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Golden Rice is part of the solution

Rice and prejudice, investigative paper published in the medical research archives journal.

On 29 February 2024, the following paper concerning Golden Rice was published by the European Society of Medicine:

“Prejudice, against GMO crops and Golden Rice, in US Academia drove unethical behaviour, with global and detrimental consequences for vitamin A deficiency alleviation”

Author: Dr A C Dubock [email protected] Executive Secretary, Golden Rice Humanitarian Board, Switzerland

In 2015, Tang et al 2012 was retracted. The paper concerned human research, relevant to public health, conducted in China in 2008. Retraction represents the most severe criticism of a scientific article. This article recounts events over a four-year period and challenges the justification for retraction based on the Committee on Publication Ethics principles.

This research focuses on analysing contemporary (2012–2015) documentary evidence, organised by key narrative participants: Greenpeace, the Chinese Government, Tufts University, the American Society for Nutrition, the US National Institutes of Health, and the US Office for Human Research Protections.

The analysis indicates that technological bias within a university and a learned society, which is also a publisher, led to unethical behaviour and the subsequent retraction. In the USA, oversight of an Institutional Review Board falls under the Office for Human Research Protections. Despite being the principal funder, the NIH's reliance on this office for the retracted paper's research to be publicly available, suggests ineffective oversight.

The retracted paper detailed a crucial nutritional study relevant to combating vitamin A deficiency, a significant cause of child mortality and blindness in low- and middle-income countries. The retraction likely heightened suspicion around this vital public health intervention.

Recommendations are made which are designed to partially ameliorate the injustices perpetrated.

The full paper can be accessed by clicking the following link: Dubock article .

And the documents referenced in the text can be accessed by clicking the following link: Support material .

The Recommendations

• The retraction of Tang et al 2012 should be rescinded by ASN for the same reasons given by KMK Vice President for Publications, ASN when she threatened Dr Tang and her co-authors on December 5 2013 (Online Resource 6): “to maintain the ethical standards of AJCN and to ensure the integrity of the scientific record.”
• Tufts University should repay Dr Tang the salary not paid in 2014, should properly consider her application for promotion to Professor withheld in 2013 and back date her pay due between then and now, and compensate her for unfair dismissal associated with this case.
• The Chinese Center for Disease Control and Prevention should reinstate the professional status of Yin Shi'an and repay lost income, and compensate for unfair treatment
• The Zhejiang Academy of Medical Sciences should reinstate the professional status of Wang Yin and repay lost income, and compensate for unfair treatment
• The Hunan Provincial Center for Disease Control and Prevention should reinstate the professional status of Hu Yuming and repay lost income, and compensate for unfair treatment
• To prevent further miscarriages of justice, Human Health Services of the US National Institutes of Health, should review their Office for Human Research Protections processes used to review supportable challenges to Institutional Review Board decisions.

Comments on the Recommendations by the responsible institutions

A few hours after publication on Friday March 1st the abstract and links above were e mailed individually to the President and to the Chief Executive of the American Society for Nutrition (ASN), to the Editor in Chief of the American Journal of Clinical Nutrition (AJCN) , and to another senior editor of the AJCN (who was the Editor in Chief during 2012- 2015), The President of Tufts University, and also to the immediate Past President of Tufts University (who was President of the University 2012- 2015), to the Chief of the Chinese Centre for Disease Control and Prevention (CCDCP), and to the Director of the US National Institutes of Health (NIH) .

On Wednesday March 5 - 4 days ago at the time of writing - comments were invited from:

• ASN (copied to both AJCN editors) relating to Recommendation 1,
• Tufts University President (copied to the Past President), relating to Recommendation 2
• The CCDCP chief relating to Recommendations 3, 4 and 5
• The Director of NIH relating to Recommendation 6.

Comments were requested by Friday March 8 midnight (CET). We said that we would record these requests and also post ‘verbatim’ any comments received on this, our website. (Comments were requested to be less than 210 words – the same as the Abstract.)

No comments have been received.

Allow Golden Rice to save lives

An Opinion paper by Felicia Wu and colleagues, published in the Proceedings of the National Academy of Sciences USA (PNAS) in December 2021, notes that 20 years after Golden Rice was first obtained by Ingo Potrykus and Peter Beyer, the tragedy we face is that this brilliant scientific success is opaqued by regulatory delays that have only led to a perpetuation of immense grief and huge losses in terms of preventable deaths, with no reported apparent benefits to consumers or the environment brought about by the overprecautionary stance of the authorities involved in the decision-making process. The urgency of getting Golden Rice approved has become more apparent, and even more urgent, with the ongoing pandemic, which has made access to healthcare services more difficult in vulnerable populations worldwide.

The World Bank recommends that micronutrient biofortification of staple crops, including specifically Golden Rice, should be the norm and not the exception in crop breeding. Golden Rice can effectively control vitamin A deficiency (VAD) and its deadly consequences, especially for children. Delaying the uptake of a genetically modified product shown to have clear health benefits has and will cost numerous lives, frequently of the most vulnerable individuals. VAD has cost more lives than the current pandemic already! Policymakers must find ways to overcome this resistance and accelerate the introduction and adoption of Golden Rice.

Link to opinion paper at PNAS (or as PDF )

In the 1990s, between 23–34% of children under 5 deaths in the world were VAD related. Progress against the UN Millennium Development Goals brought down this number to about 2% of all deaths attributed to VAD. This was achieved by a combination of mass vaccination programs against measles, better access to clean water, and vitamin A supplementation, along with economic development and education about diet reducing food insecurity. With community health programs having been adversely affected by the pandemic there is an imminent danger that VAD related deaths might climb again toward 1990s levels. It is under such circumstances that adoption of biofortified crops like Golden Rice can show their greatest potential as a safe, culturally simple and economically sustainable amelioration due to the simple facts that smallholders can grow and multiply their own biofortied crops and that such crops can vastly reduce the need for supplementation campaigns requiring recurring assembly of a costly labour and travel infrastructure to reach all those in need in the most remote areas. This current situation and possible solutions are discussed in another article by Dubock et al entitled “Golden Rice, VAD, Covid and Public Health: Saving Lives and Money” (Link to publisher IntechOpen or to PDF ).

Massive production of 'Golden Rice' seeds to start this year

Biotechnology to contribute to agriculture in the philippines.

The Presidential Communications office in the Philippines has announced that 2022 marks the start of massive production of Golden Rice seed, as well as Golden Rice for consumption, focusing on the vitamin A deficient provinces. Thus, Golden Rice spearheads the country's regional leadership in recognising biotechnology as a “powerful force to feed the future”, thereby establishing leadership in nutritional security, sustainability, agricultural productivity and economic growth. Golden Rice will be promoted as part of the Philippines Plan of Action for Nutrition. Golden Rice also featured prominently in the recent opening of the Crop Biotechnology Centre of the Philippines, by Department of Agriculture Secretary, Dr William Dar. The Philippines National Seed Industry Council has adopted a unified policy for the varietal registration of all genetically modified crops, saving the costs and time of unnecessary duplication of development work.

Commercial Propagation Permit for Golden Rice signed off in the Philippines

Philippines becomes first country to approve golden rice for planting.

Note: Commercial in this context means that rice carrying the Golden Rice trait for provitamin A production may be sold freely, which does not imply that there will be any extra cost attached to the trait itself, as this is prohibited by the agreement under which such varieties are licensed, according to the terms of donation by the creators of Golden Rice. Also, smallholders will be allowed to produce their seed without restrictions.

As of 21 July 2021, the Director of the Philippines Department of Agriculture's Bureau of Plant Industries (DA-BPI) signed off on the Commercial Propagation Permit for Golden Rice in the Philippines.

You can read about this widely publicised event in many local and international news releases:

International Rice Research Institute (IRRI) : "Philippines becomes first country to approve nutrient-enriched Golden Rice for planting" . Filipino farmers will become the first in the world to be able to cultivate a variety of rice enriched with nutrients to help reduce childhood malnutrition, after receiving the green light from regulators. Golden Rice was developed by the Department of Agriculture-Philippine Rice Research Institute (DA-PhilRice) in partnership with the International Rice Research Institute (IRRI) to contain additional levels of beta-carotene, which the body converts into vitamin A. […]

PhilRice (Philippine Rice Research Institute, Department of Agriculture): “Filipinos soon to plant and eat Golden Rice” . Filipino rice consumers are close to benefiting from a Vitamin A-infused rice with the approval of its commercial propagation permit.Dr. John C. de Leon, executive director of the Department of Agriculture-Philippine Rice Research Institute (DA-PhilRice), announced that a biosafety permit for propagating the Golden Rice has been issued on July 21, 2021. […]

Zeit online (Germany): "Philippinen genehmigen gentechnisch veränderten goldenen Reis". Die Reissorte enthält Beta-Carotin – eine Vorstufe des Vitamins A. Ein Mangel daran ist in Entwicklungsländern oft Grund für Erblindungen bei Kindern. […]

Dhaka Tribune (Bangladesh): “Philippines becomes first country to approve Golden Rice for planting” . The Philippines on Friday approved commercial cultivation of vitamin A-rich Golden Rice, long touted as a partial remedy for childhood malnutrition. It comes at a time when scientists in Bangladesh expressed deep frustration over regulators’ delay in approving the variety in the country for nearly four years. […]

The Daily Star (Bangladesh): "Philippines’ approval of Vitamin-A enriched Golden Rice a positive for Bangladesh too" . The Department of Agriculture in Philippines has approved the release of Vitamin A-enriched "Golden Rice", clearing the way for it to be cultivated commercially in the country. […]

Statement by the National Academy of Science and Technology of the Philippines

On the occasion of the approval of bt eggplant and golden rice.

The Department of Agriculture of the Philippines approved Bt eggplant for food, feed and processing, and Golden Rice for commercial propagation. On this auspicious occasion, the Academy of Science of the Philippines has congratulated the Institute of Plant Breeding, UP Los Baños, for its work on Bt eggplant, and PhilRice, the Department of Agriculture, and the International Rice Research Institute (IRRI) for their work on Golden Rice. The also congratulated the government regulatory system for the rigorous work of ensuring that these products were properly evaluated.

Golden Rice Biosafety Assessments Published

Bangladesh and the philippines leading the pack.

Golden Rice, created 20 years ago and intended as an additional intervention to combat vitamin A deficiency, is closer to being released for cultivation and human consumption in the Philippines and Bangladesh. Assessments of environmental and consumer safety, following detailed research over many years, have been submitted in applications to the corresponding authorities (more in the Regulatory section) . Separately, the efficiency of conversion of the beta-carotene provitamin A in Golden Rice to circulating vitamin A has been reported from human studies, proving that Golden Rice is an effective source of Vitamin A (Tang et al., 2009) .

Biosafety assessments involve the molecular characterisation of the introduced gene constructs and the biochemical characterisation of the improved crop plant, including a comparative compositional analysis of the biofortified Golden Rice against conventional white rice grains. The molecular characterisation involves analysing the integrity and stability of the inserted gene construct. The DNA sequence of the gene construct is also used to exclude the unintended creation of any novel gene products, including any potential allergens or toxins. Digestibility and heat stability of the gene products (proteins) determines the dietary exposure and allergenic potential of each. In our Publications section you will find four recent publications describing regulatory data generated (Swamy et al, 2019 & 2021; Biswas et al, 2021; Oliva et al, 2020) . Food and feed safety, agronomic performance and environmental interactions are reported. The reports involved collaboration of 30 scientist authors from four countries and six research institutions: the Bangladesh Agricultural University (BAU), the Bangladesh Rice Research Institute (BRRI), the Donald Danforth Plant Science Center in the USA, the International Rice Research Institute (IRRI), the Philippine Rice Research Institute (PhilRice), and the University of Freiburg in Germany.

Golden Rice event GR2E* has been crossbred with local rice varieties preferred by growers and consumers in Bangladesh and the Philippines. This gene construct, together with its surrounding DNA, is passed on from generation to generation through breeding programs, ensuring that its structure remains intact. The resulting breeding lines have been tested in multiple locations. The Golden Rice program’s objective is, following consumption, to increase circulating vitamin A levels in the blood to counteract vitamin A deficiency, thereby boosting immunity to common diseases and significantly reducing childhood blindness, of which vitamin A deficiency is the leading cause.

An important finding from the reported research is that beta-carotene levels were around 11 micrograms per gram of grain, which is sufficient to deliver between 80 and 110 per cent of the recommended daily intake of vitamin A for children and women, depending on their average rice consumption.

As a result of the donation of the technology from its creators Professors Potrykus and Beyer, and their agreements with the Government research institutes involved, the additional nutrition in Golden Rice is free of cost to growers or consumers: Golden Rice will cost no more than white rice.

*Many transformation events were produced ( Paine et al, 2005 ) from which event GR2E was selected based on molecular structure and insertion in the rice genome, together with agronomic performance. GR2E is the basis of the regulatory data generated and is the only form of Golden Rice which is offered for approval and use.

Attitudes and Influences relevant to Golden Rice’s potential use in the Philippines

Focussed group discussions and results from four different agro-economic zones of the philippines to understand attitudes and influences relevant to the adoption and use of golden rice conducted by aim students.

In late July 2008 Adrian Dubock approached The Asian Institute of Management, (‘AIM’) Manila, Philippines in connection with some Golden Rice marketing research planned for 2009. The idea was to involve Golden Rice Humanitarian Board member and Professor of Marketing, ‘JP’ Jeannet in providing a seminar for MBA students, at AIM, in consumer field research including focus group management and analysis, followed by about a month’s engagement for the trained students in conducting the focus groups and reporting back. Prof Ricardo Lim, Associate Dean of the W. Sycip Graduate School of Business at AIM kindly undertook to facilitate the request, which could form a component of course work and experience for the MBA students involved. Raul Boncodin of IRRI, and other IRRI colleagues, were closely involved in the subsequent organisation and management.

Nutrition and health go together

“……investing in the health and nutrition of vulnerable populations could lower the mortality rate of diseases such as covid-19 — as nutritional level and mortality rates are intricately linked.”.

Prof Fan Shenggen, former Director General of the International Food Policy Research Institute (IFPRI) and Chair of China Agricultural University proposes that to ensure food security in the Face of Covid-19, urgent action is needed:

• Governments need to strengthen market regulation to avoid panic, and guide growers to make rational planting decisions.
• National and international feed supply chains need to function normally, while allowing person-person contact to be minimised.
• Context specific cash or in-kind transfers, are urgently needed from Government, to protect the most vulnerable population members, and these need to continue for post-epidemic reconstruction efforts to be successful. Health and Nutrition officials need to increase their influence: improved health and nutrition of vulnerable populations could lower the mortality rate of diseases such as COVID-19, as nutritional level and mortality rates are intricately linked. [Golden Rice is a nutritional source of vitamin A. Vitamin A improves human immune response to disease – Editor]
• Contagious diseases such as Covid-19, Ebola, SARS, and Avian Flu do not respect national borders. Investment is needed in resilient food systems to allow all countries to prevent or contain the impact of food security crises they cause.
• Many, or all, of the above diseases originated in wildlife and jumped to humans. Regulation of meat, seafood and wildlife markets is essential.
• The smooth international trade in food products must continue uninterrupted by trade protectionist policies of any kind. Such uninterrupted food trade provides a safety buffer against localised shortages.

Adapted from Platform for African – European Partnership in Agricultural Research for Development (PAEPARD) , based on the original China Daily source .

Filipinos are First!

The philippines is the first asian country to approve golden rice for direct use, by adrian dubock, peter beyer & ingo potrykus.

December 2019

In a victory for science-based regulatory decision-making, the Government of the Philippines has, on 10th December 2019, authorised the direct use of GR2E Golden Rice in food, feed, and for processing. The regulatory data were submitted by the Philippine Rice Research Institute (PhilRice) and the International Rice Research Institute (IRRI) in the spring of 2017 and were scrutinized by several regulatory committees representing agriculture, environment, health, science and technology, and local governments. This decision is huge, representing the first food approval for Golden Rice in a country where rice is the staple and vitamin A deficiency a significant public health problem. Those involved in the authorisation are to be praised for their scientific integrity and courage in the face of stiff activist opposition.

In taking their decision, the Philippine Government has joined Australia, Canada, New Zealand, and the USA in affirming that Golden Rice is perfectly safe.

Unlike the industrialised countries, the Philippines is a country where rice is so important, that Pinoys (the Filipino people) do not consider any food to be a meal unless it is accompanied by rice. In 2018, per capita white rice consumption in the Philippines was 115 kg per annum —or 315 g daily (454 g = 1 lb), or more than 15-fold higher than in the USA.

Since the 1940s, the Philippine Government, at all levels, has pursued policies to deliver better health for its citizens. Nevertheless, the Philippines is a country where vitamin A deficiency (VAD) —which is globally the leading cause of child mortality and irreversible blindness— remains a significant public health problem.

The World Health Organization lists Philippine mothers as being moderately vitamin A deficient, and children less than 5 years old as being severely vitamin A deficient. This is despite, as reported in 2014, 85 percent of children consuming a vitamin-A rich food in the past day, and 76 percent of children receiving a vitamin A supplement in the past 6 months . Supplementation via Vitamin A capsule distribution in the Philippines has been in place since the early 1990s. Initially, the use of capsules was highly controversial. Globally, over the past 20 years, about 10 billion vitamin A capsules have been distributed to preschool children at a cost of about US$10 billion. In the Philippines, increasing standards of living, in combination with supplementation, reduced VAD incidence among preschool children from 40 percent in 2003 to 15 percent in 2008. By 2013, however, VAD incidence had increased again to 20 percent of preschool children, and 28 percent of children between 6 and 12 months old.

A universal source of vitamin A will reduce child mortality by 23–34 percent, and up to 50 percent in cases of measles, thanks to the immune-system-boosting effects of vitamin A. It is expected that adoption of Golden Rice —the golden colour beta-carotene is a source of vitamin A— into the regular diet will continue to reduce the incidence of VAD, and very sustainably: there is no extra cost for the additional nutrition, and no limitations on what small farmers can do with the seed. In the last month, a New Scientist article about Golden Rice commented: What shocks me is that some activists continue to misrepresent the truth about the rice. The cynic in me expects profit-driven multinationals to behave unethically, but I want to think that those voluntarily campaigning on issues they care about have higher standards .

Consistent with its commitment to public health, the Philippine authorities have ignored the misrepresentations and hyperbole around Golden Rice. Instead, they used their regulatory system and internationally accepted risk assessment principles (and their experience in assessing the safety of gmo crops , which are widely used in the Philippines) to carefully, and impartially, consider the data submitted by PhilRice and IRRI.

Children and women are dying and going blind as a result of vitamin A deficiency, despite existing interventions, and Golden Rice can assist. Even partial substitution of white rice consumption with Golden Rice — all grown in the Philippines by Philippine farmers — will combat VAD, and with no possibility of overdosing.

Before Golden Rice can be adopted by Filipino farmers, it will have to be approved for wide-scale propagation and receive varietal registration. Golden Rice field trials, already completed in both the Philippines and in Bangladesh —which share similar agro-ecosystems— have shown no cause for concern, so the outlook is very positive. Only following adoption of the publicly owned Golden Rice varieties, developed by PhilRice, into daily consumption, can Golden Rice start saving sight and lives , exactly as it was designed to do almost a quarter of a century ago.

Would you be deeply saddened if an airliner full of children crashed into the ground today?

How about two.

The equivalent of 13 jumbo jets full of children crashes into the ground every day and kills them all, because of vitamin A deficiency!!! Golden Rice has the potential to prevent all those deaths. Yet, Golden Rice lines developed by national scientists in countries where vitamin A is endemic are not given a green light by local authorities to be grown by those who would benefit most from those varieties, i.e., the poor families to which those dying children belong. And why is that the case? Simply because authorities are not prepared to face controversy generated by ill-guided activists and because the deaths of poor children do not seem to cause as much controversy, if any.

A recent opinion essay authored by the inventors and promoters of Golden Rice in Leapsmag reminds us of the senseless controversy that has stood in the way of Golden Rice helping reduce one of the main causes of children mortality on a global scale and brings us up to date regarding some positive developments on this front.

The essay, entitled "We pioneered a technology to save millions of poor children, but a worldwide smear campaign has blocked it" (click on the title to follow a link to the essay and the magazine).

Leapsmag is an editorially independent, award-winning online magazine that aims to foster a society-wide conversation about the impact of groundbreaking advances in the life sciences and related fields. Leapsmag publishes reported feature articles, commentary, personal essays, and interviews with innovators whose work stands to affect us all.

Golden Rice Named Among Project Management Institute’s Most Influential Projects of the Last 50 Years

Golden Rice is the first purposefully created biofortified food. Biofortified foods are increasingly being used to address global health issues. And are recommended as standard by the World Bank. Golden Rice, a source of vitamin A, is an additional intervention, and a disruptive technology, for use against vitamin A deficiency, a major public health issue and the most significant cause of child mortality and blindness globally.

7 October 2019 – The Golden Rice humanitarian project, announced today that it has been recognized in the top-10 Biotech Projects , as one of the most influential projects of the past 50 years by Project Management Institute (PMI) in its 2019 Most Influential Projects list. Golden Rice is the only plant-based biotech project listed, although it shares its health applications with the other nine in the list.

Additionally, PMI has released lists of the top 10 most influential projects across 14 categories in a variety of regions and industries, including a broadly-based biotechnology category. The final selections, made by PMI’s thought-leadership team, provide an inspirational reflection on what project work has enabled and the central role it has played in creating our present.

The lists are extremely eclectic, and it is gratifying to see Golden Rice recognised, in a process which the project had no input into.

The technology behind Golden Rice was donated to assist the resource poor of the world in 2000, by its inventors Professors Ingo Potrykus and Peter Beyer. Golden Rice is a not-for-profit project: no individual, nor organisation involved with its development, has any financial interest in the outcome. And, as a result of the terms of the donation by its inventors, and collaborations with Governments of countries where rice is the staple food and vitamin A deficiency endemic, Golden Rice will cost no more that the white rice variety into which the nutritional trait has been introduced.

Ingo Potrykus commented: “When starting this project in the early 90’s I was 56. Around 4,500 children a day die as a result of the ‘nutritionally acquired immune deficiency syndrome’ which is Vitamin A deficiency. Many more become blind. Now I am approaching my 86th birthday and Golden Rice is still not in the hands of those who need it so badly.

Now, though, everything is in place. The need for Golden Rice is clear, and it is registered as safe in Australia, Canada, New Zealand and USA. It is very clear it can make a huge contribution as an additional intervention for vitamin A deficiency, at no cost to growers or consumers. And it can contribute to attainment of Sustainable Development Goals 1,2,3,4,5 & 7.

Regulatory dossiers have been submitted in key developing countries. All that is now needed is for Public Health Professionals to overcome any scepticism caused by the anti-gmo activists’ activities over the past three decades and embrace Golden Rice.

Hopefully in my lifetime, you, and I, will start to see Golden Rice saving the sight and lives of some of the 3.5 billion people, half the world’s population, who consume rice, and often little else, every day.”

“This recognition reflects the incredible progress we have made in the project management profession and demonstrates how the fabric of our world has been shaped, and continues to be shaped, by the hard work of bringing ideas to life,” said Sunil Parashara, President and CEO of Project Management Institute. “This list demonstrates PMI’s vision of how excellence in project execution will be critical in meeting the challenges and opportunities of tomorrow.”

The list is part of PMI’s 50th anniversary celebration that includes various activities to recognize the important role project management has played over the past five decades and celebrate where the profession is going.

The complete list of projects honoured can be found at this PMI link

The list of Honourees of the Golden Rice project recognised for their contributions in the PMI Award

About Project Management Institute (PMI)

Project Management Institute (PMI) is the world's leading association for those who consider project, program or portfolio management their profession. Founded in 1969, PMI delivers value for more than three million professionals working in nearly every country in the world through global advocacy, collaboration, education and research. We advance careers, improve organizational success and further mature the project management profession through globally-recognized standards, certifications, communities, resources, tools, academic research, publications, professional development courses and networking opportunities. As part of the PMI family, ProjectManagement.com creates online global communities that deliver more resources, better tools, larger networks and broader perspectives. Visit us at PMI or Project Management , Facebook , and on Twitter @PMInstitute.

For more information about Golden Rice please refer to: Potrykus I (2014) From the concept of totipotency to biofortified cereals. Annual Review of Plant Biology 66(1):1-22 Dubock A (2019) Golden Rice: To Combat Vitamin A Deficiency for Public Health . DOI: 10.5772/intechopen.84445

Golden Rice: To Combat Vitamin A Deficiency for Public Health

Article by dr adrian dubock, member of the golden rice humanitarian board.

Vitamin A deficiency (VAD) has been recognised as a significant public health problem continuously for more than 30 years, despite current interventions. The problem is particularly severe in populations where rice is the staple food and diversity of diet is limited, as white rice contains no micronutrients. Golden Rice is a public-sector product designed as an additional intervention for VAD. There will be no charge for the nutritional trait, which has been donated by its inventors for use in public-sector rice varieties to assist the resource poor, and no limitations on what small farmers can do with the crop—saving and replanting seed, selling seed and selling grain are all possible. Because Golden Rice had to be created by introducing two new genes—one from maize and the other from a very commonly ingested soil bacterium—it has taken a long time to get from the laboratory to the field. Now it has been formally registered as safe as food, feed, or in processed form by four industrialised countries, and applications are pending in developing countries. The data are summarised here, and criticisms addressed, for a public health professional audience: is it needed, will it work, is it safe and is it economic? Adoption of Golden Rice, the next step after in-country registration, requires strategic and tactical cooperation across professions, non-governmental organisations (NGOs) and government departments often not used to working together. Public health professionals need to play a prominent role.

The full article can be accessed following this link to IntechOpen (From the Edited Volume «Vitamin A"» [Working title] Edited by Prof Leila Queiroz Zepka, Dr Eduardo Jacob-Lopes and Dr Veridiana Vera De Rosso; DOI: 10.5772/intechopen.84445)

Golden Rice: The Imperiled Birth of a GMO Superfood

A book by ed regis.

Supporters claim that the twenty-year delay in Golden Rice's introduction is an unconscionable crime against humanity. Critics have countered that the rice is a "hoax," that it is "fool's gold" and "propaganda for the genetic engineering industry." Here, science writer Ed Regis argues that Golden Rice is the world's most controversial, maligned, and misunderstood GMO. Regis tells the story of how the development, growth, and distribution of Golden Rice was delayed and repeatedly derailed by a complex but outdated set of operational guidelines and regulations imposed by the governments and sabotaged by anti-GMO activists in the very nations where the rice is most needed.

Writing in a conversational style, Regis separates hyperbole from facts, overturning the myths, distortions, and urban legends about this uniquely promising superfood. Anyone interested in GMOs, social justice, or world hunger will find Golden Rice a compelling, sad, and maddening true-life science tale.

Available from Amazon

ISBN-13: 978-1421433035 ISBN-10: 1421433036

And this is what Ingo Potrykus, one of the creators of Golden Rice had to tell to the author of the book: “I am half way through your book and I can’t wait to the end to tell you, how excited I am. It is simply excellent !!! Wonderful that you have devoted your talent and efforts to tell the public in such a clear presentation, what stands in the way of an important humanitarian project just because it is a GMO project.”

Lindau Nobel Laureate Meeting

Sir richard roberts talks about gm crops.

Sir Rich Roberts, FRS, organized an open letter from fellow Nobel Laureates to Greenpeace, the UN and the Governments of the World, decrying their unscientific treatment of GMO-crops. Two years later, in June 2018, Dr Roberts talked about his views at the 68th Lindau Nobel Laureate Meeting with Young Scientists, Germany.

Sir Rich: “At the meeting I described the Nobel Laureates campaign in favor of GMOs. Examples of the benefits of the new GM technology for citizens of the developing world include Golden Rice and halting both Banana Wilt and the Fall Army Worm.

For biofortification alone GMO technology can deliver high folate rice (mothers’ dietary deficiency causes birth defects), high zinc and high iron rice (dietary deficiency impedes mental development). Similarly, GMO Golden Rice provides a source of vitamin A. Vitamin A deficiency is an immune deficiency syndrome, so children die of common infections. It is also the main cause of irreversible childhood blindness. Golden Rice has been accepted as safe for consumption by the Governments of Australia, Canada, New Zealand and USA, and registrations have been applied for in Philippines and Bangladesh. Yet, significantly due to rejection of science by activists, Golden Rice is not yet available to farmers and their communities as an additional intervention for vitamin A deficiency. And neither high folate rice, nor high iron rice, nor high zinc rice, nor Golden Rice could be developed without the use of GMO-technology.

Millions of people can benefit from the use of GMO-technology in plant breeding, it is hard to comprehend how the anti-GMO movement can sleep at night.”

See the video (50%/50% presentation/discussion) here (about 40 min):

Three of the slides which are slightly difficult to read on a small screen can be seen as large pictures when clicking on the thumbnails below:

You too can sign the letter here: http://supportprecisionagriculture.org/join-us_rjr.html

Golden Rice

An update by adrian dubock, executive secretary, golden rice humanitarian board.

In early 2001, the International Rice Research Institute (IRRI) in the Philippines became the first licensee of Professors Ingo Potrykus and Peter Beyer for what became known as Golden Rice.

IRRI agreed to develop Golden Rice to fulfil the inventors' vision: to make the nutritional benefits of Golden Rice available as an additional intervention for vitamin A deficiency (VAD), without any additional cost compared to white rice, in developing countries to governments, small farmers or consumers. Except for commercial export, no restrictions were imposed on what the farmers could do with the seed. Golden Rice was designed by its inventors, and the technology donated by them, to help the ‘resource poor’.

In the same year, I was fortunate to accompany Ingo and Peter to deliver to IRRI the first 600 seeds, and six 2.5mL tubes of the genes necessary to turn any white rice into a biosynthetic factory for beta-carotene. Beta-carotene, from any source, is converted by the human body into vitamin A. It is vitamin A which is essential for a functional immune system, allowing children and their mothers to fight infection and to prevent the childhood blindness often associated with VAD. Later research confirmed that the beta-carotene in Golden Rice is converted very efficiently into vitamin A. As a source of vitamin A Golden Rice can be as effective as milk, eggs or butter. Only 40 grams consumed daily is expected to prevent death and blindness, with no possibility of overdosing, as the human body only converts the beta-carotene it needs to vitamin A and excretes the rest unchanged.

Shortly after Ingo and Peter had published their initial ‘Proof of Concept’ research in 2000, they elicited the help of Syngenta. In return for Syngenta committing to assist the inventor’s humanitarian project, Syngenta acquired the commercial rights to the inventor’s core technology. In 2004 Syngenta renounced its commercial interest in favour of more profitable opportunities. But not before its scientists had made significant improvements to the technology. As they were obligated to, Syngenta passed the technology rights and the improvements, as seed, to the inventor’s licensees, including IRRI, in 2006, so that IRRI could continue to fulfil their licence obligations to the inventors.

Meanwhile, extensive data sets have been generated —the data files alone total 32 megabytes— proving that Golden Rice differs from white rice only by the presence of beta-carotene, is safe to consume, and cannot cause allergies. It is direct descendants of one of those seeds, known as GR2E, delivered to IRRI in 2006, multiplied and introduced into Asian varieties of rice by conventional breeding, which have provided that data.

Although it is hard to imagine that such golden grains of polished rice could be included in commercial shipments of white rice by accident, in the modern world any such inclusion could be damaging to international trade. To prevent even such an unlikely situation, the regulatory data has been made available not only to countries where VAD remains a very significant public health problem, but also to other countries which import rice. Independent regulators have confirmed Golden Rice’s safety.

The inventors vision, expressed in Time magazine’s headline in July 2000, is getting closer. Despite the protesters' beliefs.

For more detailed information please refer to: http://rdcu.be/wwud ; http://rdcu.be/wwui ; http://rdcu.be/wwub

And the 2016 World Food Prize goes to ... Biofortified Sweet Potatoes

Biofortification: empowering and self-sustaining.

The 2016 World Food Prize has been awarded to the group of scientists who have tirelessly worked on breeding and introducing orange-fleshed sweet potatoes to Africa and thus benefitting millions of people, especially children, who are most susceptible to a lack of provitamin A. The World Food Prize thus once again recognises efforts to increase the quality and quantity of available food to the most vulnerable populations in the world.

Three of the 2016 laureates - Drs Maria Andrade, Robert Mwanga and Jan Low are from the CGIAR International Potato Center (CIP). The fourth winner, Dr Howard Bouis, is the founder of HarvestPlus at the CGIAR International Food Policy Research Institute (IFPRI), and is being recognised for his work over 25 years to ensure biofortification was developed into an international plant breeding strategy across more than 40 countries.

Vitamin A deficiency (VAD) is considered to be one of the most harmful forms of malnutrition in the developing world. It can cause blindness, limit growth, and weaken the body's immune system, thereby increasing morbidity and mortality. The condition affects more than 140 million pre-school children in 118 nations, and more than seven million pregnant women. It is probably the leading cause of child blindness in developing countries.

Biofortification seeks to improve nutritional quality of food crops through agronomic practices, conventional plant breeding, or modern biotechnology, as in the case of Golden Rice. The approach of providing farmers with biofortified crops, indepedently of the technology used to achieve it, is thus the most efficient way of creating a self-sustaining and virtuous cycle of nutritional independence and life quality improvement.

In the case of sweet potatoes, breeders utilise the fact that varieties producing and storing high levels of beta-carotene (=provitamin A) are available in the Andean region of South America and thus can use these for breeding purposes and create new orange-fleshed varieties acceptable to regional taste preferences in Africa. Unfortunately, such genetic variability is not available for every crop, thus requiring the use of laternative approaches to generate the new, desirable trait.

Before the introduction of orange-fleshed varieties people in Africa had a preference for white-fleshed varieties, something which is changing thanks to the work of the WFP 2016 laureates and their colleagues at various international organizations. That goes once more to prove that preferences can evolve, especially when consumers can be convinced of the benefits to their children.

And more than that, the example of the orange-fleshed sweet potato has proven that the matrix of biofortified crops are perfectly suited as a conduit to carry the much needed micronutrient, in this case is provitamin A. The outcome of this project calls for rapid introduction and adoption of a number of biofortified crops, like Golden Rice, biofortified bananas, cassava, sorghum, and other crops rich in other micronutrients like iron and zinc, which would address other major, widely spread nutritional deficiencies.

150 Nobel laureates (updated Oct 2019) have signed letter blasting Greenpeace over GMOs

From the washington post - june 2016.

More than 100 Nobel laureates have signed a letter urging Greenpeace to end its opposition to genetically modified organisms (GMOs). The letter asks Greenpeace to cease its efforts to block introduction of a genetically engineered strain of rice that supporters say could reduce Vitamin-A deficiencies causing blindness and death in children in the developing world.

By all standards, Nobel Prize laureates are usually considered the finest intellects that humanity has to offer, notwithstanding the fact that tens of thousands of other fine scientific minds and many other serious thinkers are supportive of biotechnology in agriculture. Add to that the simple fact that we all have been eating the biotechnology-derived products for the last twenty years without a single case of adverse effects linked to the biotechnological intervention as such, and non-experts should be able to arrive at the same conclusions that these fine minds have arrived at. And that is that biotechnology has already become part of the standard toolset used in plant breeding in combination with all other technologies developed and used since the inception of agriculture as we know it.

Here's a link to the press briefing by Sir Richard Roberts FRS and two other Nobel Laureates on the topic: Nobel Laureates Press Conference - 30 June 2016

You may also want to read Adrian Dubock's (Executive Secretary, Golden Rice Humanitarian Board) comments on how Greenpeace and other GMO critics misrepresent the Golden Rice Humanitarian Project at the Genetic Literacy Project site: "Disembedding grain: Golden Rice, The Green Revolution, and heirloom seeds in the Philippines"

Are you aware of the very important Support Precision Agriculture Initiative ? If you're interested in reading about the pro GMO campaign and learn more about agricultural biotechnology follow the link provided with the initiative's name, and if you like and agree with the content please Please sign on at the following page: Join Us! and do share with your colleagues"

The Golden Rice project wins the Patents for Humanity Award 2015

Patents for Humanity is a USPTO program that recognizes patent owners and licensees working to improve global health and living standards for underserved populations. The program advances the President's global development agenda by recognizing private sector leaders who bring life-saving technologies to those in need, while showing how patents are an integral part of tackling the world's challenges.

Back in 2001, in a ground-breaking humanitarian licensing arrangement , the three applicants (with Dubock then working for Syngenta) arranged in a cashless transaction for the defined commercial rights in US patent US 7,838,749 (and related patents) to be transferred to Syngenta. The inventors retained rights to the carefully and generously defined humanitarian applications. Syngenta, in return for its commercial options acquired, became obligated to support the humanitarian and non-profit vision of the inventors, and the inventors’ public sector licensees, rights to exploit any improvement, including as exemplified by patent application US20120042417 A1. Syngenta stated in 2004 that it had no continuing interest in commercial exploitation of the technology. Nevertheless, Syngenta’s obligations to support the inventors and their Golden Rice humanitarian project remain in place.

Dr Adrian Dubock (front left) collected the award at the White House on 20 April 2015 together with Prof Rob Russell, Golden Rice Humanitarian Board member(rear left).

These arrangements demonstrate that patents have a very useful role, even for projects involving developing countries, where the protection of intellectual property rights may be less well developed. Without the inventors having applied for patents, it would not have been possible to discuss and develop the above mutually beneficial arrangements between the private and public sectors. Moreover, having the Golden Rice patent in place was crucial to obtaining access to the supporting technology package from other inventors.

GOLDEN RICE NOW!

Showing the dark side of the anti gm campaigners.

This initiative, led by Dr Patrick Moore, co-founder and 15 years leader of Greenpeace and longtime adviser to government and industries on sustainability and the environment, conducts protests and forums with the aim to end the active blocking of Golden Rice by environmental organizations who claim that it is either of no value or that it is a detriment to human health and the environment. The ALLOW GOLDEN RICE NOW! Society plans to achieve this through direct public action, media communications and coalition-building.

Visit the ALLOW GOLDEN RICE NOW! Society website to find out more about dates, locations and activities.

BBC Interview with Prof Hans-Jörg Jacobsen and Vandana Shiva, 20 April 2015. Your browser does not support the audio element.

People Pope Blesses Golden Rice

Aspb news | volume 41, number 1.

BY TYRONE SPADY ASPB Legislative and Public Affairs Director

On November 7, 2013, Pope Francis gave his personal blessing to Golden Rice (GR). Why is this significant? Vitamin A deficiency (VAD) is responsible for 500,000 cases of irreversible blindness and up to 2 million deaths each year. Particularly susceptible are pregnant women and children. Across the globe, an estimated 19 million pregnant women and 190 million children suffer from the condition. The good news, however, is that dietary supplementation of vitamin A can eliminate VAD. One way that holds particular promise is the administration via GR, which had been engineered to produce large amounts of vitamin A. A 2012 study by Tang et al. published (retracted for political reasons, not because of its content) in the American Journal of Clinical Nutrition found that 100-150 g of cooked GR provided 60% of the Chinese Recommended Intake of vitamin A. Estimates suggest that supplementing GR for 20% of the diet of children and 10% for pregnant women and mothers will be enough to combat the effects of VAD.

Unfortunately, public misconceptions about genetically modified (GM) organisms have prevented GR from being available to the countries most affected by VAD. One such country is the Philippines, where more than 80% of the population identifies as Roman Catholic and field trials of GR are nearing completion. An official blessing of the church, therefore, could do a great deal to build support, allowing the Philippines to serve as a model for many of its neighbors on the potential health impacts of widespread availability and consumption of the golden grain.

Regrettably, the church did not provide an official endorsement. It turns out that there is quite a distinction between the pope's personal blessing and an official statement of support from the Vatican. To understand the nature of that distinction, we turned to the person who elicited the blessing, GR coinventor and ASPB member Ingo Potrykus. At the time of the blessing, Ingo, a member of the Pontifical Academy of Sciences, had been attending a meeting at the Vatican on the interaction of nutrition and brain development. At the end of the meeting, he was able to meet Pope Francis and took the opportunity to share a packet of GR. In response, the pope offered his personal blessing. (If an official blessing of the Holy See was given, it would come from the Pontifical Council for Justice and Peace.) From Ingo's perspective, the pope is concerned that genetic modification technology primarily benefits big business and not the poor.

The most immediate hurdle to the usage of GR, according to Ingo, is the impending deregulation by the Philippine Department of Agriculture. Although no damage has been reported from the recent typhoon (Haiyan) that struck this part of the world, the fields had already been harvested. Philippine officials have been following GR development and field trials for several years, and Ingo believes that the government will ultimately give "the green light." He expects that deregulation will occur in two phases: first consumption, then planting. The consumption phase will require a two-year study of the impacts of GR consumption on VAD in Philippine children. The study will be conducted by the Helen Keller Foundation for Research and Education (http://bit. ly/1bXh9AX), which has expertise in VAD and blindness. Only after the study will farmers be allowed to plant GR, said Ingo.

GR distribution will be carried out by existing small-scale operations. Further, it will be sold at the same price as conventional cultivars. It is believed that this will help to facilitate adoption. In addition to vitamin A production, Ingo believes that other agronomic improvements, such as increased pest resistance and yield, will further increase the attractiveness of GR to farmers.

While not a full-throated endorsement of GR or GM, the pope's blessing is a step in the right direction. It is also an important indicator of slowly shifting global attitudes regarding the role that GM foods will play in the world's long-term food security.

Copyright for this article lies with ASPB News

Biofortified rice as a contribution to the alleviation of life-threatening micronutrient deficiencies in developing countries

A good start is a food start.

Dietary micronutrient deficiencies, such as the lack of vitamin A, iodine, iron or zinc, are a major source of morbidity (increased susceptibility to disease) and mortality worldwide. These deficiencies affect particularly children, impairing their immune system and normal development, causing disease and ultimately death. The best way to avoid micronutrient deficiencies is by way of a varied diet, rich in vegetables, fruits and animal products.

The second best approach, especially for those who cannot afford a balanced diet, is by way of nutrient-dense staple crops. Sweet potatoes, for example, are available as varieties that are either rich or poor in provitamin A. Those producing and accumulating provitamin A (orange-fleshed sweetpotatoes) are called biofortified ,* as opposed to the white-fleshed sweet potatoes, which do not accumulate provitamin A. In this case, what needs to be done is to introduce the biofortified varieties to people used to the white-fleshed varieties, as is happening at present in southern Africa by introducing South American varieties of orange-fleshed sweetpotatoes.

Unfortunately, there are no natural provitamin A-containing rice varieties. In rice-based societies, the absence of β-carotene in rice grains manifests itself in a marked incidence of blindness and susceptibility to disease, leading to an increased incidence of premature death of small children, the weakest link in the chain.

Rice plants produce β-carotene (provitamin A) in green tissues but not in the endosperm (the edible part of the seed). The outer coat of the dehusked grains—the so-called aleurone layer—contains a number of valuable nutrients, e.g. vitamin B and nutritious fats, but no provitamin A. These nutrients are lost with the bran fraction in the process of milling and polishing. While it would be desirable to keep those nutrients with the grain, the fatty components are affected by oxidative processes that make the grain turn rancid when exposed to air. Thus, unprocessed rice—also known as brown rice—is not apt for long-term storage.

Even though all required genes to produce provitamin A are present in the grain, some of them are turned off during development. This is where the ingenuity of the Golden Rice inventors, Profs Ingo Potrykus (formerly ETH Zurich) and Peter Beyer (University of Freiburg) comes into play. They figured out how to turn on this complex pathway again with a minor intervention.

The shocking fact is that, far from reaching the envisaged Millenium Development Goals, more than 10 million children under the age of five are still dying every year. A high proportion of those children die victims of common diseases that could be prevented through a better nutrition. This number has been equated with a ‘Nutritional Holocaust’ . It is unfortunate that the world is not embracing more readily a number of approaches wih the potential to substantially reduce the number of deaths. It has been calculated that the life of 25 percent of those children could be spared by providing them with diets that included crops biofortified with provitamin A (beta-carotene) and zinc. Golden Rice is such a biofortified crop. Those involved in the project are hopeful that in a near future Golden Rice will be growing in farmers' fields and helping to improve the diets of millions of people.

Golden Rice grains are easily recognisable by their yellow to orange colour. The stronger the colour the more β-carotene. While a yellow rice is still unfamiliar to most of us, it is hoped that the pleasant colour will help promote its adoption. Would you believe that once upon a time carrots were white or purple? Orange-coloured carrots are the product of a mutation selected by a Dutch horticulturist a few hundred years ago, because it was the colour of the Dutch Royal House of Orange-Nassau!

*Welch RM and Graham RD (2004) Breeding for micronutrients in staple food crops from a human nutrition perspective. J Exp Bot 55:353-364.

Quantum leap:

Golden rice accumulates provitamin a (β-carotene) in the grain.

Rice produces β-carotene in the leaves but not in the grain, where the biosynthetic pathway is turned off during plant development. In Golden Rice two genes have been inserted into the rice genome by genetic engineering, to restart the carotenoid biosynthetic pathway leading to the production and accumulation of β-carotene in the grains. Both genes are naturally involved in carotene biosynthesis. The difference here is that the reconstructed pathway is not subject to downregulation, as usually happens in the grain.

Since a prototype of Golden Rice was developed in the year 2000, new lines with higher β-carotene content have been generated. The intensity of the golden colour is a visual indicator of the concentration of β-carotene in the endosperm.Our goal is to make sure that people living in rice-based societies get a full complement of provitamin A from their traditional diets. This would apply to countries such as India, Vietnam, Bangladesh. the Philippines, and Indonesia. Golden Rice could still be a valuable complement to children's diets in many countries by contributing to the reduction of clinical and sub-clinical vitamin A deficiency-related diseases.

Many people are aware that vitamin A has something to do with vision, especially at night. But many are not aware of the central role it plays in maintaining the integrity of the immune system. According to the World Health Organization, dietary vitamin A deficiency (VAD) compromises the immune systems of approximately 40 percent of children under the age of five in the developing world, greatly increasing the risk of severe illnesses from common childhood infections, thus causing hundreds of thousands of unnecessary deaths among them.

In remote rural areas Golden Rice could constitute a major contribution towards sustainable vitamin A delivery. To achieve this goal a strong, concerted, and interdisciplinary effort is needed. This effort must include scientists, breeders, farmers, regulators, policy-makers, and extensionists. The latter will play a central role in educating farmers and consumers as to their available options. While the most desirable option woud be a varied and adequate diet, this goal is not always achievable, at least not in the short term. The reasons are manifold, ranging from tradition to geographical and economical limitations. Golden Rice is a step in the right direction in that it does not create new dependencies or displace traditional foodstuff.

Golden Rice , the real thing

Who is behind golden rice, helen keller international, golden rice will reach those who need it at no additional cost, growers will be able to reuse their seed as they please.

Those most in need of this new seed-based technology are those who can least afford buying an adequate diet, rich in essential nutrients. This has been taken into consideration by the creators of Golden Rice , Profs Peter Beyer and Ingo Portrykus, and the crop protection company Syngenta, who have worked together to make the latest, improved version of Golden Rice available for humanitarian use in developing countries, free of charge.

The Golden Rice Humanitarian Board encourages further research to determine how the technology may play a part in the ongoing global effort to fight Vitamin A Deficiency in poor countries. While Golden Rice is an exciting development, it is important to keep in mind that malnutrition is to a great extent rooted in political, economic and cultural issues that will not be solved by a technical fix. Yet Golden Rice offers people in developing countries a valuable and affordable choice in the fight against the scourge of malnutrition.

This site is maintained by the Golden Rice Humanitarian Board for the purpose of providing information on the background and progress of the Golden Rice Humanitarian Project.

Eat orange! We really mean it!

Eat orange! A motto promoted by HarvestPlus

158 Nobel Laureates praise Philippines move

Richard J Roberts, 1993 Nobel Prize Winner in Physiology or Medicine, on behalf of the 157 Nobel Prize winners and 13,292 co-signers supporting GMOs, have expressed their delight with the recent announcement of the move by the Philippine Department of Agriculture to authorize the direct use of Golden Rice as food and feed or for Processing. Visit Support Precision Agriculture.

Supplementation not sustainable

Pandemic affects supplementation programs.

According to the United Nations Children's Fund (UNICEF) , in 2020, the first year of the Covid-19 pandemic, despite the potential benefits of this key child survival intervention, only two out of five children in need received the life-saving benefits of vitamin A supplementation.

Colour Blindness

Art to remind us of the insensitivity of senseless opposition, regulatory status, golden rice vs white rice, ... and the difference is ....

A study carried out by IRRI, PRRI and the Danforth Center scientists and published in June 2019 shows that the only noticeable differences between Golden Rice and its non-transgenic counterpart are the elevated levels of beta-carotene and related carotenes. For more detail continue reading here.

Vitamin A boosts the immune system

Strong reduction of mortality in measles-affected children.

“The number of measles cases reported globally from January to March has tripled since last year, says the World Health Organisation. Africa saw a 700% surge. Since only 10% of all cases of the potentially fatal disease are reported, the trend could be even stronger than these initial indications. The main cause appears to be failure to immunise enough children.” Economist Espresso 16 April 2019

Many children in countries where VAD is endemic are not immunised. WHO states, with respect to vitamin A capsules: “For deficient children, the periodic supply of high-dose vitamin A in swift, simple, low-cost, high-benefit interventions has also produced remarkable results, reducing mortality by 23% overall and by up to 50% for acute measles sufferers.”

Doesn’t that make you wonder what a difference a biofortified food like Golden Rice could do for those children?

Another reason the world needs Golden Rice

Tb continued infectious disease.

Tuberculosis is a neglected disease, according to a newly published report in the Lancet, a medical journal. The experts’ plan is to end it within a generation. That is ambitious, even by the lofty measure of such proclamations. In 2017 tuberculosis killed 1.6m people, more than any other infectious disease. A quarter of the world’s population have latent TB infections, almost all in developing countries. Of them, 5-15% will develop the disease, mostly those whose immune systems are weakened by HIV, malnourishment or smoking . The plan calls for new drugs, vaccines and diagnostic tests, as well as doubling annual spending. Treating those who fall ill is crucial to preventing its spread. Yet currently more than a third of them go untreated. And nearly half a million new cases are resistant to several tuberculosis drugs. There seems a good chance the next generation will still be living with TB’s scourge.

From Economist Espresso 22 March 2019

Golden Rice is an effective source of vitamin A

β-carotene in golden rice is as good as β-carotene in oil at providing vitamin a to children.

August 2012. Researchers from USDA (Boston and Houston), Chinese instituions in Hunan, Beijing, and Hangzhou, and NIH (Bethesda), have determined that the β-carotene in Golden Rice is as effective as pure β-carotene in oil and better than that in spinach at providing vitamin A to children. A bowl of ∼100 to 150 g cooked Golden Rice (50 g dry weight) can provide ∼60% of the Chinese Recommended Nutrient Intake of vitamin A for 6-8-year-old children. The paper, with data based on a registered clinal trial, has been published by the American Journal of Clinical Nutrition . And there is good reason to conduct these studies in China, considering the low vitamin A status of a great proportion of Chinese children (see Nutrition and Health Status Report) .

Golden Rice has got what it takes

Back in 2009, researchers were able to demonstrate that Golden Rice was an effective source of vitamin A. This investigation was done with a group of healthy adult volunteers in the USA. The study showed that the β-carotene contained in Golden Rice was highly available and easily taken up into the bloodstream by the human digestive system. While foodstuffs of plant origin are the major contributors of β-carotene in the diet, these are often absent from the diet, for customary and economic reasons. And moreover, conversion of the provitamin A carotenoids contained in them is generally inefficient. Conversion factors for provitamin A carotenoids from various fruits is in the range of 13:1 for sweet potato, 15:1 for carrots, and between 10:1 and 28:1 for green leafy vegetables. With a conversion factor of 4:1 Golden Rice displays a comparatively very favourable conversion ratio. This study was published in the American Journal of Clinical Nutrition in 2009 .

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Golden Rice

• Nutrition & Food Security

The International Rice Research Institute (IRRI) and its national research partners have developed Golden Rice to complement existing interventions to address vitamin A deficiency (VAD). VAD is a serious public health problem affecting millions of children and pregnant women globally.

In South and Southeast Asian countries, where at least half of daily caloric intake is obtained from rice, Golden Rice can help in the fight against VAD, particularly among the people who depend mostly on rice for nourishment.

Golden Rice is intended to be used in combination with existing approaches to overcome VAD, including eating foods that are naturally high in vitamin A or beta-carotene, eating foods fortified with vitamin A, taking vitamin A supplements, and optimal breastfeeding practices.

Golden Rice has been assessed to be as safe as ordinary rice with the added benefit of beta-carotene in the grains by  Food Standards Australia New Zealand (22 February 2018) , Health Canada (16 March 2018) , the  United States Food and Drug Administration (24 May 2018)  and Department of Agriculture-Bureau of Plant Industry (19 December 2019) .

In July 2021, the Philippines became the first country in the world to approve Golden Rice for commercial propagation .

Updates on the Golden Rice Project

As of 2022, Golden Rice has begun pilot-scale deployment in the Philippines. It is still under regulatory review in Bangladesh.

Biosafety approval is a prerequisite for inclusion in the rice variety listing of the National Seed Board (NSB) of Bangladesh.  To complete the biosafety review process, the Bangladesh Rice Research Institute (BRRI) lodged an application to the National Technical Committee on Crop Biotechnology (NTCCB) at the Ministry of Agriculture on November 26, 2017, who forwarded the application to the National Committee on Biosafety (NCB) at the Ministry of Environment on December 4, 2017.

PHILIPPINES

DA-PhilRice is leading pilot deployment in the Philippines , with the first batch of Golden Rice seeds distributed for planting in selected provinces during the 2022 wet season planting. Golden Rice is registered with the National Seed Industry Council as NSIC 2022 Rc682GR2E, or Malusog 1, hence the naming shift from Golden Rice to Malusog Rice in the country.

Golden Rice was assessed through the Joint Department Circular (JDC) No. 1 series of 2016 , which comprises three regulatory review processes: for direct use as food and feed, or for processing (FFP); for field trial; and for commercial propagation. Regulatory applications assessed through this process underwent approval through five different government agencies – the Department of Agriculture (DA), Department of Science and Technology (DOST), Department of Environment and Natural Resources (DENR), Department of Health (DOH), and Department of Interior and Local Government (DILG)-- as well as by a panel of independent scientific, socio-cultural, and economic experts.

The biosafety permit for the commercial propagation of GR2E Golden Rice was issued by the DA-BPI on 21 July 2021.

On 18 December 2019, the FFP permit was issued by the DA-BPI, approving GR2E Golden Rice for direct use as food and feed, or for processing in the Philippines.

The biosafety permit for field trial was released by DA-BPI on 20 May 2019. The field trial--conducted in DA-PhilRice stations in Munoz, Nueva Ecija, and San Mateo,Isabela--was completed in October 2019.

IRRI’s work with Golden Rice

IRRI works with its national research partners to develop Golden Rice as a complementary food-based approach to improve vitamin A status, using popular local inbred varieties from each country. IRRI’s work will support and strengthen the:

Development of Golden Rice varieties suitable for smallholder farmers in partner countries Breeders at the Philippine Department of Agriculture - Philippine Rice Research Institute ( DA-PhilRice ), the Bangladesh Rice Research Institute ( BRRI ), and the Indonesian Center for Rice Research ( ICRR ) are developing Golden Rice versions of existing rice varieties that are popular with their local farmers, retaining the same yield, pest resistance, and grain qualities. Golden Rice seeds are expected to cost farmers the same as other rice varieties. Once PhilRice, BRRI, and ICRR are able to secure an approval from their respective regulatory agencies, cooking and taste tests will be done to make sure that Golden Rice meets consumers' needs.

Safety assessment of Golden Rice The environmental safety of Golden Rice is assessed through field tests and other evaluations in each partner country. Golden Rice is analyzed according to internationally accepted guidelines for food safety.

Research and development of Golden Rice adhere to scientific principles developed over the last 20 years by international organizations such as the World Health Organization (WHO) , the Food and Agriculture Organization of the United Nations (FAO) , the Organization for Economic Co-operation and Development (OECD) and the Codex Alimentarius Commission . These are the same principles that inform the safety assessments of national regulatory agencies, such as FSANZ, Health Canada, and the US FDA, which have already assessed Golden Rice as safe to plant and safe to eat.

Nutrition evaluation by an independent organization After obtaining the necessary permits and approvals, an independent community nutrition study will be conducted to evaluate the contribution of Golden Rice to Vitamin A status of target communities.

Deployment of Golden Rice in priority areas IRRI supports its national partners in developing pilot-scale deployment strategies to ensure that Golden Rice reaches the farmers and consumers that need it the most. A sustainable delivery program co-designed by IRRI and its national counterparts will also be implemented to ensure that Golden Rice is affordable, acceptable, and accessible in vitamin A deficient communities.

For more information on Golden Rice, visit the Golden Rice FAQs .

Can we engineer life? 4.6 Golden rice

Golden rice symbolizes the protracted trench war between proponents and opponents of modern biotechnology. The Swiss Ingo Potrykus of ETH Zürich is the golden man behind the golden rice. With great ardour he defends this controversial rice, and attacks the opponents of gene technology. With his colleague Peter Beyer of Freiburg University he developed the first version of this provitamin-A reinforced rice, between 1991 and 1999, the year in which he retired. But still, at the age of 85, he battles to get access to ‘his rice’ for children who have serious vitamin-A deficiencies.

Hans Tramper is professor emeritus in Bioprocess Technology at Wageningen University and reflects on the development of his subject in a series of essays. His pieces were published so far on 18 June , 30 June , 11 July , 22 July , 19 August , 10 September , 21 September , 30 September , 10 October , 31 October , 8 November , 2 December and 26 December 2018.

Controversial rice

In Essay 1 of this series I write: ‘The most extensively discussed transgene vegetal case, almost to the point of boredom, is that of golden rice.’ Googling ‘golden rice’ will produce hundreds of millions of hits. Yet, we need to discuss it here, because of the importance of this provitamin-A enriched rice. Moreover, we may witness at last, after much hassle, the first commercial production in Bangladesh and the Philippines. Easily accessible sources that treat golden rice extensively are a report by Charles M. Rader from 2001, and a booklet of VIB , the Flemish Institute for Biotechnology, from 2016.

Vitamin A deficiency

WHO, the World Health Organization, estimates that 250 million people suffer from vitamin A deficiency. In developing countries, 40 percent of children suffering from this condition is under five. UNICEF statistics show that each year, 1-2 million people die prematurely because of vitamin-A deficiency. It causes the immune system to weaken, and babies and children are especially at risk. Each year, about two million people become blind and in 60 percent of those incidents in India, China and sub-Saharan Africa, this is caused by vitamin A deficiency. Worldwide, it causes the loss of eyesight in a quarter to half a million children; half of them die with a year of becoming blind. This could be counteracted by health education, vitamin supply, gardening, food programs, and the use of genetically modified rice with an enhanced beta carotene (provitamin-A) content. The UN Standing Committee on Nutrition stresses the need for an integrated program in the battle against food shortages. I fully support this, and underlined it in my book Modern Biotechnology: a new Pandora’s Box? published in 2011. Likewise I treat the golden rice problem in other books, columns etc., like We need genetic engineering on this website.

Ingo Potrykus

Golden rice and Ingo Protykus (1933) are inextricably bound up with each other. He is the driving force behind the Golden Rice Project . In 2015, he published his autobiography as a scientist, a moving story; Potrykus is 82 then. The genetically modified rice, called golden rice because of its yellow colour, has been developed with the aim of enrichment with provitamin-A (beta carotene). Together with the transgene tomato Fravr Savr (see Essay 4.1), golden rice is looked upon as the first two transgene crops with a direct positive effect on consumers. In 2001 Ingo Protykus publishes a review article titled The ‘Golden Rice’ Tale. He says: ‘Golden rice’ fulfills all the wishes the GMO opposition had earlier expressed in their criticism of the use of the technology, and it thus nullifies all the arguments against genetic engineering with plants in this specific example. – Golden Rice has not been developed by and for industry. – It fulfills an urgent need by complementing traditional interventions. – It presents a sustainable, cost-free solution, not requiring other resources. – It avoids the unfortunate negative side effects of the Green Revolution. – Industry does not benefit from it. – Those who benefit are the poor and disadvantaged. – It is given free of charge and restrictions to subsistence farmers. – It does not create any new dependencies. – It will be grown without any additional inputs. – It does not create advantages to rich landowners. – It can be resewn every year from the saved harvest. – It does not reduce agricultural biodiversity. – It does not affect natural biodiversity. – There is, so far, no conceptual negative effect on the environment. – There is, so far, no conceivable risk to consumer health. It was not possible to develop the trait with traditional methods, etc. Optimists might, therefore, have expected that the GMO opposition would welcome this case. As the contrary is the case, and GMO opposition is doing everything to prevent golden rice reaching the subsistence farmer, we have learned that GMO opposition has a hidden, political agenda.’

Fourteen years later, in the autobiography mentioned above, he poses the next question: ‘Why?’ and answers simply: ‘Golden rice is a GMO!’ Implicitly, he admits that it takes a lot of trouble to get transgene crops to the market. The shortcomings of the first version have now been overcome and the safety is warranted. Yet, opposition has become fiercer – simply because it is a genetically modified crop. The scientific evidence that underpins the products of gene technology in general and golden rice in particular has been strengthened to such an extent now, that it is amoral not to make use of these achievements.

Opponents of gene technology, particularly Greenpeace, are concerned about effectiveness, the environment, social acceptance and patents, if golden rice should be introduced in Asia. On February 9, 2001, Greenpeace launches the effective campaign Fool’s gold , in which they claim that an adult, let alone a young child, would have to eat impossible amounts of golden rice in order to ingest enough vitamin A. As for the environment, they fear that recombinant genes will jump to traditional and wild rice varieties, which could have far-reaching consequences in Asia, the region of origin of rice. They suggest that golden rice may not be socially and culturally acceptable because of its yellow colour. Golden rice is royalty free, but a number of international patents on it is in the hands of multinational companies. A British NGO, ExoNexus , publishes a critical report on this issue in 2003. The patent holders have agreed that poor farmers in developing countries will not have to pay royalties if they earn less than 10,000 American dollars and do not export the rice – but how will that work in practice? Both terms will be difficult for poor farmers to prove, simply because they lack the instruments to do so. ExoNexus also states that no environmental and food safety tests have been done on golden rice, and that an assessment of any socio-economic effects is missing. They end their report stating: ‘Meanwhile, new breeds of vitamin-A-rich grains have been announced…. Even so, as with Golden Rice, those grains cannot answer the problem of hidden hunger and malnutrition, which need a far more integrated solution.’ With the latter statement, I wholeheartedly agree, as stated above.

New developments

Meanwhile, in 2018, we have moved fifteen years onwards, and a new transgene rice variety has been developed and tested. The old version contains two daffodil genes and a gene of the bacterium Erwinia uredovora . The new variety has one bacterial gene and one gene from maize instead of the two daffodil genes. This new golden rice variety produces 23 times more provitamin-A than the old one, and therefore it will make sense to continue with this one in the battle against vitamin-A deficiency. All the more because it has been proven that a bowl with 100-150 grams of this golden rice (prepared) can ensure an intake in children between 6 and 8 of 60 percent of the recommended daily intake. A result that is corroborated by other research . Greenpeace did not succeed in discrediting these results. Nevertheless, they adhere to their 2001 website.

  As for socio-economic aspects, I highlight a few points from the VIB booklet cited above. Generally, it is expected that the provitamin-A property will not have a negative environmental impact. This property is abundantly present in nature and does not carry with it any selective advantage or disadvantage for wild varieties, in case of any crossbreeding with them. Which is not likely anyway, because rice is self-pollinating? Yet, GMO regulations require extensive environmental impact assessments before a transgene crop is allowed to be grown on a commercial scale. I will return to this in the next paragraph.

We also need to go into economic consequences. Blindness causes economic inactivity of part of the population. If golden rice could have health benefits for those people, economic participation and national wealth might increase. In China for instance, it has been estimated that national income could rise by 2% if golden rice were grown.

Varieties implementation of transgene rice crop boils down to a lot of classical enrichment activity in order to introduce the provitamin-A production system into different local rice varieties. It is important to ensure sufficient crop diversity; also, varieties should be adapted to cultural traditions, and comply with local growth conditions. In Bangladesh, the Philippines, India, Indonesia and Vietnam, major steps towards these goals are taken. There, the golden rice property is crossbred into as many local varieties as possible. Particularly Bangladesh and the Philippines are close to a commercial introduction; beneath we will go into the progress in these two countries.

Golden rice in the Philippines

In 2006, the Healthier Rice Program of the International Rice Research Institute (IRRI) at the Philippine Rice Research Institute (PhilRice) starts; it makes use of the new version of golden rice. End January 2008, Robert Zeigler, general manager of IRRI, announces that the first field trials will take place that year, and that farmers might have access to golden rice in 2011 already. He speeches at the occasion of a grant of 20 million dollars, awarded by the Bill & Melinda Gates Foundation. IRRI thinks that it could supply 18 million households, mainly in Asia and sub-Saharan Africa, with improved genetically modified rice varieties. In 2012 and 2013, they plan field trials with local golden rice varieties on five different locations in the Philippines. In accordance with existing regulation, these locations are indicated approximately. For the same reason, the fields are protected with high fences and round-the-clock patrols. Nevertheless, the most accessible location is found and destroyed on August 8, 2013. The vandals are identified and prosecuted. An editorial in Science on this event causes a storm of protest among scientists all over the world, and some 6500 scientists sign the protest.

In 2014, IRRI reports that the open field trials show a lower yield for the then best golden rice variety (GR2R) compared to the corresponding conventional local variety. The content of beta carotene on the other hand is not lower. They therefore continue the breeding program. Confined field trials between October 2014 and July 2017 show that the new variety GR2E has the same yield and the same quality as the usual local variety, and is much enriched in beta carotene. At the time of writing, open field trials are going on. If they have the same result, commercial application is just one step away, unless opponents block developments once again. Safety cannot be the issue. On December 20, 2017, the food safety authority of Australia and New Zealand (FSANZ) approves GR2E rice consumption. They judge this variety to be as safe as ordinary rice varieties. The conclusion of their safety analysis reads: ‘No potential public health and safety concerns have been identified in the assessment of GR2E. On the basis of the data provided in the present Application, and other available information, food derived from GR2E is considered to be as safe for human consumption as food derived from conventional rice varieties.’ Beginning 2018, the American FDA and Health Canada endorse this judgment.

Golden rice in Bangladesh

In Essay 4 Part 2 , I describe Bangladesh’s good results with Bt-eggplants. In 2018, some 27,000 out of 150,000 Bengalese eggplant farmers grow this transgene eggplant and… reap its benefits. This success triggers attempts to grow genetically modified crops on a larger scale. ‘We will be guided by the science-based information, not by the non-scientific whispering of a section of people,’ says the Bengali minister of agriculture. In a greenhouse in Joydebpur, in the outskirts of Dhaka that house the Bangladesh Rice Research Institute (BRRI), they work now on an important next project. They grow sprouts of a special rice variety with yellow grains, i.e. golden rice. They started with a variety supplied by the Filipino sister institute. Recently, the gene has been crossbred into three of the most common rice varieties in Bangladesh, where still millions are at risk of vitamin A deficiency. There are programs for distribution of capsules to children, but it is hard to reach those in remote areas. Moreover, they are costly. Rice on the other hand is a staple food and is consumed by everyone. To have rice or not to have rice is a matter of life and death here. If the transgene rice varieties do well on the trial fields, they will soon appear in the countryside, like Bt-eggplants. Bangladesh’s rice fields will colour yellow in a few years, at least that is the hope… and the expectation. Fortunately, opponents do not get a foothold here because of this progressive policy of the present minister for agriculture.

In the next part, we will treat a number of products of new plant breeding technologies.

Interesting? Then also read: Genetically modified food Rudy Rabbinge: genetic modification has much potential but also entails risks for world food supply and the environment We need genetic engineering

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From disagreements to dialogue: unpacking the Golden Rice debate

Annika j. kettenburg.

1 Faculty of Sustainability, Leuphana University of Lüneburg, Universitätsallee 1, 21335 Lüneburg, Germany

2 Lund University Centre for Sustainability Studies (LUCSUS), Lund University, Box 170, 22100 Lund, Sweden

Jan Hanspach

David j. abson, joern fischer, associated data.

Transgenic Golden Rice has been hailed as a practical solution to vitamin A deficiency, but has also been heavily criticized. To facilitate a balanced view on this polarized debate, we investigated existing arguments for and against Golden Rice from a sustainability science perspective. In a structured literature review of peer-reviewed publications on Golden Rice, we assessed to what extent 64 articles addressed 70 questions covering different aspects of sustainability. Using cluster analysis, we grouped the literature into two major branches, containing two clusters each. These clusters differed in the range and nature of the sustainability aspects addressed, disciplinary affiliation and overall evaluation of Golden Rice. The ‘biotechnological’ branch (clusters: ‘technical effectiveness’ and ‘advocacy’) was dominated by the natural sciences, focused on biophysical plant-consumer interactions, and evaluated Golden Rice positively. In contrast, the ‘socio-systemic’ branch (clusters: ‘economic efficiency’ and ‘equity and holism’) was primarily comprised of social sciences, addressed a wider variety of sustainability aspects including participation, equity, ethics and biodiversity, and more often pointed to the shortcomings of Golden Rice. There were little to no integration efforts between the two branches, and highly polarized positions arose in the clusters on ‘advocacy’ and ‘equity and holism’. To explore this divide, we investigated the influences of disciplinary affiliations and personal values on the respective problem framings. We conclude that to move beyond a polarized debate, it may be fruitful to ground the Golden Rice discourse in facets and methods of sustainability science, with an emphasis on participation and integration of diverging interests.

Electronic supplementary material

The online version of this article (10.1007/s11625-018-0577-y) contains supplementary material, which is available to authorized users.

Introduction

Sustainability is a contested and highly normative concept (Dobson 1999 ; Christen and Schmidt 2012 ). The solution-oriented field of sustainability science (Miller et al. 2014 ) has to address both the normative goals of sustainability itself and the, often implicit, assumptions that underpin different scientific traditions (Schumpeter 1954 ; Funtowicz and Ravetz 1993 ; Lélé and Norgaard 2005 ). Such normativity, especially when not explicitly addressed, often leads to conflicting, even polarized, discourses regarding what represents an appropriate intervention for a given sustainability problem. For example, polarized narratives in research addressing the intersecting goals of food security and biodiversity conservation are driven by the underpinning conceptualization of the problem as either technical or socio-political (Loos et al. 2014 ; Glamann et al. 2015 ). Similarly, the narrative explaining food insecurity as a result of insufficient production and population growth contrasts with explanations based on unequal distribution of social power as well as economic and physical resources (Sen 1981 ; Legwegoh and Fraser 2015 ). In the agricultural biosciences, calls for gene patenting, corporate funding of public institutions and public–private partnerships conflict with arguments that seeds should be regarded as public goods (Scoones 2002 ; Stone 2015 ). Such polarization presents serious challenges for sustainability science, not simply in terms of conflicting policy prescriptions, but also in the perceived legitimacy of the science itself (Bäckstrand 2003 ).

In this paper we use the example of the scientific discourse around “Golden Rice”—itself a microcosm of the broader debate surrounding the role of genetically modified organisms (GMOs) in agricultural sustainability—as a particularly emotive example of a polarized discourse in sustainability science. Through a systematic, quantitative (cluster) analysis of the scientific literature, we classify and describe the polarized positions within the Golden Rice debate. By viewing this discourse through an explicit sustainability lens we seek to shed light on the role of problem framing in shaping the Golden Rice discourse, and suggest ways of shifting from such polarized debates towards more constructive dialogues. In particular, we highlight the importance of understanding and acknowledging the sources of such polarization, to move beyond ‘siloed’ disagreements towards shared understandings and meaningful solutions.

The severity of conflicts around the use of GMOs in agriculture has been likened to that of a war (Lang and Heasman 2004 ; Waltz 2009 ; Stone 2015 ). Proponents see in genetically modified (GM) crops powerful tools to increase yields (Carpenter 2010 ), improve crop quality, decrease pesticide use (Christou et al. 2006 ), fight micronutrient deficiencies, adapt plants to climate change and facilitate economic growth (Phillips 2002 ). Opponents voice doubts over the long-term effectiveness of genetically modified crops in face of accelerated formation of resistances to glyphosate (Gilbert 2013 ) and to Bt toxins (Tabashnik et al. 2013 ), over the nutritional equality to non-GM crops (Bøhn et al. 2014 ), and even over their adequateness as food and feed (Séralini et al. 2014 ). Some call into question genetic engineering’s theoretical foundation on reductionist models that disregards insights from systems biology (e.g., McAfee 2003 ; Perret and Longo 2016 ). Often these GMO specific issues are entangled with political concerns regarding the role of GMOs in reinforcing corporate power (Walters 2005 ), or the promotion of monocultures and homogenization of diets and landscapes (Scrinis 2007 ).

The case of Golden Rice exemplifies many of the conflicts surrounding GMOs as a potentially sustainable solution for issues ranging from food security to biodiversity conservation. Golden Rice is a genetically modified cultivar that synthesizes beta-carotene, which in turn is metabolized into vitamin A in the human body. Some communities in the Global South show high rates of xerophthalmia, the clinical manifestation of vitamin A deficiency (Thylefors et al. 1995 ). Xerophthalmia leads to corneal ulceration and ultimately blindness. An estimated 250,000–500,000 vitamin A-deficient children became blind every year in the period 1995–2005, half of them dying within 12 months of losing their sight (WHO 2017 ). However, data are largely outdated; only 27 countries reported estimates since 2006 (Wirth et al. 2017 ). Vulnerability to xerophthalmia depends on a number of factors including eating habits such as a varied diet accompanied by fats, social determinants such as poverty or lack of education, health conditions such as parasitic infestations, and access, influenced by seasonality of vitamin A rich vegetables/fruits, land entitlements, and crisis such as famine or flight (Egana 2003 ; Oyunga et al. 2016 ). Current strategies to address vitamin A deficiency include supplementation, (bio-)fortification and dietary diversification (Ruel 2001 ; WHO 2017 ).

Golden Rice was developed by Potrykus and Beyer in Zürich and Freiburg during the 1990s in response to a call from the Rockefeller Foundation for a plant breeding solution to vitamin A deficiency. This work resulted in the first novel rice variety, which contained 1.6 µg/g carotenoids in the endosperm. The second generation of Golden Rice, created in partnership with Syngenta, contains up to 37 µg/g carotenoids, sufficient to fulfil half of daily vitamin A requirements with 60 g of uncooked rice (Paine et al. 2005 ). 1 Syngenta agreed on free licenses for famers in the Global South with incomes less than $10,000 annually. These farmers may reseed Golden Rice after every harvest. Currently, Golden Rice is still under development, with the intention that once all safety assessments are completed and it is approved by national regulators, it will be distributed accompanied by information campaigns (Potrykus 2001 ; Mayer and Potrykus 2011 ; Zeigler 2014 ).

Golden Rice promises a positive impact on human health while ensuring economic independence of smallholder farmers from large agri-business. Nevertheless divergent views regarding the benefits and sustainability of Golden Rice persist (e.g., Small 2014 ). Here, we present a systematic, quantitative assessment of the narratives in the peer-reviewed discourse on Golden Rice. Our objectives were to (1) identify and characterize narrative-based groups of articles on Golden Rice, (2) point out the scope of themes relevant to sustainability addressed by each group, and (3) propose explanations for the revealed patterns. Drawing on this, we provide ideas to facilitate a more fruitful dialogue within and beyond the scientific community regarding Golden Rice. Although our case study is specifically on Golden Rice, our approach to understanding and resolving this contentious scientific debate may also help to inform the analysis of other polarized discourses on pathways to sustainability.

Literature selection

We conducted a literature review in Scopus of English language, peer-reviewed articles and book chapters using the keyword “Golden Rice” in title–abs–key in July 2016. An article was included in the analysis if it met the following criteria. Criterion one: Golden Rice was discussed as both a biotechnological project and a health intervention, either as the focus of the paper or within a broader framing. Just mentioning Golden Rice as an illustrative example led to exclusion [e.g., in Weil (2005) “Are genetically modified plants useful and safe?”]. Criterion two: articles focusing on biophysical and technical matters only were excluded (e.g., Al-Babili et al. 2006 ). Criterion three: the article addressed three or more questions of our coding protocol. This minimum level was set to guarantee the validity of quantitative results. In contrast to the first two criteria, criterion three was applied after coding of the article.

Identification of sustainability themes

Our intention was to offer a sustainability perspective on the Golden Rice debate: which sustainability themes do the different strands of Golden Rice literature address? We defined sustainability as an ideal of human well-being within planetary boundaries across generations (Gibson 2006 ; Rockström et al. 2009 ). To operationalize this definition, we identified 16 themes, which, arguably, ought to be considered in discussions about Golden Rice from a sustainability perspective (Table  1 ). In a second step, we used an inductive approach to identify specific questions (sub-themes) under each sustainability theme that emerged from the reviewed articles. A grounded theory-based, inductive formulation and adjustment of questions during the coding process (Corbin and Strauss 1990 ) provided a higher thematic coverage of sustainability sub-themes, adding new questions and dismissing unaddressed ones. The final coding protocol resulted in 70 questions/sub-themes (Table S1 in the supplementary material). Those 70 sub-themes were coded for text analysis in MAXQDA 12 (VERBI Software 2016 ).

Description of the sustainability ‘themes’ and their relevance for the Golden Rice discourse

These themes formed the basis for the selection of the 70 sub-themes used in the coding protocol (Table S1 in supplementary material)

We summarized the coding results quantitatively in a table. A paper scored 0 if it did not address a question at all and 1 if it addressed it. The intention of this quantitative coding was to differentiate alternative discourses related to the 16 key sustainability themes (Table  1 ) and the related 70 sub-themes (supplementary materials).

Data analysis

After coding we used agglomerative hierarchical cluster analysis, a method commonly employed to recognize subsets in multivariate data. Agglomerative clustering begins with discontinuous single objects (i.e., articles) and groups these into ever larger clusters. Euclidean distance was chosen as an association measure for the clusters due to our homogeneous scale, the limited number of double zeros in pairwise comparisons (i.e., no answers) and the clear interpretation of the resulting patterns. We employed Ward’s minimum variance method for grouping. It minimizes the within-cluster sum of squared errors, that is the sum of the squared distances among cluster members divided by the number of articles per cluster, thereby usually producing clear and evenly sized clusters (Borcard et al. 2011 ). Importantly, cluster analysis is an exploratory method that is able to uncover (dis-)similarities between papers and thus to empirically show different schools of thought in the assessed literature. Therefore, unlike in the use of inferential statistics the potential lack of independence of data points (e.g., papers written by the same authors are not independent) does not invalidate or bias our analysis. Results of the cluster analysis were visualized in a dendrogram. Furthermore, we conducted an indicator analysis that listed the questions of central importance for each cluster, facilitating cluster characterization and differentiation (Dufrene and Legendre 1997 ). Analyses were performed in R Version 3.3.2 (R Core Team 2016 ).

The literature search returned 98 articles (after removing duplicates, non-peer-reviewed and inaccessible publications), of those 64 passed our inclusion criteria. Of the 34 excluded articles, 56% focused on a different topic (criterion one), 29% were entirely biochemical or technical (criterion two) and 15% addressed two or less questions (criterion three). Of the 64 reviewed articles most were authored by plant scientists including biologists and chemists (44%), followed by social scientists (25%) and economists 2 (19%; Table  2 ). Forty-two percent of the articles were authored by members of the Golden Rice Humanitarian Board and affiliated research institutes, or by employees of Syngenta or Monsanto. Seventy-seven percent of all articles were in favour of Golden Rice whereas 14% voiced doubts or opposed it, and 9% abstained from judgement. Evaluation of Golden Rice (Table  2 ) ranged from outright rejection (opposing), through pointing to serious concerns (doubtful), arguing that there is insufficient data to draw conclusions (cautious), ascribing great potential to Golden Rice if confounding factors can be overcome (optimistic), to arguing for the immediate use of Golden Rice to avoid preventable deaths (passionate). The first paper was published in 2001 and numbers of publications per year were relatively steady (mean 4, SD 2.2).

Information on message, discipline, affiliation and origin of selected articles ( n  = 64)

Cluster analysis led to two major branches containing two clusters each (Fig.  1 ). The clusters differed significantly among each other not only in the range of sustainability sub-themes addressed (Table  3 ), but also in their subjective evaluations of Golden Rice. Moreover, there was a clear disciplinary divide between the clusters. The clusters were named to reflect their thematic focus: the biotechnological branch consisted of clusters on ‘technical effectiveness’ and ‘advocacy’, whereas the socio-systemic branch included clusters on ‘economic efficiency’ and ‘equity and holism’. In the following, we describe the branches and clusters in detail (see Table S2 for illustrative quotes and Table S3 for a list of each cluster’s articles in the supplementary material). Here, it is important to note that neither the cluster names, nor the following detailed descriptions perfectly capture the approaches or emphasis of every article in a given cluster. Rather, they provide general characteristics of the different sustainability-focused narratives that have emerged within the scientific literature on Golden Rice.

Cluster analysis demonstrated two major branches of research on Golden Rice, each consisting of two clusters—a biotechnical branch (red ‘technical effectiveness’, black ‘advocacy’) versus a socio-systemic branch (blue ‘economic efficiency’ and green ‘equity and holism’). For full citations see Table S3 in the supplementary material

Indicator analysis demonstrated which aspects constituted the character of a certain cluster (no values for the ‘technical effectiveness’ cluster)

For detailed information on each abbreviated indicator corresponding to one of our 70 sub-themes see Table S1 in the supplementary material

Biotechnological branch ( n  = 40)

Articles in the biotechnological branch ( n  = 40) were predominantly authored by plant scientists (65%) and economists (15%); only one paper was written by a social scientist. All held a positive attitude towards Golden Rice, except one with a narrow focus on the potential dangers of beta-carotene engineered plants (Schubert 2008 ). The overall approach of articles in this branch was to present Golden Rice as an engineering solution to vitamin A deficiency and to argue for broad scale usage of Golden Rice based on measurements of its efficacy in producing beta-carotene. Within the biotechnological branch there were distinct clusters of articles focusing on ‘technical effectiveness’ (red cluster; Fig.  1 ) and ‘advocacy’ (black cluster; Fig.  1 ).

Technical effectiveness ( n  = 23)

“Another exciting field of modern plant biotechnology is represented by the enhancement of crop nutritional properties through genetic modification [ref.]. There are multiple nutritional advances underway and this review focuses on two representative examples that illustrate the potential impact of this technology.” (Schwember 2008 )

As in this quote, articles primarily focused on Golden Rice as an exciting achievement in the reduction of vitamin A deficiency through genetic engineering. Topics addressed in these articles included general overviews of biofortification or GM crops (nine articles), narrow foci on biotechnical processes and resulting effectiveness (six articles), the management of the Golden Rice project itself (five articles) and economic valuation methods for assessing the benefits of GMOs (three articles). Indicator analysis did not result in any predictive sustainability sub-themes within the cluster (no significant indicator values; Table  3 ). Articles in this cluster generally stated the effectiveness of Golden Rice in regard to the target of producing beta-carotene in the rice endosperm and concluded it would increase the vitamin A status of populations at risk. Considerations of issues such as changes of diets, livelihood strategies or politics were rare.

Advocacy ( n  = 17)

“The consequence [of GMO opposition]: millions of avoidable blind and dead children. The author considers those who are responsible for this avoidable suffering of many innocent children (and mothers at childbirth) a crime to humanity […]. There is a wealth of scientific information and broad consensus that GMO-technology is at least as safe as any other technology involved in any context with our food or our environment […]. Our ‘enlightenment’ and science-based successful European culture is on the verge of being replaced by unreason-based failure and lack of culture.” (Potrykus 2013 )

As the quote reveals, the tone in this cluster was often extremely emotive, including one author who called the delay of Golden Rice’s implementation a “silent holocaust” (Chassy 2010 : 543). This cluster focused on the consequences of regulation GM crops and delayed release of Golden Rice. The unconditional safety of Golden Rice was often stated, invocations of the precautionary principle—that, in the absence of scientific consensus, there is burden of proof for proponents of new products or policies to show that such products or policies are not harmful to humans or the environment (O’Riordan 1994 )—were argued as unjustified, and the behaviour of those opposed to agricultural GMOs often framed as irrational. Articles argued for Golden Rice’s implementation with reference to preventable deaths, while not mentioning any concerns related to Golden Rice. The prevailing argumentation built upon the notion of a consensus on Golden Rice’s effectiveness and the absence of any GMO-related risks. The sub-theme ‘message: passionate’ was strongly associated with this cluster in the indicator analysis (indicator value 0.90; Table  3 ).

Ten articles within this cluster were authored by members of the Golden Rice Humanitarian Board, one by an employee of Monsanto and three other authors have in the past written articles on Golden Rice with members of the board or Monsanto/ Syngenta (totalling in 14 out of 17 articles). 3

Socio-systemic branch ( n  = 24)

In comparison to the biotechnological branch, the socio-systemic branch focused less on technical achievements than on the socio-political components/contexts of food systems and vitamin A deficiency. The authors were social scientists (14 articles), economists (6 articles), biologists (2 articles) and interdisciplinary teams (2 articles). Within the branch there were two clusters: ‘economic efficiency’ (blue cluster; Fig.  1 ) and ‘equity and holism’ (green cluster; Fig.  1 ).

Economic efficiency ( n  = 10)

“We develop a methodology for comprehensive ex ante evaluation […]. We use a truly interdisciplinary approach, integrating epidemiological and nutrition details, as well as socioeconomic and policy factors. In particular, we determine the current public disease burden of VAD in a country with an important rice-eating population, and simulate to what extent this burden could be reduced through GR […]. Finally, we assess the cost-effectiveness of GR…” (Stein et al. 2008 )

The commonality of the ‘economic efficiency’ cluster was the focus on cost-benefit calculations, taking into account a wide range of variables, including political and cultural influences. Sixty percent of the articles we authored by economists. Moreover, the significance of sub-themes such as ‘acceptance’ and ‘marketing campaigns’ (Table  3 ) demonstrated a consumer oriented perspective. In comparison to the clusters in the biotechnological branch, articles in this cluster used multi-factor models in their assessment of Golden Rice and were more likely to consider alternative interventions for addressing vitamin A deficiency (e.g., Zimmermann and Qaim 2004 ; Stein et al. 2008 ). Articles tended to give a positive appraisal of Golden Rice, because it was said to be compatible with the current food system and, therefore, more realistic to implement than other interventions, especially due to its cost-effectiveness (e.g., Stein et al. 2008 ).

Equity and holism ( n  = 14)

“Addressing the most immediate and fundamental problems of food insecurity and undernutrition such as micronutrient deficiency, while essential, can only succeed in the long run by proceeding in balance with environmental, sociocultural, political, economic, behavioral and biomedical perspectives.” (Johns and Eyzaguirre 2007 )

The ‘equity and holism’ cluster took into account a variety of themes such as participation, equity, biodiversity, water and soil conservation, resilience and system thinking, values and philosophy. The number of statistically significant sub-themes in the indicator analysis exceeded that of other clusters, demonstrating the diversity of the issues and topics that defined this cluster (Table  3 ). In contrast to other clusters, only one article was in favour of Golden Rice, ascribing it the potential to “play the positive role of technological fixes […]—providing policy-makers with more options and additional means for addressing social problems” (Scott 2011 : 225). There was a particular focus on delineating and defining societal goals relating to or intersecting with the potential use of Golden Rice as an intervention for addressing vitamin A deficiency. In line with this prioritization of societal goals, most articles raised concerns over Golden Rice’s adequateness as solution to vitamin A deficiency (e.g., Lorch 2001 ; Nestle 2001 ; Small 2014 ). Articles called into question Golden Rice’s ‘real world’ nutritious efficacy in relation to contextual factors such as diet, the presence of other infectious diseases (Egana 2003 ) or storage losses (Stone and Glover 2016 ). There was also consideration of environmental and social consequences of a continued reliance on ‘mega-crops’ (Small 2014 ), the ‘placelessness’ of Golden Rice and a lack of transferability to local food systems (Stone and Glover 2016 ) as well as the disregard of indigenous knowledge (Johns and Eyzaguirre 2007 ). These concerns highlighted the need to consider complex preconditions or confounding factors in the successful use of Golden Rice as a mitigation strategy, rather than seeking to dismiss the benefits of Golden Rice itself.

Representation of sustainability themes

While the focus on sustainability themes and sub-themes varied widely between clusters (Table  3 ), there were certain sub-themes that received less attention than others. Issues that were addressed in less than 15% of the reviewed articles (largely the socio-economic branch) concerned: local interests, life quality, dignity, empowerment, autonomy, poverty alleviation, climate change, alternative farming methods, systems thinking, system dynamics across scales, details on Golden Rice’s distribution and on its monitoring, trade policies, governance and ethics. The least addressed issues such as dignity, food sovereignty, alternative farming methods and climate change (five, three, three and two mentions respectively) were largely absent across all four clusters.

The conclusions to be drawn from our results are threefold: (1) the body of literature on Golden Rice can be grouped into clusters whose range of sustainability themes correlated with the articles’ evaluation of Golden Rice and the authors’ discipline; (2) the biotechnological branch represented the dominant narrative in terms of quantity of articles, yet lacked a focus on crucial sustainability themes; and (3) there was little integration or overlap between the thematic foci or broader perspectives of the two branches, and particularly polarized positions arose in the clusters on ‘advocacy’ (e.g., Potrykus 2013 ) and ‘equity and holism’ (e.g., Small 2014 ). Such polarized debates are useful for identifying the initial differences in visions, goals and values that shaped these discourses. However, it is vital to understand the sources of such disagreements to move beyond polarization towards dialogue and mutual benefit. To inform and facilitate this process, it is useful to consider what paradigms 4 underpin the two branches of the literature, and their influences on respective problem framings and narratives surrounding Golden Rice.

Paradigms underpinning branches

The observed homogenous composition of either social or natural scientists (and their corresponding evaluations of Golden Rice) in each branch suggest the existence of a disciplinary dichotomy. This divide between scientific cultures presents a major challenge to integration (Tress et al. 2005 ), although it is not the only factor leading to polarized positions. For example, Legwegoh and Fraser ( 2015 ) argue opportunism and political economy have led to a similar case of diverging narratives in the context of the food security discourse.

The biotechnological branch’s arguments are built on premises often shared by the natural science community. A core assumption is the existence of an objective reality that can be investigated, described and to a certain extent predicted based on generalized, reductionist theories (Becher 1994 ; Moon and Blackman 2014 ). In the case of Golden Rice, this approach helps shape the observed focus on theoretical assessment of the effectiveness of plants in producing vitamin A from a purely biophysical perspective. Following positivist logic, the effectiveness in biophysical terms would translate into a successful mitigation of vitamin A deficiency in the ‘real world’. More profoundly, the idea of engineering a plant to contain beta-carotene might be traced to a framing of the problem that is characteristic to natural sciences. The nature of the problem, that is the cause of malnutrition, was perceived to be related directly to proximate biophysical factors (the lack of beta-carotene in rice plants), and not on less proximate factors such as poverty. This relatively narrowly framed problem definition naturally lends itself to a technical solution, and one aligned to the authors’ own expertise. In contrast, the existence of resource poor farmers was regarded as given and, therefore, not as target of scientific efforts (Scott 2011 ). Such bounded scientific enquiry results in generic notions of effectiveness, suggesting universal applicability directed at “the poor” (e.g., Chassy 2010 ) as a homogenous group, in “developing countries” (e.g., Zimmermann and Qaim 2004 )—an unspecified global space (Stone and Glover 2016 ).

In the socio-systemic branch the object of research shifted from the natural environment to human behaviour. Principles from the social sciences were applied, such as relativism and intersubjectivity (Moon and Blackman 2014 ), and informed a perception of the world as a multidimensional and interconnected system, whose variables cannot be understood in isolation (Loos et al. 2014 ). As result, the research centred on power and justice, systems thinking, participation and ethics within a specific place-based case, as typified by the ‘equity and holism’ cluster (Fig.  1 ; Table  3 ).

The ‘economic efficiency’ cluster only partially fits within this characterization, because most economists carry a distinct set of assumptions, rooted in rationalistic-individualistic, neoclassical, utilitarian paradigms (Becher and Trowler 2001 ; Etzioni 2010 ). These assumptions tended to favour models for assessing Golden Rice’s cost-effectiveness based on explicit assumptions informed by the study of the causal links between systems components. Quantitative methods and generalized models prevailed in this branch. This focus on generalizable models, as legitimate approaches for addressing context dependent real world problems, provides a clear link with biotechnological branch of Golden Rice research, despite the considerable differences in thematic foci between the biotechnological and socio-systemic approaches.

Distinct strengths as impetus for integration

The two branches of literature presented different strengths in their approaches to conceptualizing and solving the problem of vitamin A deficiency. The biotechnological branch offered a focused, generalizable, quickly transferable, one-time intervention, effective in regard to its target—the provision of a beta-carotene producing rice variety (e.g., Potrykus 2001 ). This rice might act as positive example of a technological fix as suggested by Scott ( 2011 ). However, the lack of focus on confounding factors that are likely to influence the success of Golden Rice, limits the nuanced understanding of how such broad brush interventions will play out in what are inevitably complex, context specific socio-political contexts.

In contrast, the socio-systemic branch’s broader, more contextualized understanding of vitamin A deficiency promoted strategies embracing the traditional approaches of supplementation and fortification along with capacity building in agroecology and education on nutrition, hygiene and health. This approach aims at synergies (e.g., by combing health checks with education measures) and at a broad notion of well-being, taking into account various aspects of health and sustainable livelihood strategies, not just vitamin A deficiency (e.g., Johns and Eyzaguirre 2007 ). Such contextual approaches face major barriers though, often not being compatible with the status quo of a targeted food system or current politics (Scoones 2002 ; Scott 2011 ). Moreover, the strong focus on the importance of context in sustainability problem framing may diminish the potential positive contribution that general, broad-brush technical solutions can have if contextual issues are addressed.

A constructive dialogue on strengths and limitations of both approaches might serve to draw a more nuanced picture of Golden Rice and to eventually inform better research outcomes by aligning both technical depth (e.g., how to design optimal seeds and growing conditions) and thematical breadth (e.g., which components of the food system influence vitamin A deficiency to what extent).

Values and vested interests in science

Despite the high promises of cooperation between the socio-systemic and the biotechnological branch, our results indicated profound differences in their respective problem framings. These problem framings required different research methods and team constellations and concluded in diverging solutions, working on different scales and time frames (short term changes in plant metabolism versus long term food system transformation). It is necessary not just to contend with the way different disciplinary traditions shape sustainability problem framing, but also with diverging values and worldviews among researchers personally (Garnett 2013 ). Moreover, there is a need to address the feedbacks between personal values and disciplinary traditions. Personal values influence individuals’ choice of discipline, and those—self-selecting—scholarly communities tend to reinforce particular worldviews. Nevertheless, the disciplinary divide seems inadequate as an explanation for the high extent of polarization between the clusters within the academic literature. Are GM crops safe, beneficial to biodiversity and a key to food security? Is it legitimate to base GMO regulations on the precautionary principle? Is our current agricultural system in crisis or at a historical peak? Polarized positions on these questions regardless of affiliations point to the influence of values and deeply held worldviews on framing the research (Fischer et al. 2014 ) and on interpreting results (Devos et al. 2014 ). These normative propositions of researchers were often obscured in the Golden Rice discourse by the assumed objectivity of scientific research. Accordingly, authors often made dichotomous policy recommendations, either supporting or rejecting Golden Rice and thus portraying the case as a formal objective problem, solvable within the realm of deduction (Levidow and Marris 2001 ; Herrick 2004 ), rather than as a normative and value-laden issue.

Moreover, in some instances vested interests have led scientists to ally with corporate representatives or activists, thereby increasing the divergence between positions through the self-amplifying process of “social bonding against a common enemy” (Stone 2017 : 590). Despite the strong influences of disciplinary affiliations, profound ideological divides, and entanglements between science and policy, our analysis showed little recognition of how these topics shape and polarize the Golden Rice discourse.

Pathways to sustainability

Offering a sustainability science perspective on how to move the debate forward, we suggest a reframing of the question and its research methodology, by prioritizing human well-being and local involvement. To transcend the reductionism of regarding rice as mere nutrient provider, neglecting its place in the eco- and cultural system (Hayes-Conroy and Sweet 2014 ), and of describing vitamin A-deficient populations as passive victims (‘the poor’) in unspecified geographic and social positions, we propose to reframe the question: from ‘how do we create a rice plant producing beta-carotene?’ or ‘how do we most efficiently raise the vitamin A status of populations at risk?’ to ‘how do we foster the well-being of people affected by malnutrition, both in short and long terms?’. Such a reframing of putting people first automatically aligns health and nutrition with equality, secure livelihoods and environmental integrity (Bennett 2017 ). Most importantly, to understand what well-being means to the people in question, there is no way around asking. This necessary physical proximity creates room for participation, for joint agenda setting, for mutual learning, for producing ‘socially-robust’ knowledge (Gibbons 1999 ), in short: for the aims and rationales of transdisciplinarity (Lang et al. 2012 ). 5

Indeed participation of non-scientists in both problem framing and solution formation was largely overlooked in the Golden Rice literature, with a notable lack of focus on sub-themes such as local culture, participation, dignity and empowerment in the articles reviewed here. This lack of participation exists despite the obstructive mistrust towards Golden Rice, witnessed both in the Global North (Baggott 2006 ) and South (Cabanilla 2007 ). Despite challenges in praxis (Brandt et al. 2013 ; Kenny et al. 2015 ) and a limited number of evaluation studies (Bath and Wakerman 2015 ), there is a growing recognition of the feasibility and potential success of transdisciplinarity, for example, in the field of health policy and systems research (Sheikh et al. 2014 ) or agricultural research (Hoffmann et al. 2007 ; Neef and Neubert 2011 ). Such approaches potentially allow for socio-technical solutions that can be adapted to specific socio-political or socio-ecological contexts and that acknowledge that multiple interventions are often required to fix what at first glance might seem like relatively simple problems (such as a vitamin A-deficient diets).

More broadly, polarized discourses regarding solutions to pressing sustainability problems may be avoided, or at least diminished, by attempts to develop shared problem definitions (both across different scientific disciplines and in conjunction with those who are impacted by the proposed solutions). This requires greater focus on exploring the way sustainability science is shaped by disciplinary traditions, underpinning assumptions, values and world-views. Furthermore, we argue that the seeming adversarial perspectives on sustainability problems that arise from more technical or socio-political perspectives may actually be complementary. The development of socio-technical solutions that seek to bridge the divide between overgeneralized technical solutions and deeply contextualized socio-political approaches with limited transferability would increase the applicability and legitimacy of sustainability science. For this to occur rather than competing narratives developed in tandem, what is required is genuine dialogue that acknowledges the underpinning factors (including problem framing) that can lead to such fractured discourses in sustainability science. Dialogue and mutual understanding denote a starting point for deeper-level institutional changes that are necessary to facilitate and mainstream inter- and transdisciplinary research projects. Currently existing institutions in science pose structural constraints to greater inclusivity. These institutional constraints include reward mechanisms that incentivise specialisation and a lack of regard for specific outputs of inter- and transdisciplinary research such as knowledge co-creation (National Research Council 2004 ; Schneidewind 2010 ). As Ostrom ( 1990 ) described, changes must happen not only at the level of operational and collective rule-making, but also at the constitutional level—in this case, the organising principles and power relationships of the institutions of science.

The findings of normative science, including in sustainability science, can be highly polarized (e.g., Fischer et al. 2014 ). Using the Golden Rice as an exemplar of such polarized debates we found that the obstacles to integration of knowledge arose from diverging problem framings, here explained as connected to disciplinary affiliation and personal values. To overcome these obstacles to shared understanding we proposed three steps: (1) to explicitly recognize why a situation is framed as a problem and what criteria constitute sustainability in the particular context; (2) to work in transdisciplinary ways, based on mutual respect, by prioritizing well-being and systems thinking; and (3) to reflect on the potentials and limitations of academia’s current institutions in facilitating inter- and transdisciplinarity. These steps may facilitate the overall aim of addressing root causes of unsustainability in, by and through science.

Below is the link to the electronic supplementary material.

Acknowledgements

JF acknowledges a Consolidator Grant by the European Research Council (ERC). Synergies with the ERC-related work helped to support this paper. DJA was supported by the VolkswagenStiftung and the Niedersächsisches Ministerium für Wissenschaft und Kultur funded project ‘Leverage Points for Sustainable Transformations: Institutions, People and Knowledge’ (Grant number A112269).

1 In comparison, raw sweet potato contains 85.1 µg/g beta-carotene (USDA 2016 ). It is the only of the 10 most produced staple crops exceeding second generation Golden Rice (FAOSTAT 2016 ).

2 Because economists tend to have a very specific approach to the normative issues addressed here, we classified them separately from other social scientists.

3 Technically seen, Monsanto is not involved in the project, having donated patents for a gene that has already been replaced, yet Stone and Glover stated the company “has been eager to take credit for Golden Rice” (2016). Further authors ascribed Golden Rice a marketing value as it facilitates consumer acceptance and eventual market penetration of Syngenta and Monsanto (Nestle 2001 ; Brooks 2011 ). Thus, we conclude the Golden Rice Humanitarian Board and biotech corporations share the interest of bringing Golden Rice to the fields.

4 The notion of paradigms is used to describe “a set of assumptions that structure the approach to research” (Moon and Blackman 2014 : 1173).

5 Following Lang et al. ( 2012 ), we define transdisciplinary as scientific principle of knowledge co-creation in teams of different disciplines and actor groups targeted at mitigating societal problems.

Handled by Osamu Saito, United Nations University Institute for the Advanced Study of Sustainability, Japan.

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• Published: 28 January 2021

Development and characterization of GR2E Golden rice introgression lines

• B. P. Mallikarjuna Swamy 1 ,
• Severino Marundan Jr. 1 ,
• Mercy Samia 1 ,
• Reynante L. Ordonio 2 ,
• Democrito B. Rebong 2 ,
• Ronalyn Miranda 2 ,
• Anielyn Alibuyog 2 ,
• Anna Theresa Rebong 2 ,
• Ma. Angela Tabil 2 ,
• Roel R. Suralta 2 ,
• Antonio A. Alfonso 2 ,
• Partha Sarathi Biswas 3 ,
• Md. Abdul Kader 3 ,
• Russell F. Reinke 1 ,
• Raul Boncodin 1 &
• Donald J. MacKenzie 4  

Scientific Reports volume  11 , Article number:  2496 ( 2021 ) Cite this article

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• Biotechnology
• Plant sciences

Golden Rice with β-carotene in the grain helps to address the problem of vitamin A deficiency. Prior to commercialize Golden Rice, several performance and regulatory checkpoints must be achieved. We report results of marker assisted backcross breeding of the GR2E trait into three popular rice varieties followed by a series of confined field tests of event GR2E introgression lines to assess their agronomic performance and carotenoid expression. Results from confined tests in the Philippines and Bangladesh have shown that GR2E introgression lines matched the performance of the recurrent parents for agronomic and yield performance, and the key components of grain quality. Moreover, no differences were observed in terms of pest and disease reaction. The best performing lines identified in each genetic background had significant amounts of carotenoids in the milled grains. These lines can supply 30–50% of the estimated average requirements of vitamin A.

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Introduction

Rice ( Oryza sativa ) is the major source of energy and nutrition for more than half the world’s population 1 . However, rice supplies minimal micronutrients in its milled form and completely lacks β-carotene which is the precursor for vitamin A. Thus, resource-poor people primarily dependent on rice with little access to diverse diets suffer from micronutrient deficiencies, also termed hidden hunger 2 , 3 . Even though efforts are being made to address micronutrient deficiencies by supplementation, fortification, and dietary diversification, the problem still persists globally. Biofortification of major staple crops has been recognized as one of the sustainable means to tackle micronutrient deficiencies especially in the vulnerable target groups in rural areas 4 .

Vitamin A is essential for various functions in the human body such as development and functioning of the visual system, differentiation and maintenance of cells, epithelial membrane integrity, and production of red blood cells, immune system, reproduction, and iron metabolism 5 , 6 . An estimated 190 million children and 19 million pregnant women have vitamin A deficiency (VAD), and almost a million children go blind every year 7 . In the Philippines, VAD ranges between 19.6 to 27.9% in infants and preschool children 8 , while in Bangladesh, over half of the preschool age (56.3%) and school age children (53.3%) at the national level were found to exhibit at least a mild grade of VAD 9 .

Several crops such as maize, cassava, and sweet potato have been successfully biofortified with elevated levels of provitamin A 10 , 11 . However, there is no naturally-occurring variation for provitamin A in grains in rice germplasm, so this has been achieved by using genetic engineering approaches. The genetic modification was made by the addition of two genes, phytoene synthase ( Zmpsy1 ) from Zea mays and carotene desaturase ( crtI ) gene from the common soil bacterium, Pantoea ananatis (syn. Erwinia uredovora ) into a temperate japonica rice variety, Kaybonnet, from the USA. This completed the carotenoid pathway in the grain and resulted in the accumulation of β-carotene in the endosperm 12 , 13 . However, the transfer of this golden rice trait from Kaybonnet into additional locally-adapted and widely-grown rice varieties is required for the successful release and adoption of golden rice in Asia.

Among the six second-generation Golden Rice (GR2) events received by the International Rice Research Institute (IRRI), event GR2E was found to contain a single intact copy of the inserted DNA integrated at a single site within the rice genome, giving rise to agronomically desirable progeny with suitable grain carotenoid content. This event has been transferred into Asian rice varieties through marker-assisted backcrossing (MABC). MABC has been successfully used to transfer high value genes/QTLs for disease resistance, submergence and drought tolerance traits into popular rice varieties without altering their desirable traits 14 , 15 , 16 .

Development of stable golden rice breeding lines with nutritionally relevant levels of provitamin A and without trait-associated yield, grain quality, or disease resistance penalties relative to the recipient parental varieties is essential for the successful adoption of golden rice. Introgression of the GR2E locus from GR2E Kaybonnet into PSBRc82, IR64, and BRRI dhan29 (BR29) was performed at IRRI through MABC along with selections for desirable agronomic and grain quality traits. The phenotypic evaluation was conducted under screen house conditions. Selection of homozygous plants and lines were carried out under field conditions at IRRI. Agronomic evaluations of selected lines were carried out under field conditions in a series of confined field tests (CTs) at IRRI, PhilRice and BRRI.

The main objectives of the present work were to: develop agronomically desirable lines of provitamin A enriched GR2E golden rice in the genetic backgrounds of popular rice varieties from Asia; to understand the effects of genetic background and environment on carotenoid expression, and to identify stable and productive lines of GR2E golden rice for varietal evaluation.

Introgression of event GR2E into multiple genetic backgrounds

A series of five backcrosses of event GR2E Kaybonnet into three widely-grown rice varieties, IR64, PSBRc82, and BR29, resulted in the identification of introgression lines that were agronomically similar to their respective recipient parents. The stability and inheritance of the GR2E locus was confirmed using event-specific PCR in every generation, where it was found to segregate without distortion in a typical 1:1 Mendelian ratio in all the backcross generations (BC 1 to BC 5 ) and genetic backgrounds. All seeds containing the GR2E event showed the typical golden yellow color, indicating the expression of the provitamin A trait in the endosperm. Hemizygous (It is a condition in a diploid organism, where only one copy of the locus is present) plants phenotypically similar to their respective recipient parents were identified, backcrossed and advanced up to BC 5 F 1 , and with each successive backcross there was a progressive increase in similarity of the progenies to their respective recurrent (recipient) parents (Fig.  1 ). A total of 400, 190, and 94 BC 5 F 1 plants of IR64, PSBRc82, and BR29, respectively, were phenotyped and genotyped by event-specific PCR. Yellow BC 5 F 2 seeds were selected and analyzed for total carotenoid content, which ranged from 3.6–6.2 ppm in IR64, 3.1–6.4 ppm in PSBRc82, and 3.2–8.0 ppm in BR29. The BC 5 F 2 plants were closer to respective recipient parents for key agronomic traits with average days to flowering (DTF), plant height (PH) and number of panicles (NP) of the selected BC 5 progenies were 71.5 days, 108 cm and 15 for IR64, 82.5 days, 122.3 cm and 15.4 for PSBRc82 and 83 days, 117 cm and 17 for BR29 respectively. The final set of BC 5 F 3 selected lines had background recovery of more than 98%. Agro-morphological traits, panicle characteristics, and grain parameters were similar to the recipient parents and no unintended, unexpected, effects due to the presence of the GR2E event were observed throughout the backcross breeding program. Based on the overall agronomic performance, carotenoid levels, and genetic background recovery, 40 BC 5 F 1 plants in the IR64 background, and 20 BC 5 F 1 plants in each of the PSBRc82 and BR29 backgrounds were selected. The BC 5 F 2 seeds produced by each of these plants were further evaluated under field conditions in confined tests and plants homozygous for the GR2E locus were selected.

( a – c ) GR2E introgression lines.

Selection of homozygous and agronomically acceptable GR2E lines

The first confined field test of GR2E breeding lines was carried out during the 2015WS at IRRI to make individual homozygous plant selections. From among 8000 BC 5 F 2 plants tested, a total of 602, 439, and 471 plants homozygous for the GR2E locus were identified in IR64, PSBRc82, and BR29, respectively (Fig S1 ). Efforts were focused on the lines homozygous for GR2E; however, hemizygous and null plants were also phenotyped to determine the impact of the presence of the GR2E locus on agronomic traits. The pair-wise t-tests were conducted between families derived from single BC 5 F 1 plants within each of the three genetic backgrounds. Significant differences between families for total carotenoids were noted in a number of the possible pair-wise comparisons (data not shown). The mean comparisons between homozygous, hemizygous and null GR2E plants within each of the three populations did not show any abnormal deviations for key agronomic traits (Fig S2 ). The mean PH of lines carrying GR2E were marginally shorter than the respective recipient parent. For the remaining traits there were no clear differences between plants carrying GR2E and the respective parent variety. A total of 70 BC 5 F 3 ILs similar to their respective parents and having higher levels of carotenoids were selected for IR64 and PSBRc82 genetic backgrounds.

Evaluation of GR2E introgression lines in multi-location replicated confined tests

Agronomic performance of GR2E Introgression Lines (ILs) and their respective control varieties were assessed in a series of CTs at IRRI (2015WS, 2016DS and 2016WS), PhilRice (2015WS and 2016DS) and BRRI in Bangladesh (2016 Boro). A total of 70 ILs similar to their respective parents in agronomic performance and having the greatest levels of carotenoids were selected from each of IR64 and PSBRc82 backgrounds. A total of 14 agronomic, yield and yield-related traits and carotenoid content were measured from the different confined tests. Among the 70 ILs tested during the 2015WS at IRRI, PSBRc82 GR2E ILs showed small but statistically significant differences from non-transgenic PSBRc82 for eight traits including days to flowering (DTF), plant height (PH), Flag leaf length (FL), flag leaf width (FW), filled spikelets (FS), total number of spikelets per plant (TSP), grain length (GL) and hundred seed weight (HSW) (Table 1 ). However, in successive CTs conducted using 32 GR2E PSBRc82 ILs at IRRI and PhilRice, only FL, GL and HSW (2016DS), and GL, HSW and plot yield (PY) (2016WS; IRRI) showed significant differences. On the other hand, no significant differences were observed during the 2016DS and only GL and HSW showed significant differences at PhilRice in 2016WS (Table 2 ). Similarly GR2E IR64 ILs showed small but significant differences to the recipient parent for FL, TSP, GL, GW and HSW in 2015DS and for FW, FS, spikelet fertility (SF) and PY in 2016DS, while only GL showed significant difference in 2016WS. For the CT conducted with GR2E BR29 ILs in Bangladesh in the 2016 Boro season there were no significant differences from BR29 for all the traits measured (Table 3 ). Significant variations in total carotenoids among different families were observed in all backgrounds. The highest concentration of total carotenoids was observed in the BR29 background, followed by the PSBRc82 background, while the IR64 background had the lowest concentration of total caroteneoids (Tables 1 , 2 , 3 ). The grain samples of GR2E ILs along with recipient parents are shown in Fig.  2 . Grain quality traits amylose content (AC), gel consistency (GC) and alkali spreading value (ASV) were measured for PSBRc82, IR64 and BR29 (Tables 1 , 2 , 3 ). There were no significant differences for AC between GR2E PSBRc82 ILs and PSBRc82 in all the trials. There were no significant differences in ASV and AC between GR2E IR64 ILs and the IR64 parent, while for BR29 there were no differences between the transgenic and the control except for AC. The background recovery of final set of selected BC 5 F 3 ILs showed more than 98% recipient genome in all the three genetic backgrounds (Fig S3 – S5 ). There was no significant difference in AC except in BR29, similarly for GC some minor significant differences were observed in PSBRc82 and IR64 in some seasons.

Grain samples of GR2E golden rice and respective recipient parents.

Correlation between yield, yield related traits and carotenoid content

The correlation among yield and yield related traits; and with total carotenoid content is presented in the Figs S6 – S8 . Over all there was no specific trend in correlations among different yield and yield related traits. Except in one environment carotenoid content was negatively but non-significantly associated with PY in all the three genetic backgrounds. The correlation analysis of carotenoid content between different seasons showed highly significant correlation in all the three genetic backgrounds.

Effect of genetic background and environment on expression of carotenoids

The combined analysis of variance for carotenoid content at two months after harvest showed that there were significant genotypic, seasonal and location effects on the expression of carotenoid content. However, there were no significant genotype and environmental interactions (G × E) for carotenoid content except CT2 PR vs CT4 (Table 4 ). However, among the three genetic backgrounds, expression of carotenoids was higher in GR2E BR29 ILs followed by PSBRc82 and lowest in GR2EIR64 ILs (Fig.  3 , Fig S9 ). There were very highly positive significant correlations for carotenoid content estimated in different locations both within and between seasons (Figs S10 – S12 ). In general carotenoids expression was bit higher in WS than in DS, but also among most of the CTs no significant G × E interaction was observed (Table 4 ).

Carotenoid levels in different genetic backgrounds.

Identification of superior GR2E NILs for multi-location evaluation

We selected five GR2E introgression lines each for PSBRc82 and IR64, for BR29 eight lines were selected from the CTs. These lines will be further evaluated in multi-location field testing in the Philippines and Bangladesh respectively. The list of selected lines and their corresponding agronomic performance is provided in Table 5 . The ILs were similar to the respective recipient parents in all the agronomic, yield and yield traits measured, and the total carotenoids ranged from 3.8 to 5.5 ppm in the DS and 4.1 to 6.1 in the WS. Among the eight selected GR2E BR29 ILs no significant variation was observed in any trait except yield, with an advantage of 12.8% over BR29.

Most of the dietary vitamin A is of plant origin in the form of provitamin A that is converted to vitamin A in the body 17 . VAD is persistent in most of the rice eating countries in Asia, Africa and Latin America 18 , 19 . Therefore, enriching rice with provitamin A through biofortification is a viable and complementary intervention to tackle the VAD. The provitamin A trait was introduced into the rice variety Kaybonnet through genetic engineering 13 , which has a temperate japonica genetic background and is not well adapted to the tropical conditions in most rice growing Asian countries. We developed GR2E event introgressed golden rice ILs in the genetic backgrounds of IR64, PSBRc82 and BR29.

Introgression of the GR2E produced agronomically superior plants

Golden rice GR2E is genetically stable and molecularly clean event useful for breeding ( https://www.dropbox.com/sh/qpiz0cftefcaceq/AAByIpj_HED3zgqH7ufW7A-ta?dl=0 ; https://www.foodstandards.gov.au/code/applications/Documents/A1138%20Application_Redacted.pdf ). The breeding process to develop GR2E introgression lines did not show any abnormal plant phenotypes both in homozygous and hemizygous conditions indicating the genetic stability of the GR2E gene and trait expression. Both the phenotypic and genotypic based segregation analysis showed typical Mendelian segregation ratio in different segregating generations. GR2E advance backcross progenies were phenotypically very similar to their respective recipient parents. Transgenic events with single copy, clean integration and showing normal Mendelian segregation are considered ideal for research and breeding purposes, as they do not alter the host plant genome 20 , 21 , 22 .

Agronomic performance at field level and G × E studies showed that the GR2E gene did not alter any of the traits of the recipient parents in all its zygosity conditions. Overall plant performance was better during DS and among the genetic backgrounds the GR2EPSBRc82 lines performed better than the GR2EIR64 lines. Morphological traits such as panicle type, panicle exertion, grain shape, flag leaf length and width were similar for the GR2E ILs. Many lines performed equally similar to the respective recurrent parents, allowing the selection of advanced lines in all backgrounds for further testing in multi-location trials. The results showed that back cross process recovered almost all the desirable agronomic, yield and grain quality traits of the respective parents with significant expression of vitamin A. Despite many typhoons, heavy rains and high winds during the trials. There were no severe lodging incidences observed. Insects and diseases incidences were monitored during the two growing seasons at two different plant growth stages: maximum tillering stage (vegetative stage) and 50% flowering. Generally, crop stand was good with manageable level of insect pests and diseases during the growing seasons. Insects observed (both pest and beneficial insects) were found to be present in both test materials. We did not notice any difference between GR2E introgression lines and their respective recipient parents for the pest or diseases pressure on the crop across the confined field tests.

Woodfield and White 23 , and Badenhorst et al . 24 opined that development of transgenic product is not limited only to transformation, but also includes breeding through further backcrossing of transgenes with recipient parents and selection for desired traits of interest, in order to expedite commercial product development. For commercial deployment of any new variety with one or more introduced new trait(s) of a staple crop, in parallel to yield and other key agronomic traits, the newly developed variety should have essentially similar or better performance against biotic and abiotic stresses and grain quality traits compared to recipient variety; the introduced trait(s) should not alter these traits of the recipient variety 25 , 26 .

Grain quality and proximate composition of GR is similar to recipient rice varieties

Furthermore, different cooking and eating quality traits like, AC and ASV did not show any significant difference between the ILs and their respective recipient parents in any CTs. The golden rice breeding lines with significant amount of provitamin A accumulated in the grains helps to tackle VAD in high risk countries such as Bangladesh and the Philippines. However, it is a requirement to assess the composition of genetically modified crops to see if any significant changes in grain quality, nutrients and anti-nutrients contents in comparison to traditional counterpart and to assess the safety of the intended or unintended changes 27 , 28 . The compositional analysis of golden rice showed that all the compounds measured are within the biologically acceptable range and does not pose any risk to human health 29 . Earlier reports on transgenic products for insect and herbicide tolerance have also shown that little biologically meaningful changes in grain quality, nutrient and anti-nutrient composition 30 . There was a clear environmental effect, even though total carotenoids varied with environments, the genotypes with high carotenoids were always the best in all the locations. Such variations in trait expression due to environmental and agronomic factors and genetic basis have been well explained 31 , 32 .

Genetic background and environment influences carotenoid expression

Stable trait expression and minimal G × E for any trait of importance, especially for grain micronutrients and vitamins is essential for varietal release as well as for their successful adoption 4 , 33 , 34 . Total carotenoids were well correlated across the sites and generations; and expressed stably across the environments but there is a genetic background effect. Carotenoids expression varied even within segregating lines of different generations in each of the genetic backgrounds. So targeted breeding and careful selection of progenies with carotenoids test in each generation is necessary for advancing the lines. Mapping background QTLs and genes and using them in MAB can provide opportunity for precise development of GR lines with highest expression. The carotenoid levels were found to vary across the genetic backgrounds, locations and seasons but there were no significant G × E interactions. The highest expression of carotenoids was observed in BR29 background and the lowest in IR64 background. Several earlier attempts to develop golden rice events and introgression lines had to face the genetic background effects. Transgenic events developed in the indica backgrounds of IR64 and BR29 reported lower expression of GR genes in IR64 and higher expression in BR29 transformants, even ILs developed in IR64 showed lesser expression 35 . Moreover, ILs did not show any significant difference in yield when expressing the genes in the carotenoid pathway 36 . In our study also lowest expression was noticed in IR64. Simultaneously efforts are being made to develop next generation golden rice events with elevated levels of carotenoids with longer stability 37 , 38 , 39 . However, a genetic background effect is still a major bottle neck for introgression of carotenoid trait. Background effect on the expression of introduced traits was reported in rice for submergence tolerance, yield and related traits, disease resistance and drought tolerance 15 , 16 , 40 , 41 .

The variation in carotenoid concentration in grains might be due to variations in sunlight exposure and intensity across the locations and seasons 42 . Differential accumulation of β-carotene due to variation in exposure period and intensity of sunlight was also observed in algae, carrots, pumpkin and maize 43 , 44 , 45 , 46 . Moreover, like other carotenoids containing crops the carotenoid concentration in the grains of golden rice degrades over time after harvest. The degradation rate is very high at first few weeks after harvest and it becomes very slow after 6–8 weeks (data not shown). The carotenoids degradation rate is highly influenced by the storage temperature, moisture and exposure to light of the storage environment 22 , 47 . So, development of golden rice varieties with stable carotenoids expression is essential to achieve the impact 37 . However, there might be genotypic effect on the retention ability for carotenoids in rice grain. Understanding background effect and standardization of post-harvest handling is needed to achieve desired level of carotenoids in the introgression lines of multiple backgrounds.

Superior introgression lines were identified for multi-location trials

The five back crosses of GR2E gene into three genetic backgrounds resulted in identification of ILs similar to respective recipient parents. Adoption by the farmers and preference by the consumers for a specific crop variety particularly rice introduced with a new trait largely depends on its yield, grain quality and eating quality parameters. The introduced trait should be stable over locations and seasons to expedite the adoption level. Considering the present levels of carotenoids and per capita consumption in these target countries, the resulting ILs would be able to supply 30–50% of the EAR for vitamin A for the high risk population group if GR2E rice is consumed regularly.

Materials and methods

Development of gr2e near isogenic lines.

Kaybonnet is a high yielding japonica rice variety with blast resistance and excellent milling quality commercially cultivated in the USA. The genetic modification was made by the addition of two genes, phytoene synthase (Zmpsy1) from Zea mays and carotene desaturase (crtI) gene from the common soil bacterium, Pantoea ananatis (syn. Erwinia uredovora ). The GR2E Kaybonnet was crossed with the popular high yielding and adopted rice varieties such as IR64, PSBRc82, and BR29. IR64 is popular in most of the Asian countries, PSBRc82 in the Philippines, and BR29 in Bangladesh. In each generation, segregating materials were genotyped using GR2E event specific molecular marker. Plants containing the GR2E event and phenotypically similar to respective recipients were selected and backcrossed in each backcross generation to advance the materials to BC 5 F 2 . Background selections were performed using 100 randomly selected SSR markers in BC 1 and BC 2 , while selected plants from BC 3 , BC 4 and BC 5 were genotyped using the 6 K SNPs set at Genotyping Service Laboratory, IRRI. Only yellow-colored BC 5 F 2 seeds were separated and analyzed for total carotenoid content. A total of 40 BC 5 F 2 families for IR64 and 20 families each for PSBRc82 and BR29 were selected for evaluation in the confined test at IRRI. We have provided details of MAB scheme and evaluation of introgression lines in the Fig.  4 .

Development and evaluation of GR2E introgression lines.

Experimental materials used in the confined tests

A total of 8000 individual plants comprised of 4000 BC 5 F 2 plants from GR2E IR64, 2000 plants each from GR2E PSBRc82 and GR2E BR29 were included in a CT in the dry season of 2015 (2015DS). Plants were genotyped using GR2E specific markers and homozygous plants were selected. Selected BC 5 F 3 homozygous plants from each genetic background along with the respective recipient and donor parents were evaluated in a series of CTs at IRRI and PhilRice in the Philippines and at BRRI in Bangladesh. The list of GR2E materials evaluated and the details of the CTs is provided in the Supplementary Table S1 . Three CTs were conducted for GR2E IR64 and GR2E PSBRc82 at IRRI, while the selected lines of GR2E PSBRc82 were evaluated for two seasons at PhilRice. Further, BC 5 F 3 seeds of GR2E BR29 were sent to Bangladesh, multiplied in the screen house, and further evaluated in a CT at BRRI, Gazipur, for one season in 2016.

Crop management and observations

Seeds of the selected plants of GR2E introgression lines, recipient and donor parents were seeded in trays. Seedlings were transplanted at 21 days after sowing with a standard spacing of 20 × 20 cm. Details of the experimental design and layout are provided in Tables S1 and S2 . Standard agronomic practices were followed to raise a good crop, including the application of need-based plant protection measures to protect the crop from diseases and insect pests. Data were gathered on key agronomic, yield and yield-related traits; and total carotenoid content was measured two months after harvest. Grain quality data were generated from the selected lines of CT2 and from all lines included in CT3 and CT4. Insect pest infestations and disease incidences were recorded at maximum tillering and at 50% flowering. Agronomic traits were measured on five random plants from each entry. Days to 50% flowering was recorded on a whole plot basis. At maturity, five selected plants were harvested from individual plots and the remaining inner plants were harvested in bulk. Final plot yield was adjusted to a uniform grain moisture content of 14%.

DNA was extracted using fresh leaf samples and following a modified cetyl trimethylammonium bromide (CTAB) protocol 48 . Nanopore was used to check the quality and quantity of the DNA extracted. The DNA samples were diluted with distilled water into an equal concentration of 25 ng/µl. Amplification of event specific markers using polymerase chain reaction (PCR) was carried out with a 10 µl reaction mixture that contained 1.5 µl of DNA template, 1.0 µl of 10 × PCR buffer with MgCl 2 , 0.5 µl each of forward and reverse primers, 0.2 µl of 1 mM dNTP and 0.1 µl of Taq DNA polymerase and 5.7 µl distilled water. The amplification reaction was carried out in a 96-well PCR plate in a thermocycler using the following temperature profile: denaturation, 95 °C for 5 min; 35 cycles of denaturation at 95 °C for 45 s, annealing at 55 °C for 45 s and extension at 72 °C for 45 s; and final extension at 72 °C for 8 min and long-term storage at 10 °C. Amplification products were separated by gel electrophoresis on 1.2% agarose (0.5 × TBE; 160 V for 45 min) and visualized using SYBR Safe DNA stain and imaging using an AlphaImager HP (Protein Simple, San Jose, CA) gel documentation system. The GR2E specific primer sequences as follows.

ZD-E1-P1 5′-GCTTAAACCGGGTGAATCAGCGTTT-3′

ZD-E1-P2 5′-CGAGAGGAAGGGAAGAGAGGCCACCAA-3′

ZD-E1-P3 5′-CTCCCTCACTGGATTCCTGCTACCCATAGTAT-3′

Grain quality analysis

Grain quality analysis was carried out at the Analytical Service Laboratory (ASL) of IRRI. We measured/analyzed grain length and width, amylose content, alkali spreading value and gel consistency, using standard protocols 49 . Similar analyses were performed at BRRI on grain samples of GR2E BR29.

Amylose content

Amylose content (AC) was determined on milled rice extracts using a segmented flow analyzer. Rice samples were ground to a fine powder using a cyclone mill. Sodium Hydroxide and Ethanol were added to a test portion of the sample and heated in a boiling bath for 10 min. Acetic acid and Iodine solution was mixed with the aliquot of the test solution to form a blue starch iodine complex and its absorbance was measured at 620 nm using a colorimeter 49 . The result of the analysis was reported as apparent amylose to take into account the contribution of amylopectin present in the rice, which also forms a blue color starch iodine complex.

Gelatinization temperature

Rice starch gelatinization temperature (GT) was estimated by determining the alkali spreading value (ASV) of milled rice grains in potassium hydroxide solution. Six kernels of whole milled rice were incubated with 10 ml of 1.7% KOH for 23 h at ambient temperature (25 °C). The appearance and disintegration of the endosperm was visually rated depending on the intensity of spreading and swelling. ASV of 1–2 was classified as high GT, 3 for intermediate to high GT, 4–5 for intermediate GT and 6–7 for low GT.

Gel consistency

Samples of milled rice were ground to a fine powder, placed in a culture tube and suspended in a mixture of ethanol and 0.2 N KOH containing thymol blue and incubated in a boiling water bath for 15 min, followed by cooling to room temperature (15 min) and placing in an ice bath (20 min). Gel consistency of the rice paste (4.4% w/v) was determined by measuring the length of the cold gel in the culture tube after placing horizontally for 1 h. Rice was differentiated into three consistency types—soft (61 to 100 mm), medium (41 to 60 mm) and hard (27 to 40 mm).

Carotenoid concentrations

Total carotenoid concentration was estimated following the protocol developed by Gemmecker et al . 50 . Dehulled and polished rice seeds were ground to a fine powder using a modified paint shaker and accurately weighed amounts (ca. 500 mg) were dispensed into 15-ml Falcon tubes, mixed by sonication with 2 ml distilled water and incubated for 10 min at 60 °C. Cooled samples were centrifuged (3000 g , 5 min) and the supernatant fractions were transferred to new 15-ml tubes. Acetone (2 ml) and 100 μl of the lipophilic metallo organic dye, VIS682A (20 μg/ml; QCR Solutions Corp.), as an internal standard were added to each sample followed by mixing with short pulses of sonication and centrifugation (3000 g , 5 min). Supernatants were transferred to 15-ml tubes and the pellets were re-extracted twice more with 2-ml volumes of acetone and the resulting supernatant fractions were combined. Two ml petroleum ether (PE): di-ethyl ether (DE) (2:1 v/v) was added to each combined supernatant fraction (ca. 8 ml) and volumes were adjusted to 14 ml with distilled water. After vortexing, phase separation was achieved by centrifugation (3000 g , 5 min). The organic phase was recovered by pipetting out and transferred into a 2 ml graduated Eppendorf tube and the remaining aqueous phase was re-extracted with another 2 ml PE:DE (2:1 v/v), followed by centrifugation (3000 g , 5 min). The combined organic phases were dried using a vacuum-concentrator (Eppendorf concentrator 5301) and re-dissolved in 1 ml acetone. Maximum absorbance of sample extract at 450 nm and maximum absorbance of internal standard at 680 nm was determined using DU730 Beckman Coulter UV/VIS spectrophotometer. Concentrations of total carotenoids were determined from A450 nm assuming an average E450  nm = 142, 180 l mol −1  cm −1 in acetone using the Beer-Lambert law corrected for sample dilution and normalized to the internal standard.

Statistical analysis

All statistical analyses were performed as a linear mixed model using R 51 and PB Tools v1.0 52 .

Mixed model for single site analysis:

where µi denotes the mean of the ith entry (fixed effect), bj denotes the effect of the jth block, and eij denotes the residual error.

Mixed model for multiple site analysis:

where µi denotes the mean of the ith entry (fixed effect), lk denotes the effect of the kth site, bj(k) denotes the effect of the jth block within the kth site, (µl)ik denotes the interaction between the entries and sites (random effect), and eijk denotes the residual error.

Mean comparison and correlation analysis

The differences in least square (LS)-mean values between GR2E rice and the control rice were tested at first step followed by significant difference (p < 0.05) was identified in the multi-year combined-sites analysis 53 . Correlation among different traits from all the replicated trials was carried out using R Program 51 .

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Acknowledgements

This work was made possible through support of grants provided from Bill and Melinda Gates Foundation and US Agency for International Development.

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B.P.M.S. designed the study, conducted experiments, analyzed data, prepared manuscript, S.M. conducted experiments, data analysis, draft preparation, M.S. involved in genotyping, data collection, carotenoid analysis, R.L.O., D.B.R., R.M., R.R.S., A.A., A.T.R., M.A.T., A.A.A., P.S.B. and M.A.K. conducted field experiments, reviewed manuscript; R.F.R., R.B. and D.J.M. involved in experimental design and edited the manuscript.

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Mallikarjuna Swamy, B.P., Marundan, S., Samia, M. et al. Development and characterization of GR2E Golden rice introgression lines. Sci Rep 11 , 2496 (2021). https://doi.org/10.1038/s41598-021-82001-0

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Received : 29 November 2019

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DOI : https://doi.org/10.1038/s41598-021-82001-0

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