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The Golden Rice Project

Biofortification to complement traditional interventions

In developing countries 500,000 people, mainly children, become blind every year, 50% of whom die within a year of becoming blind. Nearly nine million children die of malnutrion every year. Vitamin A deficiency (VAD) severely affects their immune system, hence it is involved in many of these children's deaths in the guise of multiple diseases. Malaria deaths in children under five years of age has been linked with deficiencies in the intake of protein, vitamin A and zinc. Various public and international programs for supplementation, fortification and diet diversification have achieved substantial improvements but have difficulty in attaining full coverage and above all, sustainability. Biofortification, involving conventional breeding of genetically improved basic staple crops, offers an opportunity to attain a more inclusive coverage, especially of the poorest sectors of society.

The major micronutrient deficiencies in the world are iron, zinc, and vitamin A. VAD is prevalent among the poor who depend mainly on rice for their daily energy uptake, because rice grains do not contain any β-carotene (provitamin A), which our body could in turn convert into vitamin A. Dependence on rice as the predominant food source, therefore, necessarily leads to VAD, which has the most severe effects on children and pregnant women. For the 400 million rice-consuming poor the medical consequences are severe: impaired vision, all the way to irreversible blindness, impaired epithelial integrity, exposing the affected individuals to infections, reduced immune response, impaired haematopoiesis (blood cell formation) and impaired skeletal growth, among other debilitating afflictions. Rice containing provitamin A could substantially alleviate the problem. But this can only be achieved using genetic engineering because, although there is provitamin A in the leaves of the rice plant, there is none the endosperm, which is the starch storage tissue of the seed. No variability for this trait has been detected in the world's most important rice germplasm collections.

Scientific breakthrough

Golden Rice has been engineered to contain the genes necessary to make up the biochemical pathway for pro-vitamin A production. Moreover, the genetic construct was designed to be expressed exclusively in the rice endosperm, ie in the edible part of the seed. The intensity of the golden colour is an indicator of the concentration of beta-carotene in the grain. A number of lines with different concentrations of β-carotene resulted from the work done in the laboratories of Ingo Potrykus and Peter Beyer, in Switzerland and Germany, respectively, with newer developments coming from the company Syngenta. The target of the research was to produce enough β-carotene in these rice lines to cover the recommended daily requirements, which has been now been achieved.

The transgenes leading to the production of β-carotene could be introduced into many different rice varieties at the same time, but because of the stringent, over-the-top regulatory requirements, in the end only one "regulatory clean" event will be used as a parenta line in multiple breeding programs.

Reaching out

Golden Rice will be made available to developing countries as part of a coordinated humanitarian project. This was, from the beginning, a public research project designed to reduce malnutrition in developing countries. Thanks to strong support from the private sector and free licences for humanitarian use, the hurdle of extensive intellectual property rights attached to the technologies used in the production of Golden Rice were easily overcome. This opened the way to collaborations with public rice research institutions in developing countries, with freedom-to-operate to develop locally adapted Golden Rice varieties.

Once locally developed varieties containing the Golden trait have cleared the regulatory hurdles at national level, they will be made available to subsistence farmers free of charge. The seed will become their property and they will also be able to use part of their harvest to sow their next crop, free of cost. Golden Rice is compatible with farmers using traditional farming systems, without the need for additional agronomic inputs. Therefore, no new dependencies are created.

The progress achieved since the initial scientific breakthrough in 1999 would not have been possible without an innovative public-private-partnership (PPP). Thanks to an agreement with Syngenta and other agbiotech industry technology donors, Golden Rice is royalty-free for humanitarian use, defined as an annual income below US$10,000 from farming, while income beyond that value would require a commercial licence from Syngenta. Royalty-free humanitarian sublicences are granted by the Golden Rice Humanitarian Board to public rice research institutions. These sublicence agreements ensure that the material is handled according to established biosafety guidelines and regulations, and that the target population receive the material free of charge for the provitamin A trait.

Tailored for local consumption

Development of locally adapted Golden Rice varieties and applications to national bioregulatory authorities for field testing and regulatory approval is in the hands of national and international public rice research institutions. To date, the Humanitarian Golden Rice Network includes 16 national institutions in Bangladesh, China, India, Indonesia, South Africa, The Philippines, and Vietnam. The Network is under the strategic guidance of the Golden Rice Humanitarian Board, managed by a network coordinator based at the International Rice Research Institute (IRRI) in the Philippines. The Humanitarian Board, an honorary body, benefits from the expertise of international authorities, such as Dr Gurdev Khush, retired rice breeder from IRRI (rice breeding); Prof Robert Russell, Laboratory for Human Nutrition, Tufts University Boston (vitamin A malnutrition); Dr Howarth Bouis, Director of HarvestPlus, International Food Policy Research Institute (IFPRI) Washington (biofortification); Dr Gary Toenniessen, The Rockefeller Foundation (food security in developing countries); Dr Robert Bertram, USAID Washington (development in Third World agriculture); Prof Matin Qaim, Professor and Chair in "International Food Economics and Rural Development" at the University of Göttingen, Germany (socioeconomic aspects); Dr Robert Zeigler, Director General, IRRI; Dr Sunkeswari R Rao Dept of Biotechnology, India (national cooperation in rice research); Prof Jean Pierre Jeannet, Babson College, Massachussets (global marketing); Professor Ingo Potrykus (co-inventor of Golden Rice ), professor emeritus from ETH Zurich, chairman (public relations and information); Prof Peter Beyer (co-inventor) Univ of Freiburg (scientificic advancement in the areas of biofortification for pro-vitamin A and other micronutrients); Dr Adrian Dubock, Golden Rice Project Manager (industry perspective and intellectual property rights); and the ex officio member Dr Gerard Barry, IRRI (Golden Rice Network Coordinator).

Golden Rice in the field

Biofortified seeds, a sustainable solution

Biofortification, i.e. introducing into crops the capacity to accumulate micronutrients by conventional breeding or biotecnology, is probably the most sustainable and cost-effective approach to reduce micronutrient malnutrition among the poor in developing countries. Golden Rice represents the first crop where such an enrichment was achieved in a crop plant by applying a biotechnological approach. Investment in this project has been relatively modest so far; in the nine years leading to 2008 only US$2.4 million had been spent. Product development, however, is time-consuming and requires substantial additional funding.

While expenses increase even more dramatically when it comes to biosafety assessment, as required for deregulation purposes, once a novel, biofortified variety has been deregulated and handed over to farmers, the system can develop its full potential. From this point on, the technology is built into each and every harvested seed, and does not require any additional investment. Let's consider the potential of a single Golden Rice seed: a single plant will produce in the order of 1,000 seeds; within four generations or less than two years, that one plant will have generated more than 1012 seeds. This represents up to 28-thousand metric tons of rice, which would be already sufficient to feed 100-thousand poor people for one year, and if they were eating Golden Rice they would have been automatically supplemented with provitamin A, reducing VAD. This gained protection is cost-free and sustainable. All a farmer needs –to benefit from the technology– is contained in one single seed!

Ignoring the benefits

It took 10 years —from 1980 to 1990— to develop the necessary technology to introduce genes into rice. It took another nine years—from 1990 to 1999—to introduce the genes that make up the pathway for provitamin A biosynthesis into the seed. And it took five more years—from 1999 to 2004—to develop Golden Rice. It is taking several more years to advance the first Golden Rice product through the deregulatory process. Considering that Golden Rice could substantially reduce blindness (500,000 children per year) and deaths (2-3 million per year) 20 years is a very long period of time. If it were possible to shorten the time it takes to get to the deregulated product, we could prevent blindness for hundreds of thousands of children!

Notwithstanding the fact that during the last 20 years a vast knowledge base has been accumulated in regards of the production and commercialisation of transgenic plants, the next years will have to be spent conducting the required biosafety assessments to exclude any putative harm for the environment and the consumer stemming from Golden Rice, and that despite the fact that experts cannot come up with such a harm scenario and that accumulated experience from hundreds of milions of hectares of transgenic plants are proof of what the experts are saying.

In a number of countries, the present regulatory practice is based on an overzealous interpretation of the precautionary principle, with little room left for risk management. The position at present is that even the slightest hypothetical risk must be tested and might lead to rejection of a registration application. At the same time, potential benefits are being blatantly disregarded. Recognised ecologists, including opposers of the technology, have not been able to come up with a realistic hypothetical risk to any agricultural or natural environment stemming from the production and accumulation of β-carotene in the endosperm of plants which otherwise produce high amounts of the same compound in other parts of the plant, and thus will not provide any additional selective advantage to the crop. This shows a substantial level of irrationality in the present system of environmental risk assessment. Despite this fact, the first Golden Rice field trial took place in the USA, and not in Southeast Asia, where it should have taken place, the reason being red tape imposed by a misguided precautionary principle.

An unbearable financial burden

What are those regulatory requirements? First of all, the application should be for a carefully selected, "regulatory clean" transgenic event. Criteria are not necessarily based on scientifc grounds. They include a number of requirements that are biologically irrelevant, e.g. the inserted DNA fragment should not have undergone multiple integrations or rearrangements, there should be no read-through across T-DNA borders nor microbial origins of replication and ballast DNA. This in turn requires the production of many hundreds of transgenic events using the same DNA construct, from which the regulatory clean event is then selected. The makeup of the construct itself must have been conceived taking into account the requirements imposed by the regulatory authorities. The carefully selected event can then be used to start a series of required biosafety assessment experiments expected to prove or disprove any putative biosafety hazard. The consequence of this approach is that nearly 99% of transgenic events, and often those with the highest levels of expression, must be discarded. Already this first step to mass-produce many hundreds of similar events, and the subsequent destruction of most of them, is beyond reach for most public research institutions, in developing as well as in developed countries, and funding agencies are not prepared to take over the costs.

The biosafety assessment starts with event-independent studies. These are related to the introduced genes and their function, and are valid for all events produced using these genes. These studies are followed by exposure evaluation tests for the novel trait, its intended use and bioavailability, as would be the case for a product like provitamin A. This study alone takes about three years, because without a field trial permit the material has to be produced in dedicated plant growth chambers. Next in line are protein production and equivalence analyses for the proteins encoded by the introduced genes. For this purpose the proteins have to be isolated from the plant, biochemically characterised, and their function confirmed. Further studies include, demonstration of lack of homology to known toxins and allergens, gastric degradation studies, heat lability, acute toxicity tests in a rodent feeding experiment. Screening for putative allergens and toxins is assumed to ensure that no unintended toxin or allergen will be consumed with Golden Rice.

This all would seem reasonable if it were not for the fact that most people have been eating these genes and their products from a number of other food sources throughout their lives. At one point it was even proposed to analyse whether known daffodil toxins had been introduced into Golden Rice along with the daffodil gene involved in provitamin A biosynthesis, which totally lacks a scientific basis: what has been transferred is one defined piece of DNA which is analogous to genes in other organisms, performing the same function, and these bear no relationship to any toxin or allergen. These studies take at least two years of intensive work in a well-equipped biochemistry laboratory.

The event-dependent studies are even more cumbersome; they include:

  • Molecular characterization and genetic stability: data on single-copy effect; marker gene at same locus; simple integration; Mendelian inheritance, including phenotypic and biochemical evidence for stability over at least three generations; no potential gene disruption; no unknown open reading frames; no DNA transfer beyond the T-DNA borders; no antibiotic resistance gene or origin of replication; insert size limited to the minimum necessary; insert plus flanking regions sequenced.
  • Expression profiling: gene expression levels at key growth stages; evidence for seed-specific expression.
  • Phenotypic analysis: field performance, typical agronomic traits, yield compared to isogenic lines; pest and disease status must be same as parent.
  • Compositional analysis: data from growing the event over two seasons at six locations in three replicates on proximates, macro and micronutrients, antinutrients, toxins, allergens; data must be generated on modified and isogenic background.
  • Environmental risk assessment: this type of analysis takes 4-5 years of work by an entire research team.

It is obvious that no scientist nor scientific institution in the public domain has the potential, funding or motivation to perform such lengthy and expensive biosafety experiments. It comes as no surprise then, that virtually all transgenic events that have been carried through the deregulatory process are—directly or indirectly—in the private sector and are restricted to high-value crops. Humanitarian projects do not fall into this category, even though millions of people would benefit from them. There is a lot of goodwill in the public and in the private sectors worldwide to exploit the potential of green biotechnology for the benefit of the poor. However, without a realistic, science-based risk assessment approach, public research funding is incapable of taking up the price tag. For this reason, scientific progress has become to a certain degree detached from product development, and the population at large is suffering the consequences.

No justification for extreme precaution

There are historic reasons for the present regulatory framework. In the 1970s, when gene technology was still incipient, taking a precautionary approach was sensible, and it was the scientists themselves, who at the time were not working with plants but with pathogenic micro-organisms, the ones responsible for establishing regulations based on the premise that the technology could lead to unpredictable genome alterations. More than 20 years of accumulated experience with transgenic plants, their widespread use on over 180 million hectares planted in an increasing number of countries in 2011, and many thousands of carefully conducted biosafety experiments carried out by prestigious institutions, have led to the conclusion that there is no specific risk associated with the technology that would set it apart from traditional plant breeding or natural evolution. And yet, we are still facing unjustified calls for further moratoria.

The fact that regulation of transgenic crops has become more strict lately is counterintuitive. Some people claim that we have to go to that length to build up trust in the technology with the consumer. However, more than 10 years of experience applying this strategy in Europe and many developing countries has demonstrated that this approach doesn't work. One reason is that in the public's perception a highly stringent regulation must be associated with an inherently risky technology.

The guidelines pretend to apply a predictable, risk-based evaluation system based on objective empirical science. The precautionary principle then turns the process into a subjective framework in which the assessment is based on pretence cultural and moral values, guided to a great extent by a perceptions of fear by the consumer. The EU has embedded the precautionary principle into its biosafety regulatory framework and as a consequence it's applying overly stringent health, safety, environmental, and technical product standards that go beyond other international standards, and is then exporting those regulations and standards abroad through supply chains via international treaties, standardisation bodies, and bilateral technical capacity-building intiatives. Examples of this include biotech labeling and traceability regulations implemented by the EU under the Cartagena Protocol to the UN Biodiversity Convention and the EU REACH regulation on chemicals management.

Ideally, we should be able to free the regulatory process from all scientifically unjustified ballast and end up with a set of rational regulatory guidelines. Such a move would require the involvement of institutions and governments, which at the moment lack the will to do so or are under undue pressures —often commercial in nature— that don't allow them to proceed along these lines. The pressure, amounting often to blackmailing, is then passed on to developing countries, interfering with their freedom to decide on the adoption of transgenic technologies in accordance with their particular needs. Those countries are then caught between the urgent need to adopt this technology and the commercial implications of doing so. In essence, the EU exports the high regulatory and standardisation costs abroad, resulting in the buildup of de facto trade barriers.

Gene technology has been endorsed by international agencies, such as FAO and UNIDO, to help solve food security problems in developing countries, but yet we are threading at a very low pace. The highest price for the non-adoption of green gene technology is being paid by the voiceless persons who most need it. The great potential of gene technology to reduce hunger and malnutrition and to help protect the environment will only be attained when regulatory frameworks become based on scientific evidence and a proper risk-benefit analysis. The regulatory has become so complex and expensive that at present only crop and trait combinations with large financial returns for the private sector are making it to the farmers' fields, while a huge number of publicly funded projects don't make it beyond the experimental stages. This bottleneck, caused by the technology opposers, is often used by the same people to argue against the technology. They cite the paucity of public biotech crop as "proof" that the technology benefits only the big companies, when the reason for this is their fierce opposition.

Every year there are more countries in the developing world that don't give in to questionable arguments and who have started embracing the technology, based on hard facts. Positive, highly encouraging reports on increased harvests, reduced use of pesticides, a decrease in the number of people intoxicated from using pesticides, and an increase in the number of beneficial insects in the fields are now coming from countries like South Africa and India. In the case of Golden Rice, the World Bank has produced a particularly encouraging ex-ante analysis on its potential socioeconomic impact. Sadly, the years go by, children continue dying unnecessarily, and over-the-top regulations remain the major hurdle to progress.

Traditional genome meddling

Green gene technology has the potential to support and complement traditional plant breeding. One criticism frequently brought up in relation with genetic engineering is that the insertion of genes can lead to unpredictable genome alterations. In traditional plant breeding, agronomic traits are combined or eliminated by crossing, followed by selection. Starting materials are selected varieties and landraces of crop plants. Differences between landraces were originally identified and selected by indigenous farmers, and are based on spontaneous, unpredictable mutations. In the course of traditional breeding, which may include wild relatives of crop plants, many unpredictable genome alterations, such as recombinations, translocations, deletions, inversions and horizontal gene transfer, are combined into a new cultivar. These unpredictable, significant genome alterations accumulate at every breeding step and each new, traditionally bred variety is the result of an increasing array of such genome alterations. This statement is valid for all modern crop varieties, including those used in organic farming. Nevertheless, while none of these [genetically modified] varieties has ever been assessed for biosafety, mankind has consumed them unharmed and the environment has not been affected by them either. The fact is that actually nobody could survive without eating food from these genetically modified crops.

By comparison, the creation of Golden Rice —involving the insertion of two precisely defined genes into a genome that contains fifty-thousand-odd genes— is by several orders of magnitude more precise than traditional breeding. Why should this new rice variety, despite the fact that the modification is extremely small and highly targeted, be the subject of such unjustified scrutiny?

Accepting responsibility

Green biotechnology has the potential to provide solutions to pest and disease control, improve photosynthetic efficiency and nutritional content, furnish plants with adaptation mechanisms for heat, cold and salt tolerance, and many more things to come. The benefits of Golden Rice are clear at face value, yet opposers of the technology present themselves as saviours of humanity. The blind and the dead are not at risk, they are a sad reality. Will the technology opponents accept the responsibility for this preventable tragedy being caused by them or will they keep hiding behind their "noble&; cause, to save us from the unknown unknowns? Because that's what they are afraid of.

The Nuffield Council on Bioethics concluded that "[t]he European Union is ignoring a moral imperative to promote genetically modified crops for their great potential for helping the developing world", and "[w]e believe EU regulators have not paid enough attention to the impact of EU regulations on agriculture in developing countries."


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Rice terraces