Next biotechnological plants for addressing global challenges: The contribution of transgenesis and new breeding techniques.

The aim of this survey is to identify and characterize new products in plant biotechnology since 2015, especially in relation to the advent of New Breeding Techniques (NBTs) such as gene editing based on the CRISPR-Cas system. Transgenic (gene transfer or gene silencing) and gene edited traits which are approved or marketed in at least one country, or which have a non-regulated status in the USA, are collected, as well as related patents worldwide. In addition, to shed light on potential innovation for Africa, field trials on the continent are examined. The compiled data are classified in application categories, including agronomic improvements, industrial use and medical use, namely production of recombinant therapeutic molecules or vaccines (including against Covid-19). The data indicate that gene editing appears to be an effective complement to 'classical' transgenesis, the use of which is not declining, rather than a replacement, a trend also observed in the patenting landscape. Nevertheless, increased use of gene editing is apparent. Compared to transgenesis, gene editing has increased the proportion of some crop species and decreased others amongst approved, non-regulated or marketed products. A similar differential trend is observed for breeding traits. Gene editing has also favored the emergence of new private companies. China, and prevalently its public sector, overwhelmingly dominates the patenting landscape, but not the approved/marketed one, which is dominated by the USA. The data point in the direction that regulatory environments will favor or discourage innovation.

Global developments of genome editing in agriculture.

Genome editing, particularly using of site-directed nucleases such as the CRISPR system, has spread rapidly through the biological sciences. Genome editing in crops could significantly speed up the progress of breeding programs. It could drive the development of traits in new crops and allow improvements in yield and pest resistance, adaptation to climate change, and industrial and pharmaceutical applications. However biofortification is a key challenge to satisfy nutritional needs in vitamins for developing countries and new consumer's needs for developed countries. China and the USA lead scientific research in crop editing. Nigeria, being headquarters to numerous research consortia, is the most involved country in Africa. Genome editing in animals including pig, cattle, sheep, and carp, has not merely accelerated research but has made possible research that was previously unfeasible. It has been used to increase disease resistance, to make livestock better adapted to farming or environmental conditions, to increase fertility and growth, and to improve animal welfare. The USA, the UK and China are the most involved countries in animal genome editing. Global food production needs to increase as much as 70 per cent to support the growing population. Genome editing could contribute improving the efficiency of food distribution and reducing waste. Depending on the regulatory conditions, genome editing could open up the field to smaller companies and public labs.

The ethical concerns about transgenic crops.

It is generally accepted that transgenesis can improve our knowledge of natural processes, but also leads to agricultural, industrial or socio-economical changes which could affect human society at large and which may, consequently, require regulation. It is often stated that developing countries are most likely to benefit from plant biotechnology and are at the same time most likely to be affected by the deployment of such new technologies. Therefore, ethical questions related to such biotechnology probably also need to be addressed. We first illustrate how consequentialist and nonconsequentialist theories of ethics can be applied to the genetically modified organism debate, namely consequentialism, autonomy/consent ethics (i.e. self-determination of people regarding matters that may have an effect on these people) and virtue ethics (i.e. whether an action is in adequacy with ideal traits). We show that these approaches lead to highly conflicting views. We have then refocused on moral 'imperatives', such as freedom, justice and truth. Doing so does not resolve all conflicting views, but allows a gain in clarity in the sense that the ethical concerns are shifted from a technology (and its use) to the morality or amorality of various stakeholders of this debate.

What Will Be the Benefits of Biotech Wheat for European Agriculture?

In European countries, wheat occupies the largest crop area with high yielding production. France, a major producer and exporter in Europe, ranks the fifth producer worldwide. Biotic stresses are European farmers' major challenges (fungal and viral diseases, and insect pests) followed by abiotic ones such as drought and grain protein composition. During the last 40 years, 1136 scientific articles on biotech wheat were published by USA followed by China, Australia, Canada, and European Union with the UK. European research focuses on pests and diseases resistances using widely marker-assisted selection (MAS). Transgenesis is used in basic research to develop resistance against some fungi (Fusarium head blight) while RNA interference (RNAi) silencing is used against some fungi and virus. Transgenic plants were also transformed with genes from various species for drought tolerance. The UK (mostly with transgenesis and site-specific nucleases) and France (with no transgenic tools but with MAS and site-specific nucleases) are the main countries carrying out research programs for both biotic stress and drought tolerance. Thus, few European countries used transgenesis for gluten protein composition and RNAi-mediated silencing in celiac disease. Because of vandalism field trials of transgenics dropped since 2000. No transgenic wheat is cultivated in Europe for political reasons.

Use of CRISPR systems in plant genome editing: toward new opportunities in agriculture.

Initially discovered in bacteria and archaea, CRISPR-Cas9 is an adaptive immune system found in prokaryotes. In 2012, scientists found a way to use it as a genome editing tool. In 2013, its application in plants was successfully achieved. This breakthrough has opened up many new opportunities for researchers, including the opportunity to gain a better understanding of plant biological systems more quickly. The present study reviews agricultural applications related to the use of CRISPR systems in plants from 52 peer-reviewed articles published since 2014. Based on this literature review, the main use of CRISPR systems is to achieve improved yield performance, biofortification, biotic and abiotic stress tolerance, with rice (Oryza sativa) being the most studied crop.

Next biotech plants: new traits, crops, developers and technologies for addressing global challenges.

Most of the genetically modified (GM) plants currently commercialized encompass a handful of crop species (soybean, corn, cotton and canola) with agronomic characters (traits) directed against some biotic stresses (pest resistance, herbicide tolerance or both) and created by multinational companies. The same crops with agronomic traits already on the market today will continue to be commercialized, but there will be also a wider range of species with combined traits. The timeframe anticipated for market release of the next biotech plants will not only depend on science progress in research and development (R&D) in laboratories and fields, but also primarily on how demanding regulatory requirements are in countries where marketing approvals are pending. Regulatory constraints, including environmental and health impact assessments, have increased significantly in the past decades, delaying approvals and increasing their costs. This has sometimes discouraged public research entities and small and medium size plant breeding companies from using biotechnology and given preference to other technologies, not as stringently regulated. Nevertheless, R&D programs are flourishing in developing countries, boosted by the necessity to meet the global challenges that are food security of a booming world population while mitigating climate change impacts. Biotechnology is an instrument at the service of these imperatives and a wide variety of plants are currently tested for their high yield despite biotic and abiotic stresses. Many plants with higher water or nitrogen use efficiency, tolerant to cold, salinity or water submergence are being developed. Food security is not only a question of quantity but also of quality of agricultural and food products, to be available and accessible for the ones who need it the most. Many biotech plants (especially staple food) are therefore being developed with nutritional traits, such as biofortification in vitamins and metals. The main international seed companies continue to be the largest investors in plant biotechnology R&D, and often collaborate in the developing world with public institutions, private entities and philanthropic organizations. These partnerships are particularly present in Africa. In developed countries, plant biotechnology is also used for non-food purposes, such as the pharmaceutical, biofuel, starch, paper and textile industries. For example, plants are modified to specifically produce molecules with therapeutic uses, or with an improved biomass conversion efficiency, or producing larger volumes of feedstocks for biofuels. Various plant breeding technologies are now used in the entire spectrum of plant biotechnology: transgenesis producing proteins or RNAi. Cisgenesis (transgenes isolated from a crossable donor plant) and intragenesis (transgenes originate from the same species or a crossable species), null segregants are also used. To date, the next generation precision gene editing tools are developed in basic research. They include: clustered regularly interspaced short palindromic repeats (CRISPR), oligonucleotide-directed mutagenesis (ODM), transcription activator-like effects nucleases (TALENs) and zinc-finger nuclease (ZFN).

Challenges facing European agriculture and possible biotechnological solutions.

Agriculture faces many challenges to maximize yields while it is required to operate in an environmentally sustainable manner. In the present study, we analyze the major agricultural challenges identified by European farmers (primarily related to biotic stresses) in 13 countries, namely Belgium, Bulgaria, the Czech Republic, France, Germany, Hungary, Italy, Portugal, Romania, Spain, Sweden, UK and Turkey, for nine major crops (barley, beet, grapevine, maize, oilseed rape, olive, potato, sunflower and wheat). Most biotic stresses (BSs) are related to fungi or insects, but viral diseases, bacterial diseases and even parasitic plants have an important impact on yield and harvest quality. We examine how these challenges have been addressed by public and private research sectors, using either conventional breeding, marker-assisted selection, transgenesis, cisgenesis, RNAi technology or mutagenesis. Both national surveys and scientific literature analysis followed by text mining were employed to evaluate genetic engineering (GE) and non-GE approaches. This is the first report of text mining of the scientific literature on plant breeding and agricultural biotechnology research. For the nine major crops in Europe, 128 BS challenges were identified with 40% of these addressed neither in the scientific literature nor in recent European public research programs. We found evidence that the private sector was addressing only a few of these "neglected" challenges. Consequently, there are considerable gaps between farmer's needs and current breeding and biotechnology research. We also provide evidence that the current political situation in certain European countries is an impediment to GE research in order to address these agricultural challenges in the future. This study should also contribute to the decision-making process on future pertinent international consortia to fill the identified research gaps.

Assessment of GE food safety using '-omics' techniques and long-term animal feeding studies.

Despite the fact that a thorough, lengthy and costly evaluation of genetically engineered (GE) crop plants (including compositional analysis and toxicological tests) is imposed before marketing some European citizens remain sceptical of the safety of GE food and feed. In this context, are additional tests necessary? If so, what can we learn from them? To address these questions, we examined data from 60 recent high-throughput '-omics' comparisons between GE and non-GE crop lines and 17 recent long-term animal feeding studies (longer than the classical 90-day subchronic toxicological tests), as well as 16 multigenerational studies on animals. The '-omics' comparisons revealed that the genetic modification has less impact on plant gene expression and composition than that of conventional plant breeding. Moreover, environmental factors (such as field location, sampling time, or agricultural practices) have a greater impact than transgenesis. None of these '-omics' profiling studies has raised new safety concerns about GE varieties; neither did the long-term and multigenerational studies on animals. Therefore, there is no need to perform such long-term studies in a case-by-case approach, unless reasonable doubt still exists after conducting a 90-day feeding test. In addition, plant compositional analysis and '-omics' profiling do not indicate that toxicological tests should be mandatory. We discuss what complementary fundamental studies should be performed and how to choose the most efficient experimental design to assess risks associated with new GE traits. The possible need to update the current regulatory framework is discussed.

Assessment of the health impact of GM plant diets in long-term and multigenerational animal feeding trials: a literature review.

The aim of this systematic review was to collect data concerning the effects of diets containing GM maize, potato, soybean, rice, or triticale on animal health. We examined 12 long-term studies (of more than 90 days, up to 2 years in duration) and 12 multigenerational studies (from 2 to 5 generations). We referenced the 90-day studies on GM feed for which long-term or multigenerational study data were available. Many parameters have been examined using biochemical analyses, histological examination of specific organs, hematology and the detection of transgenic DNA. The statistical findings and methods have been considered from each study. Results from all the 24 studies do not suggest any health hazards and, in general, there were no statistically significant differences within parameters observed. However, some small differences were observed, though these fell within the normal variation range of the considered parameter and thus had no biological or toxicological significance. If required, a 90-day feeding study performed in rodents, according to the OECD Test Guideline, is generally considered sufficient in order to evaluate the health effects of GM feed. The studies reviewed present evidence to show that GM plants are nutritionally equivalent to their non-GM counterparts and can be safely used in food and feed.

Is the German suspension of MON810 maize cultivation scientifically justified?

We examined the justifications invoked by the German government in April 2009 to suspend the cultivation of the genetically modified maize varieties containing the Bt insect-resistance trait MON810. We have carried out a critical examination of the alleged new data on a potential environmental impact of these varieties, namely two scientific papers describing laboratory force-feeding trials on ladybirds and daphnia, and previous data on Lepidoptera, aquatic and soil organisms. We demonstrate that the suspension is based on an incomplete list of references, ignores the widely admitted case-by-case approach, and confuses potential hazard and proven risk in the scientific procedure of risk assessment. Furthermore, we did not find any justification for this suspension in our extensive survey of the scientific literature regarding possible effects under natural field conditions on non-target animals. The vast majority of the 41 articles published in 2008 and 2009 indicate no impact on these organisms and only two articles indicate a minor effect, which is either inconsistent during the planting season or represents an indirect effect. Publications from 1996 to 2008 (376 publications) and recent meta-analyses do not allow to conclude on consistent effects either. The lower abundance of some insects concerns mainly specialized enemies of the target pest (an expected consequence of its control by Bt maize). On the contrary, Bt maize have generally a lower impact than insecticide treatment. The present review demonstrates that the available meta-knowledge on Cry1Ab expressing maize was ignored by the German government which instead used selected individual studies.

Emergence of minor pests becoming major pests in GE cotton in China: what are the reasons? What are the alternatives practices to this change of status?

A recent study in China by Lu et al.(1) shows that populations of an occasional cotton pest, mirid bugs (Heteroptera: Miridae), increased following the introduction of genetically engineered (GE) cotton plants. The GE cotton produces a delta-endotoxin from the bacteria Bacillus thuringiensis (Bt) to control the cotton bollworm. Before the introduction of Bt cotton in China, mirid bugs were usually controlled by broad-spectrum pesticide sprays targeted against the cotton bollworm, Helicoverpa armigera Hübner (Lepidoptera: Noctuidae), the most important pest of cotton in China. The effectiveness of the control of H. armigera by Bt cotton cultivation has resulted in a decrease in the amount of insecticides used on Bt cotton compared to conventional cotton. This has led to a lack of control of mirids on Bt cotton due to the reduction in broad-spectrum insecticide use and consequently to a transformation of a minor pest to a main one. We discuss the scientific evidence available in the literature of this phenomenon. We examine the reasons of the emergence of minor pests to become major pests in Bt cotton in China and possible solutions to this change of status.

[Literature review of the dispersal of transgenes from genetically modified maize]. / Revue bibliographique sur la dispersion des transgènes à partir du maïs génétiquement modifié.

This article aims at reviewing the theoretical and experimental data published in 562 publications referring to genetically modified (GM) maize dispersal. Our choice was limited to this since in the European Union (EU), GM maize is the only GM crop currently grown commercially. The pollen dispersal of transgenic maize is due to two factors: (i) pollen-mediated gene flow; (ii) seed admixture during harvest and post-harvest processes. The pollen dispersal decreases rapidly with the distance from GM plots. Climatic and topographic factors and factors of relative density between GM and non-GM maize plots impact on the pollen dispersal. The combination of both isolation distance and flowering date between source plots and sink plots limits the adventitious presence of transgenes in non-GM plots. All publications we reviewed demonstrate that the EU 0.9% threshold is technically manageable if the measures of isolation distances as well as harvesting and post harvesting processes and fully synchronous flowering are implemented.

Consultations of stakeholders on the roles of research in relation to genetically modified plants in France.

This article reports the first consultations on the roles of research in relation to genetically modified plants in France. We present a new attempt at facilitating discussion towards acceptable decisions and their results. This method consists of three steps: individual in-depth interviews of 77 French stakeholders, analysis of the interviews to identify elements that could help a constructive debate among participants, and two round-table discussions to present this analysis to stakeholders and foster discussions among them. The interviews exhibit a diversity of perceptions that are vaster than the pro or against points of view within the media. The problems raised during interviews deal with how discussions on genetically modified organisms (GMOs) are being done, how risks are taken into account, how the information is diffused, and how there is a minimal level of attention paid to social needs in GMOs' production. A series of problems more specific to the subject of the study discuss the weaknesses of the public research system. On the basis of these problems, 21 elements were identified that the stakeholders would like to see improved. One request seemed to be important for all types of stakeholders: "Raising the objectivity of the debate on GMOs." Our facilitation exercise led to a set of innovative concrete proposals for the design of an effective national debate.