Crop Biotechnology Research Articles

Reflections on Social Justice in the Promotion of Biotechnology Crops and Institutionalisation of Food Values among Smallholder Farmers in Central Uganda

The academic discourse of agricultural biotechnology carries a lot of information on how biotechnology crops are crucial in realizing food security once they are fully adopted by farmers. Biotechnology crops are seen as the solution to ameliorate the Malthusian catastrophe of population increase threatening food security. While studies on agricultural biotechnology are increasing, they mainly focus on how the new agriculture science produces crops with desirable traits in supporting food production interventions. Few studies have concerned themselves with the social justice implications among smallholder farmers, posed by institutionalised agricultural biotechnology knowledge with its sophistication. This paper addresses itself to the above lacuna. The data collection used an embedded mixed method design with focus group discussions, in-depth interviews, non-participant observation, and questionnaire survey methods. The paper shows that mechanisms that are used to promote biotechnology crops are highly institutionalized. The institutions not only develop the biotechnology crops but also engage in mechanisms that encourage smallholder farmers to adopt these crops. The major social justice concerns addressed by this study are plant diversity, equity and commercialization of biotechnology-generated seeds/crops, participation, and farming choices

An assessment of the linkages between GM crop biotechnology and climate change mitigation

ABSTRACT This article provides an analysis and evaluation of peer-reviewed evidence on the contribution of crop biotechnology to climate change mitigation and adaption. While there is a range of agricultural technologies and products that contribute to climate change mitigation, this literature landscape analysis focuses on the development of genetically modified traits, their use and adoption in major commodity crops and responsive changes in production techniques. Jointly, these technologies and products are contributing to climate change mitigation, yet the technology, the literature and evidence is still evolving as more sophisticated research methods are used with greater consistency. The literature analysis is undertaken with consideration of the consequential impact that regulatory regimes have on technology development. This assessment utilizes the Maryland Scientific Methods Scale and citation analysis, concluding that GM crops provide benefits that contribute to climate change mitigation.

RNA Interference in Insects: From a Natural Mechanism of Gene Expression Regulation to a Biotechnological Crop Protection Promise.

Insect pests rank among the major limiting factors in agricultural production worldwide. In addition to direct effect on crops, some phytophagous insects are efficient vectors for plant disease transmission. Large amounts of conventional insecticides are required to secure food production worldwide, with a high impact on the economy and environment, particularly when beneficial insects are also affected by chemicals that frequently lack the desired specificity. RNA interference (RNAi) is a natural mechanism gene expression regulation and protection against exogenous and endogenous genetic elements present in most eukaryotes, including insects. Molecules of double-stranded RNA (dsRNA) or highly structured RNA are the substrates of cellular enzymes to produce several types of small RNAs (sRNAs), which play a crucial role in targeting sequences for transcriptional or post-transcriptional gene silencing. The relatively simple rules that underlie RNAi regulation, mainly based in Watson-Crick complementarity, have facilitated biotechnological applications based on these cellular mechanisms. This includes the promise of using engineered dsRNA molecules, either endogenously produced in crop plants or exogenously synthesized and applied onto crops, as a new generation of highly specific, sustainable, and environmentally friendly insecticides. Fueled on this expectation, this article reviews current knowledge about the RNAi pathways in insects, and some other applied questions such as production and delivery of recombinant RNA, which are critical to establish RNAi as a reliable technology for insect control in crop plants.

Remembering Ronald L. Phillips: Pioneer in crop biotechnology, mentor, and humanitarian

Remembering Ronald L. Phillips: Pioneer in crop biotechnology, mentor, and humanitarian

Evaluation of an in vitro experimental platform of human polarized intestinal epithelial monolayers for the hazard assessment of insecticidal proteins

Previous work demonstrated the utility of using human-derived intestinal epithelial cell (IEC) lines cultured as polarized monolayers on Transwell® filters to differentiate between hazardous and non-hazardous proteins. The current study seeks to further resolve appropriate concentrations for evaluating proteins of unknown hazard potential using the IEC experimental platform and leverages these parameters for evaluating the potential toxicity of insecticidal proteins characteristic of those expressed in genetically modified (GM) agricultural biotechnology crops. To establish optimal test protein concentrations, effects of several known hazardous (C. perfringens epsilon toxin, Listeriolysin O, Phaseolus vulgaris erythroagglutinin, E. coli Shiga toxin 1, C. difficile Toxin B and wheat germ agglutinin) and non-hazardous (Ara-h2, β-lactoglobulin, fibronectin and Rubisco) proteins on IEC barrier integrity and cell viability were evaluated at concentration ranges. Two insecticidal proteins (AfIP-1A and AfIP–1B) were evaluated for effects in the IEC assay, a seven-day insecticidal bioassay, and assessed in a high-dose 14-day acute oral toxicity study in mice. The results obtained from the human in vitro IEC assay were consistent with results obtained from an in vivo acute oral toxicity study, both demonstrating that the combination of AfIP-1A and AfIP–1B do not exhibit any identifiable harmful impacts on mammalian cells.

Genetic dissection of drought resistance for trait improvement in crops

Reliance on agriculture for food security is a constant in all modern societies. Global climate change and population growth have put immense pressure on sustainable agriculture, exacerbating the effects of environmental stresses. Drought is one of the most pressing abiotic stresses that farmers face, presenting an annual threat to crop growth and yield. Crops have evolved extensive morphological, physiological, and molecular mechanisms to combat drought stress. Drought resistance is a polygenic trait, controlled by a complex genetic network and an array of genes working together to ensure plant survival. Many studies have aimed at dissecting the genetic mechanisms underlying drought resistance. Recent studies using linkage and association mapping have made progress in identifying genetic variations that affect drought-resistance traits. These loci may potentially be engineered by genetic transformation and genome editing aimed at developing new, stress-resistant crop cultivars. Here we summarize recent progress in elucidating the genetic basis of crop drought resistance. Molecular-breeding technologies such as marker-assisted selection, genome selection, gene transformation, and genome editing are currently employed to develop drought-resistant germplasm in a variety of crops. Recent advances in basic research and crop biotechnology covered in this review will facilitate delivery of drought-resistant crops with unprecedented efficiency.

Plant Promoters: Their Identification, Characterization, and Role in Gene Regulation.

One of the strategies to overcome diseases or abiotic stress in crops is the use of improved varieties. Genetic improvement could be accomplished through different methods, including conventional breeding, induced mutation, genetic transformation, or gene editing. The gene function and regulated expression through promoters are necessary for transgenic crops to improve specific traits. The variety of promoter sequences has increased in the generation of genetically modified crops because they could lead to the expression of the gene responsible for the improved trait in a specific manner. Therefore, the characterization of the promoter activity is necessary for the generation of biotechnological crops. That is why several analyses have focused on identifying and isolating promoters using techniques such as reverse transcriptase-polymerase chain reaction (RT-PCR), genetic libraries, cloning, and sequencing. Promoter analysis involves the plant genetic transformation method, a potent tool for determining the promoter activity and function of genes in plants, contributing to understanding gene regulation and plant development. Furthermore, the study of promoters that play a fundamental role in gene regulation is highly relevant. The study of regulation and development in transgenic organisms has made it possible to understand the benefits of directing gene expression in a temporal, spatial, and even controlled manner, confirming the great diversity of promoters discovered and developed. Therefore, promoters are a crucial tool in biotechnological processes to ensure the correct expression of a gene. This review highlights various types of promoters and their functionality in the generation of genetically modified crops.

Mini-review: Lipid flippases as putative targets for biotechnological crop improvement.

An increasing world population and drastic changes in weather conditions are challenging agricultural production. To face these challenges and ensure sustainable food production in the future, crop plants need to be improved to withstand several different biotic and abiotic stresses. Commonly, breeders select varieties that can tolerate a specific type of stress and then cross these varieties to stack beneficial traits. This strategy is time-consuming and strictly dependent on the stacked traits been genetically unlinked. Here, we revise the role of plant lipid flippases of the P4 ATPase family in stress-related responses with a special focus on the pleiotropic nature of their functions and discuss their suitability as biotechnological targets for crop improvement.

Genome editing in maize: Toward improving complex traits in a global crop.

Recent advances in genome editing have enormously enhanced the effort to develop biotechnology crops for more sustainable food production. CRISPR/Cas, the most versatile genome-editing tool, has shown the potential to create genome modifications that range from gene knockout and gene expression pattern modulations to allele-specific changes in order to design superior genotypes harboring multiple improved agronomic traits. However, a frequent bottleneck is the delivery of CRISPR/Cas to crops that are less amenable to transformation and regeneration. Several technologies have recently been proposed to overcome transformation recalcitrance, including HI-Edit/IMGE and ectopic/transient expression of genes encoding morphogenic regulators. These technologies allow the eroding of the barriers that make crops inaccessible for genome editing. In this review, we discuss the advances in genome editing in crops with a particular focus on the use of technologies to improve complex traits such as water use efficiency, drought stress, and yield in maize.

An established protocol for generating transgenic wheat for wheat functional genomics via particle bombardment.

Wheat is one of the most important food crops in the world and is considered one of the top targets in crop biotechnology. With the high-quality reference genomes of wheat and its relative species and the recent burst of genomic resources in Triticeae, demands to perform gene functional studies in wheat and genetic improvement have been rapidly increasing, requiring that production of transgenic wheat should become a routine technique. While established for more than 20 years, the particle bombardment-mediated wheat transformation has not become routine yet, with only a handful of labs being proficient in this technique. This could be due to, at least partly, the low transformation efficiency and the technical difficulties. Here, we describe the current version of this method through adaptation and optimization. We report the detailed protocol of producing transgenic wheat by the particle gun, including several critical steps, from the selection of appropriate explants (i.e., immature scutella), the preparation of DNA-coated gold particles, and several established strategies of tissue culture. More importantly, with over 20 years of experience in wheat transformation in our lab, we share the many technical details and recommendations and emphasize that the particle bombardment-mediated approach has fewer limitations in genotype dependency and vector construction when compared with the Agrobacterium-mediated methods. The particle bombardment-mediated method has been successful for over 30 wheat genotypes, from the tetraploid durum wheat to the hexaploid common wheat, from modern elite varieties to landraces. In conclusion, the particle bombardment-mediated wheat transformation has demonstrated its potential and wide applications, and the full set of protocol, experience, and successful reports in many wheat genotypes described here will further its impacts, making it a routine and robust technique in crop research labs worldwide.

THE TENDENCY OF TRANSGENIC PLANTS SPREAD IN 2019-2021

The achievement of modern biotechnology, in particular genetic engineering in crop production, provides not only the possibility of increasing production, but also more efficient use of natural resources and reduces the negative impact on the environment. Recent publications indicate the widespread distribution of genetically modified (GM) agricultural crops in the world that are currently in commercial use, research and development. All over the world, the most cultivated GM crops are cotton, soybeans, corn, and rapeseed, and the area under these biotechnological crops continues to grow every year. However, the analysis of publicly available sources of information revealed the risks of negative effects of GMOs on human and animal bodies, as well as on the environment and biodiversity.
 The article analyzes the results of research on the presence of GM ingredients in plant raw materials, processing products, and feed for productive and non-productive animals for the period 2019-2021. The study was conducted by the polymerase chain reaction method in real time (PCR-RF), according to DSTU ISO 21569:2008 (ISO 21569:2005, IDT), and DSTU ISO 21571:2008 (ISO 21571:2005, IDT). In the studied samples were detected target sequences of the promoter p35S CaMV (cauliflower mosaic virus), promoter p34S FMV, terminator NOS (T-NOS) T1 of the Agrobacterium tumefaciens plasmid, genes Pat, EPSPS, Cry 3A.
 Transgenic plants are present in circulation in the agricultural market of Ukraine, and the area under their crops is increasing.
 The largest number of GM plants was found in samples of rapeseed, soybeans, and feed for productive and non-productive animals. There was a tendency for an increase in the number of transgenic plants in tested samples. During rapeseed research in 2019, the number of positive samples was 6.5%, in 2020 – 7.4%, and in 2021 – 14.3%. When studying soybeans in 2019, the number of positive samples was 6.7%, in 2020 – 16.7%, and in 2021 – 18.2%.
 GM ingredients were also found in feed for productive and non-productive animals. In 2020 16.7% of compound feed for poultry were positive, while in 2021 there were no samples for research.
 As for feeds for non-productive animals, during 2019–2020 there were no detected positive samples, while in 2021 the number of positive samples was 12%.

Turning promise into practice: Crop biotechnology for increasing genetic diversity and climate resilience.

As climate change increasingly threatens agricultural production, expanding genetic diversity in crops is an important strategy for climate resilience in many agricultural contexts. In this Essay, we explore the potential of crop biotechnology to contribute to this diversification, especially in industrialized systems, by using historical perspectives to frame the current dialogue surrounding recent innovations in gene editing. We unearth comments about the possibility of enhancing crop diversity made by ambitious scientists in the early days of recombinant DNA and follow the implementation of this technology, which has not generated the diversification some anticipated. We then turn to recent claims about the promise of gene editing tools with respect to this same goal. We encourage researchers and other stakeholders to engage in activities beyond the laboratory if they hope to see what is technologically possible translated into practice at this critical point in agricultural transformation.

Progress and prospects of C4 trait engineering in plants.

Incorporating C4 photosynthetic traits into C3 crops is a rational approach for sustaining future demands for crop productivity. Using classical plant breeding, engineering this complex trait is unlikely to achieve its target. Therefore, it is critical and timely to implement novel biotechnological crop improvement strategies to accomplish this goal. However, a fundamental understanding of C3 , C4 , and C3 -C4 intermediate metabolism is crucial for the targeted use of biotechnological tools. This review assesses recent progress towards engineering C4 photosynthetic traits in C3 crops. We also discuss lessons learned from successes and failures of recent genetic engineering attempts in C3 crops, highlighting the pros and cons of using rice as a model plant for short-, medium- and long-term goals of genetic engineering. This review provides an integrated approach towards engineering improved photosynthetic efficiency in C3 crops for sustaining food, fibre and fuel production around the globe.

Tissue-specific enhancement of OsRNS1 with root-preferred expression is required for the increase of crop yield

IntroductionRoot development is a fundamental process that supports plant survival and crop productivity. One of the essential factors to consider when developing biotechnology crops is the selection of a promoter that can optimize the spatial–temporal expression of introduced genes. However, there are insufficient cases of suitable promoters in crop plants, including rice. ObjectivesThis study aimed to verify the usefulness of a new rice root-preferred promoter to optimize the function of a target gene with root-preferred expression in rice. Methodsosrns1 mutant had defects in root development based on T-DNA insertional mutant screening and CRISPR technology. To optimize the function of OsRNS1, we generated OsRNS1-overexpression plants under two different promoters: a whole-plant expression promoter and a novel root-preferred expression promoter. Root growth, yield-related agronomic traits, RNA-seq, and reactive oxygen species (ROS) accumulation were analyzed for comparison. ResultsOsRNS1 was found to be involved in root development through T-DNA insertional mutant analysis and gene editing mutant analysis. To understand the gain of function of OsRNS1, pUbi1::OsRNS1 was generated for the whole-plant expression, and both root growth defects and overall growth defects were found. To overcome this problem, a root-preferential overexpression line using Os1-CysPrxB promoter (Per) was generated and showed an increase in root length, plant height, and grain yield compared to wild-type (WT). RNA-seq analysis revealed that the response to oxidative stress-related genes was significantly up-regulated in both overexpression lines but was more obvious in pPer::OsRNS1. Furthermore, ROS levels in the roots were drastically decreased in pPer::OsRNS1 but were increased in the osrns1 mutants compared to WT. ConclusionThe results demonstrated that the use of a root-preferred promoter effectively optimizes the function of OsRNS1 and is a useful strategy for improving root-related agronomic traits as well as ROS regulation.

Lessons for a SECURE Future: Evaluating Diversity in Crop Biotechnology Across Regulatory Regimes.

Regulation of next-generation crops in the United States under the newly implemented “SECURE” rule promises to diversify innovation in agricultural biotechnology. Specifically, SECURE promises to expand the number of products eligible for regulatory exemption, which proponents theorize will increase the variety of traits, genes, organisms, and developers involved in developing crop biotechnology. However, few data-driven studies have looked back at the history of crop biotechnology to understand how specific regulatory pathways have affected diversity in crop biotechnology and how those patterns might change over time. In this article, we draw upon 30 years of regulatory submission data to 1) understand historical diversification trends across the landscape and history of past crop biotechnology regulatory pathways and 2) forecast how the new SECURE regulations might affect future diversification trends. Our goal is to apply an empirical approach to exploring the relationship between regulation and diversity in crop biotechnology and provide a basis for future data-driven analysis of regulatory outcomes. Based on our analysis, we suggest that diversity in crop biotechnology does not follow a single trajectory dictated by the shifts in regulation, and outcomes of SECURE might be more varied and restrictive despite the revamped exemption categories. In addition, the concept of confidential business information and its relationship to past and future biotechnology regulation is reviewed in light of our analysis.

Cytogenetics to functional genomics: six decades journey of Professor P.K. Gupta.

SummaryWe had the fortune of starting our scientific/research careers in the Molecular Biology and Crop Biotechnology Laboratory of Professor P.K. Gupta at Ch. Charan Singh University, Meerut, UP, India. Here, we describe the most important scientific contributions of our beloved mentor in the area of cytotaxonomy, cytogenetics, mutation breeding, quantitative genetics, molecular biology, crop biotechnology and plant genomics, on his 85th birthday. Important contributions made in the development and use of genomics resources including the development and use of different kinds of molecular markers, genetic and physical mapping, quantitative trait locus (QTL) interval mapping, genome‐wide association mapping and molecular breeding including marker‐assisted selection have been briefly summarized. Efforts have been also made to give readers a glimpse of important contributions in the study of cytology/apomixis of grasses, cytotaxonomic studies in asteraceae/fabaceae, nuclear/repetitive DNA content in Lolium, interspecific/intergeneric relationships involving the genus Hordeum and re‐examining taxonomy of the tribe Triticeae.

Compendium on Food Crop Plants as a Platform for Pharmaceutical Protein Production.

Tremendous advances in crop biotechnology related to the availability of molecular tools and methods developed for transformation and regeneration of specific plant species have been observed. As a consequence, the interest in plant molecular farming aimed at producing the desired therapeutic proteins has significantly increased. Since the middle of the 1980s, recombinant pharmaceuticals have transformed the treatment of many serious diseases and nowadays are used in all branches of medicine. The available systems of the synthesis include wild-type or modified mammalian cells, plants or plant cell cultures, insects, yeast, fungi, or bacteria. Undeniable benefits such as well-characterised breeding conditions, safety, and relatively low costs of production make plants an attractive yet competitive platform for biopharmaceutical production. Some of the vegetable plants that have edible tubers, fruits, leaves, or seeds may be desirable as inexpensive bioreactors because these organs can provide edible vaccines and thus omit the purification step of the final product. Some crucial facts in the development of plant-made pharmaceuticals are presented here in brief. Although crop systems do not require more strictly dedicated optimization of methodologies at any stages of the of biopharmaceutical production process, here we recall the complete framework of such a project, along with theoretical background. Thus, a brief review of the advantages and disadvantages of different systems, the principles for the selection of cis elements for the expression cassettes, and available methods of plant transformation, through to the protein recovery and purification stage, are all presented here. We also outline the achievements in the production of biopharmaceuticals in economically important crop plants and provide examples of their clinical trials and commercialization.

Performance Prediction of Durum Wheat Genotypes in Response to Drought and Heat in Climate Change Conditions.

With an approach combining crop modelling and biotechnology to assess the performance of three durum wheat cultivars (Creso, Duilio, Simeto) in a climate change context, weather and agronomic datasets over the period 1973–2004 from two sites, Benatzu and Ussana (Southern Sardinia, Itay), were used and the model responses were interpreted considering the role of DREB genes in the genotype performance with a focus on drought conditions. The CERES-Wheat crop model was calibrated and validated for grain yield, earliness and kernel weight. Forty-eight synthetic scenarios were used: 6 scenarios with increasing maximum air temperature; 6 scenarios with decreasing rainfall; 36 scenarios combining increasing temperature and decreasing rainfall. The simulated effects on yields, anthesis and kernel weights resulted in yield reduction, increasing kernel weight, and shortened growth duration in both sites. Creso (late cultivar) was the most sensitive to simulated climate conditions. Simeto and Duilio (early cultivars) showed lower simulated yield reductions and a larger anticipation of anthesis date. Observed data showed the same responses for the three cultivars in both sites. The CERES-Wheat model proved to be effective in representing reality and can be used in crop breeding programs with a molecular approach aiming at developing molecular markers for the resistance to drought stress.

Effect of African basil (Ocimum gratissimum) extracts against Water mould (Pythium aphanidermatum) in cowpea

The experiment was designed to investigate the effects of Ocimum gratissimum extracts in the control of Pythium aphanidermatum in cowpea. This was conducted in Crop Science and Biotechnology laboratory, Imo State University, Owerri between May to August 2018 using completely randomized design (C.R.D). There were 8 treatments replicated 4 times. Powder and Liquid extracts of the leaves of Ocimum gratissimum were applied at different concentrations (0,5, 10, 15) g and (0, 5, 10, 15) mls respectively, inoculated on the media, Potato Dextrose Agar (PDA) in petri dishes to check the incidence and spread of the disease. Extracts of Ocimum gratissimum were effective against Pythium aphanidermatum in all the parameters tested. On inoculation of PDA media, different forms of Pythium aphanidermatum (ring or circular and dotted) in all treatments were observed, except the control. Quantitative analyses and Infrared spectroscopy of Ocimum gratissimum leaf extracts revealed the presence of active phytochemicals (alkaloids 3.6, tannins 15.4, flavonoids, oxalate 5.8, saponins 3.1) ug/g and infrared compounds (alkenes (3201.2), alcohol (3344.5), carboxylic acid (2995.5), aldehyde (2719.5), and isocyanate (2804.4) absorbed at different wave lengths in cm-1 were identified.Different fungal growths were cleared using different levels of extracts, (15>10>5>0) mls thanpowder extracts respectively. The ability of the extracts to exert fungicidal effects was due to thepresence of active phytochemicals and infrared compounds present in the extracts.

PYL1- and PYL8-like ABA Receptors of Nicotiana benthamiana Play a Key Role in ABA Response in Seed and Vegetative Tissue.

To face the challenges of climate change and sustainable food production, it is essential to develop crop genome editing techniques to pinpoint key genes involved in abiotic stress signaling. The identification of those prevailing abscisic acid (ABA) receptors that mediate plant-environment interactions is quite challenging in polyploid plants because of the high number of genes in the PYR/PYL/RCAR ABA receptor family. Nicotiana benthamiana is a biotechnological crop amenable to genome editing, and given the importance of ABA signaling in coping with drought stress, we initiated the analysis of its 23-member family of ABA receptors through multiplex CRISPR/Cas9-mediated editing. We generated several high-order mutants impaired in NbPYL1-like and NbPYL8-like receptors, which showed certain insensitivity to ABA for inhibition of seedling establishment, growth, and development of shoot and lateral roots as well as reduced sensitivity to the PYL1-agonist cyanabactin (CB). However, in these high-order mutants, regulation of transpiration was not affected and was responsive to ABA treatment. This reveals a robust and redundant control of transpiration in this allotetraploid plant that probably reflects its origin from the extreme habitat of central Australia.