Transgenic Crops Research Articles

Worldwide study on field trials of biotechnological crops: new promises but old policy hurdles

Field trials (FTs) are a necessary step towards future commercialization of biotech crops and products thereof, whether for research and development or cultivation approval. A total of 187 FTs in 30 countries have been compiled for 2022 and 2023 using a survey and intergovernmental databases. FTs have been classified according to methods, crops and traits. Compiled FTs are mostly conducted by the public sector on eight plant species with improved stress resistance, industrial application, yield, and quality. Regarding genome editing (GenEd), 23 FTs (12% of total) are carried out in 6 countries, on 10 crops. Regulations were examined in 141 countries to discuss why in some countries FTs are not performed, although basic biotech research is carried out. The EU particularly is compared to the rest of the world. Regarding the new proposal in the EU for GenEd product classification, it was found that all recent FTs of such products fall in the category that the EU would consider as ‘equivalent to conventional plants’ (NGT-1). We also studied current cultivation approvals to highlight differences with crops tested in the field and those may be approved in the future.

Selection on synonymous codon usage in soybean (Glycine max) WRKY genes

The WRKY transcription factor gene family in soybean [Glycine max (L.) Merr.] (GmWRKY) is critical for the plant’s development and stress responses. This study examines the evolutionary dynamics of the GmWRKY gene family, focusing on its synonymous codon usage bias (CUB) in a comprehensive set of 179 coding sequences. CUB was analyzed using various indices, revealing a preference for A/T-ending codons and relatively low codon bias. Codon adaptation index (CAI) analysis suggested that these genes are optimized for efficient translation despite relatively low bias, reflecting a balance between codon diversity and translation efficiency. Neutrality and NC plots indicated that selective forces dominate over mutational forces in shaping codon usage, while selection signature analysis showed purifying selection being prevalent across the gene family. However, episodic positive selection was also detected in certain clades, highlighting potential adaptive diversification in response to environmental stress. Additionally, promoter binding site analysis uncovered correlations between codon usage and transcriptional regulation, indicating a context-dependent relationship between CUB and gene expression. Phylogenetic analysis identified 11 well-supported clades in the modern GmWRKY gene family and ancestral sequence reconstruction revealed more relaxed codon preferences and reduced selection constraints in modern GmWRKY genes, potentially linked to neofunctionalization and adaptation to environmental changes. These findings provide a framework for optimizing gene expression in transgenic soybean crops with resilience. Further functional validation of positively selected genes is recommended to elucidate their role in stress responses.

Biological performance of Bollgard II® Bt-resistant vs susceptible population of pink bollworm, Pectinophora gossypiella (Saunders) on non-toxic diet

BackgroundThe pink bollworm, Pectinophora gossypiella (Saunders), is a devastating global pest of cotton that has caused substantial economic damage to Bt Bollgard-II® cotton plants in recent years due to the evolution of Bt resistance. The associated fitness cost is assumed to be one of the factors delaying the development of resistance against Bt transgenic crops. Hence, the present study was undertaken to assess the biological performance of pink bollworms by comparing the life history and demographic parameters of a resistant (Field-R) and susceptible (Lab-S) population.ResultsProlonged larval duration (23.40 days in Field-R vs 18.80 days in Lab-S population), total life cycle (male = 50.00 vs 42.80 days; female = 53.60 vs 46.20 days), reduced fecundity (100.60 vs 154.20 eggs/female) and fertility (88.00 vs 138.00 fertile eggs/female) was observed. The demographic parameters indicated a significant reduction in the net reproductive rate (184.27 vs 276.72), innate capacity for increase in number (0.11 vs 0.15), finite rate of increase in number (1.12 vs 1.16 female progenies produced/female/day), weekly multiplication rate (2.16 vs 2.86), potential fecundity (545.06 vs 634.11 eggs), number of hypothetical F2 females (33 955.65 vs 76 572.41), but longer mean length of generation (47.54 vs 37.74 days) and population doubling time (6.30 vs 4.62 days) in Field-R compared with Lab-S population. A stage-specific life table demonstrated the differences in survival rates between susceptible and resistant populations at various life stages, with the resistant population having higher generation mortality (0.22 vs 0.19).ConclusionsThe study confirms the involvement of fitness costs associated with Bt resistance in P. gossypiella. Despite reduced reproductive fitness, the resistant population tried prolonging the larval stage as a compensatory mechanism to repair the damaged host tissues due to Bt intoxication and for accumulation of enough nutrient reserves for normal pupation and adult emergence. Presence of a high proportion of double Bt-resistant larvae in the field coupled with continued noncompliance with refug planting certainly favours the flaring up of this monophagous pest despite the observed fitness costs. The resistance cannot be effectively reversed unless suitable alternative management strategies are deployed.

Bacillus thuringiensis Cry14A family proteins as novel anthelmintics against gastrointestinal nematode parasites.

Bacillus thuringiensis crystal (Cry) proteins have been expressed in commercial transgenic crops for nearly 30 years, providing safe and effective control of insect pests and significantly reducing the application of hazardous chemical pesticides. B. thuringiensis crystal proteins have also been shown to target parasitic nematodes, including plant parasitic nematodes. Recently, transgenic soybean crops expressing Cry14Ab have been shown to provide control against the soybean cyst nematode Heterodera glycines, marking the first time a crystal protein is being commercialized in transgenic crops for control of a nematode pest. However, apart from H. glycines and the free-living nematode, Caenornabditis elegans, the breadth of nematode activity of Cry14Ab, e.g., against gastrointestinal parasitic nematodes (GINs), has not been reported. Here we study the efficacy of Cry14Ab against a wide range of gastrointestinal nematode parasites (GINs) in vitro and in vivo. We find that Cry14Ab is effective in vitro against the barber's pole worm Haemonchus contortus larvae, small strongyles cyathostomin larvae, the hookworm Ancylostoma ceylanicum adults, the roundworm Ascaris suum L4 larvae, and the whipworm Trichuris muris adults. In rodents infected with GIN parasites, Cry14Ab is effective as an in vivo anthelmintic against the hookworms A. ceylanicum and N. americanus, against the mouse parasite Heligmosomoides polygyrus bakeri, and against the roundworm A. suum. Cry14Ab also variably reduces the reproduction of the whipworm T. muris in vivo. Using optimized profile Markov Models, we looked for other putative anthelmintic Cry proteins and, within this list, identified a Bt crystal protein, GenBank accession no. MF893203, that we produced and demonstrated intoxicated GINs. This protein, with 90% amino acid identity to Cry14Ab, is active against C. elegans, A. ceylanicum adults, and A. suum L4 larvae in vitro. MF893203 was given the official designation of Cry14Ac. Cry14Ac is also an effective in vivo anthelmintic against A. ceylanicum hookworms in hamsters and intestinal A. suum in mice. Taken together, our results demonstrate that Cry14Ab and Cry14Ac have wide therapeutic utility against GINs.

Biodiversity assessment and environmental risk analysis of the single line transgenic pod borer resistant cowpea.

The discussion surrounding biological diversity has reached a critical point with the introduction of Nigeria's first transgenic food crop, the pod borer-resistant (PBR) cowpea. Questions have been raised about the potential risks of the transgenic Maruca vitrata-resistant cowpea to human health and beneficial insects. Public apprehension, coupled with social activists' calling for the removal of this crop from the nation's food market, persists. However, there is a lack of data to counter the assertion that cultivating PBR cowpea may have adverse effects on biodiversity and the overall ecological system. This research, with its multifaceted objective of examining the environmental safety of PBR cowpea and assessing its impact on biodiversity compared to its non-transgenic counterpart, IT97KN, is of utmost importance in providing the necessary data to address these concerns. Seeds for both the transgenic PBR cowpea and its isoline were obtained from the Institute for Agricultural Research (IAR) Zaria before planting at various farm sites (Addae et al., 2020). Throughout the experiment, local cultural practices were strictly followed to cultivate both transgenic and non-transgenic cowpeas. Elaborate taxonomic keys were used to identify arthropods and other non-targeted organisms. Principal component analysis (PCA) was used to evaluate potential modifications in all ecological niches of the crops. The lmer function of the R package lme4 was used to analyze diversity indices, including Shannon, Pielou, and Simpson. The Bray-Curtis index was used to analyzed potential modifications in the dissimilarities of non-targeted organisms' communities. Examination of ecological species abundance per counting week (CW) revealed no disruption in the biological properties of non-targeted species due to the cultivation of transgenic PBR cowpea. Analysis of species evenness and diversity indices indicated no significant difference between the fields of transgenic PBR cowpea and its isoline. Principal component analysis results demonstrated that planting PBR cowpea did not create an imbalance in the distribution of ecological species. All species and families observed during this study were more abundant in transgenic PBR cowpea fields than in non-transgenic cowpea fields, suggesting that the transformation of cowpea does not negatively impact non-targeted organisms and their communities. Evolution dynamics of the species community between transgenic and non-transgenic cowpea fields showed a similar trend throughout the study period, with no significant divergence induced in the community structure because of PBR cowpea planting. This study concludes that planting transgenic PBR cowpea positively influences biodiversity and the environment.

The future is only the beginning

Recent advancements in genome editing have captured the attention of scientists and policymakers, who contend that the technology has a large role to play in advancing food and climate security on the African continent. However, the modest results of earlier generations of biotechnology—such as genetically modified (GM) crops—raise questions about the sustainability of new technological interventions. This special feature examines lessons learned from previous generations of GM crops and other agricultural technologies, using them to analyze the portfolio of gene edited crops being developed for African farmers today. In this article, we introduce the 6 papers that make up the special feature by way of examining future-oriented discourses around the advancement of genome editing. Drawing on Science and Technology Studies, political ecology, and critical development studies, this introduction highlights the crucial factors that shape technology development, agricultural practice, and the politics of knowing and emphasizes the need to look toward multiple, diverse futures.

“Blue-red-purple” multicolored lateral flow immunoassay for simultaneous detection of GM crops utilizing RPA and CRISPR/Cas12a

Advanced multiplexed testing techniques should be designed and developed to ensure an accurate and reliable evaluation for unknown samples. In this study, an efficient platform coupled with the “Blue-Red-Purple” strategy based on recombinant polymerase amplification (RPA), CRISPR/Cas12a and lateral flow strip was established, which could realize the dual-target detection of CP4-EPSPS and Cry1Ab/Ac in genetically modified crops. The lateral flow immunoassay was developed using different colored microspheres to label the antibodies to realize the visualization of results and avoid cross-reactions. The proposed method exhibits high specificity, sensitivity and stability. The visual detection limits of standard plasmids and real samples reached 10 copies/μL and 0.5 %, respectively, which could be stored at 4 °C for 12 months with high detection ability. Moreover, the entire detection process could be completed within 50 min without any complex instruments or professional operators. These findings indicated that a sensitive, specific, rapid and accurate method was established for on-site detection of GM crops.

Mathematical Modelling of Spatially Inhomogeneous Non-Stationary Interaction of Pests with Transgenic and Non-Modified Crops Considering Taxis

Introduction. This paper addresses a unified spatially inhomogeneous, non-stationary model of interaction between genetically modified crop resources (corn) and the corn borer pest, which is also present on a relatively small section of non-modified corn. The model assumes that insect pests influence both types of crops and are capable of independent movement (taxis) towards the gradient of plant resources. It also considers diffusion processes in the dynamics of all components of the unified model, biomass growth, genetic characteristics of both types of plant resources, processes of crop consumption, phenomena of growth and degradation, diffusion, and mutation of pests. The model allows for predictive calculations aimed at reducing crop losses and increasing the resistance of transgenic crops to pests by slowing down the natural mutation rate of the pest. Materials and Methods. The mathematical model is an extension of Kostitsin’s model and is formulated as an initial-boundary value problem for a nonlinear system of convection-diffusion equations. These equations describe the spatiotemporal dynamics of biomass density changes in two types of crops — transgenic and non-modified — as well as the specific populations (densities) of three genotypes of pests (the corn borer) resulting from mutations. The authors linearized the convection-diffusion equations by applying a time-lag method on the time grid, with nonlinear terms from eachequation taken from the previous time layer. The terms describing taxis are presented in a symmetric form, ensuring the skew-symmetry of the corresponding continuous operator and, in the case of spatial grid approximation, the finite-difference operator. Results. A stable monotonic finite-difference scheme is developed, approximating the original problem with second-order accuracy on a uniform 2D spatial grid. Numerical solutions of model problems are provided, qualitatively corresponding to observed processes. Solutions are obtained for various ratios of modified and non-modified sections of the field. Discussion and Conclusion. The obtained results regarding pest behavior, depending on the type of taxis, could significantly extend the time for pests to acquire Bt resistance. The concentration dynamics of pests moving in the direction of the food gradient differs markedly from the concentration of pests moving towards a mate for reproduction.

Considerations for safety assessment of intractable proteins expressed in genetically modified crops

Current requirements for safety assessment by regulatory agencies necessitate the production of multi-gram quantities of functionally active proteins newly expressed in genetically modified (GM) crops. This presents challenges for so-called intractable proteins (those that are difficult to isolate in an active form). Significant experience and knowledge have been gained by both GM crop developers and global regulatory agencies regarding the safety assessment of the newly expressed proteins (NEPs). By leveraging this knowledge and scientific data that have accumulated over the last several decades, the safety assessments of NEPs can be refined and improved. In this technical review, we examine the options for characterizing intractable NEPs and producing protein test substances from different sources to support the safety assessments of novel GM crops.

Overexpressing Carbonic Anhydrase Transgenic Rice Plants Maintain the Regular Soil Functions and Microbial Activities of Soil

Background: Genetically modified (GM) crops are focused on establishing desirable traits to support the food demand of increasing populations. The effect of transgenic plants on soil diversity has been highlighted by a lot of researchers. Some of them showed the negative effect of transgenic crop on soil microbes and the food chain. So there is an urgent need to develop transgenic plants with no harmful effects on environmental health. Methods: We have examined the effects salt tolerant carbonic anhydrase overexpressing transgenic rice plants on the physiology of rhizospheric microbes and their enzymatic activities. In the present study, we have used pot experiments in the greenhouse of Centurion University of Technology and management, Bhubaneswar, Odisha, India. Result: No significant variation in the soil properties viz., pH, Eh, organic C, P, K, N, Ca, Mg, S, Na and Fe+2 were observed. The population of bacteria, fungi and nematodes were remained same in the soil. The enzymatic activities like soil dehydrogenase, nitrate reductase, urease and alkaline phosphatase were not significantly affected. Further, plant growth promotion (PGP) functions such as HCN, siderophore, salicylic acid, GA, IAA, zeatin, were, not influenced considerably. The present study ecologically pertinent of salt tolerant CA transgenic rice to usual functions of the rhizospheric organisms.

Identification, Cloning, and Characterization of Two Acupuncture-Injury-Inducing Promoters in Rice

As an important global food crop, rice is damaged by a variety of piercing–sucking pests. Identifying a broad-spectrum promoter induced by the physical signal of sucking pests and applying it to transgenic breeding to mitigate the damage caused by different sucking pests will significantly improve the efficiency of our breeding. This study compared the transcriptome changes in two rice varieties under needle-wounding stress to investigate their differential responses to mechanical damage. The results showed that the insect-susceptible variety TN1 exhibited more differentially expressed genes (DEGs) and greater changes in expression levels after needle treatment, indicating a more active internal gene regulatory network. GO and KEGG enrichment analysis further revealed that TN1 not only exhibited changes in genes related to the extracellular environment, but also mobilized more genes associated with stress response and defense. By screening the differentially expressed genes, we identified two promoters (P1 and P2) with inducible expression characteristics in both the resistant and susceptible rice varieties. These promoters were able to effectively drive the expression of the insect resistance gene OsLecRK1* and enhance the resistance of transgenic plants against the brown planthopper. This study provides promoter resources for the development of insect-resistant transgenic crops.

Tracing scientific and technological development in genetically modified crops.

Genetically Modified (GM) Organisms have been used in various domains since their introduction in the 1980s. According to ISAAA data, the use of GM crops in agriculture has also increased significantly in the past 30years. However, even after 3 decades of commercialisation, GM crops are still surrounded with controversies with different countries adopting varying approaches to their introduction in the consumer markets, owing to different stances of various stakeholders. Motivated by this multitude of opinions, and absence of knowledge mapping, this study has undertaken scientometric analysis of the publication (Web of Science) and patent (Lens.org) data about genetically modified technology use in agriculture to explore the changing knowledge patterns and technological advancements in the area. It explores both scientific and technological perspectives regarding the use of Genetically Modified Crops, by using publication as well as patent data. The findings of this study highlight the major domains of research, technology development, and leading actors in the ecosystem. These findings can be helpful in taking effective policy decisions, and furthering the research activities. It presents a composite picture using both publications and patent data. Further, it will be of utility to explore the other technologies which are replacing GM technology in agriculture in future studies.

Impact of dual Bt-transgenic maize (2A7) on soil microbial communities and enzyme activities: A comparative study with control variety Z58

The impacts of transgenic crops on soil microbiology and fertility are critical in determining their biosafety. While transgenic crops can alter soil microbes, their effects are often context-dependent; therefore, the ecological importance of these changes remains a topic of ongoing research. Using high-throughput sequencing, we investigated the effects of Bacillus thuringiensis (Bt) maize expressing the mcry1Ab and mcry2Ab genes (2A7) on soil nutrient dynamics, as well as the diversity and function of soil microbial communities, including bacteria and fungi, within different soil compartments. Our findings revealed a plant-shaped rhizosphere (RS) microbial community as a result of the selective recruitment of microorganisms from the surrounding environment. The transgene insertion had a significant impact on the RS niche, and several species eventually became associated with Z58 and 2A7 plants. For example, Neocosmospora rubicola fungal and Pantoea dispersa bacterial microorganisms were significantly decreased in the dual Bt-transgenic 2A7 rhizosphere but enriched in the Z58 rhizospheres. The activity of soil enzymes such as urease, invertase, and alkaline phosphatase was boosted by Bt-transgenic 2A7. LefSe analysis identified significant bacterial and fungal biomarker species that were responsible for the differential effects of Bt-transgenic 2A7 and control Z58 within rhizosphere soils. Mantel analysis further demonstrated that the root exudates of 2A7 altered nutrient-acquisition enzymes by influencing biomarker taxa. PICRUSt2 functional characterization revealed a significantly higher abundance of the phosphate-starvation-inducible protein in control Z58 than in Bt-transgenic 2A7. Furthermore, taxonomy, alpha (Shannon diversity), and beta diversity analyses all revealed niche-driven microbial profile differentiation. Niche partitioning also had a significant impact on N- and P-related COGs as well. Our findings suggests that Bt-transgenic 2A7 modulates rhizosphere microbial communities by affecting biomarker taxa and soil enzyme activity. These findings will promote sustainable agriculture practices by advancing our knowledge of the ecological effects of Bt crops on soil microbial communities.

Downregulation of APN1 and ABCC2 mutation in Bt Cry1Ac-resistant Trichoplusia ni are genetically independent.

The resistance to the insecticidal protein Cry1Ac from the bacterium Bacillus thuringiensis (Bt) in the cabbage looper, Trichoplusia ni, has previously been identified to be associated with a frameshift mutation in the ABC transporter ABCC2 gene and with altered expression of the aminopeptidase N (APN) genes APN1 and APN6, shown as missing of the 110-kDa APN1 (phenotype APN1¯) in larval midgut brush border membrane vesicles (BBMV). In this study, genetic linkage analysis identified that the APN1¯ phenotype and the ABCC2 mutation in Cry1Ac-resistant T. ni segregated independently, although they were always associated under Cry1Ac selection. The ABCC2 mutation and APN1¯ phenotype were separated into two T. ni strains respectively. Bioassays of the T. ni strains with Cry1Ac determined that the T. ni with the APN1¯ phenotype showed a low level resistance to Cry1Ac (3.5-fold), and the associated resistance is incompletely dominant in the background of the ABCC2 mutation. Whereas the ABCC2 mutation-associated resistance to Cry1Ac is at a moderate level, and the resistance is incompletely recessive in the genetic background of downregulated APN1. Analysis of Cry1Ac binding to larval midgut BBMV indicated that the midgut in larvae with the APN1¯ phenotype had reduced binding affinity for Cry1Ac, but the number of binding sites remained unchanged, and the midgut in larvae with the ABCC2 mutation had both reduced binding affinity and reduced number of binding sites for Cry1Ac. The reduced Cry1Ac binding to BBMV from larvae with the ABCC2 mutation or APN1¯ phenotype correlated with the lower levels of resistance.IMPORTANCEThe soil bacterium Bacillus thuringiensis (Bt) is an important insect pathogen used as a bioinsecticide for pest control. Bt genes coding for insecticidal proteins are the primary transgenes engineered into transgenic crops (Bt crops) to confer insect resistance. However, the evolution of resistance to Bt proteins in insect populations in response to exposure to Bt threatens the sustainable application of Bt biotechnology. Cry1Ac is a major insecticidal toxin utilized for insect control. Genetic mechanisms of insect resistance to Cry1Ac are complex and require to be better understood. The resistance to Cry1Ac in Trichoplusia ni is associated with a mutation in the ABCC2 gene and also associated with the APN expression phenotype APN1¯. This study identified the genetic independence of the APN1¯ phenotype from the ABCC2 mutation and isolated and analyzed the ABCC2 mutation-associated and APN1¯ phenotype-associated resistance traits in T. ni to provide new insights into the genetic mechanisms of Cry1Ac resistance in insects.

Advancements in genetically modified insect pest-resistant crops in India.

The review offers insights into the current state of research on insect pest-resistant GM crops and the regulations governing the cultivation of GM crops in India. India has a rich crop diversity of more than 160 major and minor crops through its diverse agroclimatic conditions. Insect pests alone cause around USD 36 billion in crop loss annually in India. The last two decades witnessed considerable progress in managing insect pests by adopting innovative techniques including transgenics. In research, significant advancement has been brought in insect pest-resistant transgenics in India since its inception in 2002. However, any events have not been endorsed owing to biosafety impediments, except Bt cotton reaching the commercial release stage. A landmark decision to exempt certain types of gene-edited plants from genetically modified organism (GMO) regulations offers great promise for developing novel insect-resistant crops in India. The article reviews the current research on insect pest-resistant transgenics and its regulations in India.

Identification and functional characterization of bidirectional gene pairs and their intergenic regions in cotton

BackgroundIn research to improve the quality of transgenic crops, it is often necessary to introduce multiple functionally related genes into recipient plants simultaneously to improve crop genetic traits effectively. Compared with unidirectional promoters, bidirectional promoters simultaneously regulate the expression of multiple genes and improve the efficiency of biotechnology. Therefore, in this study, bidirectional gene pairs were systematically analyzed in Gossypium hirsutum TM-1, and the structure, function and evolutionary relationships of the bidirectional genes were analyzed. The endogenous bidirectional promoters of cotton were mined, and their specific regulatory elements and biological functions were explored to provide useful promoter resources and a theoretical basis for cultivating new cotton germplasms with excellent fiber quality.ResultsUsing an improved search model, a total of 1,383 bidirectional transcript pairs were identified in the Gossypium hirsutum TM-1 genome, and their gene structure and functional annotations were systematically analyzed. Thirty bidirectional intergenic sequences were randomly screened for promoter activity analysis via a transient expression system, and 25 intergenic sequences were found to have bidirectional promoter activity. Comparative analysis of the bidirectional gene profiles of the four cotton subspecies revealed that these subspecies presented abundant bidirectional gene pairs with high homology and that the bidirectional genes in the cotton subspecies were more similar in terms of their molecular functions, cellular components and biological processes. In addition, parallel analysis of bidirectional genes in dicotyledons and monocotyledons revealed that abundant bidirectional gene pairs exist in different species. Although the total number of orthologous bidirectional genes was similar, there was a significant difference in the number of orthologous bidirectional gene pairs between dicotyledons and monocotyledons. This evolutionary analysis of the function and structure of homologous bidirectional gene pairs in different varieties and different subspecies of the same species revealed potential pathways by which these gene pairs originated, which may be necessary for the evolution of a new species.ConclusionIn this study, many bidirectional gene pairs in Gossypium hirsutum TM-1 were identified using computer programming, and systematic analysis was conducted to explore their functions and evolutionary relationships. In addition, the promoter activity of the bidirectional intergenic sequences was verified. The combination of computer programming screening, experimental validation and other methods is expected to provide preferred bidirectional promoters for transgenic breeding work via multigene cotransformation methods, and this information is valuable for genetic engineering research and applications.

Inheritance and ecological effects of exogenous genes from transgenic Brassica napus to Brassica juncea hybrids

●With the rapid development of new breeding techniques, the ecological impacts of transgenic plants receive wide concern again, particularly for potential gene flow from transgenic crops to their relatives. The transgene insertion position, number of gene copies, and flanking sequence of exogenous genes integrated into the recipient genome affect the genetic stability and fitness of offspring.●We employed hybrids F1 and F2, six backcross generations BC1-BC6 and BC1F1 from transgenic Brassica napus with Bacillus thuringiensis (Bt) cry1Ac gene and its wild relative B. juncea through hand pollination. We detected exogenous gene copies, mRNA transcription, and protein expression by ddPCR, qRT-PCR and ELISA, and the fitness of hybrid and backcross generations.●Exogenous genes followed Mendelian segregation expectations in hybrid and backcrossed generations, with stable gene copies, mRNA transcription and protein concentration of exogenous genes in offspring. Exogenous gene copies had no significant effects on plant fitness, but had positive effects on mRNA transcription and protein concentration that varied with growth stages in hybrid and backcross generations.●Our study demonstrated inheritance and ecological effects of exogenous genes from transgenic plants to wild relatives, which will help ecological risk management of biotechnological plants released in the nature.

Species-specific evolution of lepidopteran TspC5 tetraspanins associated with dominant resistance to Bacillus thuringiensis toxin Cry1Ac

Transgenic crops producing insecticidal proteins from the bacterium Bacillus thuringiensis (Bt) have proven to be highly effective in managing some key pests. However, the evolution of resistance by the target pests threatens the sustainability of Bt crops. The L31S mutation in a tetraspanin encoded by HarmTspC5 (previously known as HaTSPAN1) has been shown to confer dominant resistance to the Bt protein Cry1Ac in Helicoverpa armigera, a globally damaging lepidopteran pest. However, the broader implications of the L31S mutation in the tetraspanins of other lepidopteran species remain unclear. The evolutionary analyses in this study indicate that TspC5s have evolved in a species-specific manner among the lepidopteran insects. To investigate the role of TspC5s in conferring dominant resistance to Cry1Ac, we used the piggyBac-based transformation system to generate four transgenic H. armigera strains that express exogenous TspC5 variants from three phylogenetically close species (Helicoverpa zea, Helicoverpa assulta and Heliothis virescens) and one phylogenetically distant species (Plutella xylostella). In comparison with the background SCD strain of H. armigera, the transgenic strains expressing HzeaTspC5-L31S, HassTspC5-L31S, or HvirTspC5-L31S exhibited significant resistance to Cry1Ac (10.0-, 21.4-, and 81.1-fold, respectively), whereas the strain expressing PxylTspC5-L27S remained susceptible. Furthermore, the Cry1Ac resistant phenotypes followed an autosomal dominant inheritance pattern and were closely linked to the introduced mutant TspC5s. These findings reveal the conserved role of TspC5s from Helicoverpa and Heliothis species in mediating the dominant resistance to Cry1Ac, and they provide crucial insights for assessing resistance risks related to mutant tetraspanins and devising adaptive resistance management strategies for these major lepidopteran pests.

Environmental impacts of genetically modified crops.

Genetically modified (GM) crops have been adopted by some of the world's leading agricultural nations, but the full extent of their environmental impact remains largely unknown. Although concerns regarding the direct environmental effects of GM crops have declined, GM crops have led to indirect changes in agricultural practices, including pesticide use, agricultural expansion, and cropping patterns, with profound environmental implications. Recent studies paint a nuanced picture of these environmental impacts, with mixed effects of GM crop adoption on biodiversity, deforestation, and human health that vary with the GM trait and geographic scale. New GM or gene-edited crops with different traits would likely have different environmental and human health impacts.

The B-box transcription factor BnBBX22.A07 enhances salt stress tolerance by indirectly activating BnWRKY33.C03.

Salt stress has a detrimental impact on both plant growth and global crop yields. B-box proteins have emerged as pivotal players in plant growth and development regulation. Although the precise role of B-box proteins orchestrating salt stress responses in B. napus (Brassica napus) is not well understood in the current literature, further research and molecular explorations are required. Here, we isolated the B-box protein BnBBX22.A07 from B. napus. The overexpression of BnBBX22.A07 significantly improved the salt tolerance of Arabidopsis (Arabidopsis thaliana) and B. napus. Transcriptomic and histological analysis showed that BnBBX22.A07 enhanced the salt tolerance of B. napus by activating the expression of reactive oxygen species (ROS) scavenging-related genes and decreasing salt-induced superoxide anions and hydrogen peroxide. Moreover, BnBBX22.A07 interacted with BnHY5.C09, which specifically bound to and activated the promoter of BnWRKY33.C03. The presence of BnBBX22.A07 enhanced the activation of BnHY5.C09 on BnWRKY33.C03. Overexpression of BnHY5.C09 and BnWRKY33.C03 improved the salt tolerance of Arabidopsis. Functional analyses revealed that BnBBX22.A07-mediated salt tolerance was partly dependent on WRKY33. Taken together, we demonstrate that BnBBX22.A07 functions positively in salt responses not only by activating ROS scavenging-related genes but also by indirectly activating BnWRKY33.C03. Notably, our study offers a promising avenue for the identification of candidate genes that could be harnessed in breeding endeavours to develop salt-resistant transgenic crops.