Transgenic Varieties Research Articles

Identification of the sweet orange (Citrus sinensis) bHLH gene family and the role of CsbHLH55 and CsbHLH87 in regulating salt stress.

Salt stress is one of the primary environmental stresses limiting plant growth and production and adversely affecting the growth, development, yield, and fruit quality of Citrus sinensis. bHLH (basic helix-loop-helix) genes are involved in many bioregulatory processes in plants, including growth and development, phytohormone signaling, defense responses, and biosynthesis of specific metabolites. In this study, by bioinformatics methods, 120 CsbHLHgenes were identified, and phylogenetic analysis classified them into 18 subfamilies that were unevenly distributed on nine chromosomes. The cis-acting elements of the CsbHLH genes were mainly hormone-related cis-acting elements. Seventeen CsbHLH genes exhibited significant differences in expression under salt stress. Six CsbHLH genes with significant differences in expression were randomly selected for quantitative real-time polymerase chain reaction (qRT-PCR) validation. The qRT-PCR results showed a strong correlation with the transcriptome data. Phytohormones such as jasmonic acid (JA) are essential for biotic and abiotic stress responses in plants, and CsbHLH55 and CsbHLH87 are considered candidate target genes for sweet orange MYC2 transcription factors involved in the JA signaling pathway. These genes are the main downstream effectors in the JA signaling pathway and can be activated to participate in the JA signaling pathway. Activation of the JA signaling pathway inhibits the production of reactive oxygen species and improves the salt tolerance of sweet orange plants. The CsbHLH55 and CsbHLH87 genes could be candidate genes for breeding new transgenic salt-resistant varieties of sweet orange.

Low temperature exposure influences nitrogen metabolism resulting in decreased Cry1Ac insecticidal endotoxin content in cotton seeds

BackgroundSudden temperature drops, resulting from extreme weather events, often occur during the boll-setting period of cotton in Xinjiang, China, causing decreased expression of Bacillus thuringiensis (Bt) insecticidal proteins in cotton bolls. The precise threshold temperatures and durations that lead to significant changes in Cry1Ac endotoxin levels under low temperatures remain unclear. To address this, we investigated the effects of different temperatures and stress durations on Cry1Ac endotoxin levels in cotton bolls. In 2020–2021, two Bt transgenic cotton varieties, conventional Sikang1 and hybrid Sikang3, were selected as experimental materials. Various low temperatures (ranging from 16 to 20 °C) with different durations (12 h, 24 h and 48 h) were applied during the peak boll-setting period.ResultsAs the temperature decreased, the Cry1Ac endotoxin content in the boll shell, fiber, and seed exhibited a declining trend. Moreover, the threshold temperature which caused a significant reduction in Cry1Ac endotoxin content increased with the prolonged duration of low-temperature stress. Among the components of cotton bolls, seeds were most affected by low-temperature stress, with the threshold temperature for a significant reduction in Cry1Ac endotoxin content ranging from 17 °C to 19 °C. Correlation analysis indicated that low temperatures led to a decrease in protein synthesis capacity and an increase in degradation ability, resulting in reduced Cry1Ac endotoxin content. Pathway analysis revealed that both free amino acid and peptidase had significant negative effects on Cry1Ac endotoxin content.ConclusionIn summary, when the daily average temperature was ≤ 19 °C, implementing cultural practices to reduce free amino acid content and peptidase activity could serve as effective cold defense strategies for Bt cotton production.

Biofortification of Major Crops through Conventional and Modern Biotechnological Approaches to Fight Hidden Hunger: An Overview

Biofortification, the process of enhancing the nutritional content of crops, offers a promising strategy to combat hidden hunger—micronutrient deficiencies affecting over two billion people globally. This review article explores the biofortification of major crops, focusing on both conventional breeding techniques and modern biotechnological approaches. Conventional methods, such as selective breeding and crossbreeding, have been instrumental in increasing the levels of essential micronutrients like iron (Fe) and zinc (Zn) in staple crops such as wheat, rice, and maize. For instance, wild relatives of cultivated wheat, including Triticum dicoccoides and Aegilops tauschii, have been utilized to significantly enhance Fe and Zn content in modern cultivars. Advancements in biotechnological tools, including genetic engineering, marker-assisted selection (MAS), and genome editing (CRISPR/Cas9), have further accelerated the development of biofortified crops. These technologies enable precise modifications to increase the accumulation of micronutrients and improve nutrient bioavailability. For example, transgenic rice varieties enriched with β-carotene (Golden Rice) and enhanced Fe and Zn content through gene editing showcase the potential of biotechnology in addressing micronutrient deficiencies. The review also highlights ongoing efforts and challenges in the field, such as regulatory hurdles, public acceptance, and the need for comprehensive strategies integrating conventional and modern approaches. Furthermore, it discusses the role of international research organizations and collaborations in facilitating the development and dissemination of biofortified crops. In conclusion, combining conventional breeding with cutting-edge biotechnological innovations presents a robust approach to biofortify major crops, offering a sustainable solution to mitigate hidden hunger and improve global food security. Continued research and multi-disciplinary collaborations are essential to fully realize the potential of biofortification in enhancing human nutrition.

Discrimination of maize transgenic and non-transgenic varieties by laser induced spectroscopy (LIBS) and machine learning algorithms

BackgroundIn the last decades, the production and consumption of genetically modified agricultural products have increased markedly due to the worldwide population growth and rising demand for food and feed. Consequently, genetically modified crops have been extensively produced and consumed, which required identifying and discriminating transgenic and non-transgenic products. ResultsLaser-induced breakdown spectroscopy (LIBS) combined with chemometric methods was applied to identify and discriminate two varieties of conventional (not-genetically modified, NGM) maize from four varieties of transgenic maize (genetically modified, GM). The LIBS spectra acquired under reduced pressure (100 Torr) conditions over two ranges, i.e., 175–330 nm and 275–770 nm, were subjected to Standard Normal Variation (SNV) and multivariate methods such as Principal Component Analysis (PCA) to reduce data matrices dimensionality and spectral noise. The supervised machine learning algorithms k-nearest neighbor (k-NN) and support vector machine (SVM) have been applied to discriminate among NGM and GM maize reserving 25 % of data for external validation. The training data were employed for hyperparameter optimization of classifiers using the Leave-One-Out Cross-Validation (LOOCV) method. Considering all six maize varieties simultaneously, the highest training accuracy achieved was 90.56 %, with an external validation accuracy of 88.33 %. In an alternative approach based on pairwise combinations of one GM variety against one NGM variety, the best outcome achieved was 100 % LOOCV and external validation accuracy. ConclusionsThese results showed that LIBS supported by appropriate chemometric methods represents an alternative screening technique for identifying and discriminating transgenic from non-transgenic maize.

Impact Analysis of Biorational Insecticides under Weeded and Non Weeded Habitat on Whitefly, Bemisia tabaci Management in Cotton Agroecosystem

Transgenic Bt cotton is effective to a high degree of, with respect to target lepidoptran bollworms, however sucking pests especially whitefly posed a serious issues in absence of bollworm in genetically modified (GM) crops. Therefore, to understand the efficacy of various biorational compounds against whitefly under non-weedy as well as weedy field conditions, studies on whitefly carried out during kharif 2018, to know the changes in number of whitefly adults as well as nymphs. Transgenic hybrid cotton variety RCH 650 was sown with a spacing of 100×45 cm and replicated four times in both non-weedy and weedy habitat. Although, standard check Dimethoate 30 EC proved it’s efficacy after first and second spray in cotton crop against whitefly. Not with standing, after the application of biorational first time as spray in 29th Standard Meteoroloical Week (SMW), Nimbecidine 300 ppm was admitted as most effective in non-weedy (66.76 and 60.65% mean mortality in whitefly adult and nymph, respectively) as well as in weedy habitat that caused 56.77 and 55.54% mean mortality in adult and nymph, respectively over a period of seven days after application. Entomopathogenic fungi Verticillium lecanii found better to control whitefly adults (42.06 and 40.63%), while Metarhizium anisopliae (42.39 and 36.01%) found more effective for nymphal mortality in non-weedy as well as in weedy habitat. One and same pattern of efficacy of biorational was reconstructed against whitefly again after second application of biorational in all the treatment selected to test against whitefly.

Effect of soil pH and mineralogy on the sorption and desorption of phosphite and phosphate in Ultisols of the Southeastern Coastal Plain

AbstractThe development of transgenic crop varieties capable of utilizing phosphite (Phi) as a phosphorus (P) source is a promising strategy to increase P use efficiency while decreasing reliance on phosphate (Pi)‐based fertilizers. However, little information is available on Phi sorption and desorption in soils. We conducted batch experiments to investigate the sorption of Phi and Pi by three Ultisols from the Coastal Plain of the southeastern United States. At the soils' acidic field pH (pH 4.7–6.2), Pi had a higher affinity than Phi for soil solids, where maximum sorbed concentrations of Pi were an average of 44% greater than those of Phi. The sorption of both P species decreased when experiments were repeated adjusting soil pH to 6.5. More Phi than Pi was recovered during desorption experiments, indicating that Phi was more reversibly sorbed and, therefore, may be more plant‐available than Pi. Multiple linear regression between calculated linear distribution coefficients (KD; 10 mg P L−1 input solutions) and soil properties suggested that amorphous Al‐ and Fe‐oxides controlled Pi sorption. Alternatively, amorphous Al‐oxides and gibbsite controlled Phi sorption. Our results show that a lower affinity of Phi than Pi in Ultisols could improve P availability for plant uptake of Phi‐based fertilizers but may increase the risk of soil P buildup over time.

Bt-Modified Transgenic Rice May Shift the Composition and Diversity of Rhizosphere Microbiota.

Plants significantly shape root-associated microbiota, making rhizosphere microbes useful environmental indicator organisms for safety assessment. Here, we report the pyrosequencing of the bacterial 16S ribosomal RNA in rhizosphere soil samples collected from transgenic cry1Ab/cry1Ac Bt rice Huahui No. 1 (GM crop) and its parental counterpart, Minghui63. We identified a total of 2579 quantifiable bacterial operational taxonomic units (OTUs). Many treatment-enriched microbial OTUs were identified, including 14 NonGM-enriched OTUs and 10 GM-enriched OTUs. OTUs belonging to the phyla Proteobacteria, Actinobacteria, Acidobacteria, Firmicutes, Nitrospirae, Chlorobi and GN04 were identified as statistically different in abundance between GM and the other two treatments. Compared with the different impacts of different rice varieties on microbiota, the impact of rice planting on microbiota is more obvious. Furthermore, Huahui No. 1 transgenic Bt rice had a greater impact on the rhizosphere bacterial communities than Minghui63. Early developmental stages of the transgenic Bt rice had a significant impact on many Bacillaceae communities. Soil chemical properties were not significantly altered by the presence of transgenic Bt rice. The peak concentration level of Bt protein products was detected during the seedling stage of transgenic Bt rice, which may be an intriguing factor for bacterial diversity variations. Based on these findings, we conclude that transgenic Bt rice has a significant impact on root-associated bacteria. This information may be leveraged in future environmental safety assessments of transgenic Bt rice varieties.

Susceptibility of Fall Armyworm Field Populations to Vip3Aa/Cry Bt Maize in a Tropical Agricultural Region

Fall armyworm (FAW, Spodoptera frugiperda) is a polyphagous and migratory lepidopteran pest insect in field crops and is notoriously invasive worldwide. In large portions of the Americas, its populations are managed using transgenic maize or cotton varieties producing insecticidal proteins from Bacillus thuringiensis (Bt), primarily Vip3Aa pyramided with Cry Bt proteins. We determined the susceptibility of FAW field populations from locations pressured with such maize hybrids for over five years. We used time–mortality bioassays with F1 third-instar larvae of six geographically distinct populations collected in maize fields of a tropical agricultural region encompassing four Brazilian states. We maintained the neonate progeny from the field populations on an artificial diet until the third instar, and then determined their survival curves on the foliage of three Vip3Aa/Cry-producing Bt maize hybrids. Death of the mid-size, third-instar FAWs occurred relatively rapidly, with larval mortality rates reaching 98–100% in less than five days regardless of Bt maize hybrid. However, median survival time (ST50) for the larvae differed among the populations, with the lowest and highest ST50 values occurring for PI-Cr (42 h, 1.75 d) and PI-Ur populations (66–90 h, 2.75–3.75 h), respectively. Therefore, the F1 third-instar larvae of FAW populations were largely susceptible to Vip3Aa/Cry-producing maize foliage, and the most contrasting susceptibility occurred in the insects from Piauí state, Brazil. These results indicate that progeny of FAWs from areas highly pressured with Vip3Aa/Cry Bt maize hybrids are killed on maize foliage producing Vip3Aa and Cry Bt proteins despite field reports of increased leaf damage by the larvae in some locations. This research informs decision making for Bt-crop resistance management by producers, technicians, and researchers in local, regional, and world agriculture.

From genes to fields: Environmental compatibility of herbicide tolerant transgenic cotton

IntroductionThere are several enticing reasons to use genetic engineering to develop transgenic cotton plants with economically relevant new traits. The use of transgenic cotton to protect crops from biotic stressors like weeds may prove to be a cost-effective solution. ObjectivesThe development of herbicide-resistant cotton variants is the primary motivation for gene transfer in this crop. In the current study, we conducted a battery of experiments on the herbicide-tolerant cotton variety MNH-786 to investigate its expression patterns and environmental adaptation. Material and methodsThe 35S promoter controls the expression of the codon-optimized Cp4-EPSPS gene that is present in this variety. Several different molecular approaches, such as polymerase chain reaction (PCR), Florescent In-Situ Hybridization analysis, and enzyme-linked immunosorbent assay (ELISA), are used to confirm the presence of transgenic cotton plants. The glyphosate spray assay was performed using molecular analysis, employing a dosage of 2000 ml per acre. The suitability of transgenic and non-transgenic cotton plants in four unique environments—the Multan, Vehari, Bahawalpur, and Faisalabad Districts of Punjab province, Pakistan was assessed using a comparative comparison. A gene x environment interaction (GEI) biplot was used to analyze the correlation between genes and their environments. ResultsTransgenic cotton plants screened using PCR to target 1.467 kb regions showed effective integration into cotton genomes. Results showed that Cp4-EPSPS proteins fluctuated and associated with environmental variables across all three generations (T0, T1, and T2). In the herbicide spray assay, all transgenic cotton plants survived, but non-transgenic cotton and all weeds died. According to the "which-won-where" polygon, transgenic cotton plant Events 3, 4, 5, 6, and 7 performed better in three settings, except Faisalabad. According to correlation and heat-map yield research, transgenic cotton variety MNH-786 Events 4, 5 and 6 have a higher consistency and influence in Vehari district's natural habitats. Multan and Bahawalpur also show encouraging results. In Faisalabad, no transgenic events perform well. ConclusionWe conclude that Events 4 and 5 show a substantial abundance of genetic potential that is robust over a wide range of environments thanks to our thorough study of the outcomes obtained under these conditions. SignificanceOur study emphasises the importance of transgenic cotton, its performance, and its exposure to appropriate agroecological zones. This knowledge can be advantageous for farmers in attaining optimal conditions to maximize productivity.

Differentiation of the Pathotypes in Race 29 of cotton Bacterial Leaf Blight Pathogen Xanthomonas axonopodis pv. malvacearum (Xam) and the reaction of some transgenic ( Bt ) and non-transgenic cotton varieties grown in Vidharbha region to the prevalent Xam races and pathotypes.

Differentiation of the Pathotypes in Race 29 of cotton Bacterial Leaf Blight Pathogen Xanthomonas axonopodis pv. malvacearum (Xam) and the reaction of some transgenic ( Bt ) and non-transgenic cotton varieties grown in Vidharbha region to the prevalent Xam races and pathotypes.

The potential of cardiac xenotransplantation for management of infants with complex congenital heart disease.

Gene editing of the porcine genome has enabled the production of pigs that do not express the three known carbohydrate antigens that are associated with hyperacute rejection of a pig organ xenotransplant. In addition, it is now possible to insert a variety of human transgenes to protect against the human immune response, e.g., to protect from complement and coagulation activation. As a result, cardiac xenotransplantation of the gene-edited porcine heart is progressing towards clinical application. Many hope that it will definitively address the disparity between organ supply and demand. The role of cardiac xenotransplantation in pediatric care remains controversial but we believe there is an infant patient population with complex congenital heart disease (CHD) (not optimally managed by conventional surgical approaches) that is ideally suited to initial clinical application of this new technology. The most efficacious start would be to initiate clinical use as a short-term bridge to allotransplantation, particularly in infants with single ventricle pathology and significant risk factors for first stage Norwood palliation. Infants with end-stage heart failure after first stage palliation would represent a second target population. Infants experience unacceptably high mortality and morbidity when placed on mechanical circulatory support as a bridge to allotransplant. Effectively bridging these vulnerable populations could promote acceptance of cardiac xenotransplantation, allowing indications and use to expand, e.g., by (I) bridging patients with failed second and third stage single ventricle disease, or (II) with complex biventricular CHD, or (III) those with a restrictive or dilated cardiomyopathy. Finally, there is a reasonable expectation that the immunologic privilege of infants will allow porcine heart xenotransplantation to be destination therapy for some patients. In summary, heart allotransplantation in infants offers superior outcomes when compared to three-stage single ventricle palliation, but there is a continual shortage of deceased human donor organs. We should pursue research towards the application of xenotransplantation in patients with single ventricle pathology, in whom the results of staged palliation are likely to be suboptimal. There are many remaining issues to be resolved before cardiac xenotransplantation enters regular pediatric clinical use, but experience in this field is progressing rapidly.

Analysis of the Unintended Effects of the Bacillus thuringiensis Insecticidal Protein in Genetically Modified Rice Using Untargeted Transcriptomics

The safety and unintended effects of genetically modified (GM) crops have been the focus of public attention. Transcriptome analysis is a powerful tool to assess the potential impact of genetic modification on plant genomes. In this study, three transgenic (KMD, KF6, and TT51-1) and three non-transgenic (XS11, MH86, and MH63) rice varieties were assessed at the genomic and protein levels. The results of polymerase chain reaction (PCR) and Cry1Ab/1Ac speed test strips showed that the Bt gene was successfully expressed in transgenic rice. The results of RNA-seq analysis to analyze the unintended effects of transgenic Bt rice showed fewer differentially expressed genes (DEGs) between the transgenic and non-transgenic rice varieties than among the different varieties. Meanwhile, the results of principal component analysis and cluster analysis found no significant genetic variation between the transgenic and non-transgenic rice varieties, except for the presence of Bt in transgenic rice. There were only two co-upregulated DEGs and no co-downregulated DEGs among three comparison groups. Although there were various DEGs among the groups, the two co-upregulated DEGs were not related to any significantly enriched gene ontology (GO) term or Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway, indicating that the differences among the subgroups were more likely caused by complex environmental or genetic factors, rather than unintended effects due to Bt expression. This study provides useful information to further explore the unexpected effects and safety of GM rice.

Five-years post commercial approval monitoring of eucalyptus H421.

Eucalyptus comprises the largest planted area of cultivated production forest in Brazil. Genetic modification of eucalyptus can provide additional characteristics for increasing productivity, protecting plant yield, and potentially altering fiber for various industrial uses. With this objective, a transgenic eucalyptus variety, event H421, received regulatory approval for commercial release after 6years of approved risk assessment studies by the Brazilian National Technical Biosafety Commission (CTNBio) in 2015, becoming the first approved genetically modified (GM) eucalyptus in the world. GM event H421 enables increased plant biomass accumulation through overexpression of the Arabidopsis 1,4-β-endoglucanase Cel1, which remodels the xyloglucan-cellulose matrix of the cell wall during development to promote cell expansion and growth. As required, in that time, by the current normative from CTNBio, a post-commercial release monitoring plan for H421 was submitted, incorporating general surveillance for five consecutive years with the submission of annual reports. The monitoring plan was conducted on fields of H421 progenies, with conventional clones as comparators, cultivated in representative regions where eucalyptus is cultivated in the states of São Paulo, Bahia, and Maranhão, representing Southeast, Northeast, and Northern Brazil. Over the course of the five-year general surveillance monitoring plan for the approved GM eucalyptus H421, no adverse effect that could impact the biosafety of the commercially approved event was identified. Additionally, the GM eucalyptus exhibited behavior highly consistent with that of conventional commercial clones. Therefore, there was no need for an extra risk assessment study of a case-specific monitoring plan. The results show the importance of continuously updating the regulation norms of governmental agencies to align with scientific advances.

Biotic stress alleviating strategies in chickpea

The third-most important food legume in terms of economic importance worldwide is the chickpea (Cicer arietinum L.). Its potential production is frequently constrained by numerous biotic stressors, such as the nematodes, insects Ascochyta blight, fusarsium wilt, and botrytis grey mould are the three major fungal diseases that cause significant economic losses, while Helicoverpa armigera, Aphis craccivora, cowpea weevil are the three major pre-harvest pest of chickpea. Several biological, chemical, cultural and, agronomical practices are used to control biotic stress, apart from that few modern biotechnological approaches also developed for high yielding and biotic stress resistant varieties. This paper aims to elaborate about different biotic stresses that affect Chickpea plant, their management strategies including traditional chemicals and adaptation of transgenic varieties with their limitations and also enlightened newer ray of hope i.e., plant growth promoting rhizobacteria that holds the ability to combat against biotic stress by mitigating stress ethylene level.

Strategies for delivery of CRISPR/Cas-mediated genome editing to obtain edited plants directly without transgene integration.

Increased understanding of plant genetics and the development of powerful and easier-to-use gene editing tools over the past century have revolutionized humankind's ability to deliver precise genotypes in crops. Plant transformation techniques are well developed for making transgenic varieties in certain crops and model organisms, yet reagent delivery and plant regeneration remain key bottlenecks to applying the technology of gene editing to most crops. Typical plant transformation protocols to produce transgenic, genetically modified (GM) varieties rely on transgenes, chemical selection, and tissue culture. Typical protocols to make gene edited (GE) varieties also use transgenes, even though these may be undesirable in the final crop product. In some crops, the transgenes are routinely segregated away during meiosis by performing crosses, and thus only a minor concern. In other crops, particularly those propagated vegetatively, complex hybrids, or crops with long generation times, such crosses are impractical or impossible. This review highlights diverse strategies to deliver CRISPR/Cas gene editing reagents to regenerable plant cells and to recover edited plants without unwanted integration of transgenes. Some examples include delivering DNA-free gene editing reagents such as ribonucleoproteins or mRNA, relying on reagent expression from non-integrated DNA, using novel delivery mechanisms such as viruses or nanoparticles, using unconventional selection methods to avoid integration of transgenes, and/or avoiding tissue culture altogether. These methods are advancing rapidly and already enabling crop scientists to make use of the precision of CRISPR gene editing tools.

The current scenario and future perspectives of transgenic oilseed mustard by CRISPR-Cas9.

Production of a designer crop having added attributes is the primary goal of all plant biotechnologists. Specifically, development of a crop with a simple biotechnological approach and at a rapid pace is most desirable. Genetic engineering enables us to displace genes among species. The newly incorporated foreign gene(s) in the host genome can create a new trait(s) by regulating the genotypes and/or phenotypes. The advent of the CRISPR-Cas9 tools has enabled the modification of a plant genome easily by introducing mutation or replacing genomic fragment. Oilseed mustard varieties (e.g., Brassica juncea, Brassica nigra, Brassica napus, and Brassica carinata) are one such plants, which have been transformed with different genes isolated from the wide range of species. Current reports proved that the yield and value of oilseed mustard has been tremendously improved by the introduction of stably inherited new traits such as insect and herbicide resistance. However, the genetic transformation of oilseed mustard remains incompetent due to lack of potential plant transformation systems. To solve numerous complications involved in genetically modified oilseed mustard crop varieties regeneration procedures, scientific research is being conducted to rectify the unwanted complications. Thus, this study provides a broader overview of the present status of new traits introduced in each mentioned varieties of oilseed mustard plant by different genetical engineering tools, especially CRISPR-Cas9, which will be useful to improve the transformation system of oilseed mustard crop plants. This review presents recent improvements made in oilseed mustard genetic engineering methodologies by using CRISPR-Cas9 tools, present status of new traits introduced in oilseed mustard plant varieties. The review highlighted that the transgenic oilseed mustard production is a challenging process and the transgenic varieties of oilseed mustard provide a powerful tool for enhanced mustard yield. Over expression studies and silencing of desired genes provide functional importance of genes involved in mustard growth and development under different biotic and abiotic stress conditions. Thus, it can be expected that in near future CRISPR can contribute enormously in improving the mustard plant's architecture and develop stress resilient oilseed mustard plant species.

Plant protection from virus: a review of different approaches.

This review analyzes methods for controlling plant viral infection. The high harmfulness of viral diseases and the peculiarities of viral pathogenesis impose special requirements regarding developing methods to prevent phytoviruses. The control of viral infection is complicated by the rapid evolution, variability of viruses, and the peculiarities of their pathogenesis. Viral infection in plants is a complex interdependent process. The creation of transgenic varieties has caused much hope in the fight against viral pathogens. The disadvantages of genetically engineered approaches include the fact that the resistance gained is often highly specific and short-lived, and there are bans in many countries on the use of transgenic varieties. Modern prevention methods, diagnosis, and recovery of planting material are at the forefront of the fight against viral infection. The main techniques used for the healing of virus-infected plants include the apical meristem method, which is combined with thermotherapy and chemotherapy. These methods represent a single biotechnological complex method of plant recovery from viruses in vitro culture. It widely uses this method for obtaining non-virus planting material for various crops. The disadvantages of the tissue culture-based method of health improvement include the possibility of self-clonal variations resulting from the long-term cultivation of plants under in vitro conditions. The possibilities of increasing plant resistance by stimulating their immune system have expanded, which results from the in-depth study of the molecular and genetic bases of plant resistance toward viruses and the investigation of the mechanisms of induction of protective reactions in the plant organism. The existing methods of phytovirus control are ambiguous and require additional research. Further study of the genetic, biochemical, and physiological features of viral pathogenesis and the development of a strategy to increase plant resistance to viruses will allow a new level of phytovirus infection control to be reached.

Response of pink bollworm Pectinophora gossypiella (Saunders) to Cry1Ac and Cry2Ab toxin

BackgroundDuring field surveillance in Gujarat, India, in 2009, scientists found that pink bollworm was surviving in the first generation of Bollgard cotton, which contains a single protein. This survival was observed in four districts of Gujarat. From 2012 to 2014, surveys revealed that pink bollworm larvae had higher survival rates on Bollgard II cotton in Amreli and Bhavnagar districts. In 2016, a pink bollworm outbreak in Maharashtra caused significant losses in cotton production, leading to damage, yield losses, and increased management costs in several regions. The objective of the present study was to determine the reaction of Pectinophora gossypiella to Cry1Ac and Cry2Ab toxins.ResultsNeonates of P. gossypiella collected from six distinct locations in Vidarbha were subjected to seven different concentrations of Cry1Ac. The highest LC50 value was recorded in the population collection of Yavatmal (1.362 μg/ml diet) but the lowest LC50 value of 0.600 μg/ml diet in Buldana population. The highest MIC50 value was recorded in the population collected from Yavatmal (0.743 μg/ml diet) and the lowest MIC50 value of 0.413 μg/ml diet in the Buldana population (0.431 μg/ml diet). The highest IC50 value was recorded in the population collected from Yavatmal (0.303 μg/ml diet) but the lowest IC50 value of 0.127 μg/ml diet in the Buldana population. Neonates of P. gossypiella collected from six different locations in Vidarbha were exposed to seven different concentrations (9.0, 3.0, 1.0, 0.333, 0.111, 0.037 and 0.012 for Cry1Ac and 27.0, 9.0, 3.0, 1.0, 0.333, 0.111 and 0.037 μg/ml for Cry2Ab) which revealed that the Yavatmal population had the highest LC50 value of 94.294 μg/ml diet and the lowest LC50 value was recorded in the Amravati population (42.144 μg/ml diet). The highest MIC50 value of 27.258 μg/ml diet was recorded in Buldana population, but the lowest MIC50 value was recorded in the Amravati population (12.881 μg/ml diet). The highest IC50 value of 3.209 μg/ml diet was observed in Yavatmal population, and the lowest one was recorded in the Amravati population (1.574 μg/ml diet).ConclusionsAccording to the LC50 values, all of the P. gossypiella populations in Vidarbha were extremely insensitive and had field-evolved resistance to the Bt proteins found in the transgenic cotton varieties of Bollgard II.

Azotobacter inoculation can enhance the resistance of Bt cotton to cotton bollworm, Helicoverpa armigera.

The rising trend in the cultivation of Bacillus thuringiensis (Bt) transgenic crops may cause a destabilization of agroecosystems, thus increasing concerns about the sustainability of Bt crops as a valid pest management method. Azotobacter can be used as a biological regulator to increase environmental suitability and improve the soil nitrogen utilization efficiency of crops, especially Bt cotton. A laboratory test investigated effects on the development and food utilization of Helicoverpa armigera fed with different Cry1Ab/Cry1Ac proteins and nitrogen metabolism-related compounds from cotton (transgenic variety SCRC37 vs non-Bt cotton cv. Yu2067) inoculated with Azospirillum brasilense (Ab) and Azotobacter chroococcum (Ac). The findings indicate that inoculation with Azotobacter significantly decreased the partial development and food utilization indexes (pupal weight; pupation rate; adult longevity; fecundity; relative growth rate, RGR; efficiency of conversion of digested food, ECD; and efficiency of conversion of ingested food, ECI) of H.armigera fed on Bt cotton, but contrasting trends were found among these indexes in H.armigera fed on non-Bt cotton inoculated with Azotobacter, as a result of differences in Bt toxin production. Overall, the results showed that inoculation with Azotobacter had negative effects on the development and food utilization of H.armigera fed on Bt cotton, leading to enhanced target insect resistance. Presumably, Azotobacter inoculation can be used to stimulate plant soil nitrogen uptake to increase nitrogen metabolism-related compounds and promote plant growth for Bt and non-Bt cotton, simultaneously raising Bt protein expression and enhancing resistance efficacy against cotton bollworm in Bt cotton.

Heterologous expression of Zygophyllum xanthoxylon zinc finger protein gene (ZxZF) enhances the tolerance of poplar photosynthetic function to drought stress

The ZxZF transcription factor (TF) of Zygophyllum xanthoxylon (Bunge) Maxim, an extremely drought-resistant woody plant, is a C2H2 zinc finger protein. Studies have shown that C2H2 zinc finger proteins play important roles in activating stress-related genes and enhancing plant resistance. However, their function in regulating plant photosynthesis under drought stress is not well understood. Since poplar is an important greening and afforestation tree species, it is particularly important to cultivate excellent drought-tolerant varieties. The ZxZF transcription factor (TF) was heterogeneously expressed in Euroamerican poplar (Populus × euroameracana cl.'Bofengl’) by genetic transformation. Based on the mechanism and potential function of poplar photosynthesis regulated by ZxZF under drought stress, transcriptomic and physiological determinations were used to reveal the important role of this gene in improving the drought resistance of poplar. The results showed that the overexpression of ZxZF TF in transgenic poplars could improve the inhibition of Calvin cycle by regulating stomatal opening and increasing the concentration of intercellular CO2. The chlorophyll content, photosynthetic performance index, and photochemical efficiency of transgenic lines under drought stress were significantly higher than those of the wild type (WT). The overexpression of ZxZF TFs could alleviate the degree of photoinhibition of photosystems II and I under drought stress and maintain the efficiency of light energy capture and the photosynthetic electron transport chain. The transcriptomic data also showed that differentially expressed genes between the transgenic poplar and WT under drought stress were primarily enriched in metabolic pathways related to photosynthesis, such as photosynthesis, photosynthesis-antenna protein, porphyrin and chlorophyll metabolism, and photosynthetic carbon fixation, and the downregulation of genes related to chlorophyll synthesis, photosynthetic electron transport and Calvin cycle were alleviated. In addition, the overexpression of ZxZF TF can alleviate the inhibition of NADH dehydrogenase-like (NDH) cyclic electron flow of the poplar NDH pathway under drought stress, which plays an important role in reducing the excess pressure of electrons on the photosynthetic electron transport chain and maintaining the normal photosynthetic electron transport. In summary, the overexpression of ZxZF TFs can effectively alleviate the inhibition of drought on the assimilation of carbon in poplar and have a positive impact on light energy capture, the orderly transport of photosynthetic electron transport chain and the integrity of the photosystem, which is highly significant to acheivean in-depth understanding of the function of ZxZF TFs. This also provides an important basis for the breeding of new transgenic poplar varieties.