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Rapeseed (Brassica napus L.) is known for its high-quality seed oil and protein content. However, its use in animal feed is restricted due to antinutritional factors present in the seedcake, with sinapine being one of the main compounds that reduces palatability. Attempts to develop rapeseed germplasm with lower sinapine levels through traditional breeding methods have shown limited progress. Genetic transformation methods could create new genotypes with reduced sinapine levels by silencing key genes involved in sinapine biosynthesis, though these methods often result in transgenic or genetically modified plants. The recent development of CRISPR-Cas technology provides a precise and efficient approach to crop improvement, with the potential to generate transgene-free mutants. In this study, we targeted the BnaX.SGT.a genes for knockout using CRISPR-Cas editing. By utilizing our newly established protoplast regeneration and transfection protocol for rapeseed, we demonstrated that DNA-free CRISPR editing via protoplast-based ribonucleoprotein (RNP) delivery was highly effective. We achieved successful knockout of the BnaX.SGT.a paralogues, with an average mutation efficiency of over 30%. Sequencing results revealed a variety of mutation types, from 1 bp insertions to 10 bp deletions, with most mutants exhibiting frameshift mutations that led to premature stop codons. The mutants displayed no visible phenotypic differences in growth patterns or flowering compared to the wild type. Importantly, sinapine content was significantly reduced in all T2 generation mutants analysed, while seed weight remained comparable between mutants and the wild type. [ABSTRACT FROM AUTHOR]

The Fusarium oxysporum species complex (FOSC), comprising a multitude of soil borne fungal pathogens, represent a major group of plant invaders that cause vascular wilt diseases of crops worldwide. Although sexual reproduction is nebulous in the FOSC, horizontal gene transfer (also well known as trans-kingdom) may contribute to the observed diversity in pathogenic strains. Concerning the safety and costly process of generating genetically modified crops, a plant-disease management strategy not reliant on transgenic approaches remains the most economic and environmental-friendly option. Here, we highlight the current scenario via beginning with F. oxysporum formae speciales and races as well as caused Fusarium wilt disease in field. Subsequently, based on the mechanisms by which small RNA (sRNA) acts in gene silencing, especially in bidirectionally trans-kingdom sRNA silencing between F. oxysporum and their hosts in the development of disease, we summarize technological breakthroughs like Spray-Induced Gene Silencing (SIGS) technology in the developing and applying to protect crops from Fusarium wilt disease. Given that SIGS has been developed and recently applied in controlling crop Fusarium wilt diseases, we propose a potential prevention and control system to exploit crop Fusarium-vasculature interaction leading to further study trans-kingdom sRNA silencing. [ABSTRACT FROM AUTHOR]

Flower appearance stands as a key characteristic of flowering plants and is closely linked to their ornamental value. Phytohormone Gibberellin (GA), essential for plant growth and development are widely reported for expansion in flower. DELLA proteins are known to negatively regulate GA signaling and influences plant growth and development through the regulation of cell expansion. However, the specific biological function of DELLA proteins in the woody plant Osmanthus fragrans remains unclear. In this study, O. fragrans 'Sijigui' was utilized as the experimental material, and OfRGA was isolated using the PCR method. OfRGA is expressed in various tissues and is localized in the nucleus. A negative association was observed between OfRGA expression and petal size across four different Osmanthus fragrans cultivars. Transformation experiments in tobacco revealed that transgenic plants overexpressing OfRGA exhibited increased plant height, greater node spacing, shorter leaf length, and wider leaves during the vegetative phase. Notably, the flower organs of transgenic tobacco plants displayed noticeable alterations, including reduced petal size, shorter corolla tubes, pedicels, male and female stamens, and lighter petal color. Furthermore, a decrease in the length and area of petal and corolla tube cells was observed as well. DEGs were found in RNA-seq studies of OfRGA transgenic plants. Subsequent investigation revealed a considerable quantity of down-regulated genes were associated with cell wall synthesis genes and expansion genes, such as CesA1 , XEH , and EXPB1 , as well as genes related to anthocyanin biosynthesis. Overall, our findings suggest that OfRGA undermines tobacco petal size by influencing cell expansion. The present study offers a fundamental comprehension of the role of DELLA protein in the organ development in Osmanthus fragrans. [ABSTRACT FROM AUTHOR]

Conventional approaches like Agrobacterium-mediated transformation, viral transduction, biolistic particle bombardment, and polyethylene glycol (PEG)-facilitated delivery methods have been optimized for transporting specific genes to various plant cells. These conventional approaches in genetically modified crops are dependent on several factors like plant types, cell types, and genotype requirements, as well as numerous disadvantages such as time-consuming, untargeted distribution of genes, and high cost of cultivation. Therefore, it is suggested to develop novel techniques for the transportation of genes in crop plants using tailored nanoparticles (NPs) of manipulative and controlled high-performance features synthesized using green and chemical routes. It is observed that site-specific delivery of genes exhibits high efficacy in species-independent circumstances which leads to an increased level of productivity. Therefore, to achieve these outcomes, NPs can be utilized as gene nano-carriers for excellent delivery inside crops (i.e., cotton, tobacco, rice, wheat, okra, and maize) for desired genetic engineering modifications. As outcomes, this review provides an outline of the conventional techniques and current application of numerous nano-enabled gene delivery needed for crop gene manipulation, the benefits, and drawbacks associated with state-of-the-art techniques, which serve as a roadmap for the possible applicability of nanomaterials in plant genomic engineering as well as crop improvement in the future. [ABSTRACT FROM AUTHOR]

A multi-chromatic and multi-component lateral flow immunoassay (MCMC-LFIA) was developed for simultaneous detection of CP4 EPSPS, Bt-Cry1Ab, Bt-Cry1Ac, and PAT/bar proteins in genetically modified (GM) crops. Captured antibodies specific to these exogenous proteins were separately immobilized on a nitrocellulose membrane as test zones. Multi-colored microspheres, used as visible multi-probes, were conjugated with corresponding antibodies and sprayed on the conjugate pad. The assay results can be visually interpreted within 10 min by observing the appearance of colored bands. The MCMC-LFIA demonstrated high sensitivity, with detection of limits of 7.8 ng/mL for CP4 EPSPS and 2.5 ng/mL for Bt-Cry1Ab, Bt-Cry1Ac, and PAT/bar proteins, significantly improving the performance of previously reported LFIAs. The MCMC-LFIA exhibited excellent specificity and was validated for practical use in field-based applications. The proposed MCMC-LFIA offers a rapid, sensitive, and user-friendly tool for the on-site large-scale screening of GM materials. [ABSTRACT FROM AUTHOR]

Summary: Nitrate (NO3−) serves as the primary inorganic nitrogen source assimilated by most terrestrial plants. The acquisition of nitrate from the soil is facilitated by NITRATE TRANSPORTERS (NRTs), with NRT2.1 being the key high‐affinity nitrate transporter. The activity of NRT2.1, which has multiple potential phosphorylation sites, is intricately regulated under various physiological conditions. Here, we discovered that CALCIUM‐DEPENDENT PROTEIN KINASE 28 (CPK28) positively regulates nitrate uptake under nitrogen deprivation conditions.We found CPK28 as the kinase targeted by immunoprecipitation followed by mass spectrometry and examined the in‐planta phosphorylation status of NRT2.1 in cpk28 mutant plants by employing quantitative MS‐based phosphoproteomics. Through a combination of in vitro phosphorylation experiment and immunoblotting using phospho‐specific antibody, we successfully demonstrated that CPK28 specifically phosphorylates NRT2.1 at Ser21.Functional analysis conducted in Xenopus oocytes revealed that co‐expression of CPK28 significantly enhanced high‐affinity nitrate uptake of NRT2.1. Further investigation using transgenic plants showed that the phosphomimic variant NRT2.1S21E, but not the nonphosphorylatable variant NRT2.1S21A, fully restored high‐affinity 15NO3− uptake ability in both nrt2.1 and cpk28 mutant backgrounds.This study clarifies that the kinase activity of CPK28 is promoted during nitrogen deprivation conditions. These significant findings provide valuable insights into the intricate regulatory mechanisms that govern nitrate‐demand adaptation. [ABSTRACT FROM AUTHOR]

Sulphur limitation 1 (SLIM1), a member of ethylene‐insensitive3‐like (EIN3/EIL) protein family, is recognised as the pivotal transcription factor regulating sulphur assimilation, essential for maintaining sulphur homoeostasis in Arabidopsis. However, the function of its monocot homologues is largely unknown. In this study, we identified PvEIL3a, a homologous gene of AtSLIM1, from switchgrass (Panicum virgatum L.), a significant perennial bioenergy crop. Our results demonstrated that introducing PvEIL3a into Arabidopsis slim1 mutants significantly increased the expression of genes responsive to sulphur deficiency, and transgenic plants exhibited shortened root length and delayed development. Moreover, PvEIL3a activated the expression of AtAPR1, AtSULTR1;1 and AtBGLU30, which plays an important role in sulphur assimilation and glucosinolate metabolism. Results of transcriptome and metabonomic analysis further indicated a perturbation in the metabolic pathways of tryptophan‐dependent indole glucosinolates (IGs), camalexin and auxin. In addition, PvEIL3a conservatively regulated sulphur assimilation and the biosynthesis of tryptophan pathway‐derived secondary metabolites, which reduced the biosynthesis of indole‐3‐acetic acid (IAA) and inhibited the root elongation of transgenic Arabidopsis. In conclusion, this study highlights the functional difference of the ethylene‐insensitive 3‐like (EIL) family gene in monocot and dicot plants, thereby deepening the understanding of the specific biological roles of EIL3 in monocot plant species. Summary statement: The heterologous expression of PvEIL3a in Arabidopsis slim1 mutant altered sulphur assimilation and glucosinolate metabolism significantly.Glucosinolates and their derivatives increased in pAtSLIM1:PvEIL3a/slim1‐1 transgenic Arabidopsis plants inhibited the root elongation. [ABSTRACT FROM AUTHOR]

Transgenic soybean [Glycine max(L.) Merrill] currently covers approximately 80% of the global crop area for this species, with the majority of transgenic plants being glyphosate resistant (Roundup Ready, GR or RR). However, there is significant concern regarding the potential effects of GM crops and their byproducts on soil microbial communities. During our research, we discovered a type of semiwild soybean that emerged due to genetic drift at a transgenic test site. Nevertheless, the ecological risk to soil rhizosphere microorganisms associated with planting semiwild soybean following genetic drift remains unclear. Therefore, we conducted a field experiment and collected soil samples at various stages of plant growth. Our results indicate that the species diversity of rhizosphere bacteria in transgenic glyphosate-resistant semiwild soybean was also not significantly different from that observed in other types of soybean. Additionally, Basidiomycota had beneficial effects on rhizosphere fungi during the flowering and maturation stages in transgenic glyphosate-tolerant semiwild soybean. These findings provide valuable insights into how genetic drift in transgenic soybean may impact the soil microenvironment. [ABSTRACT FROM AUTHOR]

Soybean looper (SBL), Chrysodeixis includens (Walker 1858) (Lepidoptera: Noctuidae), is one of the most damaging insect pests of soybean, Glycine max (L.) Merr., in the mid-south region of the United States, and causes significant economic losses to cotton, sunflower, tomato, and tobacco crops in the United States, Brazil, and Argentina. Soybean production in the southern region accounted for 15.5% of the total production in the United States, and yield losses due to invertebrate pests were 5.8%, or 1.09 million metric ton, in 2022. As insecticide resistance of SBL continues to rise, the lack of alternate control strategies is a serious concern. Numerous studies have been reported on pest status, distribution, semiochemical-based attractant blends, pesticides and resistance mechanisms, host-plant resistance mechanisms, and molecular tools for controlling this pest in soybeans and other crops. However, there is no comprehensive review that summarizes and discusses these research on SBL and soybeans. The current management strategies for SBL remain heavily reliant on chemical insecticides and transgenic crops. In contrast, integrated pest management (IPM) strategies are needed to control the pest in an effective and environmentally friendly way. This review examines and synthesizes the literature on SBL as a significant pest of soybeans and other important crops, highlighting recent progress in ecological interactions, host plant defenses, and control strategies and identifying information gaps, thereby suggesting avenues for further research on this pest. [ABSTRACT FROM AUTHOR]

Societal Impact Statement Summary The European Union is in the midst of changing the current regulatory framework for new genomic techniques (NGTs) to accelerate the production of plant varieties, in order to achieve the goals of the European Green Deal. These techniques are highly contested, with divergent views on how they should be governed. So far, there has been little effort to engage citizens in this legislative reform process. By engaging with Dutch citizens, we give the public a voice in shaping the future of agriculture and the food system. By facilitating the exchange of multiple views, we allow for more effective governance arrangements. The European Commission (EC) has proposed a new regulation for plants obtained by new genomic techniques (NGTs). Currently, food crops developed with NGTs are subject to the EU Directive 2001/18/EC on the deliberate release into the environment of genetically modified organisms (GMOs). The current proposal for a new regulation differentiates between two categories of NGT plants. Category 1 NGT plants will be subject to the new regulation, whereas Category 2 plants will remain subject to the GMO legislation, although the risk assessment may be adapted. In this paper, we analyze the views of Dutch citizens on NGT crops and their governance, prior to the publication of the new proposal. We find significant reservations arising from doubts about NGT crops delivering on their promises, the likelihood of unanticipated consequences, and unnaturalness. We extrapolate our findings to anticipate citizen's response to the new proposal and reflect on ways to move forward, both for policy making, and for the plant science community. [ABSTRACT FROM AUTHOR]

Biodegradable materials are currently one of the main focuses of research and technological development. The significance of these products grows annually, particularly in the fight against climate change and environmental pollution. Utilizing artificial biopolymers offers an opportunity to shift away from petroleum-based plastics with applications spanning various sectors of the economy, from the pharmaceutical and medical industries to food packaging. This paper discusses the main groups of artificial biopolymers. It emphasizes the potential of using genetically modified plants for its production, describing the primary plant species involved in these processes and the most common genetic modifications. Additionally, the paper explores the potential applications of biobased polymers, highlighting their key advantages and disadvantages in specific context. [ABSTRACT FROM AUTHOR]

CRISPR/Cas9 is the most popular genome editing platform for investigating gene function or improving traits in plants. The specificity of gene editing has yet to be evaluated at a genome-wide scale in seed-propagated Camelina sativa (L.) Crantz (camelina) or clonally propagated Solanum tuberosum L. (potato). In this study, seven potato and nine camelina stable transgenic Cas9-edited plants were evaluated for on and off-target editing outcomes using 55x and 60x coverage whole genome shotgun sequencing data, respectively. For both potato and camelina, a prevalence of mosaic somatic edits from constitutive Cas9 expression was discovered as well as evidence of transgenerational editing in camelina. CRISPR/Cas9 editing provided negligible off-target activity compared to background variation in both species. The results from this study guide deployment and risk assessment of genome editing in commercially relevant traits in food crops. [ABSTRACT FROM AUTHOR]

Global climate change exposes crops to numerous abiotic and biotic stress factors, significantly threatening their growth, development, and productivity. Among the various abiotic stresses, salt and drought are the most detrimental, severely impacting yield stability. Late embryogenesis abundant (LEA) proteins represent a broad and varied class of polypeptides that accumulate in response to cellular dehydration across diverse organisms. Nonetheless, there has been a scarcity of studies on this protein family in the common bean (Phaseolus vulgaris L.) prior to this investigation. In this study, we identified 69 members of the LEA gene family in the common bean, characterized by conserved LEA and dehydrin domains and additional functional domains. Based on their phylogenetic relationships, these proteins are categorized into eight distinct subfamilies: LEA_1, LEA_2, LEA_3, LEA_4, LEA_5, LEA_6, dehydrin, and SMP. In this study, approximately 62% of the identified PvLEAs are classified within the LEA_2 group, characterized by their higher hydrophobicity. Notably, they are clustered on chromosomes 7 and 8, suggesting their evolutionary conservation within these chromosomal regions. Analysis of promoter regions uncovered a variety of elements, including those responsive to stress, including ARE, ABRA, WRKY, TCA element, and CGTCA motif, implying their potential role in stress response mechanisms. The PvHDN3 overexpressed transgenic lines exhibited enhanced physiological health, increased height, and better-developed roots and leaves than wild-type plants under salt and drought stresses. In addition, it was found that PvHDN3 expression in response to different abiotic stress (salt and drought) intensity was closely linked to several stress-related physiological parameters, including proline and malondialdehyde concentrations, ascorbate peroxidase, catalase, and peroxidase activity. The current study offers novel insights into the formation of LEA proteins in common beans and their roles in developmental processes under stress conditions, providing a valuable foundation for future research. [ABSTRACT FROM AUTHOR]

Rice blast, caused by the fungal pathogen Magnaporthe oryzae, is a destructive disease that affects rice (Oryzae sativa L.) on a global scale. Polyamines (PAs) play diverse roles in plant growth and development and responses to biotic and abiotic stimuli. Putrescine (PUT), spermidine (SPD), and spermine (SPM) are the primary forms of polyamines (PAs). In this study, we observed that the accumulation of apoplastic PAs significantly increased in rice plants after treatment with salt or M. oryzae. The salt-treated plants exhibited enhanced resistance to rice blast disease. RNA sequencing data indicate that S-adenosylmethionine decarboxylase (SAMDC), a key enzyme involved in the synthesis of polyamines, plays a significant role in enhancing plant resistance. Overexpression of rice SAMDC (OsSAMDC) led to a significant decrease of pathogen infection in the transgenic rice plants. Additionally, OsSAMDC overexpression plants accumulated polyamines in the cytosol and apoplast, particularly SPD and SPM. Conversely, the disease resistance and accumulation of PAs were reduced in OsSAMDC-silenced plants. Exogenous application of PAs inhibited the mycelium growth, spore germination, germ tube elongation, and appressorium formation in M. oryzae. These results demonstrated that OsSAMDC-mediated polyamine biosynthesis, especially SPD and SPM, plays an essential role in rice plants to resist biotic and abiotic stresses. [ABSTRACT FROM AUTHOR]

Phosphoenolpyruvate carboxykinase (PEPCK) is an enzyme of the lyase family utilized in the gluconeogenesis pathway in plants. It converts oxaloacetate into Phosphoenolpyruvate (PEP) and carbon dioxide. PEPCK acts as a prime decarboxylase cytosolic enzyme and may have exhibited a positive response against salinity (main abiotic stress) stress in certain plants. Transgenic plants (C3) with high-level expression of C4 enzymes PEPC or PEPCK can serve as a perfect example for increasing the crop photosynthetic capacity through genetic engineering. In this review, we have focused on the recent advances in the utilization of the PEPCK gene and its role in increasing photosynthesis as well as in abiotic stress tolerance. [ABSTRACT FROM AUTHOR]

Amino acids are the primary form of nitrogen utilization in higher plants, mainly transported by amino acid transporters. In this study, we analyzed the natural variation of amino acid transporter-like 4 (OsATL4) in rice germplasm resources, identified its spatiotemporal expression characteristics, determined its substrate transport, and validated its function using transgenic plants. We found that the promoter sequence of OsATL4 varied across 498 rice varieties. The expression level of OsATL4 was higher in japonica rice, which was negatively correlated with tiller number and grain yield. OsATL4 was highly expressed in the basal part, leaf sheath, stem, and young panicle, with its two splicing variants localized to the cell membrane. OsATL4a (the long splicing variant) had a high affinity for transporting Ser, Leu, Phe, and Thr, while OsATL4b (the short splicing variant) had a high affinity for transporting Ser, Leu, and Phe. Blocking OsATL4 promoted axillary bud outgrowth, rice tillering, and grain yield, whereas overexpression lines exhibited the opposite phenotype. Exogenous application of low concentrations of Ser promoted axillary bud outgrowth in overexpression lines, while high concentrations of Ser inhibited it. Conversely, the mutant lines showed the opposite response. Altered expression of OsATL4 might affect the expression of genes in nitrogen, auxin, and cytokinin pathways. We propose that two splicing variants of OsATL4 negatively regulate rice tillering and yield by mediating the transport of amino acids, making it a significant target for high-yield rice breeding. [ABSTRACT FROM AUTHOR]

Cytokinins are one of the main groups of plant hormones that regulate growth and development of plants. Cytokinin oxidase/dehydrogenase (CKX) is an enzyme that rapidly and irreversibly degrades cytokinins and thus directly affects their concentration and physiological effect. Genetically modified plants with reduced CKX activity in the shoot, i.e. with a higher concentration of cytokinins, showed e.g. increased tolerance to drought stress, formed larger inflorescences and had higher grain yield. For these reasons, chemical compounds capable of inhibiting the CKX activity (CKX inhibitors) were sought. First, they were identified among strong synthetic cytokinins, but their inhibitory activity was low. The trend has been to develop potent CKX inhibitors with minimal intrinsic cytokinin activity in the hope of avoiding the negative effect of cytokinins on root growth. Cloning CKX, production of key recombinant enzymes from Arabidopsis (AtCKX2) and maize (ZmCKX1 and ZmCKX4a), development of screening bioassays and progress in X-ray crystallography and synthetic organic chemistry led to extensive progress in the development of these compounds. Currently, the most suitable CKX inhibitors are seeking their application in research and the commercial sphere in two main areas -- plant tissue cultures and agriculture. The key milestones that preceded it are summarized in this review. [ABSTRACT FROM AUTHOR]

Simple Summary: The corn earworm causes persistent ear damage to corn grown in the southeastern United States. Increased levels of ear damage have been associated with mycotoxin contamination such as aflatoxin and fumonisin. Corn hybrids expressing the Bt traits Vip3Aa20 provided substantial corn earworm control and prevented kernel damage. Older Bt corn without this trait did not control corn earworm in the ear and prevent ear damage. Bt corn that prevented kernel damage had a variable effect on grain yield but may prevent yield loss. Bt corn that prevented ear damage did not suffer grain contamination from aflatoxin but did show reduced grain contamination by fumonisin. The corn earworm, Helicoverpa zea (Boddie), causes persistent ear damage to corn grown in the southeastern United States region. Increased levels of ear damage have been associated with mycotoxin contamination in addition to yield loss. Corn hybrids expressing proteins from the Bacillus thuringiensis (Bt) may provide corn earworm control. A selection of hybrids expressing various Bt traits were evaluated in field experiments across Georgia over two years to assess their efficacy for corn earworm control, grain yield and quality protection, and grain mycotoxin mitigation. Ear damage was significantly reduced only by Bt hybrids expressing the Vip3Aa20 protein. The remaining Bt hybrids expressing Cry proteins provided only marginal control. Ear damage had a variable effect on grain yield and was not correlated with grain aflatoxin contamination. In contrast, grain fumonisin contamination was positively associated with earworm damage. These results indicate Bt hybrids that effectively reduce corn earworm damage may also assist in reducing fumonisin contamination and possibly yield loss. [ABSTRACT FROM AUTHOR]

Drought stress is one of the main factors limiting the high yield and quality of potatoes. Arabinogalactan proteins (AGPs) are an important class of glycoproteins widely present in the cell walls, plasma membranes, and extracellular matrices of higher plants. Among them, fasciclin-like arabinogalactan proteins (FLAs) are involved in plant development, stress responses, and hormone signal regulation. However, little is known about the FLAs gene in potatoes. Based on transcriptome sequencing data, this study screened a drought stress-related candidate FLA gene (StFLA4) through bioinformatics and expression analysis in potatoes. qRT-PCR analysis showed that StFLA4 was induced by drought stress, and its expression decreased with the extension of stress time. Moreover, the relative expression level of StFLA4 in the drought-resistant variety "Kexin 1" was lower than in the drought-sensitive variety "Atlantic". The StFLA4 protein was located in the cell membrane and interacted with nineteen proteins, mainly related to response to environmental stimulus, cellular response to abiotic stimulus, and cell maturation. After heterologous overexpression of StFLA4 in tobacco, the transgenic plants showed more withered leaves than the wild-type tobacco under drought stress. During the drought stress period, the expression level of StFLA4 in the transgenic plants significantly decreased, and the activity of SOD and POD was significantly lower than that of WT. However, the MDA content was higher than that of WT. These results indicated that StFLA4 negatively regulates the response to drought stress. In addition, in the germination test of potato "Variety V7" tubers, it was found that the variation tendency of StFLA4 expression was along with the concentration of arabinogalactan proteins, and it may participate in the regulation of potato tuber germination. This study lays the foundation for elucidating the function and expression pattern of StFLA4 response to drought stress and tuber germination in potatoes. [ABSTRACT FROM AUTHOR]

Background/Objectives: The sucrose non-fermentation-related kinase 1 (SnRK1) protein complex in plants plays an important role in energy metabolism, anabolism, growth, and stress resistance. SnRK1 is a heterotrimeric complex. The SnRK1 complex is mainly composed of α, β, βγ, and γ subunits. Studies on plant SnRK1 have primarily focused on the functional α subunit, with the β regulatory subunit remaining relatively unexplored. The present study aimed to elucidate the evolutionary relationship, structural prediction, and interaction with the core α subunit of peach SnRK1β3 (PpSnRK1) subunit. Methods: Bioinformatics analysis of PpSnRK1 was performed through software and website. We produced transgenic tomato plants overexpressing PpSnRK1 (OEPpSnRK1). Transcriptome analysis was performed on OEPpSnRK1 tomatoes. We mainly tested the growth index and drought resistance of transgenic tomato plants. Results: The results showed that PpSnRK1 has a 354 bp encoded protein sequence (cds), which is mainly located in the nucleus and cell membrane. Phylogenetic tree analysis showed that PpSnRK1β3 has similar domains to other woody plants. Transcriptome analysis of OEPpSnRK1β3 showed that PpSnRK1β3 is widely involved in biosynthetic and metabolic processes. Functional analyses of these transgenic plants revealed prolonged growth periods, enhanced growth potential, improved photosynthetic activity, and superior drought stress tolerance. Conclusions: The study findings provide insight into the function of the PpSnRK1 subunit and its potential role in regulating plant growth and drought responses. This comprehensive analysis of PpSnRK1 will contribute to further enhancing our understanding of the plant SnRK1 protein complex. [ABSTRACT FROM AUTHOR]

We provide guidance on how to incorporate best practices around gender integration in the development of genetically improved crops by adapting a gender integration framework for conventional crop breeding to the GM product development pipeline, which places greater emphasis on the discovery and launch phases because the technical nature of the development process means fewer opportunities for farmer engagement or pivoting possibilities between these two ends of the product development spectrum. For crop innovation to be relevant to both women and men producers, during the discovery phase, developers can conduct baseline gender analysis consisting of gender-disaggregated value chain analysis, systematic learning about gender-specific crop trait preferences, and identification of varietal preferences by women and men along the value chain. The latter opportunity in the GM product development pathway for intentional gender integration is deployment, including pre-launch activities such as field demonstrations and consumer testing. We also describe ex ante and ex post gender impact assessment methods. We conclude with a number of gender integration recommendations for GM product developers: improving gender data collection and analysis to inform crop innovation efforts, investing in staffing and training of scientific teams to enhance gender expertise, and increasing accountability of product development teams with gender-intentional monitoring and evaluation systems. [ABSTRACT FROM AUTHOR]

Genetically modified (GM) crops are the most important agricultural commodities that can improve the yield of African smallholder farmers. The intricate circumstances surrounding the introduction of GM agriculture in Africa, however, underscore the importance of comprehending the moral conundrums, regulatory environments, and public sentiment that exist today. This review examines the current situation surrounding the use of GM crops in Africa, focusing on moral conundrums, regulatory frameworks, and public opinion. Only eleven of the fifty-four African countries currently cultivate GM crops due to the wide range of opinions resulting from the disparities in cultural, socioeconomic, and environmental factors. This review proposed that addressing public concerns, harmonizing regulations, and upholding ethical standards will improve the adoption of GM crops in Africa. This study offers ways to enhance the acceptability of GM crops for boosting nutrition and food security globally. [ABSTRACT FROM AUTHOR]

Maize (Zea mays L.) is the most important cereal crop in the world. Flowering period and photoperiod play important roles in the reproductive development of maize. This study, investigated ZmMADS42, a gene that is highly expressed in the shoot apical meristem. Agrobacterium infection was used to successfully obtain overexpressed ZmMADS42 plants. Fluorescence quantitative PCR revealed that the expression of the ZmMADS42 gene in the shoot apical meristem of transgenic plants was 2.8 times higher than that of the wild-type(WT). In addition, the expression of the ZmMADS42 gene in the endosperm was 2.4 times higher than that in the wild-type. The seed width of the T2 generation increased by 5.35%, whereas the seed length decreased by 7.78% compared with that of the wild-type. Dissection of the shoot tips of transgenic and wild-type plants from the 7-leaf stage to the 9-leaf stage revealed that the transgenic plants entered the differentiation stage earlier and exhibited more tassel meristems during their vegetative growth period. The mature transgenic plants were approximately 20 cm shorter in height and had a lower panicle position than the wild-type plants. Comparing the flowering period, the tasseling, powdering, and silking stages of the transgenic plants occurred 10 days earlier than those of the wild-type plants. The results showed that the ZmMADS42 gene played a significant role in regulating the flowering period and plant height of maize. [ABSTRACT FROM AUTHOR]

Since the first genetically engineered or modified crops or organisms (GMO) were approved for commercial production in 1995, no new GMO has been proven to be a hazard or cause harm to human consumers. These modifications have improved crop efficiency, reduced losses to insect pests, reduced losses to viral and microbial plant pathogens and improved drought tolerance. A few have focused on nutritional improvements producing beta carotene in Golden Rice. Regulators in the United States and countries signing the CODEX Alimentarius and Cartagena Biosafety agreements have evaluated human and animal food safety considering potential risks of allergenicity, toxicity, nutritional and anti-nutritional risks. They consider risks for non-target organisms and the environment. There are no cases where post-market surveillance has uncovered harm to consumers or the environment including potential transfer of DNA from the GMO to non-target organisms. In fact, many GMOs have helped improve production, yield and reduced risks from chemical insecticides or fungicides. Yet there are generic calls to label foods containing any genetic modification as a GMO and refusing to allow GM events to be labeled as organic. Many African countries have accepted the Cartagena Protocol as a tool to keep GM events out of their countries while facing food insecurity. The rationale for those restrictions are not rational. Other issues related to genetic diversity, seed production and environmental safety must be addressed. What can be done to increase acceptance of safe and nutritious foods as the population increases, land for cultivation is reduced and energy costs soar? [ABSTRACT FROM AUTHOR]

ve: Increasing tree productivity by genetic engineering methods is one of the main trends of forest biotechnology. A promising strategy for this is to improve the use efficiency of nitrogen, which is the main limiting factor of plant growth. For this purpose, the GS1 gene from Scots pine was transferred to downy birch (Betula pubescens) plants. This gene encodes the cytosolic form of glutamine synthetase, the main enzyme of nitrogen metabolism in plants. Methods: To assess the effects of insertion of this gene, the birch plant metabolome was analyzed using GC-MS and HPLC-MS. Results and Discussion: GC-MS analysis found 197 metabolites in birch extracts, but the metabolomes of two transgenic clones showed no statistically significant differences from the control. Using the S-plot based on the OPLS-DA model, 32 metabolite markers affecting the separation of control and transgenic birch plants were detected; 22 of them were identified. Three metabolites among them were nitrogen-containing, including γ-aminobutyric acid, the immediate precursor of which is glutamine. HPLC-MS analysis found 48 metabolites, but transgenic plants did not differ from the control. GC-MS, however, showed a decrease in the content of two phenolic compounds in transgenic plants, which is characteristic of improved nitrogen supply. Conclusions: The study shows that modification of nitrogen metabolism in birch plants does not significantly affect the biochemical composition of tree shoots. [ABSTRACT FROM AUTHOR]

Wheat (Triticum aestivum L.) is a key crop for the global food supply. Its complex polyploid genome and recalcitrance to tissue culture pose challenges for genetic transformation, especially for elite cultivars. This study aimed to improve transformation efficiency in elite Korean wheat cultivars by integrating morphogenic regulator genes (GRF4–GIF1), using an engineered Agrobacterium tumefaciens strain. By integrating speed breeding to accelerate donor plant growth, we also reduced the time required to generate transgenic plants. Modifying the resting period and incorporating GRF4–GIF1 significantly improved the outcomes, leading to a transformation efficiency of 4.46% ± 0.93% in Bobwhite cultivar. Application of these methods to Korean elite cultivars also successfully yielded transformations with Keumkang and Baekkang cultivars. Furthermore, using a T3SS AGL-1 strain expressing the type III effector AvrPto enhanced the transformation efficiency in Baekkang cultivar by 2.27 times (17.7%) compared to the standard AGL-1 strain. Using the protocol as described here, we successfully developed stable transgenic lines of the Korean wheat cultivar, marking a pioneering achievement in Korean wheat molecular breeding. The modified protocol can contribute to wheat genetic research and breeding programs in Korea. [ABSTRACT FROM AUTHOR]

AN2 was thought to be the key gene inducing the fruit color differentiation of Lycium barbarum and Lycium ruthenicum, but there was no direct evidence. In this study, AN2 from L. ruhtenicum was overexpressed in L. barbarum to conform its function in regulating anthocyanin biosynthesis. The results showed that the stems, leaves, and fruits of transgenic plants were purple and contained significantly higher anthocyanin levels compared to the wild-type plants. Transcriptome analysis of leaf and fruit identified 5,832 differentially expressed genes (DEGs) in leaves and 2,029 DEGs in fruits, with 910 genes common to both tissues. GO and KEGG pathway enrichment analyses indicated a significant impact of LrAN2 on flavonoid biosynthesis and anthocyanin biosynthesis. The structural genes involved in the anthocyanin pathway, such as 4CL, CHS, F3H, F3'5'H, DFR, ANS, and UFGT, were up-regulated in transgenic lines, while the expression of transcription factors bHLH and WD40 were remained unchanged. These findings confirm the role of LrAN2 in the color differentiation of L. barbarum and L. ruthenicum and highlight its potential for the genetic enhancement of anthocyanin content in L. barbarum and other Solanaceae species. [ABSTRACT FROM AUTHOR]

SUMMARY: Sorghum bicolor (sorghum) is a vital C4 monocotyledon crop cultivated in arid regions worldwide, valued for its significance in both human and animal nutrition. Despite its agricultural prominence, sorghum research has been hindered by low transformation frequency. In this study, we examined sorghum transformation using the pVS1‐VIR2 ternary vector system for Agrobacterium, combined with the morphogenic genes BABY BOOM and WUSCHEL2 and selection using G418. We optimized Agrobacterium‐mediated infection, targeting key parameters such as bacterial optical density, co‐cultivation time, and temperature. Additionally, an excision‐based transformation system enabled us to generate transgenic plants free of morphogenic regulators. The method yielded remarkable transformation frequencies, reaching up to 164.8% based on total isolated plantlets. The same combination of ternary vector, morphogenic genes and geneticin‐based selection also resulted in a marked increase in transformation efficiency of the Zea mays (maize) inbred line B104. The potential for genomic editing using this approach positions it as a valuable tool for the development of sorghum and maize varieties that comply with evolving European regulations. Our work marks a significant stride in sorghum biotechnology and holds promise for addressing global food security challenges in a changing climate. Significance Statement: This study achieved unprecedented transformation frequencies in Sorghum bicolor using the pVS1‐VIR2 ternary vector system combined with BABY BOOM and WUSCHEL2 morphogenic genes. These advancements promise to enhance sorghum and maize breeding, addressing global food security in a changing climate. [ABSTRACT FROM AUTHOR]

Environmental stresses are responsible for limiting soybean yield. To mitigate the impacts generated by water deficit, molecular biology tools are being used to develop genetically modified plants. Previous studies showed that two independent events (B1 and B3) of soybean transgenic plants expressing a Solanum nigrum osmotin (SnOLP) had an increment in drought tolerance. The present study aims to investigate the modulated pathways that results in the drought tolerance promoted by osmotin overexpression in soybean. Transgenic and non-transgenic (NT) plants in the vegetative stage were submitted to water deficit by irrigation suppression for seven days. Control plants were kept irrigated. Physiological variables were monitored and confirmed that the transgenic plants present better performance when compared to the NT plants. The total RNA extracted from leaves was sequenced and data was normalized by DESeq2. A total of 2044 and 1505 differentially expressed genes (DEGs) were identified in B1 and B3 events, respectively. Regarding the B1 event, 769 genes were upregulated and 1275 downregulated. For B3, 541 genes were upregulated and 964 genes were downregulated. Excluding common differentially expressed genes (DEGs) between transgenic and non-transgenic (NT) plants yielded 395 upregulated and 234 downregulated genes, which were shared by B1 and B3 events. The metabolic pathways and gene ontology categories identified are known to be involved in plant responses to drought. Hormonal, photosynthetic, carbohydrate and amino acid metabolism, reactive oxygen species, and post-translational modifications pathways were significantly modulated in transgenic plants. Altogether, the results suggest that osmotin promotes tolerance through an increment in the plant responses elicited by drought. [ABSTRACT FROM AUTHOR]

Minghuai 1 (MH1) is a yam (Dioscorea alata) cultivar characterized by its strong resistance to Cd stress. This study aims to investigate the enzymatic antioxidant system of MH1 under Cd stress. Under Cd treatment, MH1 exhibited an elevation in superoxide dismutase (SOD) activity, while the activities of ascorbate peroxidase, catalase, and peroxidase remained unchanged. A Cu/Zn-SOD gene, DaSOD1, was cloned and found to be up-regulated following Cd treatment. The DaSOD1 gene was further introduced into tobacco plants for functional analysis. Transgenic tobacco seedlings overexpressing DaSOD1 under Cd treatments exhibited increased chlorophyll contents and seed germination rate, while the levels of superoxide anion and malondialdehyde decreased compared to the wild-type plants. This suggests that DaSOD1 overexpression mitigated the negative effects of Cd stress by reducing oxidative damage in plants. Characterization of the SOD activity and its corresponding DaSOD1 gene is expected to improve our understanding of Cd-resistance mechanism in yam plants. [ABSTRACT FROM AUTHOR]

The present study explored the impact of rolB gene transformation on biological activities of Nicotiana tabacum. PCR, and RT-qPCR were used to validate the integration of rolB gene and its expression. HPLC analysis was used for phytochemical profiling, and various in-vitro and in-vivo assays were used to assess pharmacological activities of wild-type and transgenic extracts using ANOVA and Tukey's multiple comparison method for data analysis. Our findings indicated increased total phenolic content (24.4 ± 0.2 mg GAE/g DW of extract) and total flavonoid content (58.8 ± 1.1 mg QE/g DW of extract) in transgenic plants. In case of antioxidant activities, an increase in reducing power (51.82 ± 0.4 AAE mg/g), and radical scavenging (IC50 188 ± 2.7 µg/mL) was observed. Similarly, transgenic lines exhibited DNA protection against damage caused by free radicals. Moreover, we detected significant antileishmanial potential (IC50 350.7 ± 8.3 µg/mL), butyrylcholinesterase (IC50 268.6 ± 4.7 µg/mL), and acetylcholinesterase inhibitions (IC50 421.6 ± 3.7 µg/mL) in transgenic plants. Similarly, compared with the non-transgenic plants, the rolB-transgenic plants showed increased analgesic (71.4%), anti-inflammatory activities (77.8%), and 35.6% decrease in depression compared to non-transgenic plants. The improved pharmacological activities in transgenic plants have been linked to the level of phytochemicals that increase the therapeutic and curative properties of N. tabacum plants. [ABSTRACT FROM AUTHOR]

Summary: The endeavour to elevate the nutritional value of oat (Avena sativa) by altering the oil composition and content positions it as an optimal crop for fostering human health and animal feed. However, optimization of oil traits on oat through conventional breeding is challenging due to its quantitative nature and complexity of the oat genome. We introduced two constructs containing three key genes integral to lipid biosynthesis and/or regulatory pathways from Arabidopsis (AtWRI1 and AtDGAT1) and Sesame (SiOLEOSIN) into the oat cultivar 'Park' to modify the fatty acid composition. Four homozygous transgenic lines were generated with a transformation frequency of 7%. The expression of these introduced genes initiated a comprehensive transcriptional reprogramming in oat grains and leaves. Notably, endogenous DGAT, WRI1 and OLEOSIN genes experienced upregulation, while genes associated with fatty acid biosynthesis, such as KASII, SACPD and FAD2, displayed antagonistic expression patterns between oat grains and leaves. Transcriptomic analyses highlighted significant differential gene expression, particularly enriched in lipid metabolism. Comparing the transgenic oat plants with the wild type, we observed a remarkable increase of up to 34% in oleic acid content in oat grains. Furthermore, there were marked improvements in the total oil content in oat leaves, as well as primary metabolites changes in both oat grains and leaves, while maintaining homeostasis in the transgenic oat plants. These findings underscore the effectiveness of genetic engineering in manipulating oat oil composition and content, offering promising implications for human consumption and animal feeding through oat crop improvement programmes. [ABSTRACT FROM AUTHOR]

WRKY transcription factors (TF) are identified as important regulating plant proteins involved in stress response signaling pathways. Overexpression of these transcription factors in plants improved plant biotic and abiotic stress responses. In this context, we have envisaged transferring a cDNA encoding the grapevine VvWRKY2TF in potato plants. Four transgenic lines were selected (BFW2A, BFW2C, BFW2D, and BFW2F). In the present study, their response to Cadmium (Cd) stress (50, 100, 150, and 300 μM) was evaluated in vitro. Cadmium is recognized as being among the most harmful heavy metals to plants. Its accumulation in plant cells and tissues disturbs cell homeostasis and causes numerous metabolic damages that affect productivity. The wildtype (WT) plants from the BF15 potato variety and the transgenic plants overexpressing VvWRKY2TF were submitted to cadmium in vitro stress for 20 days. Plant growth and oxidative stress parameters were followed in these plants. All transgenic plants appeared more vigorous than WT. The BFW2A, BFW2C, and BFW2D lines showed better stem development rates than the WT and BFW2F lines. Malondialdehyde (MDA) production in both roots and leaves was reduced in BFW2A, BFW2C, and BFW2D plants as compared to BFW2F and WT plants. This result was associated with the best antioxidant activities of superoxide dismutase (SOD) and catalase (CAT) displayed by these genetically modified lines suggesting their better adaptation to Cd stress conditions. Cd accumulation in plant tissues was investigated, and higher levels of Cd were found in transgenic plants than in WT plants. These findings point to a functional Cd sequestration mechanism in the roots of transgenic plants expressing VvWRKY2. These findings imply that the VvWRKY2 TF is implicated in heavy metal response signaling processes. Its overexpression in plants may be an efficient strategy to reduce the negative effects of Cd stress, promoting the growth patterns and the activity of reactive oxygen species-scavenging enzymes in potato plants. [ABSTRACT FROM AUTHOR]

As the global demand for seafood surges, the expanding aquaculture industry faces a pressing need for viable aquafeed ingredients. The raw material for fish oil is limited and expensive due to unpredictable fishery resources in the fishing zones and the overexploitation of wild fisheries, underscoring the urgency of finding alternatives. This review explores diverse plant oil sources, including soybean, rapeseed, linseed, and algal oils, emphasizing their crucial role in nutritionally balanced aquafeeds. These oils support aquatic animals' growth, health, and development, influencing membrane structure, energy storage, and hormone production. Genetically modified oilseeds (GM), such as camelina and canola, offer a controlled nutrient content, enabling customized nutrient profiles. This comprehensive review provides an overview of different plant oil sources, elucidates their nutrient profiles, and assesses their potential applications in aquaculture. The discussion encompasses their impact on growth, feed efficiency, lipid profile, health, immunological status, disease resistance, and overall performance of both freshwater and marine fish. Furthermore, the review compiles relevant data on the current status of genetically modified plant oils and explores their potential integration into aquaculture practices. In summary, substituting plant oils for fish oil in aquafeed presents a promising solution to aquaculture industry challenges to meet nutritional requirements for fish. [ABSTRACT FROM AUTHOR]

BACKGROUND: Lepidopteran pest control in agriculture has become heavily dependent on cultivars that express Bacillus thuringiensis (Bt) toxins as 'plant‐incorporated protectants'. However, populations of Spodoptera frugiperda (Smith) in Brazil appear resistant to the Bt traits currently available in commercial soybean cultivars. RESULTS: This study evaluated S. frugiperda life history when feeding on three different Bt soybean cultivars. Cultivars expressing Cry1Ac + Cry1F and Cry1A.105 + Cry2Ab2 + Cry1Ac Bt toxins caused 100% larval mortality in S. frugiperda. Both non‐Bt and Cry1Ac‐expressing soybean induced transgenerational effects that increased the survival of subsequent generations. A Cry1Ac soybean diet reduced the generation time (T) of S. frugiperda relative to non‐Bt soybean, resulting in shorter generation time and more rapid population growth. CONCLUSION: The implications of these results revealed how diet can alter aspects of insect life history and biology, and have important implications for sustainable management of S. frugiperda on soybean. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Herbicides are essential tools for the phytosanitary security of agricultural areas, but their excessive use can cause problems in agricultural production systems and have negative impacts on human health and the environment. The objective of this study was to present and discuss the main causes behind the increase in herbicide commercialization in Brazil between 2010 and 2020. Data from the Brazilian pesticide database, provided by the Instituto Brasileiro do Meio Ambiente e dos Recursos Naturais Renováveis (IBAMA), were used. In 2010 and 2020, Brazil sold 157,512 and 329,697 tons of herbicide active ingredients, respectively, representing a 128.1% increase in commercialization over 11 years. Some herbicides, such as clethodim, haloxyfop-methyl, triclopyr, glufosinate, 2,4-D, diclosulam, and flumioxazin, showed increases in sales volumes between 2010 and 2020 of 2672.8%, 896.9%, 953.5%, 290.2%, 233.8%, 561.3%, and 531.6%, respectively, percentages far exceeding the expansion of Brazil's agricultural area. The primary reason for this sharp increase in herbicide sales was the worsening cases of weeds resistant and tolerant to glyphosate, with species such as Conyza spp., Amaranthus spp., Digitaria insularis, and Eleusine indica standing out. This situation created the necessity of the use of additional herbicides to achieve effective chemical control of these weed species. [ABSTRACT FROM AUTHOR]

The German debate about genetically engineered crops for human consumption (GE) has been polarized for nearly three decades. Efforts to overcome this polarization generally involve distributing information, but research indicates that information has little to no impact on GE attitudes, especially among those with extreme positions. Recent studies suggest that moral value concerns drive GE opposition. However, the specific moral values underlying both support and opposition for GE are unknown, as is whether these values differ between GE supporters and opponents. The present research addresses these questions through latent content analysis. Findings show that most arguments in the debate address moral value concerns related to loyalty, often focusing on trust issues. Trust issues are more prevalent than moral concerns about preventing harm and risks. Furthermore, moral value conflicts are likely since GE supporters emphasize concerns related to authority, fairness, and liberty, while GE opponents focus more on values related to care and purity. This paper discusses these findings and recommends steps for improved science communication. [ABSTRACT FROM AUTHOR]

CLE (CLAVATA3/ENDOSPERM SURROUNDING REGION-related) peptides are systemic regulators of legume-rhizobium symbiosis that negatively control the number of nitrogen-fixing nodules. In Medicago truncatula, the expression of the MtCLE35 gene is activated in response to rhizobial inoculation and nitrate treatment, and its overexpression systemically inhibits nodulation. However, little is known about the molecular mechanisms underlying MtCLE35-mediated inhibition of nodulation. In order to elucidate target genes regulated by the MtCLE35-induced signaling cascade, we analyzed the transcriptome of MtCLE35-overexpressing roots inoculated by rhizobia using MACE (Massive Analysis of cDNA Ends) sequencing. Totally, 1390 genes were found to be differentially expressed between MtCLE35-overexpressing (35S::MtCLE35) and control (35S::GUS) roots after rhizobial inoculation, among them 268 genes were upregulated and 1122 genes were downregulated. Among downregulated genes, many known regulators of legume-rhizobia symbiosis were found. Genes upregulated in rhizobia-inoculated MtCLE35-overexpressing roots included ones associated with defense response and cellular redox status. Furthermore, stable transgenic plants overexpressing the MtCLE35 gene were obtained in this study, and gene expression qPCR analysis of selected differentially expressed genes in rhizobia-inoculated roots was performed in such plants. Collectively, our data suggest that overexpression of the MtCLE35 gene prevents the induction of nodulation program, which is accompanied by the expression of defense-related gene in MtCLE35-overexpressing rhizobia-inoculated M. truncatula roots. [ABSTRACT FROM AUTHOR]

This research aims to determine and quantify peri-urban land-use patterns for genetically modified crops, woody and nonwoody vegetation, and noncrop areas around Argentine cities where the agro-industrial model is implemented. Through a selection process, a database of urban areas for 715 cities was obtained. Perimeter rings of 100, 500, 1,000, and 2,000 m were used to calculate the areas occupied by the different peri-urban land covers, total crops, no crops, and tree cover. As a general pattern, it is observed that as larger peripheral areas are included, the predominance of the total crop class increases, while that of no crops and tree cover decreases. In addition, it was estimated that an average of 9.7 million kilograms of pesticides are being used per crop season on the 1.8 million hectares analysed, that is, on the 2,000 peripheral meters of the 715 cities under study. [ABSTRACT FROM AUTHOR]

SUMMARY Cold stress affects the growth, development, and yield of asparagus bean (Vigna unguiculata subsp. sesquipedalis). Mediator (MED) complex subunits regulate the cold tolerance of asparagus bean, but the underlying regulatory mechanisms remain unclear. Here, VunMED2 positively responds to cold stress of asparagus beans. Under cold acclimation and freezing treatment, the survival rate, ROS scavenging activity, and expression levels of VunMED2 were increased in VunMED2 transgenic plants. Natural variation in the promoter of VunMED2 in two different cold‐tolerant asparagus beans was observed. Under cold stress, the expression of the GUS reporter gene was higher in cold‐tolerant plants than in cold‐sensitive plants, and the expression of the GUS reporter gene was tissue‐specific. VunHY5 positively influenced the expression of VunMED2 by binding to the E‐box motif, and the transcriptional activation of the promoter was stronger in the cold‐tolerant variety than in cold‐sensitive plants. VunHY5 overexpression improved plant freezing resistance by increasing the antioxidant capacity and expression of dehydrin genes. VunHY5 and VunMED2 play a synergistic role in binding to the G‐box/ABRE motif and transcriptionally activating the expression of VunERD14. VunERD14 complemented the med2 mutant, which could positively respond to plant freezing resistance by reducing membrane lipid peroxidation and improving the antioxidant capacity. Therefore, the VunHY5‐VunERD14 module and the VunHY5‐VunMED2‐VunERD14 positive cascade effect are involved in the cold signal transduction in asparagus bean. Our findings have implications for the breeding of asparagus bean varieties with improved cold tolerance. [ABSTRACT FROM AUTHOR]

The role of molecular hydrogen (H 2) in autophagy during inflammatory response is controversial in mammalian cells. Although the stimulation of H 2 production in response to osmotic stress was observed in plants, its synthetic pathway and the interrelationship between its induction and plant autophagy remain unclear. Here, the induction of autophagy was observed in Arabidopsis upon osmotic stress, assessing by the autophagosome formation and autophagy-related genes expression. Above responses were intensified by H 2 fumigation. Meanwhile, the reduction in seedling growth and roots vigor was obviously abolished, accompanied by reestablishing redox balance. These H 2 responses were markedly impaired in T-DNA knockout lines atg2 , atg5 , and atg18. Further evidence showed that the increased endogenous H 2 synthesis by genetic manipulation, not only stimulated autophagosome formation, but also triggered various plant responses toward osmotic stress. By contrast, these responses were obviously abolished by the disruption of endogenous H 2 synthesis with the addition of 2,6-dichloroindophenol sodium salt. Together, the integrated genetic and molecular evidence clearly illustrated the requirement of autophagy activation in H 2 control of plant osmotic tolerance. [Display omitted] • H 2 fumigation contributed to plant tolerance against osmotic stress. • Osmotic stress-induced autophagy was intensified by H 2 , in which ATGs acted as downstream targets. • Endogenous H 2 -mediated autophagy played important roles in promoting osmotic tolerance in Arabidopsis. • H 2 fumigation and transgenic plants were more reliable methods to explore the role of H 2 in plants. [ABSTRACT FROM AUTHOR]

Rice (Oryza sativa) produces phenolamides and diterpenoids as major phytoalexins. Although the biosynthetic pathways of phenolamides and diterpenoids in plants have been revealed, knowledge of their accumulation regulatory mechanisms remains limited, and, in particular, no co-regulatory factor has been identified to date. Here, using a combined co-expression and evolutionary analysis, we identified the basic helix–loop–helix (bHLH) transcription factor OsbHLH5 as a positive bifunctional regulator of phenolamide and diterpenoid biosynthesis in rice. Metabolomic analysis revealed that OsbHLH5 significantly increased the content of phenolamides (such as feruloyl tryptamine (Fer-Trm) and p-coumaroyl tyramine (Cou-Tyr)) and diterpenoid phytoalexins (such as momilactones A, momilactones B) in the overexpression lines, while their content was reduced in the OsbHLH5 knockout lines. Gene expression and dual-luciferase assays revealed that OsbHLH5 activates phenolamide biosynthetic genes (including putrescine hydroxycinnamoyltransferase 3 (OsPHT3), tyramine hydroxycinnamoyltransferases 1/2 (OsTHT1/2), and tryptamine benzoyltransferase 2 (OsTBT2)) as well as diterpenoid biosynthetic genes (including copalyl diphosphate synthase 4 (OsCPS4) and kaurene synthase-like 4/7/10/11 (OsKSL4/7/10/11)). Furthermore, we have demonstrated that OsbHLH5 is induced by jasmonic acid (JA), while pathogen inoculation assays indicated that the overexpression of OsbHLH5 in transgenic rice plants leads to enhanced resistance to Xanthomonas oryzae pv. oryzae (Xoo). Overall, we have identified a positive regulator of phenolamide and diterpenoid biosynthesis and have demonstrated that biotic stress induces phytoalexin accumulation partly in an OsbHLH5-dependent manner, providing new insights into the metabolic interactions involved in pathogen response and offering valuable gene resources for the development, through genetic improvement, of new rice varieties that are resistant to diseases. [ABSTRACT FROM AUTHOR]

Bracts are leaf‐like structures in flowering plants. They serve multiple functions such as attracting pollinators, aiding tolerance of abiotic stressors, and conducting photosynthesis. While previous studies extensively examine bract function, the molecular mechanisms underlying bract growth remain unknown. Here, the map‐based isolation and characterization of a crucial factor responsible for cotton bract development, identified from a mutant known as frego bract (fg), discovered by Frego in 1945 are presented. This gene, named Ghfg, encodes a CC‐NB‐ARC‐LRR (CNL) family protein. Through analysis of bract form in plants with virus‐induced gene silencing (VIGS) and transgenic plants, this gene is confirmed to be the causal gene under the fg locus. Furthermore, high‐resolution single‐cell transcriptomic landscape of cotton bracts is generated, which reveals differences related to auxin in proliferating cells from TM‐1 and T582; differences in auxin distribution and ROS accumulation are experimentally verified. These findings suggest that GhFG is in a self‐activated state in the fg mutant, and its activity leads to ROS accumulation that impacts auxin distribution and transport. Finally, an island cotton variety with the frego bract trait is developed, demonstrating a novel solution for reducing the high impurity rate caused by bract remnants. [ABSTRACT FROM AUTHOR]

SUMMARY: Radish (Raphanus sativus L.) is one of the most important root vegetable crops worldwide. However, gene function exploration and germplasm innovation still face tremendous challenges due to its extremely low transformation efficiency. Here, an economic and highly efficient genetic transformation method for radish was explored by Agrobacterium rhizogenes‐mediated transformation with the help of combining special developmental regulator (DR) genes and the visual identification reporter. Firstly, the RUBY gene, a betalain biosynthesis system, could result in a visual red‐violet color used as a convenient and effective reporter for monitoring transgenic hairy roots screening of radish. However, the hairy roots‐to‐shoots conversion system of radish still stands as a barrier to the obtainment of whole transgenic plants, although different hormone combinations and various culture conditions were tried. Following, two DR genes including Wuschel2 (Wus2) and isopentenyl transferase (ipt), as well as their combination Wus2‐ipt were introduced for the shoot regeneration capacity improvement. The results showed that the transgenic shoots could be directly generated without externally supplying any hormones in the presence of a Wus2‐ipt combination. Then, Wus2‐ipt along with the RUBY reporter was employed to establish an efficient genetic transformation system of radish. Moreover, this system was applied in generating gene‐edited radish plants and the phytoene desaturase (RsPDS) gene was effectively knockout through albino phenotype observation and sequencing analysis. These findings have the potential to be widely applied in genetic transformation and genome‐editing genetic improvement of other vegetable species. Significance Statement: We successfully explored an economic and highly efficient genetic transformation method for radish by Agrobacterium rhizogenes‐mediated transformation with the help of the Wus2‐ipt‐RUBY system, which did not need any hormone supplement for shoot regeneration and could achieve a transformation efficiency of 29.21–40.00% in less than 4 months and be used to generate knockout mutants through CRISPR/Cas9 genome editing. These findings have the potential to be widely applied in genetic transformation and genome‐editing genetic improvement of other vegetable species. [ABSTRACT FROM AUTHOR]

The heat shock transcription factor (HSF) family is one of the most widely studied transcription factor families in plants; HSFs can participate in the response to various stressors, such as heat stress, high salt, and drought stress. Based on garlic transcriptome data, we screened and identified 22 garlic HSFs. The HSF proteins of garlic and Arabidopsis can be divided into three (A, B, C) subfamilies. The phylogenetic relationship, chromosome localization, sequence characteristics, conserved motifs, and promoter analysis of the HSF family were analyzed through bioinformatics methods. RT-qPCR analysis showed that the nine selected genes had different degrees of response to heat stress. In addition, we isolated and identified a class B HSF gene, AsHSFB1, from garlic variety 'Xusuan No.6'. Subsequently, the AsHSFB1 gene was overexpressed in Arabidopsis thaliana. Under heat stress, the germination rate and growth of wild-type plants were better than that of transgenic plants. Moreover, after heat treatment, the contents of peroxidase, catalase, and chlorophyll a and b of transgenic plants were lower, but the contents of malondialdehyde (MDA) and leaf conductivity were higher. Nitroblue tetrazolium (NBT) staining showed that the stained area of transgenic plant leaves was larger than that of the wild type. Further studies showed that AsHSFB1 overexpression inhibited the expression of related reverse resistance genes. These results indicate that AsHSFB1 might play a negative regulatory role in garlic resistance under high stress. Altogether, these findings provide valuable data for revealing the function of HSF genes and lay a foundation for the subsequent selection of heat-resistant garlic varieties. [ABSTRACT FROM AUTHOR]

Barley (Hordeum vulgare L.), a major cereal crop grown in arid and semi-arid regions, faces significant yield variability due to drought and heat stresses. In this study, the HvABF2 gene, encoding an ABA-dependent transcription factor, was cloned using specific primers from water deficit-stressed barley seedlings. Gene expression analysis revealed high HvABF2 expression in developing caryopses and inflorescences, with significant induction under stress conditions. The HvABF2 coding sequence was utilized to generate transgenic barley plants with both stress-inducible and constitutive expression, driven by the rice SNAC1 and maize Ubiquitin promoters, respectively. Selected transgenic barley lines, along with control lines, were subjected to water deficit-stress experiments at seedling and flag leaf stages under controlled and greenhouse conditions. The transgenic lines exhibited higher relative water content and stomatal resistance under stress compared to control plants. However, constitutive overexpression of HvABF2 led to growth retardation under well-watered conditions, resulting in reduced plant height, grain weight, and grain number. In contrast, stress-inducible expression mitigated these effects, demonstrating improved drought tolerance without adverse growth impacts. This study highlights that the stress-inducible expression of HvABF2, using the SNAC1 promoter, effectively improves drought tolerance while avoiding the negative pleiotropic effects observed with constitutive expression. [ABSTRACT FROM AUTHOR]