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Abstract The current reports on phytopathogens multidrug resistance have become a significant issue for plant health and global food security. Antimicrobial peptides (AMPs) have recently gained generous attention as potential alternatives to prevent plant disease resistance because of their potent, multifarious antimicrobial activity. AMPs are low-weight protein molecules. Living organisms secrete a wide range of AMPs, with some synthesised by canonical gene expression, known as ribosomal AMPs, and non-ribosomal AMPs, synthesised by modular enzyme-generating systems. Plants produce an array of AMPs, yet they are still unknown to many infection processes of causal agents. Plant-derived AMPs have a wide range of structures and functions, and they induce an innate immune system in plants. The biologically active AMPs in plants mainly depend on direct and indirect interactions with membrane lipids. Transgenic plants have expressed several AMPs, the basis for the model of new synthetic analogues, to provide support against diseases. These peptides have shown significant ability to manage plant diseases and can provide an eco-friendly alternative to hazardous conventional methods. Here, we have a comprehensive study on AMPs to identify their role in plant pathogen stress suppression activities and their mode of action. This would surely facilitate a bottomless insight into AMPs' mode of action against pathogen infections. An improved understanding of the mechanism will facilitate the development of the next generation of antimicrobial peptides, potentially employing a multitargeted approach.

Abstract As a typical bulb flower, lily is widely cultivated worldwide because of its high ornamental, medicinal and edible value. Although breeding efforts evolved over the last 10000 years, there are still many problems in the face of increasing consumer demand. The approach of biotechnological methods would help to solve this problem and incorporate traits impossible by conventional breeding. Target traits are dormancy, development, color, floral fragrance and resistances against various biotic and abiotic stresses, so as to improve the quality of bulbs and cut flowers in planting, cultivation, postharvest, plant protection and marketing. Genetic transformation technology is an important method for varietal improvement and has become the foundation and core of plant functional genomics research, greatly assisting various plant improvement programs. However, achieving stable and efficient genetic transformation of lily has been difficult worldwide. Many gene function verification studies depend on the use of model plants, which greatly limits the pace of directed breeding and germplasm improvement in lily. Although significant progress has been made in the development and optimization of genetic transformation systems, shortcomings remain. Agrobacterium-mediated genetic transformation has been widely used in lily. However, severe genotypic dependence is the main bottleneck limiting the genetic transformation of lily. This review will summarizes the research progress in the genetic transformation of lily over the past 30 years to generate the material including a section how genome engineering using stable genetic transformation system, and give an overview about recent and future applications of lily transformation. The information provided in this paper includes ideas for optimizing and improving the efficiency of existing genetic transformation methods and for innovation, provides technical support for mining and identifying regulatory genes for key traits, and lays a foundation for genetic improvement and innovative germplasm development in lily.

Abstract Background γ-Hexachlorocyclohexane (γ-HCH), an organochlorine insecticide of anthropogenic origin, is a persistent organic pollutant (POP) that causes environmental pollution concerns worldwide. Although many γ-HCH-degrading bacterial strains are available, inoculating them directly into γ-HCH-contaminated soil is ineffective because of the low survival rate of the exogenous bacteria. Another strategy for the bioremediation of γ-HCH involves the use of transgenic plants expressing bacterial enzyme for γ-HCH degradation through phytoremediation. Results We generated transgenic Arabidopsis thaliana expressing γ-HCH dehydrochlroninase LinA from bacterium Sphingobium japonicum strain UT26. Among the transgenic Arabidopsis T2 lines, we obtained one line (A5) that expressed and accumulated LinA well. The A5-derived T3 plants showed higher tolerance to γ-HCH than the non-transformant control plants, indicating that γ-HCH is toxic for Arabidopsis thaliana and that this effect is relieved by LinA expression. The crude extract of the A5 plants showed γ-HCH degradation activity, and metabolites of γ-HCH produced by the LinA reaction were detected in the assay solution, indicating that the A5 plants accumulated the active LinA protein. In some A5 lines, the whole plant absorbed and degraded more than 99% of γ-HCH (10 ppm) in the liquid medium within 36 h. Conclusion The transgenic Arabidopsis expressing active LinA absorbed and degraded γ-HCH in the liquid medium, indicating the high potential of LinA-expressing transgenic plants for the phytoremediation of environmental γ-HCH. This study marks a crucial step toward the practical use of transgenic plants for the phytoremediation of POPs.

Abstract Peptide deformylase can catalyse the removal of formyl groups from the N-terminal formyl methionine of the primary polypeptide chain. The peptide deformylase genes of a few herbaceous plants have been studied to some extent, but the peptide deformylase genes of woody plants have not been studied. In this study, we isolated EuPDF1B from Eucommia ulmoides Oliv. The full-length sequence of EuPDF1B is 1176 bp long with a poly-A tail and contains an open reading frame of 831 bp that encodes a protein of 276 amino acids. EuPDF1B was localized to the chloroplast. qRT‒PCR analysis revealed that this gene was expressed in almost all tissues tested but mainly in mature leaves. Moreover, the expression of EuPDF1B was enhanced by ABA, MeJA and GA and inhibited by shading treatment. The expression pattern of EuPDF1B was further confirmed in EuPDF1Bp: GUS transgenic tobacco plants. Among all the transgenic tobacco plants, EuPDF1Bp-3 showed the highest GUS histochemical staining and activity in different tissues. This difference may be related to the presence of enhancer elements in the region from − 891 bp to − 236 bp of the EuPDF1B promoter. In addition, the expression of the chloroplast gene psbA and the net photosynthetic rate, fresh weight and height of tobacco plants overexpressing EuPDF1B were greater than those of the wild-type tobacco plants, suggesting that EuPDF1B may promote the growth of transgenic tobacco plants. This is the first time that PDF and its promoter have been cloned from woody plants, laying a foundation for further analysis of the function of PDF and the regulation of its expression.

Improving the nutritional value of grain sorghum, a drought- and heat-tolerant grain crop, is an important task in the context of global warming. One of the reasons for the low nutritional value of sorghum grain is the resistance of its storage proteins (kafirins) to proteolytic digestion, which is due, among other things, to the structural organization of protein bodies, in which γ-kafirin, the most resistant to proteases, is located on the periphery, encapsulating more easily digested α-kafirins. The introduction of genetic constructs capable of inducing RNA silencing of the γ-kafirin (gKAF1) gene opens up prospects for solving this problem. Using Agrobacterium-mediated genetic transformation of immature embryos of the grain sorghum cv. Avans we have obtained a mutant with improved digestibility of endosperm proteins (up to 92 %) carrying a genetic construct for RNA silencing of the gKAF1 gene. The goal of this work was to study the stability of inheritance of the introduced genetic construct in T2–T4 generations, to identify the number of its copies, as well as to trace the manifestation of agronomically valuable traits in the offspring of the mutant. The mutant lines were grown in experimental plots in three randomized blocks. The studied lines were characterized by improved digestibility of kafirins, a modified type of endosperm, completely or partially devoid of the vitreous layer, an increased percentage of lysine (by 75 %), reduced plant height, peduncle length, 1000-grains weight, and grain yield from the panicle. In T2, a line with monogenic control of GA resistance was selected. qPCR analysis showed that in different T3 and T4 plants, the genetic construct was present in 2–4 copies. In T3, a line with a high digestibility of endosperm proteins (81 %) and a minimal decrease in agronomically valuable traits (by 5–7 %) was selected.

ABSTRACTMinghuai 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.

Wheat is a strategic cereal crop of global importance and plays a leading role in the food supply of mankind. Despite the general trend to increase in its production, climatic changes leading to significant temperature changes, unpredictable precipitation or droughts and the appearance of new races of pathogens and pests significantly affect its yield. In order to prevent the negative impact of changes in climatic conditions on the productivity of this crop, it is necessary to develop innovative technologies for improving the resistance of wheat to environmental stresses. RNA interference (RNAi) represents a new potential tool for wheat breeding by introducing small non-coding RNA sequences with the ability to silence gene expression in a sequence-specific manner. A decrease in the expression of a certain gene determines the acquisition of a new characteristic through the elimination or accumulation of certain plant traits, which leads to biochemical or phenotypic changes that the original plants do not have. This literature review describes the progress achieved over the past decades in the application of RNAi to create wheat plants with improved economic and valuable traits. The main stages of the gene silencing mechanism mediated by short interfering RNA (siRNA) and microRNA (miRNA), features of their biogenesis, modes of action and distribution are presented. Examples of the use of various biotechnological approaches to wheat improvement using gene transformation, endogenous and exogenous double-stranded RNA molecules (dsRNA) are given. The possibility of using RNAi technology to increase the nutritional value and quality of grain, remove toxic compounds and allergens is highlighted. Considerable attention is paid to the practical results of various applications of RNAi to increase the resistance of wheat to biotic stress factors, in particular, viruses, bacteria, fungi, insect pests, and nematodes. Examples of the use of siRNA-mediated RNAi and the role of miRNA in improving wheat tolerance to abiotic stresses are summarized.

Under changing climatic scenarios, grassland conservation and development have become imperative to impart functional sustainability to their ecosystem services. These goals could be effectively and efficiently achieved with targeted genetic improvement of native grass species. To the best of our literature search, very scant research findings are available pertaining to gene editing of non-cultivated grass species (switch grass, wild sugarcane, Prairie cordgrass, Bermuda grass, Chinese silver grass, etc.) prevalent in natural and semi-natural grasslands. Thus, to explore this novel research aspect, this study purposes that gene editing techniques employed for improvement of cultivated grasses especially sugarcane might be used for non-cultivated grasses as well. Our hypothesis behind suggesting sugarcane as a model crop for genetic improvement of non-cultivated grasses is the intricacy of gene editing owing to polyploidy and aneuploidy compared to other cultivated grasses (rice, wheat, barley, maize, etc.). Another reason is that genome editing protocols in sugarcane (x = 10–13) have been developed and optimized, taking into consideration the high level of genetic redundancy. Thus, as per our knowledge, this review is the first study that objectively evaluates the concept and functioning of the CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 technique in sugarcane regarding high versatility, target specificity, efficiency, design simplicity, and multiplexing capacity in order to explore novel research perspectives for gene editing of non-cultivated grasses against biotic and abiotic stresses. Additionally, pronounced challenges confronting sugarcane gene editing have resulted in the development of different variants (Cas9, Cas12a, Cas12b, and SpRY) of the CRISPR tool, whose technicalities have also been critically assessed. Moreover, different limitations of this technique that could emerge during gene editing of non-cultivated grass species have also been highlighted.

The sensitivity of crops to ultraviolet B (UVB, 280–315 nm) radiation varies significantly. Plants' sensitivity to UVB is heavily influenced by the activity of the enzyme cyclobutane pyrimidine dimer (CPD) photolyase, which fixes UVB-induced CPDs. Crops grown in tropical areas with high level of UVB radiation, like O. glaberrima from Africa and O. sativa ssp. indica rice from Bengal, are more sensitive to UVB radiation and could suffer more as a result of rising UVB levels on the earth's surface. Therefore, creating crops that can withstand high UVB is crucial in tropical regions. There is, however, little information on current techniques for breeding UVB-resistant plants. The most recent techniques for producing UVB-resistant crops are presented in this review. The use of DNA methylation, boosting the antioxidant system, regulating the expression of micro-RNA396, and overexpressing CPD photolyase in transgenic plants are some of the methods that are discussed. CPD photolyase overexpression in transgenic plants is the most popular technique for producing UVB-resistant rice. The study also offers several strategies for creating UVB-resistant plants using gene editing techniques. To feed the world's rapidly expanding population, researchers can use the information from this study to improve food production.

Abstract Remote sensing of vegetation by spectroscopy is increasingly used to characterize trait distributions in plant communities. How leaves interact with electromagnetic radiation is determined by their structure and contents of pigments, water, and abundant dry matter constituents like lignins, phenolics, and proteins. High-resolution (“hyperspectral”) spectroscopy can characterize trait variation at finer scales, and may help to reveal underlying genetic variation—information important for assessing the potential of populations to adapt to global change. Here, we use a set of 360 inbred genotypes of the wild coyote tobacco Nicotiana attenuata: wild accessions, recombinant inbred lines (RILs), and transgenic lines (TLs) with targeted changes to gene expression, to dissect genetic versus non-genetic influences on variation in leaf spectra across three experiments. We calculated leaf reflectance from hand-held field spectroradiometer measurements covering visible to short-wave infrared wavelengths of electromagnetic radiation (400–2500 nm) using a standard radiation source and backgrounds, resulting in a small and quantifiable measurement uncertainty. Plants were grown in more controlled (glasshouse) or more natural (field) environments, and leaves were measured both on- and off-plant with the measurement set-up thus also in more to less controlled environmental conditions. Entire spectra varied across genotypes and environments. We found that the greatest variance in leaf reflectance was explained by between-experiment and non-genetic between-sample differences, with subtler and more specific variation distinguishing groups of genotypes. The visible spectral region was most variable, distinguishing experimental settings as well as groups of genotypes within experiments, whereas parts of the short-wave infrared may vary more specifically with genotype. Overall, more genetically variable plant populations also showed more varied leaf spectra. We highlight key considerations for the application of field spectroscopy to assess genetic variation in plant populations.

The amino acid proline accumulates in plants during abiotic stresses such as drought and salinity and is considered a reliable marker of environmental stress. While its accumulation is well established, its precise role in stress tolerance and its underlying molecular mechanism remain less clear. To address these issues, we performed a meta-analysis—a robust statistical technique that synthesizes results from multiple independent studies while accounting for experimental differences. We focused on 16 physiological and morphological parameters affected by drought and salt stress in transgenic plants expressing proline metabolic genes. For each parameter, we calculated the effect size as the response ratio (RR), which represents the logarithm of the mean value in the transgenic group over the mean value of the control group (lnRR). Under stress, most parameters exhibited significantly higher response ratios in the transgenic group, confirming the beneficial effects of proline during drought and salt stress. Surprisingly, under non-stressed conditions, most stress markers showed no significant differences between transgenic and non-transgenic plants, despite elevated proline levels in the former. These results suggest that the benefits of proline may be related to proline catabolism or may only become apparent during stress, possibly due to interactions with reactive oxygen species (ROS), which accumulate predominantly under stress conditions.

Abstract Background C-lignin is a homopolymer of caffeyl alcohol present in the seed coats of a variety of plant species including vanilla orchid, various cacti, and the ornamental plant Cleome hassleriana. Because of its unique chemical and physical properties, there is considerable interest in engineering C-lignin into the cell walls of bioenergy crops as a high-value co-product of bioprocessing. We have used information from a transcriptomic analysis of developing C. hassleriana seed coats to suggest strategies for engineering C-lignin in a heterologous system, using hairy roots of the model legume Medicago truncatula. Results We systematically tested strategies for C-lignin engineering using a combination of gene overexpression and RNAi-mediated knockdown in the caffeic acid/5-hydroxy coniferaldehyde 3/5-O-methyltransferase (comt) mutant background, monitoring the outcomes by analysis of lignin composition and profiling of monolignol pathway metabolites. In all cases, C-lignin accumulation required strong down-regulation of caffeoyl CoA 3-O-methyltransferase (CCoAOMT) paired with loss of function of COMT. Overexpression of the Selaginella moellendorffii ferulate 5-hydroxylase (SmF5H) gene in comt mutant hairy roots resulted in lines that unexpectedly accumulated high levels of S-lignin. Conclusion C-Lignin accumulation of up to 15% of total lignin in lines with the greatest reduction in CCoAOMT expression required the strong down-regulation of both COMT and CCoAOMT, but did not require expression of a heterologous laccase, cinnamyl alcohol dehydrogenase (CAD) or cinnamoyl CoA reductase (CCR) with preference for 3,4-dihydroxy-substituted substrates in M. truncatula hairy roots. Cell wall fractionation studies suggested that the engineered C-units are not present in a heteropolymer with the bulk of the G-lignin.

Four popular genotypes of Brassica napus, cv. BARI Sarisha-8, BARI Sarisha-13, Binasarisha-4, and Binasarisha-8 were subjected to cadmium (Cd) and lead (Pb) stress to identify a genotype with enhanced tolerance of Cd and Pb. The goal was to use the selected genotype for genetic engineering, thereby improving traits related to heavy metal tolerance and accumulation to remediate polluted agricultural soils. The genotype, BARI Sarisha-8 with superior tolerance to both Cd and Pb stresses, was chosen for genetic engineering with Arabidopsis thaliana ABC transporter of the mitochondrion 3 (AtATM3) gene. The transformation process involved coculturing the cotyledon explants with Agrobacterium strain GV3101 carrying a binary vector containing the hygromycin phosphotransferase (HPT) gene as a selectable marker and the AtATM3 gene. Transformation efficiency was significantly enhanced with a two-day co-cultivation period on shoot induction medium composed of MS medium, α-naphthaleneacetic acid (0.5 mg/L), and 6-benzyladenine (3 mg/L), supplemented with acetosyringone (20 mg/L), along with a four-day delay in exposing the explants to the selective agent hygromycin. Hygromycin-resistant shoots were obtained by employing a three-step selection process. The refined protocol resulted in a transformation efficiency of 15.38%. Polymerase chain reaction (PCR) analysis confirmed the integration of AtATM3 and HPT genes into the host genome in all recombinant plants. Transgenic B. napus plants expressing the AtATM3 gene exhibited notable improvements in tolerance, demonstrating a 1.4- to 1.7fold increase in Cd tolerance and a 1.3- to 1.5-fold increase in Pb tolerance compared to the wild type (nontransformed) under both Cd and Pb stress conditions.

Calcium ions function as key messengers in the context of intracellular signal transduction. The ability of plants to respond to biotic and abiotic stressors is highly dependent on the calcineurin B-like protein (CBL) and CBL-interacting protein kinase (CIPK) signaling network. Here, a comprehensive effort was made to identify all members of the soybean CBL gene family, leading to the identification of 15 total genes distributed randomly across nine chromosomes, including 13 segmental duplicates. All the GmCBL gene subfamilies presented with similar gene structures and conserved motifs. Analyses of the expression of these genes in different tissues revealed that the majority of these GmCBLs were predominantly expressed in the roots. Significant GmCBL expression and activity increases were also observed in response to a range of stress-related treatments, including salt stress, alkaline stress, osmotic stress, or exposure to salicylic acid, brassinosteroids, or abscisic acid. Striking increases in GmCBL1 expression were observed in response to alkaline and salt stress. Subsequent analyses revealed that GmCBL1 was capable of enhancing soybean salt and alkali tolerance through the regulation of redox reactions. These results offer new insight into the complex mechanisms through which the soybean CBL gene family regulates the responses of these plants to environmental stressors, highlighting promising targets for efforts aimed at enhancing soybean stress tolerance.

Radish (Raphanus sativus L.) is a globally significant vegetable and relies on cytoplasmic male sterile (CMS) lines for hybrid seed production. The NWB CMS type is favored over Ogura CMS for its ease in maintainer screening. Despite its varied mitochondrial configurations and unvalidated sterile gene, we re-sequenced the mitochondrial genome of NWB CMS Tibet A and verified the function of the sterility gene via genetic transformation of Arabidopsis thaliana. The mitochondrial genomes of Tibet A could be assembled into circular DNA molecules, with a mitochondrial genome size of 239,184 bp. Our analysis indicated that the specific orf463a was the CMS-associated gene in Tibet A, sharing sequence consistency with the CMS gene in DCGMS and NWB CMS YB-A. Collinearity analysis showed that the mitochondrial genomes of NWB CMS Tibet A, DCGMS, and NWB CMS YB-A share the same mitotype, with structural variations due to recombination via a 9731 bp long repeat sequence and a 508 bp short repeat sequence. Driven by the Ap3 promoter, transgenic Arabidopsis with orf463a exhibited male sterility, confirming the gene’s potential role in CMS. In this study, we assembled a new isomeric form of NWB CMS mitochondrial genome and proved the function of the candidate sterile gene.

CRISPR-Cas9, the “genetic scissors”, is being presaged as a revolutionary technology, having tremendous potential to create designer crops by introducing precise and targeted modifications in the genome to achieve global food security in the face of climate change and increasing population. Traditional genetic engineering relies on random and unpredictable insertion of isolated genes or foreign DNA elements into the plant genome. However, CRISPR-Cas based gene editing does not necessarily involve inserting a foreign DNA element into the plant genome from different species but introducing new traits by precisely altering the existing genes. CRISPR edited crops are touching markets, however, the world community is divided over whether these crops should be considered genetically modified (GM) or non-GM. Classification of CRISPR edited crops, especially transgene free crops as traditional GM crops, will significantly affect their future and public acceptance in some regions. Therefore, the future of the CRISPR edited crops is depending upon their regulation as GM or non-GMs, and their public perception. Here we briefly discuss how CRISPR edited crops are different from traditional genetically modified crops. In addition, we discuss different CRISPR reagents and their delivery tools to produce transgene-free CRISPR edited crops. Moreover, we also summarize the regulatory classification of CRISPR modifications and how different countries are regulating CRISPR edited crops. We summarize that the controversy of CRISPR-edited plants as GM or non-GM will continue until a universal, transparent, and scalable regulatory framework for CRISPR-edited plants will be introduced worldwide, with increased public awareness by involving all stakeholders.

Abstract Background Studies have indicated that long non-coding RNAs (lncRNAs) play important regulatory roles in many biological processes. However, the regulation of seed oil biosynthesis by lncRNAs remains largely unknown. Results We comprehensively identified and characterized the lncRNAs from seeds in three developing stages in two accessions of Brassica napus (B. napus), ZS11 (high oil content) and WH5557 (low oil content). Finally, 8094 expressed lncRNAs were identified. LncRNAs MSTRG.22563 and MSTRG.86004 were predicted to be related to seed oil accumulation. Experimental results show that the seed oil content is decreased by 3.1–3.9% in MSTRG.22563 overexpression plants, while increased about 2% in MSTRG.86004, compared to WT. Further study showed that most genes related to lipid metabolism had much lower expression, and the content of some metabolites in the processes of respiration and TCA (tricarboxylic acid) cycle was reduced in MSTRG.22563 transgenic seeds. The expression of genes involved in fatty acid synthesis and seed embryonic development (e.g., LEC1) was increased, but genes related to TAG assembly was decreased in MSTRG.86004 transgenic seeds. Conclusion Our results suggest that MSTRG.22563 might impact seed oil content by affecting the respiration and TCA cycle, while MSTRG.86004 plays a role in prolonging the seed developmental time to increase seed oil accumulation.

The global climate changes and the consequent increase in the number of soil and air droughts during the vegetation period of grain crops require the development of new strategies to adapt plants to those yield-decreasing stressors. A relevant way of increasing drought-tolerance of cereals is the use of biotechnological methods, particularly RNA interference, which can down-regulate the activity of plants’ genes and increase concentration of stress metabolites that perform osmoprotective functions during drought. We studied the tolerance to soil moisture shortage in transgenic plants of winter wheat with partial suppression of the proline dehydrogenase gene, obtained using the technology of short interfering RNAs. We analyzed physiological and biochemical parameters and structural elements of yield productivity of 4 wild genotypes and their transgenic lines with reduced activity of proline dehydrogenase in the conditions of 7-day drought during the late booting–ear emergence. We determined that the presence of double-stranded RNA suppressor of the proline dehydrogenase gene in transgenic lines led to increase in the level of accumulation of free proline in flag leaves. At the same time, its concentration in transgenic lines was higher than in untransformed plants of the wild genotypes in both drought conditions and conditions of sufficient moisture. We found that against the background of water deficiency, the total chlorophyll content in leaves of plants of transgenic lines was significantly higher, and the ratio of carotenoids to chlorophyll was lower than in plants of the wild genotypes, suggesting mitigation of the negative impact of drought on the plants of transgenic lines. Lacking soil moisture, genetically altered lines of wheat had significantly higher parameters of the structure of grain yield compared with untransformed genotypes. At the same time, we observed genotypic difference according to grain productivity in biotechnological plants. Therefore, the results we obtained confirm the perspectives of using the technology of short interfering RNAs to increase tolerance of winter wheat to water deficiency.

Uncontrolled distribution of goods produced by genetically modified plants should be prevented by the state in order to secure food safety in the Russian Federation and to minimize health risks for consumers. We analyzed foods of plant origin for children to identify components of genetically modified organisms in them. It was done to ensure safety of such foods. The highest specific weight among the analyzed foods belonged to nectars (40.0 %) and juice-containing drinks (36.0 %). Juices and fruit drinks accounted for 12 % each. Genetically modified organisms were determined in foods by identifying regulatory sequences (35S promoter, FMV promoter and NOS terminator) that are widely used in constructions of genetically modified plants. Occurrence of regulatory genetic elements specific for genetically modified organisms was checked in juice products for children by the polymerase chain reaction in real-time mode with hybridization-fluorescent detection of amplification products and with the use of the “AmpliSens GM Plant-1-FL” and “AmpliKvant GM soya-FL” test systems. The results of this study showed that no analyzed foods of plant origin contained any regulatory sequences (35S, NOS и FMV) indicating presence of genetically modified organisms. Fluorescence through the FAM, Cy5 and ROX channels did not exceed its threshold value. Therefore, we did not detect any violations of the established requirements to occurrence of genetically modified organisms in foods for children. Further investigation that would involve examining a more extensive material is required to ensure proper assessment and control of food contamination with genetically modified organisms in order to ensure food safety.

Currently, several species of the genus Gossypium are cultivated to produce fiber. Cotton has been grown for a long time, however, many aspects of its cultivation and processing are still researched. When talking about the agronomy of cotton, some fundamental problems should be mentioned. For example, the amounts of pesticides used in the cultivation of cotton are greater than for any other crop. Chemicals sprayed on cotton are washed away from the fields and pollute fresh water sources, causing significant damage to the environment. Fortunately, such challenges can be overcome by switching to the cultivation of transgenic cotton. The introduction of transgenic cotton has already brought many important environmental, social and economic benefits, including a reduction in the use of pesticides, indirect influence on the increase in yields, minimization of environmental pollution, reduction of economic costs and labor for cultivating the crop. Until today, the main methods of obtaining transgenic cotton lines are still agrobacterial transformation and biolistics. In recent years, however, innovative methods of transformation have also been developed. For example, the pollen tube-mediated introduction of genetic material for obtaining commercial transgenic cotton is actively used in China. Although transgenic lines with resistance to diseases and abiotic stresses, and with improved fiber quality have been obtained in recent decades, the market of transgenic cotton is still dominated by insect- and herbicide-resistant lines. All the above indicates an insufficient integration between institutes as sources of advanced developments and agricultural industry. The present review collected and summarized the results of research on the cultivation and genetic modification of cotton. The main methods of genetic transformation of cultivated representatives of the genus Gossypium, both actively used at present and still under development, were considered. Also, the most remarkable transgenic lines were also described, among which are those that have already been adopted by the agricultural industry and those that have been obtained only recently. Thus, the reader will be able to get a general idea of the current achievements in the field of cotton genetic modification.

MADS-box is a large transcription factor family in plants and plays a crucial role in various plant developmental processes; however, it has not been systematically analyzed in kiwifruit. In the present study, 74 AcMADS genes were identified in the Red5 kiwifruit genome, including 17 type-I and 57 type-II members according to the conserved domains. The AcMADS genes were randomly distributed across 25 chromosomes and were predicted to be mostly located in the nucleus. A total of 33 fragmental duplications were detected in the AcMADS genes, which might be the main force driving the family expansion. Many hormone-associated cis-acting elements were detected in the promoter region. Expression profile analysis showed that AcMADS members had tissue specificity and different responses to dark, low temperature, drought, and salt stress. Two genes in the AG group, AcMADS32 and AcMADS48, had high expression levels during fruit development, and the role of AcMADS32 was further verified by stable overexpression in kiwifruit seedlings. The content of α-carotene and the ratio of zeaxanthin/β-carotene was increased in transgenic kiwifruit seedlings, and the expression level of AcBCH1/2 was significantly increased, suggesting that AcMADS32 plays an important role in regulating carotenoid accumulation. These results have enriched our understanding of the MADS-box gene family and laid a foundation for further research of the functions of its members during kiwifruit development.

The rapid advancement of genetic technologies has made it possible to modify various plants through both genetic transformation and gene editing techniques. Poplar, with its rapid in vitro growth and regeneration enabling high rates of micropropagation, has emerged as a model system for the genetic transformation of woody plants. In this study, Populus × berolinensis K. Koch. (Berlin poplar) was chosen as the model organism due to its narrow leaves and spindle-shaped crown, which make it highly suitable for in vitro manipulations. Various protocols for the Agrobacterium-mediated transformation of poplar species have been developed to date. However, the genetic transformation procedures are often constrained by the complexity of the nutrient media used for plant regeneration and growth, which could potentially be simplified. Our study presents a cheaper, simplified, and relatively fast protocol for the Agrobacterium-mediated transformation of Berlin poplar. The protocol involved using internode sections without axillary buds as explants, which were co-cultivated in 10 µL droplets of bacterial suspension directly on the surface of a solid agar-based medium without rinsing and sterile paper drying after inoculation. We used only one regeneration Murashige and Skoogbased medium supplemented with BA (0.2 mg·L−1), TDZ (0.02 mg·L−1), and NAA (0.01 mg·L−1). Acetosyringone was not used as an induction agent for vir genes during the genetic transformation. Applying our protocol and using the binary plasmid pBI121 carrying the nptII selective and uidA reporter genes, we obtained the six transgenic lines of poplar. Transgenesis was confirmed through a PCR-based screening of kanamycin-selected regenerants for the presence of both mentioned genes, Sanger sequencing, and tests for detecting the maintained activity of both genes. The transformation efficiency, considering the 100 explants taken originally, was 6%.

The development and improvement of molecular biology methods have led to the creation of new technologies that make it possible to modify plant genomes by transferring and integrating into the genomes’ heterologous genes from various expression systems (genetic engineering), as well as inducing knockouts of one or more target genes of interest (genomic editing). The development of genome-editing methods is a new milestone in the development of modern breeding methods and certainly relies on the knowledge and technologies developed for transgenesis. This review will discuss issues related to the advantages and disadvantages of both technologies for improving the economically valuable traits of important crops.

The development of legume–rhizobial symbiosis results in the formation of nitrogen-fixing root nodules. In response to rhizobial molecules, Nod factors, signal transduction is mediated by the interaction of activated receptors with downstream signaling proteins. Previously, some new regulators of the signal pathway, such as phospholipases D, which regulate the level of phosphatidic acid (PA), as well as mitogen-activated protein kinases (MAPKs), have been identified in legumes. Since PA is an important signal messenger, we tested the hypothesis that increasing the level of proteins involved in the reversible binding of PA in plant tissues may have a positive effect on symbiosis. Our findings showed that overexpression of MtSPHK1-PA, encoding the PA-binding domain of sphingosine kinase 1 (SPHK1), stimulated plant growth and nodule development in legume plants. Furthermore, the influence of MAPK6 on the development of symbiosis was studied. Using genetic engineering methods, we increased MAPK6 transcriptional activity in transgenic roots, leading to an increase in the number of nodules and the biomass of pea plants. Therefore, new approaches to obtain plants with an increased efficiency of symbiosis were tested. We report here that both genes that encode signaling proteins may be used as potential targets for future modification using biotechnological approaches.

This article is devoted to the study of the phenotypic manifestations of transformation induced by the Agrobacterium rhizogenes strain K599 in Nicotiana tabacum L., a model plant sensitive to agrobacteria. For this purpose, we carried out a genetic transformation of tobacco leaf disks that resulted in spontaneous callus and shoot formation on hormone-free media. On average, 6 regenerated shoots per leaf explant were obtained. Overall, 2 out of 87 transplanted shoots spontaneously formed roots, and 21 shoots died. In the remaining shoots, rooting was observed only when the growth regulator IAA was added to the medium. Using the above protocol, a total of 39 transgenic plantlets adapted to the given soil conditions were produced from spontaneous points of regeneration. The transformation also initiated the formation of hairy roots, but there were very few of them – 0.77 roots per explant on average. The isolated cultures of tobacco hairy roots generated using strain K599 did not phenotypically differ from those induced by strains A4 and 15834. On the hairy roots, 10 shoots transgenic for rol genes were regenerated by the induction of shoots using growth regulators, 8 of them were later rooted and acclimated to the given soil conditions. The PCR analysis showed the presence of rol genes in all transgenic plants that regenerated spontaneously or were induced on roots by growth regulators. Thus, strain K599, unlike other strains of A. rhizogenes, has a potential to be used to grow transgenic tobacco plants on hormone-free nutrient media by direct regeneration of shoots on explants.

Background Tannins are the main bottlenecks restricting the transformation efficiency of plants. Hongyingzi is a special tannin-containing sorghum cultivar used in brewing. Methods In this study, a highly efficient microprojectile transformation system for tannin-containing sorghum was successfully exploited using immature embryos (IEs) of Hongyingzi as explants. Results Hongyingzi presented two types of calli. Type II calli were found to be the most suitable and effective explants for transformation. After optimization of the geneticin (G418) concentration and tissue culture medium, an average transformation frequency of 27% was achieved. Molecular analyzis showed that all transgenic plants were positive and showed transgenes expression. The inheritance analyzis confirmed that the transgenes could be inherited into the next generation. Thus, we successfully established an efficient transformation system for tannin-containing sorghum and demonstrated the possibility of breaking the restriction imposed by tannins in plants.

Transgenic Arabidopsis plants expressing a potato D200 gene encoding a hypothetical protein were subjected to salinity stress and assessed for their tolerance. The D200 Arabidopsis lines exhibited increased chlorophyll content, improved stomatal conductance, less electrolyte leakage, lower accumulation of malondialdehyde (MDA), and a higher amount of proline compared to the wild type (WT) plants under salinity stress. The gene expression analysis revealed that D200 plants accumulated a significantly higher amount of mRNA transcripts of genes encoding three major antioxidant enzymes ascorbate peroxidase (APX), catalase (CAT), and superoxide dismutase (SOD). Chlorophyll a fluorescence kinetics analyses showed the D200 plants were more efficient in terms of primary photochemistry of photosystem II and performance indices. Furthermore, the quantum yields and efficiencies that represent the critical steps of photosynthetic light reactions were analyzed and it was found that D200 plants were photosynthetically more active than the WT plants under salt stress conditions. Overall, these findings suggest that the D200 gene is a potential candidate gene for developing stress-resilient crops in future.

Rice (Oryzae sativa cv. dongjin) is a cornerstone of global food security; however, Burkholderia glumae BGR1, which is responsible for bacterial panicle blight (BPB), threatens its productive output, with dire consequences for rice and other crops. BPB is primarily caused by toxoflavin, a potent phytotoxin that disrupts plant growth at various developmental stages. Therefore, understanding the mechanisms through which toxoflavin and BPB affect rice plants is critical. Toxoflavin biosynthesis in B. glumae BGR1 relies on the toxABCDE operon, with ToxA playing a central role. In response to this threat, our study explores a metagenome-derived toxoflavin-degrading enzyme, TxeA, as a potential defense mechanism against toxoflavin’s destructive impact. TxeA-induced degradation of toxoflavin represents a potential strategy to mitigate crop damage. We introduce a groundbreaking approach: engineering transgenic rice plants to produce toxoflavin-degrading enzymes. These genetically modified plants, armed with TxeA, hold significant potential for combating toxoflavin-related crop losses. However, removal of toxoflavin, a major virulence factor in B. glumae BGR1, does not completely inhibit virulence. This innovative perspective offers a new shift from pathogen eradication to leveraging transgenic plants’ power, offering a beacon of hope for crop protection and disease management. Our study offers insights into the intricate interplay between toxoflavin, BPB, and TxeA, providing a promising avenue to safeguard rice crops, ensure food security, and potentially enhance the resilience of various agricultural crops to B. glumae BGR1-induced diseases.

Transgenic tobacco plants overexpressing the choline oxidase gene from A. globiformis showed an increase in resistance at the level of primary and secondary biosynthesis of metabolites, removing the damage characteristic of salinity and stabilizing the condition of plants. We used 200 mM NaCl, which inhibits the growth of tobacco plants at all stages of development. Leaves of transgenic and wild-type (WT) plants Nicotiána tabácum were used for biochemical, cytological and molecular biological analysis. However, for transgenic lines cultivated under normal conditions (without salinity), we noted juvenile characteristics, delay in flowering, and slowing down of development, including the photosynthetic apparatus. This caused changes in the amount of chlorophyll, a delay in the plastid grana development with the preservation of prolamellar bodies. It also caused changes in the amount of sugars and indirectly downstream processes. A significant change in the activity of antioxidant enzymes and a change in metabolism is probably compensated by the regulation of a number of genes, the expression level of which was also changed. Thus, the tolerance of transgenic tobacco plants to salinity, which manifested itself as a result of the constitutive expression of codA, demonstrates an advantage over WT plants, but in the absence of salinity, transgenic plants did not have such advantages due to juvenilization.

Heat shock transcription factors (Hsfs) participate in a variety of plant physiological processes including the regulation of transcription factors associated with thermotolerance. Here, a Hsf gene DcHsfA1d was identified from carnation (Dianthus caryophyllus L.). The open reading frame (ORF) of DcHsfA1d was 1 368 bp and encoded a protein of 455 amino acids with a molecular mass of 51.039 kDa and an isoelectric point of 4.94. Sequence domain prediction revealed that DcHsfA1d protein exhibited five typical functional features and motifs. The transcription of DcHsfA1d was significantly up-regulated under heat stress or ABA treatment. Yeast two-hybrid experiment indicated that DcHsfA1d and DcHsp70 physically interact with each other. Overexpression of DcHsfA1d in Arabidopsis ecotype Columbia enhanced seedling thermotolerance by increasing the activities of catalase, peroxidase, and superoxide dismutase while reducing relative electrolyte leakage, malondialdehyde content, accumulation of O2- and H2O2 and by initiating transcriptional regulation of thermal protective gene expression under heat stress. Furthermore, under salt stress, the root length and fresh mass of Arabidopsis ectopically expressing DcHsfA1d were significantly higher than those of wild type, which indicated that the salt tolerance of transgenic Arabidopsis was improved to a certain extent. In summary, DcHsfA1d was demonstrated to play a positive regulatory role in heat stress response and it might be a candidate gene for salt tolerance using genetic modification.

The physicochemical properties of starches from different botanical origin, such as viscosity, gelatinization temperature, solubility and degradability depend on the amylose/amylopectin ratio and the length and frequency of the α-1,6-glycosidic bonds in amylopectin. These branches depend on the action of the starch branching enzymes (SBEs), which are highly structurally conserved in plants. We recently identified a novel gene from Ostreococcus tauri (OsttaSBEIII) which codes for a protein showing starch branching activity (OsttaSBEIII) and with a different structure than other known SBEs from plants, containing two in-tandem carbohydrate binding modules (CBM41-CBM48) at its N-terminus. OsttaSBEIII overexpression in A. thaliana plants resulted in a higher starch content and smaller granules with an increased degradability. OEOsttaSBE lines showed also an increase in the expression and activity of starch degradative enzymes and a higher content of glucose and inorganic phosphate, which suggests a remodeling of the granule structure in response to the expression of OsttaSBEIII. These results allow us to propose the use of OsttaSBEIII as a new strategy to obtain starches with greater degradability that would be useful for different biotechnological applications.

The Lemnaceae or duckweed family comprises 37 species of smallest and simplest flowering plants. Duckweeds have a fast growth rate, can survive under a wide range of temperature and pH conditions and are easy to maintain and harvest which makes them an excellent candidate for bioremediation of wastewaters. The main objective of the present review is to extend an appreciation for the potential of living and non-living biomass of duckweed in remediating waters contaminated with heavy metals. Along with showing the detailed mechanism of phytoremediation by duckweed, the paper also discusses the enhancement of duckweed phytoremediation by the integration of transgenic technology. Furthermore, the paper explores other applications of duckweed specifically as fuel, animal feed, in human nutrition, in medicine and as a life support system. Apart from this, various disposal mechanisms for harvested duckweed have been analysed. Current understanding of removal efficiencies of several contaminants by employing duckweed is limited mainly to laboratory experiments. More concentrated and persistent efforts to develop efficient approaches for the genetic transformation of duckweeds can expand the development and utilization of duckweeds.

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

Recombinant proteins are of paramount importance for research, industrial and medical use. Numerous expression chassis are available for recombinant protein production, and while bacterial and mammalian cell cultures are the most widely used, recent developments have positioned transgenic plant chassis as viable and often preferential options. Plant chassis are easily maintained at low cost, are hugely scalable, and capable of producing large quantities of protein bearing complex post-translational modification. Several protein targets, including antibodies and vaccines against human disease, have been successfully produced in plants, highlighting the significant potential of plant chassis. The aim of this review is to act as a guide to producing recombinant protein in plants, discussing recent progress in the field and summarising the factors that must be considered when utilising plants as recombinant protein expression systems, with a focus on optimising recombinant protein expression at the genetic level, and the subsequent extraction and purification of target proteins, which can lead to substantial improvements in protein stability, yield and purity.

The need for studies on the nutritional value of plants depends on their composition. The first generation genetically modified Plants (GMPs) have the same composition as their near-isogenic lines. Therefore, they have the same nutritional value, and most of the animal feeding studies have found no significant differences in the production and health parameters of animals that consumed first-generation GMPs in comparison to non-GMPs. Due to the recent production of transgenic plants with specific nutritional properties (second generation GMPs), their use as feed for animals is viewed with skepticism in very many countries. In this critical review, it is concluded that most of these nutritionally improved plants have not shown adverse effects on the performance of various animals compared to their near-isogenic lines and can therefore be considered in the animal feed industry. However, most of the experiments were conducted on laboratory animals. There is a need to conduct them with animals that are mainly consumed by humans, such as ruminants. There is also a need to feed the whole plant to these animals and not just certain parts of the plant to get a clear picture of its overall safety. In addition, there is a need to determine a suitable long-term nutritional and toxicological approach assessment.

Abiotic stresses are major limiting factors that pose severe threats to agricultural production. Conventional breeding has significantly improved crop productivity in the last century, but traditional breeding has reached its maximum capacity due to the multigenic nature of abiotic stresses. Alternatively, biotechnological approaches could provide new opportunities for producing crops that can adapt to the fast-changing environment and still produce high yields under severe environmental stress conditions. Many stress-related genes have been identified and manipulated to generate stress-tolerant plants in the past decades, which could lead to further increase in food production in most countries of the world. This review focuses on the recent progress in using transgenic technology and gene editing technology to improve abiotic stress tolerance in plants, and highlights the potential of using genetic engineering to secure food and fiber supply in a world with an increasing population yet decreasing land and water availability for food production and fast-changing climate that will be largely hostile to agriculture.

Abiotic stresses create an unfavourable environment for plant growth, increasing the possibilities of low yield and associated economic loss. Several steps have been taken to address this problem. During the last twenty years, techniques of genetic engineering/transgenic breeding have made significant advances in gene manipulation for inciting desirable traits in transgenic plants. Transgenic techniques allow us to identify potential genes, transcription factors (TFs) and miRNAs, engaged in certain processes in plants, allowing us to gain a comprehensive understanding of the processes at molecular and physiological levels which determine plant resilience and production. The reliability and specificity of this approach ensure that future plant enhancements will be a huge success. As a result, transgenic breeding has determined to be a viable strategy in improving crop abiotic stress tolerance. The approach of CRISPR/Cas gene-editing technique to create stress-tolerant plant variants is gaining popularity right now. The researchers like this user-friendly technology because of its versatility. In the gene-editing process, the DNA sequence "CRISPR" and the endonuclease "Cas" collaborate under the supervision of specific guide RNA. In a variety of plant species, the CRISPR/Cas system is being utilized. In the majority of situations, Cas9 is employed. Various reports have surfaced which demonstrate the utilization of CRISPR/Cas9 technology to improve abiotic stress tolerance of plants. The focus of this review is on the promising and effective applications of transgenic plant breeding for enhancing environmental stress tolerance and crop productivity, as well as its recent developments.

Soybean is one of the most widely grown oilseed crops worldwide. Several unfavorable factors, including salt and salt–alkali stress caused by soil salinization, affect soybean yield and quality. Therefore, exploring the molecular basis of salt tolerance in plants and developing genetic resources for genetic breeding is important. Sucrose non-fermentable protein kinase 1 (SnRK1) belongs to a class of Ser/Thr protein kinases that are evolutionarily highly conserved direct homologs of yeast SNF1 and animal AMPKs and are involved in various abiotic stresses in plants. The GmPKS4 gene was experimentally shown to be involved with salinity tolerance. First, using the yeast two-hybrid technique and bimolecular fluorescence complementation (BiFC) technique, the GmSNF1 protein was shown to interact with the GmPKS4 protein. Second, the GmSNF1 gene responded positively to salt and salt–alkali stress according to qRT-PCR analysis, and the GmSNF1 protein was localized in the nucleus and cytoplasm using subcellular localization assay. The GmSNF1 gene was then heterologously expressed in yeast, and the GmSNF1 gene was tentatively identified as having salt and salt–alkali tolerance function. Finally, the salt–alkali tolerance function of the GmSNF1 gene was demonstrated by transgenic Arabidopsis thaliana, soybean hairy root complex plants overexpressing GmSNF1 and GmSNF1 gene-silenced soybean using VIGS. These results indicated that GmSNF1 might be useful in genetic engineering to improve plant salt and salt–alkali tolerance.

Decades after the production of recombinant vaccines, the production of large scale and commercial use of such vaccines has become an important issue in the academic community. Newcastle disease is an infectious and highly contagious viral disease that causes diseases with different virulence and high infectivity in birds, especially chickens. Newcastle disease imposes severe economic losses on the poultry industry, and as a result, tackling it is always a priority for all countries of the world. In this regard, many vaccines have been produced, some of which are commercialized and some of which are in the testing phase. Given the economic importance of controlling Newcastle disease, the production of recombinant vaccines against the disease has been one of the hottest areas in the history of recombinant vaccines. Over the past three decades, many laboratory studies have been conducted to produce recombinant Newcastle disease vaccines on various platforms, which in many cases have yielded promising results. This article reviews the literature on the production of recombinant Newcastle disease vaccines. In this regard, while introducing Newcastle disease and its causative agent, the basics of producing recombinant vaccines and production platforms are explained. The following are some studies that have shown promising results for the production of the recombinant Newcastle disease vaccine. At the end of this article, while summarizing, areas for future research are introduced.

Tartary buckwheat (TB) is a pseudocereal rich in flavonoids, mainly including flavonols and anthocyanins. The flavonoid 3′-hydroxylase (F3′H) is a key enzyme in flavonoid biosynthesis and is encoded by two copies in TB genome. However, its biological function and effects on flavonol and anthocyanin synthesis in TB have not been well validated yet. In this study, we cloned the full-length FtF3′H1 gene highly expressed in all tissues (compared with FtF3′H2) according to TB flowering transcriptome data. The corresponding FtF3′H1 protein contains 534 amino acids with the molecular properties of the typical plant F3′H and belongs to the CYP75B family. During the flowering stage, the FtF3′H1 expression was highest in flowers, and its expression pattern showed a significant and positive correlation with the total flavonoids (R2 > 0.95). The overexpression of FtF3′H1 in Arabidopsis thaliana, Nicotiana tabacum and TB hairy roots resulted in a significant increase in anthocyanin contents (p

Sugar beet is vulnerable to rhizomania as the most destructive viral disease. Two selected events of transgenic sugar beet carrying cassettes inducing RNA silencing mechanism, 219-T3:S3-13.2 (S3) and 6018-T3:S6-44 (S6), were shown to inhibit propagation of Beet Necrotic Yellow Vein Virus, the causative agent. As a method for signifying the substantial equivalence, we analyzed the levels of some metabolites through LC-MS in order to demonstrate possible unintended changes in the leaves of the transgenic events. There was no significant difference in the concentrations of examined key metabolites but cis-aconitate and fructose-1,6-bisphosphatase which were decreased in S3. Also, ATP was reduced in both genetically modified sugar beets. Among free amino acids, only glycine level in S6 was increased compared to the wild plant, while the production levels of 5 and 12 ones were increased in S3 compared to S6 event and the wild type plants, respectively.

Drought stress is one of the most destructive abiotic stresses. Expansins are one of the cell wall expandable proteins that can expand the cell wall under a pH-dependent pathway. Stomatal opening rate Due to the expression of AtEXPA1 gene in transgenic plants has been observed. In this study, second generation (T2) of three transgenic tobacco lines with AtEXPA1 gene were studied for some physiological and biochemical traits under drought stress. Transgenic lines includes 2, 4 and 7 and Non-transgenic commercial cultivar Samson as control were cultivated. RWCL and leaf chlorophyll content has been reduced under drought stress, and ELIL, Lipid peroxidation as MDAL and leaf proline content has been increase. The activity of antioxidant enzymes such as, catalase (CAT), guaiacol peroxidase (GPOX), ascorbate peroxidase (APOX) and polyphenol oxidase (PPO) was increased in leaves under drought stress. In this study, according to biochemical traits Line 2 and Line 7 and according to physiological traits Line4 in Compared to the control cultivar, could cope with stress and introduce as tolerate cultivars.

Oral immune tolerance is a physiological process to achieve tolerance against autoimmunity by oral ingestion of self-antigen(s) or other therapeutics. At the cellular level, oral tolerance suppresses autoimmune diseases by activating FoxP-positive and -negative regulatory T cells (Tregs) and/or causing clonal anergy or deletion of autoreactive T cells, affecting B cell tolerance. However, oral delivery of antigens/biologics is challenging due to their instability in the harsh environment of the gastrointestinal (GI) tract. Several antigen/drug delivery tools and approaches, including micro/nanoparticles and transgenic plant-based delivery systems, have been explored to demonstrate oral immune tolerance for different autoimmune diseases successfully. However, despite the effectiveness, variation in results, dose optimization, and undesirable immune system activation are the limitations of the oral approach to further advancement. From this perspective, the current review discusses the oral tolerance phenomenon, cellular mechanisms, antigen delivery tools and strategies, and its challenges.

Ficus lyrata is a popular ornamental foliage plant with unique violin- or guitar-shaped green leaves. In our previous study, a grapevine gene VvMybA1 was introduced into F. lyrata via Agrobacterium-mediated transformation, which resulted in the availability of purple-leaved plants. Since VvMybA1 is a transcription factor, the regeneration of transgenic purple-leaved plants might potentially produce variants with multicolored leaves. The objective of this study was to establish a method for regenerating purple-leaved F. lyrata and determine if variants with different coloration or variegation could be isolated from regenerated populations. Leaf explants derived from a completely purple-leaved transgenic plant were cultured on Murashige and Skoog (MS) basal medium supplemented with different concentrations of 6-benzyladenine (BA) and α-naphthalene acetic acid (NAA). Callogenesis occurred in leaf explants, and a subculture of callus-borne explants on the same medium resulted in callus proliferation and the occurrence of somatic embryos. Somatic embryos were more effectively induced from callus pieces cultured on MS medium supplemented with 8.88 μM of BA and 0.27 μM of NAA. More than 30 embryos were induced per callus piece, and the embryos matured and converted to plantlets. MS medium supplemented with 4.92 μM of indolyl-3-butanoic acid (IBA) greatly improved root development. Plantlets were transplanted into soilless substrate and grown in a shaded greenhouse for morphological evaluation. Nine variants with different degrees of coloration and variegation were isolated from the regenerated populations. Our results suggest that the regeneration of transgenic plants that harbor a transcription factor, such as VvMybA1, could be an additional way of isolating novel variants for the development of new cultivars of ornamental plants.

The development of recombinant DNA technology during the past thirty years has enabled scientists to isolate, characterize, and manipulate a myriad of different animal, bacterial, and plant genes. This has, in turn, led to the commercialization of hundreds of useful products that have significantly improved human health and well-being. Commercially, these products have been mostly produced in bacterial, fungal, or animal cells grown in culture. More recently, scientists have begun to develop a wide range of transgenic plants that produce numerous useful compounds. The perceived advantage of producing foreign compounds in plants is that compared to other methods of producing these compounds, plants seemingly provide a much less expensive means of production. A few plant-produced compounds are already commercially available; however, many more are in the production pipeline.

Cysteine proteases (CPs) are vital proteolytic enzymes that play critical roles in various plant processes. However, the particular functions of CPs in maize remain largely unknown. We recently identified a pollen-specific CP (named PCP), which highly accumulated on the surface of maize pollen. Here, we reported that PCP played an important role in pollen germination and drought response in maize. Overexpression of PCP inhibited pollen germination, while mutation of PCP promoted pollen germination to some extent. Furthermore, we observed that germinal apertures of pollen grains in the PCP-overexpression transgenic lines were excessively covered, whereas this phenomenon was not observed in the wild type (WT), suggesting that PCP regulated pollen germination by affecting the germinal aperture structure. In addition, overexpression of PCP enhanced drought tolerance in maize plants, along with the increased activities of the antioxidant enzymes and the decreased numbers of the root cortical cells. Conversely, mutation of PCP significantly impaired drought tolerance. These results may aid in clarifying the precise functions of CPs in maize and contribute to the development of drought-tolerant maize materials.

Genetically modified plants and crops can contribute to remarkable increase in global food supply, with improved yield and resistance to plant diseases or insect pests. The development of biotechnology introducing exogenous nucleic acids in transgenic plants is important for plant health management. Different genetic engineering methods for DNA delivery, such as biolistic methods, Agrobacterium tumefaciens-mediated transformation, and other physicochemical methods have been developed to improve translocation across the plasma membrane and cell wall in plants. Recently, the peptide-based gene delivery system, mediated by cell-penetrating peptides (CPPs), has been regarded as a promising non-viral tool for efficient and stable gene transfection into both animal and plant cells. CPPs are short peptides with diverse sequences and functionalities, capable of agitating plasma membrane and entering cells. Here, we highlight recent research and ideas on diverse types of CPPs, which have been applied in DNA delivery in plants. Various basic, amphipathic, cyclic, and branched CPPs were designed, and modifications of functional groups were performed to enhance DNA interaction and stabilization in transgenesis. CPPs were able to carry cargoes in either a covalent or noncovalent manner and to internalize CPP/cargo complexes into cells by either direct membrane translocation or endocytosis. Importantly, subcellular targets of CPP-mediated nucleic acid delivery were reviewed. CPPs offer transfection strategies and influence transgene expression at subcellular localizations, such as in plastids, mitochondria, and the nucleus. In summary, the technology of CPP-mediated gene delivery provides a potent and useful tool to genetically modified plants and crops of the future.