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Salinity of cultivable land is a growing global concern that has been affecting the yields of major crops worldwide such as pigeon pea. In the current study, transgenic pigeon pea plants were developed using an in-planta Agrobacterium-mediated genetic transformation method wherein OsRuvB, a rice DNA helicase gene, was incorporated to induce salt to...
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Background: Pigeonpea (Cajanus cajan (L.) Millsp.; 2n = 2× = 22) is a drought-tolerant perennial grain legume commonly cultivated in India's rain-fed and dry land zones, and it is a remarkable natural source of minerals and protein worldwide. Despite decades of research, the constraints linked to tissue culture continues to hinder the genetic impro...
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... The first DNA helicase was discovered in Escherichia coli in the mid-1970s (Abdel-Monem et al. 1976). Following this, extensive research was undertaken on these proteins and their involvement in stress tolerance (Gorbalenya et al. 1989;Tuteja et al. 2009;Tuteja et al. 2015;Singh et al. 2020). ...
Recombination UVB (sensitivity) like (RuvBL) helicase genes represent a conserved family of genes, which are known to be involved in providing tolerance against abiotic stresses like heat and drought. We identified nine wheat RuvBL genes, one each on nine different chromosomes, belonging to homoeologous groups 2, 3, and 4. The lengths of genes ranged from 1647 to 2197 bp and exhibited synteny with corresponding genes in related species including Ae. tauschii, Z. mays, O. sativa, H. vulgare, and B. distachyon. The gene sequences were associated with regulatory cis-elements and transposable elements. Two genes, namely TaRuvBL1a-4A and TaRuvBL1a-4B, also carried targets for a widely known miRNA, tae-miR164. Gene ontology revealed that these genes were closely associated with ATP-dependent formation of histone acetyltransferase complex. Analysis of the structure and function of RuvBL proteins revealed that the proteins were localized mainly in the cytoplasm. A representative gene, namely TaRuvBL1a-4A, was also shown to be involved in protein-protein interactions with ten other proteins. On the basis of phylogeny, RuvBL proteins were placed in two sub-divisions, namely RuvBL1 and RuvBL2, which were further classified into clusters and sub-clusters. In silico studies suggested that these genes were differentially expressed under heat/drought. The qRT-PCR analysis confirmed that expression of TaRuvBL genes differed among wheat cultivars, which differed in the level of thermotolerance. The present study advances our understanding of the biological role of wheat RuvBL genes and should help in planning future studies on RuvBL genes in wheat including use of RuvBL genes in breeding thermotolerant wheat cultivars.
... The rst DNA helicase was discovered in Escherichia coli in mid-1970s (Abdel-Monem et al. 1976). Following this, extensive research was undertaken on these proteins and their involvement in stress tolerance (Gorbalenya et al. 1989;Tuteja et al. 2009;Tuteja et al. 2015;Singh et al. 2020 Liu et al. 2020). Among these families of helicase genes, the genes belonging to RuvBL family encode proteins, which belong to AAA+ (ATPases associated with diverse cellular activities) superfamily of proteins. ...
RuvBL helicase genes represent a conserved family of genes, which are known to be involved in providing tolerance against abiotic stresses like heat and drought in plants. We identified nine wheat RuvBL genes on nine different chromosomes, belonging to homoeologous groups 2, 3, and 4. Analysis of the structure and function of these genes revealed that the (i) length of genes ranged from 1647 to 2197 bp; (ii) genes exhibit synteny with corresponding genes in related species including Ae. tauschii , Z. mays , O. sativa , H. vulgare and B. distachyon ; (iii) gene sequences were associated with cis-elements and transposable elements; (iv) the genes TaRuvBL1a-4A and TaRuvBL1a-4B also carried targets for a widely known miRNA, tae-miR164. Gene ontology revealed that these genes were closely associated with ATP-dependent formation of histone acetyltransferase complex. Analysis of the structure and function of RuvBL proteins revealed that (i) proteins were localized mainly in the cytoplasm; (ii) the protein encoded by the representative gene TaRuvBL1a-4A was shown to be involved in protein-protein interactions with ten other proteins; (iii) on the basis of phylogeny, RuvBL proteins were placed in two sub-divisions, namely RuvBL1 and RuvBL2, which were further classified into clusters and sub-clusters. In-silico expression analysis suggested that these genes were differentially expressed under heat/drought. The qRT-PCR analysis confirmed that expression of TaRuvBL genes differed among wheat cultivars with varying degrees of thermotolerance. This study advances our understanding of the biological role of wheat RuvBL genes and should help in planning future studies on RuvBL genes in wheat.
... For developing other transgenic lines mentioned in Table 2 [17][18][19][20][21][22][23][24][25][26], the seeds were directly transferred to potted soil after co-cultivation, omitting the step of germinating in the presence of cefotaxime. For wheat and rice transformation, 200 µM acetosyringone was added to the bacterial suspension during co-cultivation. ...
... The presence of a single band observed by Southern hybridization analysis revealed that all the transgenic lines developed in various crops reported here (Table 2) carried a single copy of the transgene (Figure 2), except transgenic pigeon pea lines separately carrying the OsRuvB gene and the OsLecRLK gene, where one of line each was observed to carry two copies (last lane in Figure 2B; third lane in Figure 2G) as two bands in each of these lines were detected. Similar results were observed by real time-PCR analysis [17][18][19][20][21][22][23][24][25][26]. ...
... As an example, the inheritance pattern of the OsRuvB gene in transgenic pigeon pea is presented here [20]. The inheritance pattern of the OsRuvB gene in the T 1 generation was assessed by the presence of a transgene detected through direct PCR amplification with gene-specific primers. ...
Crop improvement under changing climatic conditions is required to feed the growing global population. The development of transgenic crops is an attractive and conceivably the most effective approach for crop improvement with desired traits in varying climatic situations. Here, we describe a simple, efficient and robust in planta Agrobacterium-mediated genetic transformation method that can be used in most crops, including rice, wheat and cotton, and particularly in tissue culture recalcitrant crops, such as chickpea and pigeon pea. The protocol was successfully used for the development of transgenic chickpea and pigeon pea lines for resistance against pod borer. Transgenic lines in chickpea, pigeon pea and wheat were also developed for salt stress tolerance. These lines exhibited improved salt tolerance in terms of various physio-biochemical parameters studied. Since the protocol is rapid, as no tissue culture step is involved, it will significantly contribute to the improvement of most crops and will be of interest for plant biologists working with genetic engineering or genome editing.
... A number of traits have been improved by transformation in pigeon peas also, e.g., insect resistance (Krishna et al. 2011;Kaur et al. 2016;Ghosh et al. 2017), increased salt tolerance (Singh et al. 2020) and seed nutritional quality (Thu et al. 2007). Insect resistance (Solleti et al. 2008;Grazziotin et al. 2020) and virus resistance (Cruz & Aragão 2013;Kumar et al. 2017) were induced by transformation methods in the cowpea also. ...
This review recapitulates the history, important milestones, the current status, and the perspectives of the pea (Pisum sativum L.) transformation as a tool for pea crop breeding. It summarises the developments of the pea transformation from the first methodological experiments to achieving the complete transformation and regeneration of genetically modified (GM) plants, transformation with the first genes of interest (GOI), to recent techniques of targeted genome editing. We show how recent biotechnological methods and genetic engineering may contribute to pea breeding in order to speed up the breeding process and for the creation of new pea cultivars. The focus is laid on genetic engineering which represents an excellent technology to enhance the pea gene pool with genes of interest which are not naturally present in the pea genome. Different methods of pea transformation are mentioned, as well as various GOI that have been used for pea transformation to date, all aimed at improving transgenic pea traits. Tolerance to herbicides or resistance to viruses, fungal pathogens, and insect pests belong, among others, to the pea traits that have already been modulated by methods of genetic engineering. The production of phytopharmaceuticals is also an important chapter in the use of genetically modified peas. We compare different methods of introducing transgenes to peas and also the usage of different selective and reporter genes. The transformation of other major legumes (soybeans, beans) is marginally mentioned. The effect of genetically modified (GM) peas on animal health (feeding tests, allergenicity) is summarised, the potential risks and benefits of pea modification are evaluated and also the prime expectations of GM peas and the real current state of this technology are compared. Unfortunately, this technology or, more precisely, the products created by this technology are under strict (albeit not scientifically-based) legislative control in the European Union.
... The first impacts of salinity cause a decrease in early plant development owing to a lack of water. Simultaneously, salt stress causes a nutritional imbalance in plants, resulting in a decrease in plant nutrient absorption and development (Singh et al., 2020). The results of this study show that salt stress reduces root and shoot length in all linseed genotypes. ...
Salinity, which is one of the abiotic stress factors, severely restricts plant production as a result of the negative effects of plants in different growth and development periods. Therefore, it is extremely important to determine the tolerance limits of plants to salinity in order to eliminate the limiting effect in terms of plant growth. Flax is an industrial plant that is used for multiple purposes and has commercial importance in the world. This research was carried out in controlled laboratory conditions in 2021 to determine the effects of salinity on the germination of flax seeds.
In the study, germination rate, root length, root fresh weight, shoot length and shoot fresh weight were evaluated. The result showed that significant differences between different NaCl solutions for all evaluated characters. Although the highest value was obtained in the control group in Mures variety, the highest value was obtained in 25 nM NaCl concentration in all other characters except for the germination rate in Dakota variety. The highest germination rate of 93.3% was obtained from the control application (0 mM NaCl) in both varieties. On the other hand, there was no germination in both varieties in 200 mM application.
... Kwapata et al. [157] created a drought-tolerant common bean crop using Hordeum vulgare's late embryogenesis abundant (LEA) protein HVA1. Likewise, Singh et al. utilized the rice DNA helicase (OsRuvB) gene to confer salinity tolerance in pigeonpea [158]. Similarly, Hanafy et al. heterologously expressed the potato gene PR10a in faba bean to enhance its salinity and drought tolerance [159]. ...
Legumes are a better source of proteins and are richer in diverse micronutrients over the nutritional profile of widely consumed cereals. However, when exposed to a diverse range of abiotic stresses, their overall productivity and quality are hugely impacted. Our limited understanding of genetic determinants and novel variants associated with the abiotic stress response in food legume crops restricts its amelioration. Therefore, it is imperative to understand different molecular approaches in food legume crops that can be utilized in crop improvement programs to minimize the economic loss. ‘Omics’-based molecular breeding provides better opportunities over conventional breeding for diversifying the natural germplasm together with improving yield and quality parameters. Due to molecular advancements, the technique is now equipped with novel ‘omics’ approaches such as ionomics, epigenomics, fluxomics, RNomics, glycomics, glycoproteomics, phosphoproteomics, lipidomics, regulomics, and secretomics. Pan-omics—which utilizes the molecular bases of the stress response to identify genes (genomics), mRNAs (transcriptomics), proteins (proteomics), and biomolecules (metabolomics) associated with stress regulation—has been widely used for abiotic stress amelioration in food legume crops. Integration of pan-omics with novel omics approaches will fast-track legume breeding programs. Moreover, artificial intelligence (AI)-based algorithms can be utilized for simulating crop yield under changing environments, which can help in predicting the genetic gain beforehand. Application of machine learning (ML) in quantitative trait loci (QTL) mining will further help in determining the genetic determinants of abiotic stress tolerance in pulses.
... Low soil water availability to plants is the main limiting environmental factor to the growth of plants and, consequently, to agricultural productivity (Campos et al., 2014;Kardile et al., 2018). Water deficit in cultivated areas occurs worldwide and it is the main limiting abiotic stress (Singh and Reddy, 2011;Singh et al., 2020). The available water to plants can be reduced due to the decrease in soil water content and increasing salt concentrations in the solution (Sheldon et al., 2017). ...
Drought and soil salinity are the main abiotic stresses in semiarid regions of the world. This study aims to evaluate the effect of water tensions generated by the reduction of soil moisture and salt on the leaf water potential of cowpea (Vigna unguiculata L. Walp). The experiments were conducted in a randomized complete block design, with a 6 × 2 factorial arrangement consisted of six soil water tensions (0.025, 0.265, 0.485, 0.705, 0.925, and 1.145 MPa) and two tension sources (water deficit and salt), with four replications. Two experiments were performed with the same environmental conditions to evaluate the influence of the tensions on vegetative and reproductive stages. Water and osmotic potentials, relative water content, leaf succulence, and shoot biomass yield were evaluated. Soil water tension was not the main factor of changes on water and osmotic potentials of V. unguiculata plants; the water deficit treatments at soil water tensions of up to 1.145 MPa did not reduce the water and osmotic potentials either at the vegetative or flowering phenological stages; high correlations were found between shoot biomass yield and the leaf water potential at seven days after stress. The osmotic potential was the main indicator of stress in plants at the vegetative and flowering stages subjected to water deficit by the presence of salts in the soil solution.
Plant transformation remains a major bottleneck to the improvement of plant science, both on fundamental and practical levels. The recalcitrant nature of most commercial and minor crops to genetic transformation slows scientific progress for a large range of crops that are essential for food security on a global scale. Over the years, novel stable transformation strategies loosely grouped under the term “in planta” have been proposed and validated in a large number of model (e.g. Arabidopsis and rice), major (e.g. wheat and soybean) and minor (e.g. chickpea and lablab bean) species. The in planta approach is revolutionary as it is considered genotype-independent, technically simple (i.e. devoid of or with minimal tissue culture steps), affordable, and easy to implement in a broad range of experimental settings. In this article, we reviewed and categorized over 300 research articles, patents, theses, and videos demonstrating the applicability of different in planta transformation strategies in 105 different genera across 139 plant species. To support this review process, we propose a classification system for the in planta techniques based on five categories and a new nomenclature for more than 30 different in planta techniques. In complement to this, we clarified some grey areas regarding the in planta conceptual framework and provided insights regarding the past, current, and future scientific impacts of these techniques. To support the diffusion of this concept across the community, this review article will serve as an introductory point for an online compendium about in planta transformation strategies that will be available to all scientists. By expanding our knowledge about in planta transformation, we can find innovative approaches to unlock the full potential of plants, support the growth of scientific knowledge, and stimulate an equitable development of plant research in all countries and institutions.
Worldwide, abiotic stresses including heat and drought are the major obstructions that threaten the agricultural production. Development of climate-resilient cultivars is the easy and economical way to combat drought and heat stress with limited resources. Plants do follow adaptation strategies to mitigate the impact of stress and lead to alteration in some of the morphological traits such as leaf rolling, leaf angle, cuticular wax content, stomatal conductance, deep root system, altered signalling and metabolic pathways. Targeting such traits along with the economical yield will help to identify suitable genotypes which perform better under stress environment. The basic step is to explore the available physiological trait variation among the cultivars, germplasm set and wild relatives to main stream alleles of importance to breeding material from the donor parent. Conventional and advanced breeding strategies can be implemented to develop climate-resilient cultivars with the suitable breeding and screening methods. As a key factor hybridization and selection along with the implication of advanced breeding methods like MABB, MARS, GS and transgenic approach make it easy and accurate to develop varieties in less time. Linkage, QTL and genome-wide association mapping helps to identify the genomic region of interest to target during marker-aided breeding approaches. A cocktail of breeding methods from conventional to transgenic may help in the development of high-yielding climate-resilient varieties which can help to serve farmers to escape from glitch of crop loss due to dry spell during cropping season. The recent advancement and methodologies regarding drought and heat tolerance breeding in wheat are discussed in this chapter along with the difficulties posed.KeywordsDroughtHeatWheatPhysiological breedingMolecular markers
Pulses are climate-smart grain legumes important to nutritional security and sustainable agriculture. Abiotic stresses take a heavy toll in pulse production, and genetic engineering offers a solution to add adaptive traits in the germplasm. Abiotic stresses being mostly polygenic are difficult to manipulate and require a thorough understanding of the underlying mechanism. Impact of abiotic stresses in eight different pulses, genetic mechanism involved, and transgenics approach adopted for enhancing the stress tolerance in those pulses are discussed. Traits engineered in chickpea (drought and salt tolerance), pigeon pea (salt tolerance), mung bean (salt and cold tolerance), urdbean (salt and drought tolerance and aluminum toxicity), cowpea (salt tolerance), field pea (salt, frost, and heat tolerance), common bean (drought tolerance), and lentil (cold and freezing tolerance) and resulting phenotypes are also discussed. Currently, only two transgenic pulses for biotic stress (insect resistance for cowpea and golden mosaic virus in common bean) are commercialized. Climate change poses various challenges, and genetic engineering and emerging genome editing techniques for abiotic stress-adaptive traits shall play a crucial role in abiotic stress management.
