Unlocking the nutritional potential of chickpea: strategies for biofortification and enhanced multinutrient quality.

Chickpea (Cicer arietinum L.) is a vital grain legume, offering an excellent balance of protein, carbohydrates, fats, fiber, essential micronutrients, and vitamins that can contribute to addressing the global population's increasing food and nutritional demands. Chickpea protein offers a balanced source of amino acids with high bioavailability. Moreover, due to its balanced nutrients and affordable price, chickpea is an excellent alternative to animal protein, offering a formidable tool for combating hidden hunger and malnutrition, particularly prevalent in low-income countries. This review examines chickpea's nutritional profile, encompassing protein, amino acids, carbohydrates, fatty acids, micronutrients, vitamins, antioxidant properties, and bioactive compounds of significance in health and pharmaceutical domains. Emphasis is placed on incorporating chickpeas into diets for their myriad health benefits and nutritional richness, aimed at enhancing human protein and micronutrient nutrition. We discuss advances in plant breeding and genomics that have facilitated the discovery of diverse genotypes and key genomic variants/regions/quantitative trait loci contributing to enhanced macro- and micronutrient contents and other quality parameters. Furthermore, we explore the potential of innovative breeding tools such as CRISPR/Cas9 in enhancing chickpea's nutritional profile. Envisioning chickpea as a nutritionally smart crop, we endeavor to safeguard food security, combat hunger and malnutrition, and promote dietary diversity within sustainable agrifood systems.

Delineation of genes for a major QTL governing heat stress tolerance in chickpea.

Chickpea (Cicer arietinum) is a cool season grain legume experiencing severe yield loss during heat stress due to the intensifying climate changes and its associated gradual increase of mean temperature. Hence, understanding the genetic architecture regulating heat stress tolerance has emerged as an important trait to be addressed for enhancing yield and productivity of chickpea under heat stress. The present study is intended to identify the major genomic region(s) governing heat stress tolerance in chickpea. For this, an integrated genomics-assisted breeding strategy involving NGS-based high-resolution QTL-seq assay, QTL region-specific association analysis and molecular haplotyping was deployed in a population of 206 mapping individuals and a diversity panel of 217 germplasm accessions of chickpea. This combinatorial strategy delineated a major 156.8 kb QTL genomic region, which was subsequently narrowed-down to a functional candidate gene CaHSFA5 and its natural alleles associated strongly with heat stress tolerance in chickpea. Superior natural alleles and haplotypes delineated from the CaHSFA5 gene have functional significance in regulating heat stress tolerance in chickpea. Histochemical staining, interaction studies along with differential expression profiling of CaHSFA5 and ROS scavenging genes suggest a cross talk between CaHSFA5 with ROS homeostasis pertaining to heat stress tolerance in chickpea. Heterologous gene expression followed by heat stress screening further validated the functional significance of CaHSFA5 for heat stress tolerance. The salient outcomes obtained here can have potential to accelerate multiple translational genomic analysis including marker-assisted breeding and gene editing in order to develop high-yielding heat stress tolerant chickpea varieties.

Non-coding RNAs (ncRNAs) in plant: Master regulators for adapting to extreme temperature conditions.

Unusual daily temperature fluctuations caused by climate change and climate variability adversely impact agricultural crop production. Since plants are immobile and constantly receive external environmental signals, such as extreme high (heat) and low (cold) temperatures, they have developed complex molecular regulatory mechanisms to cope with stressful situations to sustain their natural growth and development. Among these mechanisms, non-coding RNAs (ncRNAs), particularly microRNAs (miRNAs), small-interfering RNAs (siRNAs), and long-non-coding RNAs (lncRNAs), play a significant role in enhancing heat and cold stress tolerance. This review explores the pivotal findings related to miRNAs, siRNAs, and lncRNAs, elucidating how they functionally regulate plant adaptation to extreme temperatures. In addition, this review addresses the challenges associated with uncovering these non-coding RNAs and understanding their roles in orchestrating heat and cold tolerance in plants.

Investigating the influence of elevated temperature on nutritional and yield characteristics of mung bean (Vigna radiata L.) genotypes during seed filling in a controlled environment.

Rising temperatures impact different developmental stages of summer crops like mung bean, particularly during the crucial seed-filling stage. This study focused on two mung bean genotypes, categorized as heat-tolerant [HT] or heat-sensitive [HS]. These genotypes were grown in pots in an outdoor natural environment (average day/night temperature 36°C/24.3°C) until the onset of podding (40 days after sowing) and subsequently relocated to controlled-environment walk-in growth chambers for exposure to heat stress (42°C/30°C) or control conditions (35°C/25°C) until maturity. For all measured attributes, heat stress had a more pronounced effect on the HS genotype than on the HT genotype. Heat-stressed plants exhibited severe leaf damage, including membrane damage, reduced chlorophyll content, diminished chlorophyll fluorescence, and decreased leaf water content. Heat stress impeded the seed-filling rate and duration, decreasing starch, protein, fat, and mineral contents, with a notable decline in storage proteins. Heat stress disrupted the activities of several seed enzymes, inhibiting starch and sucrose accumulation and consequently decreasing individual seed weights and seed weight plant-1. This study revealed that heat stress during seed filling severely impaired mung bean seed yield and nutritional quality due to its impact on various stress-related traits in leaves and enzyme activities in seeds. Moreover, this research identified potential mechanisms related to heat tolerance in genotypes with contrasting heat sensitivity.

Understanding the effect of heat stress during seed filling on nutritional composition and seed yield in chickpea (Cicer arietinum L.).

Increasing temperature affects all food crops, thereby reducing their yield potential. Chickpea is a cool-season food legume vital for its nutritive value, but it is sensitive to high temperatures (> 32/20 °C maximum/minimum) during its reproductive and seed-filling stages. This study evaluated the effects of heat stress on yield and qualitative traits of chickpea seeds in a controlled environment. Chickpea genotypes differing in heat sensitivity [two heat-tolerant (HT) and two heat-sensitive (HS)] were raised in pots, initially in an outdoor environment (average 23.5/9.9 °C maximum/minimum), until the beginning of pod set (107-110 days after sowing). At this stage, the plants were moved to a controlled environment in the growth chamber to impose heat stress (32/20 °C) at the seed-filling stage, while maintaining a set of control plants at 25/15 °C. The leaves of heat-stressed plants of the HT and HS genotypes showed considerable membrane damage, altered stomatal conductance, and reduced leaf water content, chlorophyll content, chlorophyll fluorescence, and photosynthetic ability (RuBisCo, sucrose phosphate synthase, and sucrose activities) relative to their corresponding controls. Seed filling duration and seed rate drastically decreased in heat-stressed plants of the HT and HS genotypes, severely reducing seed weight plant-1 and single seed weight, especially in the HS genotypes. Yield-related traits, such as pod number, seed number, and harvest index, noticeably decreased in heat-stressed plants and more so in the HS genotypes. Seed components, such as starch, proteins, fats, minerals (Ca, P, and Fe), and storage proteins (albumin, globulins, glutelin, and prolamins), drastically declined, resulting in poor-quality seeds, particularly in the HS genotypes. These findings revealed that heat stress significantly reduced leaf sucrose production, affecting the accumulation of various seed constituents, and leading to poor nutritional quality. The HT genotypes were less affected than the HS genotypes because of the greater stability of their leaf water status and photosynthetic ability, contributing to better yield and seed quality traits in a heat-stressed environment.

Major viral diseases in grain legumes: designing disease resistant legumes from plant breeding and OMICS integration.

Grain legumes play a crucial role in human nutrition and as a staple crop for low-income farmers in developing and underdeveloped nations, contributing to overall food security and agroecosystem services. Viral diseases are major biotic stresses that severely challenge global grain legume production. In this review, we discuss how exploring naturally resistant grain legume genotypes within germplasm, landraces, and crop wild relatives could be used as promising, economically viable, and eco-environmentally friendly solution to reduce yield losses. Studies based on Mendelian and classical genetics have enhanced our understanding of key genetic determinants that govern resistance to various viral diseases in grain legumes. Recent advances in molecular marker technology and genomic resources have enabled us to identify genomic regions controlling viral disease resistance in various grain legumes using techniques such as QTL mapping, genome-wide association studies, whole-genome resequencing, pangenome and 'omics' approaches. These comprehensive genomic resources have expedited the adoption of genomics-assisted breeding for developing virus-resistant grain legumes. Concurrently, progress in functional genomics, especially transcriptomics, has helped unravel underlying candidate gene(s) and their roles in viral disease resistance in legumes. This review also examines the progress in genetic engineering-based strategies, including RNA interference, and the potential of synthetic biology techniques, such as synthetic promoters and synthetic transcription factors, for creating viral-resistant grain legumes. It also elaborates on the prospects and limitations of cutting-edge breeding technologies and emerging biotechnological tools (e.g., genomic selection, rapid generation advances, and CRISPR/Cas9-based genome editing tool) in developing virus-disease-resistant grain legumes to ensure global food security.

Drought priming triggers diverse metabolic adjustments and induces chilling tolerance in chickpea (Cicer arietinum L.).

Chickpea (Cicer arietinum L.) suffers from chilling stress at the reproductive stage (<15 °C) which leads to significant yield loss. This study presents a comprehensive plant response to drought priming and its effect on chilling tolerance during the reproductive stage in two chickpea cultivars PBG1 and PBG5. Lipidome profiling (Fatty acid methyl esters analysis), metabolome profiling (GC-MS based untargeted analysis), fatty acid desaturases and antioxidative gene expression (qRT-PCR) were analyzed to monitor physiological and biochemical events after priming during flowering, podding and seed filling stages. Drought priming alleviated membrane damage and chlorophyll degradation by increasing membrane unsaturated fatty acids (18:3) along with up-regulation of various fatty acid desaturases (CaFADs) genes and antioxidative machinery during flowering and improved seed yield in PBG5. PCA, HCA, and KEGG pathway analysis of 87 identified metabolites showed that metabolites were regulated differently in both cultivars under non-primed and primed conditions. The plant response was more apparent at flowering and podding stages which coincided with chilling temperature (<15 °C). Drought priming stimulated many important genes, especially FADs, antioxidative proteins and accumulation of key metabolites (proline and TCA intermediates) required for defense especially in PBG5. This explains that plant's response to drought priming not only depends on developmental stage, and temperature regime (<15 °C) but also on the genotypic-specificity.

Assessing the heat sensitivity of Urdbean (Vigna mungo L. Hepper) genotypes involving physiological, reproductive and yield traits under field and controlled environment.

The rising temperatures are seriously impacting the food crops, including urdbean; hence efforts are needed to identify the sources of heat tolerance in such crops to ensure global food security. In the present study, urdbean genotypes were evaluated for heat tolerance under natural outdoor for two consecutive years (2018, 2019) and subsequently in the controlled environment of the growth chamber to identify high temperature tolerant lines. The genotypes were assessed involving few physiological traits (membrane damage, chlorophyll, photosynthetic efficiency, stomatal conductance, lipid peroxidation), reproductive traits (pollen germination % and pollen viability %) and yield related traits (total number of pods plant-1, total seeds plant-1, single seed weight and seed yield plant-1). Based upon these tested traits, PantU31, Mash114, UTTARA and IPU18-04 genotypes were identified as promising genotypes for both years under heat stress condition. Further confirming heat tolerance, all these four tolerant and four sensitive genotypes were tested under controlled environment under growth chamber condition. All these four genotypes PantU31, Mash114, UTTARA and IPU18-04 showed high chlorophyll content, photosynthetic efficiency, stomatal conductance, leaf area, pods plant-1, total seeds plant-1 and low reduction in pollen germination % and pollen viability under stress heat stress condition. Moreover, yield and yield related traits viz., pods plant-1, seeds plant-1, single seed weight and seed yield plant-1 showed very strong positive correlation with pollen germination and pollen viability except electrolyte leakage and malondialdehyde content. Thus, these genotypes could be potentially used as donors for transferring heat tolerance trait to the elite yet heat-sensitive urdbean cultivars.

Legume Pangenome: Status and Scope for Crop Improvement.

In the last decade, legume genomics research has seen a paradigm shift due to advances in genome sequencing technologies, assembly algorithms, and computational genomics that enabled the construction of high-quality reference genome assemblies of major legume crops. These advances have certainly facilitated the identification of novel genetic variants underlying the traits of agronomic importance in many legume crops. Furthermore, these robust sequencing technologies have allowed us to study structural variations across the whole genome in multiple individuals and at the species level using 'pangenome analysis.' This review updates the progress of constructing pangenome assemblies for various legume crops and discusses the prospects for these pangenomes and how to harness the information to improve various traits of economic importance through molecular breeding to increase genetic gain in legumes and tackle the increasing global food crisis.

Phenotypic evaluation of agronomic and root related traits for drought tolerance in recombinant inbred line population derived from a chickpea cultivar (C. arietinum L.) and its wild relative (C. reticulatum).

Drought is a major abiotic stress that drastically reduces chickpea yields. The present study was aimed to identify drought-responsive traits in chickpea by screening a recombinant inbred line population derived from an inter-specific cross between drought cultivar of GPF2 (C. arietinum L.) and drought sensitive accession of ILWC292 (C. reticulatum), at two locations in India. Twenty-one traits, including twelve morphological and physiological traits and nine root-related traits were measured under rainfed (drought-stress) and irrigated conditions (no-stress). High genotypic variation was observed among RILs for yield and root traits indicated that selection in these germplasms would be useful in achieving genetic progress. Both correlation and principal component analysis revealed that plant height, number of pods per plant, biomass, 100-seed weight, harvest index, membrane permeability index, and relative leaf water content were significantly correlated with yield under both irrigated and drought stress environments. Root length had significant positive correlations with all root-related traits except root length density in drought-stressed plants. Path analysis and multiple and stepwise regression analyses showed that number of pods per plant, biomass, and harvest index were major contributors to yield under drought stress conditions. Thus, a holistic approach across these analyses identified number of pods per plant, biomass, harvest index, and root length as key traits for improving chickpea yield through indirect selection for developing drought-tolerant cultivars. Overall, on the basis of yield components morphological and root traits, a total of 15 promising RILs were identified for their use in chickpea breeding programs for developing drought tolerant cultivars. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-022-01218-z.

Drought priming induces chilling tolerance and improves reproductive functioning in chickpea (Cicer arietinum L.).

KEY MESSAGE: Priming alleviates membrane damage, chlorophyll degradation along with cryoprotectants accumulation during chilling stress that leads to improved reproductive functioning and increased seed yield. Chilling temperatures below 15 °C have severe implications on the reproductive growth and development of chickpea. The abnormal reproductive development and subsequent reproductive failure lead to substantial yield loss. We exposed five chickpea cultivars (PBG1, GPF2, PDG3, PDG4, and PBG5) to drought stress (Priming) during the vegetative stage and analyzed for chilling tolerance during the reproductive stage. These varieties were raised in the fields in two sets: one set of plants were subjected to drought stress at the vegetative stage for 30 days (priming) and the second set of plants were irrigated regularly (non-primed). The leaf samples were harvested at the flowering, podding, and seed filling stage and analyzed for membrane damage, water status, chlorophyll content, cellular respiration, and certain cryoprotective solutes. The reproductive development was analyzed by accessing pollen viability, in vivo and in vitro germination, pollen load, and in vivo pollen tube growth. Principal component analysis (PCA) revealed that priming improved membrane damage, chlorophyll b degradation, and accumulation of cryoprotectants in GPF2, PDG3, and PBG5 at the flowering stage (< 15 °C). Pearson's correlation analysis showed a negative correlation with the accumulation of proline and carbohydrates with flower, pod, and seed abortion. Only, PBG5 responded best to priming while PBG1 was worst. In PBG5, priming resulted in reduced membrane damage and lipid peroxidation, improved water content, reduced chlorophyll degradation, and enhanced cryoprotective solutes accumulation, which led to increased reproductive functioning and finally improved seed yield and harvest index. Lastly, the priming response is variable and cultivar-specific but overall improve plant tolerance.

Physiological and Molecular Approaches for Developing Thermotolerance in Vegetable Crops: A Growth, Yield and Sustenance Perspective.

Vegetables are a distinct collection of plant-based foods that vary in nutritional diversity and form an important part of the healthy diet of the human being. Besides providing basic nutrition, they have great potential for boosting human health. The balanced consumption of vegetables is highly recommended for supplementing the human body with better nutrition density, dietary fiber, minerals, vitamins, and bioactive compounds. However, the production and quality of fresh vegetables are influenced directly or indirectly by exposure to high temperatures or heat stress (HS). A decline in quality traits and harvestable yield are the most common effects of HS among vegetable crops. Heat-induced morphological damage, such as poor vegetative growth, leaf tip burning, and rib discoloration in leafy vegetables and sunburn, decreased fruit size, fruit/pod abortion, and unfilled fruit/pods in beans, are common, often rendering vegetable cultivation unprofitable. Further studies to trace down the possible physiological and biochemical effects associated with crop failure reveal that the key factors include membrane damage, photosynthetic inhibition, oxidative stress, and damage to reproductive tissues, which may be the key factors governing heat-induced crop failure. The reproductive stage of plants has extensively been studied for HS-induced abnormalities. Plant reproduction is more sensitive to HS than the vegetative stages, and affects various reproductive processes like pollen germination, pollen load, pollen tube growth, stigma receptivity, ovule fertility and, seed filling, resulting in poorer yields. Hence, sound and robust adaptation and mitigation strategies are needed to overcome the adverse impacts of HS at the morphological, physiological, and biochemical levels to ensure the productivity and quality of vegetable crops. Physiological traits such as the stay-green trait, canopy temperature depression, cell membrane thermostability, chlorophyll fluorescence, relative water content, increased reproductive fertility, fruit numbers, and fruit size are important for developing better yielding heat-tolerant varieties/cultivars. Moreover, various molecular approaches such as omics, molecular breeding, and transgenics, have been proved to be useful in enhancing/incorporating tolerance and can be potential tools for developing heat-tolerant varieties/cultivars. Further, these approaches will provide insights into the physiological and molecular mechanisms that govern thermotolerance and pave the way for engineering "designer" vegetable crops for better health and nutritional security. Besides these approaches, agronomic methods are also important for adaptation, escape and mitigation of HS protect and improve yields.

Ensuring Global Food Security by Improving Protein Content in Major Grain Legumes Using Breeding and 'Omics' Tools.

Grain legumes are a rich source of dietary protein for millions of people globally and thus a key driver for securing global food security. Legume plant-based 'dietary protein' biofortification is an economic strategy for alleviating the menace of rising malnutrition-related problems and hidden hunger. Malnutrition from protein deficiency is predominant in human populations with an insufficient daily intake of animal protein/dietary protein due to economic limitations, especially in developing countries. Therefore, enhancing grain legume protein content will help eradicate protein-related malnutrition problems in low-income and underprivileged countries. Here, we review the exploitable genetic variability for grain protein content in various major grain legumes for improving the protein content of high-yielding, low-protein genotypes. We highlight classical genetics-based inheritance of protein content in various legumes and discuss advances in molecular marker technology that have enabled us to underpin various quantitative trait loci controlling seed protein content (SPC) in biparental-based mapping populations and genome-wide association studies. We also review the progress of functional genomics in deciphering the underlying candidate gene(s) controlling SPC in various grain legumes and the role of proteomics and metabolomics in shedding light on the accumulation of various novel proteins and metabolites in high-protein legume genotypes. Lastly, we detail the scope of genomic selection, high-throughput phenotyping, emerging genome editing tools, and speed breeding protocols for enhancing SPC in grain legumes to achieve legume-based dietary protein security and thus reduce the global hunger risk.

Response of Physiological, Reproductive Function and Yield Traits in Cultivated Chickpea (Cicer arietinum L.) Under Heat Stress.

Under global climate change, high-temperature stress is becoming a major threat to crop yields, adversely affecting plant growth, and ultimately resulting in significant yield losses in various crops, including chickpea. Thus, identifying crop genotypes with increased heat stress (HS) tolerance is becoming a priority for chickpea research. Here, we assessed the response of seven physiological traits and four yield and yield-related traits in 39 chickpea genotypes grown in normal-sown and late-sown environments [to expose plants to HS (>32/20°C) at the reproductive stage] for two consecutive years (2017-2018 and 2018-2019). Significant genetic variability for the tested traits occurred under normal and HS conditions in both years. Based on the tested physiological parameters and yield-related traits, GNG2171, GNG1969, GNG1488, PantG186, CSJ515, RSG888, RSG945, RVG202, and GNG469 were identified as promising genotypes under HS. Further, ten heat-tolerant and ten heat-sensitive lines from the set of 39 genotypes were validated for their heat tolerance (32/20°C from flowering to maturity) in a controlled environment of a growth chamber. Of the ten heat-tolerant genotypes, GNG1969, GNG1488, PantG186, RSG888, CSJ315, and GNG1499 exhibited high heat tolerance evidenced by small reductions in pollen viability, pollen germination, and pod set %, high seed yield plant-1 and less damage to membranes, photosynthetic ability, leaf water status, and oxidative processes. In growth chamber, chlorophyll, photosynthetic efficiency, pollen germination, and pollen viability correlated strongly with yield traits. Thus, GNG1969, GNG1488, PantG186, RSG888, CSJ315, and GNG1499 genotypes could be used as candidate donors for transferring heat tolerance traits to high-yielding heat-sensitive varieties to develop heat-resilient chickpea cultivars.

Progress of Genomics-Driven Approaches for Sustaining Underutilized Legume Crops in the Post-Genomic Era.

Legume crops, belonging to the Fabaceae family, are of immense importance for sustaining global food security. Many legumes are profitable crops for smallholder farmers due to their unique ability to fix atmospheric nitrogen and their intrinsic ability to thrive on marginal land with minimum inputs and low cultivation costs. Recent progress in genomics shows promise for future genetic gains in major grain legumes. Still it remains limited in minor legumes/underutilized legumes, including adzuki bean, cluster bean, horse gram, lathyrus, red clover, urd bean, and winged bean. In the last decade, unprecedented progress in completing genome assemblies of various legume crops and resequencing efforts of large germplasm collections has helped to identify the underlying gene(s) for various traits of breeding importance for enhancing genetic gain and contributing to developing climate-resilient cultivars. This review discusses the progress of genomic resource development, including genome-wide molecular markers, key breakthroughs in genome sequencing, genetic linkage maps, and trait mapping for facilitating yield improvement in underutilized legumes. We focus on 1) the progress in genomic-assisted breeding, 2) the role of whole-genome resequencing, pangenomes for underpinning the novel genomic variants underlying trait gene(s), 3) how adaptive traits of wild underutilized legumes could be harnessed to develop climate-resilient cultivars, 4) the progress and status of functional genomics resources, deciphering the underlying trait candidate genes with putative function in underutilized legumes 5) and prospects of novel breeding technologies, such as speed breeding, genomic selection, and genome editing. We conclude the review by discussing the scope for genomic resources developed in underutilized legumes to enhance their production and play a critical role in achieving the "zero hunger" sustainable development goal by 2030 set by the United Nations.

Breeding and Genomics Interventions for Developing Ascochyta Blight Resistant Grain Legumes.

Grain legumes are a key food source for ensuring global food security and sustaining agriculture. However, grain legume production is challenged by growing disease incidence due to global climate change. Ascochyta blight (AB) is a major disease, causing substantial yield losses in grain legumes worldwide. Harnessing the untapped reserve of global grain legume germplasm, landraces, and crop wild relatives (CWRs) could help minimize yield losses caused by AB infection in grain legumes. Several genetic determinants controlling AB resistance in various grain legumes have been identified following classical genetic and conventional breeding approaches. However, the advent of molecular markers, biparental quantitative trait loci (QTL) mapping, genome-wide association studies, genomic resources developed from various genome sequence assemblies, and whole-genome resequencing of global germplasm has revealed AB-resistant gene(s)/QTL/genomic regions/haplotypes on various linkage groups. These genomics resources allow plant breeders to embrace genomics-assisted selection for developing/transferring AB-resistant genomic regions to elite cultivars with great precision. Likewise, advances in functional genomics, especially transcriptomics and proteomics, have assisted in discovering possible candidate gene(s) and proteins and the underlying molecular mechanisms of AB resistance in various grain legumes. We discuss how emerging cutting-edge next-generation breeding tools, such as rapid generation advancement, field-based high-throughput phenotyping tools, genomic selection, and CRISPR/Cas9, could be used for fast-tracking AB-resistant grain legumes to meet the increasing demand for grain legume-based protein diets and thus ensuring global food security.

'Omics' approaches in developing combined drought and heat tolerance in food crops.

Global climate change will significantly increase the intensity and frequency of hot, dry days. The simultaneous occurrence of drought and heat stress is also likely to increase, influencing various agronomic characteristics, such as biomass and other growth traits, phenology, and yield-contributing traits, of various crops. At the same time, vital physiological traits will be seriously disrupted, including leaf water content, canopy temperature depression, membrane stability, photosynthesis, and related attributes such as chlorophyll content, stomatal conductance, and chlorophyll fluorescence. Several metabolic processes contributing to general growth and development will be restricted, along with the production of reactive oxygen species (ROS) that negatively affect cellular homeostasis. Plants have adaptive defense strategies, such as ROS-scavenging mechanisms, osmolyte production, secondary metabolite modulation, and different phytohormones, which can help distinguish tolerant crop genotypes. Understanding plant responses to combined drought/heat stress at various organizational levels is vital for developing stress-resilient crops. Elucidating the genomic, proteomic, and metabolic responses of various crops, particularly tolerant genotypes, to identify tolerance mechanisms will markedly enhance the continuing efforts to introduce combined drought/heat stress tolerance. Besides agronomic management, genetic engineering and molecular breeding approaches have great potential in this direction.

Nitric oxide secures reproductive efficiency in heat-stressed lentil (Lens culinaris Medik.) plants by enhancing the photosynthetic ability to improve yield traits.

Rising temperatures, globally and locally, would be detrimental for cool- and summer-season food legumes, such as lentil (Lens culinaris Medik.). Lentil is highly sensitive to supra-optimal temperatures (> 30 °C), particularly during reproductive growth, resulting in flower and pod losses. Thus, suitable strategies are needed to introduce heat tolerance in this legume. Here, we evaluated the efficacy of nitric oxide (NO)-applied as foliar treatment of 1 mM sodium nitroprusside (SNP), twice (one day before final exposure to high temperature, and again five days later)-on heat-stressed (32/20 °C) lentil genotypes, differing in heat sensitivity. As a result of heat stress, endogenous NO increased significantly in heat-tolerant genotypes (46-62% in leaves and 66-68% in anthers, relative to the respective controls), while it decreased in heat-sensitive (HS) genotypes (27-30% in leaves and 28-33% in anthers, relative to the respective controls). Foliar supplementation with SNP markedly increased endogenous NO in leaves and anthers of both the control and heat-treated plants. Heat stress significantly accelerated phenology, damaged membranes, chlorophyll, chlorophyll fluorescence, cellular viability, and decreased leaf water status, carbon fixing and assimilating ability, less so in plants treated with SNP. Heat stress plus SNP significantly improved carbon fixation (as RuBisCo activity) and assimilation ability, (as sucrose concentration (in leaves and anthers), sucrose synthase and vacuolar acid invertase activity, reducing sugars), as well as osmolyte accumulation (proline and glycine betaine) in leaves and anthers. Moreover, SNP-treated plants had significantly less oxidative damage-measured as malondialdehyde and hydrogen peroxide concentrations-in leaves and anthers, relative to the respective control. Reproductive function-assessed as pollen grain germination and viability, stigma receptivity, and ovular viability-decreased markedly in plants exposed to heat stress alone, more so in HS genotypes, but increased significantly with SNP treatment as a consequence of improved leaf and anther function, to significantly increase the pod and seed numbers in heat-stressed lentil plants, relative to heat-stress alone.

Cold Tolerance during the Reproductive Phase in Chickpea (Cicer arietinum L.) Is Associated with Superior Cold Acclimation Ability Involving Antioxidants and Cryoprotective Solutes in Anthers and Ovules.

Chickpea is sensitive to cold stress, especially at reproductive stage, resulting in flower and pod abortion that significantly reduces seed yield. In the present study, we evaluated (a) whether cold acclimation imparts reproductive cold tolerance in chickpea; (b) how genotypes with contrasting sensitivity respond to cold acclimation; and (c) the involvement of cryoprotective solutes and antioxidants in anthers and ovules in cold acclimation. Four chickpea genotypes with contrasting cold sensitivity (cold-tolerant: ICC 17258, ICC 16349; cold-sensitive: ICC 15567, GPF 2) were grown in an outdoor environment for 40 days in November (average maximum/minimum temperature 24.9/15.9 °C) before being subjected to cold stress (13/7 °C), with or without cold acclimation in a controlled environment of walk-in-growth chambers. The 42-d cold acclimation involved 7 d exposure at each temperature beginning with 23/15 °C, 21/13 °C, 20/12 °C, 20/10 °C, 18/8 °C, 15/8 °C (12 h/12 h day/night), prior to exposing the plants to cold stress (13/7 °C, 12 h/12 h day/night; 700 µmol m-2 s-1 light intensity; 65-70% relative humidity). Cold acclimation remarkably reduced low temperature-induced leaf damage (as membrane integrity, leaf water status, stomatal conductance, photosynthetic pigments, and chlorophyll fluorescence) under cold stress in all four genotypes. It only reduced anther and ovule damage in cold-tolerant genotypes due to improved antioxidative ability, measured as enzymatic (superoxide dismutase, catalase, ascorbate peroxidase, and glutathione reductase) and non-enzymatic (ascorbate and reduced glutathione), solutes (particularly sucrose and γ-aminobutyric acid) leading to improving reproductive function and yield traits, whereas cold-sensitive genotypes were not responsive. The study concluded that cold tolerance in chickpea appears to be related to the better ability of anthers and ovules to acclimate, involving various antioxidants and cryoprotective solutes. This information will be useful in directing efforts toward increasing cold tolerance in chickpea.

Non-Coding RNAs in Legumes: Their Emerging Roles in Regulating Biotic/Abiotic Stress Responses and Plant Growth and Development.

Noncoding RNAs, including microRNAs (miRNAs), small interference RNAs (siRNAs), circular RNA (circRNA), and long noncoding RNAs (lncRNAs), control gene expression at the transcription, post-transcription, and translation levels. Apart from protein-coding genes, accumulating evidence supports ncRNAs playing a critical role in shaping plant growth and development and biotic and abiotic stress responses in various species, including legume crops. Noncoding RNAs (ncRNAs) interact with DNA, RNA, and proteins, modulating their target genes. However, the regulatory mechanisms controlling these cellular processes are not well understood. Here, we discuss the features of various ncRNAs, including their emerging role in contributing to biotic/abiotic stress response and plant growth and development, in addition to the molecular mechanisms involved, focusing on legume crops. Unravelling the underlying molecular mechanisms and functional implications of ncRNAs will enhance our understanding of the coordinated regulation of plant defences against various biotic and abiotic stresses and for key growth and development processes to better design various legume crops for global food security.