KASP: a high-throughput genotyping system and its applications in major crop plants for biotic and abiotic stress tolerance.

Advances in plant molecular breeding have resulted in the development of new varieties with superior traits, thus improving the crop germplasm. Breeders can screen a large number of accessions without rigorous and time-consuming phenotyping by marker-assisted selection (MAS). Molecular markers are one of the most imperative tools in plant breeding programmes for MAS to develop new cultivars possessing multiple superior traits. Single nucleotide polymorphisms (SNPs) are ideal for MAS due to their low cost, low genotyping error rates, and reproducibility. Kompetitive Allele Specific PCR (KASP) is a globally recognized technology for SNP genotyping. KASP is an allele-specific oligo extension-based PCR assay that uses fluorescence resonance energy transfer (FRET) to detect genetic variations such as SNPs and insertions/deletions (InDels) at a specific locus. Additionally, KASP allows greater flexibility in assay design, which leads to a higher success rate and the capability to genotype a large population. Its versatility and ease of use make it a valuable tool in various fields, including genetics, agriculture, and medical research. KASP has been extensively used in various plant-breeding applications, such as the identification of germplasm resources, quality control (QC) analysis, allele mining, linkage mapping, quantitative trait locus (QTL) mapping, genetic map construction, trait-specific marker development, and MAS. This review provides an overview of the KASP assay and emphasizes its validation in crop improvement related to various biotic and abiotic stress tolerance and quality traits.

A comprehensive and conceptual overview of omics-based approaches for enhancing the resilience of vegetable crops against abiotic stresses.

MAIN CONCLUSION: Abiotic stresses adversely affect the productivity and production of vegetable crops. The increasing number of crop genomes that have been sequenced or re-sequenced provides a set of computationally anticipated abiotic stress-related responsive genes on which further research may be focused. Knowledge of omics approaches and other advanced molecular tools have all been employed to understand the complex biology of these abiotic stresses. A vegetable can be defined as any component of a plant that is eaten for food. These plant parts may be celery stems, spinach leaves, radish roots, potato tubers, garlic bulbs, immature cauliflower flowers, cucumber fruits, and pea seeds. Abiotic stresses, such as deficient or excessive water, high temperature, cold, salinity, oxidative, heavy metals, and osmotic stress, are responsible for the adverse activity in plants and, ultimately major concern for decreasing yield in many vegetable crops. At the morphological level, altered leaf, shoot and root growth, altered life cycle duration and fewer or smaller organs can be observed. Likewise different physiological and biochemical/molecular processes are also affected in response to these abiotic stresses. In order to adapt and survive in a variety of stressful situations, plants have evolved physiological, biochemical, and molecular response mechanisms. A comprehensive understanding of the vegetable's response to different abiotic stresses and the identification of tolerant genotypes are essential to strengthening each vegetable's breeding program. The advances in genomics and next-generation sequencing have enabled the sequencing of many plant genomes over the last twenty years. A combination of modern genomics (MAS, GWAS, genomic selection, transgenic breeding, and gene editing), transcriptomics, and proteomics along with next-generation sequencing provides an array of new powerful approaches to the study of vegetable crops. This review examines the overall impact of major abiotic stresses on vegetables, adaptive mechanisms and functional genomic, transcriptomic, and proteomic processes used by researchers to minimize these challenges. The current status of genomics technologies for developing adaptable vegetable cultivars that will perform better in future climates is also examined.

Mechanistic insight on boron-mediated toxicity in plant vis-a-vis its mitigation strategies: a review.

Boron (B) is an essential micronutrient, crucial for the growth and development of crop plants. However, the essential to a toxic range of B in the plant is exceptionally narrow, and symptoms develop with a slight change in its concentration in soil. The morphological and anatomical response, such as leaf chlorosis, stunted growth, and impairment in the xylem and phloem development occurs under B-toxicity. The transport of B in the plant occurs via transpiration stream with the involvement of B-channels and transporter in the roots. The higher accumulation of B in source and sink tissue tends to have lower photosynthetic, chlorophyll content, infertility, failure of pollen tube formation and germination, impairment of cell wall formation, and disruption of membrane systems. Excess B in the plant hinders the uptake of other micronutrients, hormone transport, and metabolite partitioning. B-mediated reactive oxygen species production leads to the synthesis of antioxidant enzymes which help to scavenge these molecules and prevent the plant from further oxidative damage. This review highlights morpho-anatomical, physiological, biochemical, and molecular responses of the plant under B toxicity and thereby might help the researchers to understand the related mechanism and design strategies to develop B tolerant cultivars.

The physio-biochemical and molecular responses and mechanism of B uptake under its toxic condition have been illustrated. The spatial distribution of boron under its toxic condition and its accumulation in the plant might be regulated with sugar alcohols (polyols). This review throws light on the elevated level of B in the soil-plant system and provides management strategies for alleviating B toxicity in the plant.

Insight into melatonin-mediated response and signaling in the regulation of plant defense under biotic stress.

KEY MESSAGE: Melatonin plays a crucial role in the mitigation of plant biotic stress through induced defense responses and pathogen attenuation. Utilizing the current knowledge of signaling and associated mechanism of this phytoprotectant will be invaluable in sustainable plant disease management. Biotic stress in plants involves complex regulatory networks of various sensory and signaling molecules. In this context, the polyfunctional, ubiquitous-signaling molecule melatonin has shown a regulatory role in biotic stress mitigation in plants. The present review conceptualized the current knowledge concerning the melatonin-mediated activation of the defense signaling network that leads to the resistant or tolerant phenotype of the infected plants. Fundamentals of signaling networks involved in melatonin-induced reactive oxygen species (ROS) or reactive nitrogen species (RNS) scavenging through enzymatic and non-enzymatic antioxidants have also been discussed. Increasing evidence has suggested that melatonin acts upstream of mitogen-activated proteinase kinases in activation of defense-related genes and heat shock proteins that provide immunity against pathogen attack. Besides, the direct application of melatonin on virulent fungi and bacteria showed disrupted spore morphology, destabilization of cell ultrastructure, reduced biofilm formation, and enhanced mortality that led to attenuate disease symptoms on melatonin-treated plants. The transcriptome analysis has revealed the down-regulation of pathogenicity genes, metabolism-related genes, and up-regulation of fungicide susceptibility genes in melatonin-treated pathogens. The activation of melatonin-mediated systemic acquired resistance (SAR) through cross-talk with salicylic acid (SA), jasmonic acid (JA) has been essential for viral disease management. The high endogenous melatonin concentration has also been correlated with the up-regulation of genes involved in pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI) and effector-triggered immunity (ETI). The present review highlights the versatile functions of melatonin towards direct inhibition of pathogen propagule along with active participation in mediating oxidative burst and simulating PTI, ETI and SAR responses. The hormonal cross-talk involving melatonin mediated biotic stress tolerance through defense signaling network suggests its suitability in a sustainable plant protection system.

Physiological and molecular insights on wheat responses to heat stress.

Increasing temperature is a key component of global climate change, affecting crop growth and productivity worldwide. Wheat is a major cereal crop grown in various parts of the globe, which is affected severely by heat stress. The morphological parameters affected include germination, seedling establishment, source-sink activity, leaf area, shoot and root growth. The physiological parameters such as photosynthesis, respiration, leaf senescence, water and nutrient relation are also affected by heat. At the cellular level, heat stress leads to the generation of reactive oxygen species that disrupt the membrane system of thylakoid, chloroplast and plasma membrane. The deactivation of the photosystem, reduction in photosynthesis and inactivation of rubisco affect the production of photoassimilates and their allocation. This ultimately affects anthesis, grain filling, size, number and maturity of wheat grains, which hamper crop productivity. The interplay of various systems comprising antioxidants and hormones plays a crucial role in imparting heat stress tolerance in wheat. Thus, implementation of various omics technologies could foster in-depth insights on heat stress effects, eventually devising heat stress mitigation strategies by conventional and modern breeding to develop heat-tolerant wheat varieties. This review provides an integrative view of heat stress responses in wheat and also discusses approaches to develop heat-tolerant wheat varieties.

Salt stress influences the proliferation of Fusarium solani and enhances the severity of wilt disease in potato.

Soil salinity has emerged as a critical abiotic stress in potato production, whereas wilt disease, caused by Fusarium solani, is the significant biotic stress. An experiment was performed to decipher the occurrence of wilt incidence by F. solani FJ1 under the influence of salinity in both in vitroand pot culture conditions. High salt concentration negatively influenced root and shoot development in the variety "Kufri Jyoti" but positively affected the mycelial growth and sporulation behaviours of F. solani FJ1. There was abundant whitish mycelial growth with enhanced biomass and high sporulation (microconidia production) in F. solani FJ1 cultured on salt-supplemented media. Moreover, under high salinity conditions (EC 2-8 dS m-1), severe wilting and rotting of vascular bundles were observed in plants artificially inoculated with F. solani FJ1. The mortality rate of potato plants was significantly higher under individual and combined stresses as compared to control. The wilt index of individual and combined stressed plants was also substantially higher compared to the control. Additionally, compared to the control, there was a significant decrease in total chlorophyll content and membrane stability index of the leaves under combined stress. However, the total phenols were increased under stress conditions. The total sugar content of potato plants decreased in infected plants, but increased when exposed to salt stress or a combination of salt stress and pathogen infection. F. solani infection also increased the activity of peroxidase (POX) and decreased the activity of phenylalanine ammonia-lyase (PAL) and catalase (CAT). These results suggest that Fusarium wilt and dry rot will be a more severe disease for potato cultivation in saline soils.

Development of near homozygous lines for diploid hybrid TPS breeding in potatoes.

Diploid inbred-based F1 hybrid True Potato Seed (DHTPS) breeding is a novel technique to transform potato breeding and cultivation across the globe. Significant efforts are being made to identify elite diploids, dihaploids and develop diploid inbred lines for heterosis exploitation in potatoes. Self-incompatibility is the first obstacle for developing inbred lines in diploid potatoes, which necessitates the introgression of a dominant S locus inhibitor gene (Sli) for switching self-incompatibility to self-compatibility. We evaluated a set of 357 diploid clones in different selfing generations for self-compatibility and degree of homozygosity using Kompetitive Allele Specific PCR (KASP) Single Nucleotide Polymorphism (SNP) markers. A subset of 10 KASP markers of the Sli candidate region on chromosome 12 showed an association with the phenotype for self-compatibility. The results revealed that the selected 10 KASP markers for the Sli gene genotype could be deployed for high throughput rapid screening of self-compatibility in diploid populations and to identify new sources of self-compatibility. The homozygosity assessed through 99 KASP markers distributed across all the chromosomes of the potato genome was 20-78 % in founder diploid clones, while different selfing generations, i.e., S0, S1, S2 and S3 observed 36.1-80.4, 56.9-82.8, 59.5-85.4 and 73.7-87.8 % average homozygosity, respectively. The diploid plants with ∼80 % homozygosity were also observed in the first selfing generation, which inferred that homozygosity assessment in the early generations itself could identify the best plants with high homozygosity to speed up the generation of diploid inbred lines.

Digitalization of potato breeding program: Improving data collection and management.

A plant breeding program involves hundreds of experiments, each having number of entries, genealogy information, linked experimental design, lists of treatments, observed traits, and data analysis. The traditional method of arranging breeding program information and data recording and maintenance is not centralized and is always scattered in different file systems which is inconvenient for retrieving breeding information resulting in poor data management and the loss of crucial data. Data administration requires a significant amount of manpower and resources to maintain nurseries, trials, germplasm lines, and pedigree records. Further, data transcription in scattered spreadsheets and files leads to nomenclature and typing mistakes, which affects data analysis and selection decisions in breeding programs. The accurate data recording and management tools could improve the efficiency of breeding programs. Recent interventions in data management using computer-based breeding databases and informatics applications and tools have made the breeder's life easier. Because of its digital nature, the data obtained is improved even further, allowing for the acquisition of images, voice recording and other specific data kinds. Public breeding programs are far behind the industry in the use of data management tools and softwares. In this article, we have compiled the information on available data recording tools and breeding data management softwares with major emphasis on potato breeding data management.

CRISPR/Cas9-mediated editing of phytoene desaturase (PDS) gene in an important staple crop, potato.

The gene editing using the CRISPR/Cas9 system has become an important biotechnological tool for studying gene function and improving crops. In this study, we have used CRISPR/Cas9 system for editing the phytoene desaturase gene (PDS) in popular Indian potato cultivar Kufri Chipsona-I. A construct (pHSE401) carrying two target gRNAs with glycine tRNA processing system under the control of Arabidopsis U6 promoter and the Cas9 protein was constructed and transformed in potato plants using Agrobacterium-mediated genetic transformations. The regeneration efficiency of 45% was observed in regenerated plants, out of which 81% of the putative transformants shoot lines exhibited mutant or bleached phenotype (albinism). The deletion mutations were detected within the StPDS gene in the genotyped plants and a mutation efficiency of 72% for gRNA1 and gRNA2 has been detected using Sanger sequencing. Hence, we set up a CRISPR/Cas9-mediated genome editing protocol which is efficient and generates mutations (deletions) within StPDS gene in potato. The bleached phenotype is easily detectable after only few weeks after Agrobacterium-mediated transformation. This is the first report as a proof of concept for CRISPR/Cas9-based editing of PDS gene in Indian potato cv. Kufri Chipsona-I. This study demonstrates that CRISPR/Cas9 can be used to edit genes at high frequency within the genome of the potato for various traits. Therefore, this study will aid in creating important mutants for modifying molecular mechanisms controlling traits of agronomic importance.

Functional Fermented Probiotics, Prebiotics, and Synbiotics from Non-Dairy Products: A Perspective from Nutraceutical.

The current trend of health-conscious consumers and healthy food habits prompts researchers to explore developing food products with synbiotic benefits. Synbiotic foods have gained popularity in recent years due to their functional, nutritional, physiological, and therapeutic characteristics. Lactose intolerance, dyslipidemia, and allergic milk proteins become the barriers in the development of dairy probiotics. The present scenario of an increase in the demand for vegetarian products leads to a rise in the consumption of non-dairy probiotics. Prebiotics like, resistant starch, inulin, and polyphenols are selectively used by gut microbiota to enhance the selection and colonization of probiotics bacteria. Probiotic's action mechanisms include the production of bacteriocins, peptides, short-chain fatty acids, amino acids, vitamins, and other metabolites. Therefore, this review article explores the alternative sources of probiotics so it will help to an understanding of non-dairy based functional fermented foods for both pro and prebiotics. Dietary fibers in vegetables, fruits, and cereals are one of prospective prebiotics and highlighted the various methods for making non-dairy synbiotics based on dietary fibers, such as microencapsulation, freeze-drying, and spray drying is also addressed.

Characterizing the Potato Growing Regions in India Using Meteorological Parameters.

Currently, the multi-location testing of advanced hybrids in India is carried out at 25 centers under the All India Co-ordinated Research Project on Potato (AICRP-P), which is spread across the country. These centres have been chosen to represent different potato growing regions based on soil and agronomic features. However, the reliable deployment of the newly bred varieties in different regions requires a scientific delineation of potato growing zones with homogenous climates. The present study was undertaken to develop homogenous zones in the Indian sub-continent based on the environmental parameters of the potato growing season. A total of 1253 locations were identified across the country as having a plausible potato growing season of at least 70 days with suitable thermal limits. Six variables including five meteorological parameters including Physiological days (P days), Growing degree days (GDD), Mean daily temperature, Mean night temperature and Mean daily incident solar radiation, together with altitude as the sixth variable, were used for Agglomerative Hierarchical Clustering (AHC) and the Principal Component Analysis by Multidimensional Scaling (MDS) technique to derive identical classes. The thematic map of the classes was overlaid on potato growing districts of India using ArcGIS 9.1 software. The study clearly depicted that the clustering technique can effectively delineate the target population of environments (TPE) for potato genotypes performing well at different testing environments in India. The study also identifies target locations for future focus on breeding strategies, especially the high night temperature class having a large expanse in India. This is also vital in view of the impending climate change situation.

Tolerance and adaptation mechanism of Solanaceous crops under salinity stress.

Solanaceous crops act as a source of food, nutrition and medicine for humans. Soil salinity is a damaging environmental stress, causing significant reductions in cultivated land area, crop productivity and quality, especially under climate change. Solanaceous crops are extremely vulnerable to salinity stress due to high water requirements during the reproductive stage and the succulent nature of fruits and tubers. Salinity stress impedes morphological and anatomical development, which ultimately affect the production and productivity of the economic part of these crops. The morpho-physiological parameters such as root-to-shoot ratio, leaf area, biomass production, photosynthesis, hormonal balance, leaf water content are disturbed under salinity stress in Solanaceous crops. Moreover, the synthesis and signalling of reactive oxygen species, reactive nitrogen species, accumulation of compatible solutes, and osmoprotectant are significant under salinity stress which might be responsible for providing tolerance in these crops. The regulation at the molecular level is mediated by different genes, transcription factors, and proteins, which are vital in the tolerance mechanism. The present review aims to redraw the attention of the researchers to explore the mechanistic understanding and potential mitigation strategies against salinity stress in Solanaceous crops, which is an often-neglected commodity.

Salinity Stress in Potato: Understanding Physiological, Biochemical and Molecular Responses.

Among abiotic stresses, salinity is a major global threat to agriculture, causing severe damage to crop production and productivity. Potato (Solanum tuberosum) is regarded as a future food crop by FAO to ensure food security, which is severely affected by salinity. The growth of the potato plant is inhibited under salt stress due to osmotic stress-induced ion toxicity. Salinity-mediated osmotic stress leads to physiological changes in the plant, including nutrient imbalance, impairment in detoxifying reactive oxygen species (ROS), membrane damage, and reduced photosynthetic activities. Several physiological and biochemical phenomena, such as the maintenance of plant water status, transpiration, respiration, water use efficiency, hormonal balance, leaf area, germination, and antioxidants production are adversely affected. The ROS under salinity stress leads to the increased plasma membrane permeability and extravasations of substances, which causes water imbalance and plasmolysis. However, potato plants cope with salinity mediated oxidative stress conditions by enhancing both enzymatic and non-enzymatic antioxidant activities. The osmoprotectants, such as proline, polyols (sorbitol, mannitol, xylitol, lactitol, and maltitol), and quaternary ammonium compound (glycine betaine) are synthesized to overcome the adverse effect of salinity. The salinity response and tolerance include complex and multifaceted mechanisms that are controlled by multiple proteins and their interactions. This review aims to redraw the attention of researchers to explore the current physiological, biochemical and molecular responses and subsequently develop potential mitigation strategies against salt stress in potatoes.