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Not AvailableThe rise of the most appealing genome editing technology i.e. CRISPR/Cas (clustered regularly interspaced short palindromic repeat/CRISPR-associated protein) had stimulated researchers to exploit it for its wide implementation in several eukaryotic organisms along with crop plants. The technology has proven to be highly efficient, economical, easy to use and efficient in precise genome editing. Rice being a primary food crop worldwide has a lot of scope for improvement in terms of its productivity, quality and resistance to environmental stress. The various abiotic and biotic stresses pose a potential threat and are constantly diminishing rice yields globally. After rice genome sequencing, the researchers were engaged in decoding and reading genomes and since then there has been considerable advancement in the improvement of this crop. Moreover, the recent development in different CRISPR/Cas systems has widened its applicability in crops and the research community is very keen on rescripting and editing genomes for a better change. This chapter entails the strategies and protocols for CRISPR mediated genome editing in rice, refined CRISPR systems, recent development and progress of this technology in rice system, all aspects of transgenic development and complications associated with it and its future scope in the improvement of rice crop.Not Availabl

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Not AvailableRice yield is affected by various biotic stresses including fungi, bacteria, viruses, parasites, nematodes, weeds and insects, posing a major threat to global food security. Therefore, one of the major objectives of rice breeders is to develop rice cultivars resistant to biotic stresses and it has been achieved to large extent through traditional and molecular breeding approaches. However, frequent breakdown of resistance to these biotic stresses is a challenging issue and therefore, continuous efforts are needed to develop cultivars with durable resistance. Recently, genetic engineering technologies like transgenics and RNAi have enabled breeders to develop such durable resistance in rice against number of bacterial, fungal and viral diseases by utilizing the genes conferring resistance to trait and isolated from various organisms like plants, animals, microbes, etc. Genetic engineering is more preferred in some of the cases as it has advantages like requirement of lesser duration, no linkage drag and no crossing barrier compared to molecular breeding. Although varieties developed through genetic engineering require prior regulation before commercialization, it has the enormous potential to develop plants resistant/tolerant to biotic stresses. Present status of use of genetic engineering for developing biotic stress resistance in rice is briefly described in this chapter.Not Availabl

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Not AvailableUnder changing climatic conditions, high temperature stress is the most severe problem for the whole agriculture. Identification and utilization of crop plants which can sustain and yield better under high temperature conditions is need of the day. In this study, we established finger millet as thermotolerant crop. For this, we characterized thermotolerant cotton, thermosensitive wheat along with finger millet by MDA accumulation after heat stress and shown that finger millet is even better than cotton. Further, using seed germination test and growing seedlings at higher temperature, it was observed that finger millet was least affected at 42 oC whereas germination percent and fresh weight reduced at 47 oC. With biochemical assay, it was shown that finger millet had very less difference at 42 oC as compared to 37 oC, however there is significant reduction at 47 oC in chlorophyll and carotenoid content and relative water content (RWC) percent whereas increase in electrolyte leakage (%) and H2O2 and O2 concentration. Still finger millet plants can tolerate temperature of 47 oC. Overall, the present study strongly identified finger millet as thermotolerant crop and can be utilized for allele mining of known genes and prospecting of novel genes for crop improvement for high temperature stress.Not Availabl

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Not AvailableRubus ellipticus Smith. commonly known as ‘Yellow Himalayan raspberry’ is an important member of Rosaceae family with high medicinal importance having antioxidant and antibacterial properties. This study was done with the objective of characterizing 21 R. ellipticus collections from different regions of India, based on morphological descriptors and expressed sequence tags-simple sequence repeat (EST-SSR) markers. Broad phenotypic variability among the R. ellipticus collections was detected by using morphological descriptors. The Euclidian distance matrix discriminated two groups, based on morphological characters. A set of 68 EST-SSR markers were developed based on 7777 unigenes generated by us from ‘Kumarhatti-1’ collection of R. ellipticus using assembled pair—end sequences from Illumina NextSeq 500. The ESTs containing simple sequence repeat motifs were extracted using Microsatellite identification tool and primers were designed. Out of 68 EST-SSRs polymorphism was detected using 61 (89.7%) primer pairs, indicating high genetic variability in the R. ellipticus collections. Effective multiplex ratio, marker index and polymorphism information content were 0.43, 0.24 and 0.50, respectively. The unweighted pair group method with arithmetic mean coincided with the bootstrap analysis and two groups were formed, separating the collections Uttrakhand from the other. Morphological descriptors along with EST-SSR markers proved efficient in detecting the levels of genetic variability among the collections maintained in the field. These results can be used as an additional source of exploitation in R. ellipticus breeding programs.Not Availabl

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Not AvailableBasic leucine zipper (bZIP) transcription factors comprise one of the largest gene families in plants. They play a key role in almost every aspect of plant growth and development and also in biotic and abiotic stress tolerance. In this study, we report isolation and characterization of EcbZIP17, a group B bZIP transcription factor from a climate smart cereal, finger millet (Eleusine coracana L.). The genomic sequence of EcbZIP17 is 2662 bp long encompassing two exons and one intron with ORF of 1722 bp and peptide length of 573 aa. This gene is homologous to AtbZIP17 (Arabidopsis), ZmbZIP17 (maize) and OsbZIP60 (rice) which play a key role in endoplasmic reticulum (ER) stress pathway. In silico analysis confirmed the presence of basic leucine zipper (bZIP) and transmembrane (TM) domains in the EcbZIP17 protein. Allele mining of this gene in 16 different genotypes by Sanger sequencing revealed no variation in nucleotide sequence, including the 618 bp long intron. Expression analysis of EcbZIP17 under heat stress exhibited similar pattern of expression in all the genotypes across time intervals with highest upregulation after 4 h.Not Availabl

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Not AvailableReactive oxygen species (ROS) are byproducts of metabolic processes such as respiration and photosynthesis in plants. Production of ROS leads to rapid cell damage, and plants developed a complex system of enzymatic and non-enzymatic antioxidants to scavenge these ROS. Catalase is an important enzyme, which plays a key role in elimination of toxic effects of hydrogen peroxide and plays a major role as an antioxidant. When characterizing heat responsive genes in finger millet (Eleusine coracana L.) using a suppression subtractive hybridization (SSH) library, we isolated two catalase genes and named them as EcCATA1 and EcCATB1. The lengths of the EcCATA1 and EcCATB1 open reading frames were 1 482 and 1 426 bp, respectively. We characterized these genes under different abiotic stresses and in different tissues. The tissue wise expression revealed that EcCATA1 expression was higher in leaves whereas EcCATB1 expression was higher in roots than in other organs. Under stress conditions, the expression of EcCATA1 was highest under salt stress followed by mannitol treatment. In the case of EcCATB1, the highest expression was observed under mannitol treatment followed by cold and dehydration. We also studied expression of both the genes under heat stress in different finger millet genotypes and observed that expressions of these genes can be correlated with heat tolerance. For both the genes, a detailed computational investigation was also performed for understanding their structural properties and physicochemical characteristics. Overall, this is the first study to identify and characterize catalase genes from climate resilient finger millet crop.Not Availabl

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Not AvailableOver various conventional techniques, a CRISPR/Cas system of genome editing has revolutionized research through targeted and precise changes at the genome level. A great number of applications of this system in vegetable crops affirms its multifaceted nature in functional genomics. In combination with other breeding techniques, it may speed up the strengthening of genetic resources, which results in improved vegetable crops. In vegetables, the majority of modifications have been done in tomato and potato plants. Lack of enough genomic resources and efficient delivery methods in a large number of vegetables limits the use of CRISPR/Cas in these crops. CRISPR/Cas targets can be coding sequence (results in null alleles), regulatory elements of promoter region (results in novel alleles) and upstream ORFs to modify at the protein level. This editing system has been applied in vegetable crops for functional studies of genes as well as for the improvement of important traits such as abiotic and biotic stress tolerance, quality and yield, which shows the relevance of the technique. This editing system has been successfully used to modify the plant architecture along with inflorescence patterns. Improved tolerance to stress is highly fascinating since unfavourable conditions greatly affect vegetable crops. To cope with this situation, highly materialized cultivation system is required to grow vegetables, which make higher cost. To overcome this situation, knowledge on vegetable crops will guide us to modify the endogenous genetic traits along with the addition of new beneficial traits, which leads to obtain ‘environmentally fitted’ plants that will help to achieve food and nutrition security for growing human population.Not Availabl

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Not AvailableThe occurrence of various stresses, as the outcome of global climate change, results in the yield losses of crop plants. Prospecting of genes in stress tolerant plant species, may help to protect and improve their agronomic performance. Finger millet (Eleusine coracana L.) is a valuable source of superior genes and alleles for stress tolerance. In this study, we isolated a novel endoplasmic reticulum (ER) membrane tethered bZIP transcription factor from finger millet, EcbZIP17. Transgenic tobacco plants overexpressing this gene showed better vegetative growth and seed yield compared with WT plants under optimal growth conditions and confirmed upregulation of brassinosteroid signalling genes. Under various abiotic stresses, such as 250 mM NaCl, 10% PEG6000, 400 mM mannitol, water withdrawal, and heat stress, the transgenic plants showed higher germination rate, biomass, primary and secondary root formation, and recovery rate, compared with the wild type (WT) plants. The transgenic plants exposed to an ER stress inducer resulted in greater leaf diameter and plant height as well as higher expression of the ER stress-responsive genes BiP, PDIL, and CRT1. Overall, our results indicated that EcbZIP17 improves plant growth at optimal conditions through brassinosteroid signalling and provide tolerance to various environmental stresses via ER signalling pathways.Not Availabl

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Not AvailableFinger millet (Eleusine coracana L.) commonly known as ragi is an orphan cereal crop having significant climate-smart stress adaptation system. Apart from various abiotic stresses worldwide, heat stress is the major limiting constraint for crop yield and the loss of crop yield due to heat stress is increasing every year. Hence, to generate better heat tolerant crop plants there is a need to mine superior alleles. In this regard, efforts were made to isolate and characterize role of small heat shock protein gene HSP17.8 from finger millet. Here, we obtained an EST of having Hsp20 domain based on the expression analysis under heat stress and identified complete gene using homology-based analysis. The obtained gene was named as EcHSP17.8 based on its molecular weight. The complete Open Reading Frame (ORF) of EcHSP17.8 is of 489 bp with peptide length of 162 aa. Expression analysis of this gene in finger millet revealed its maximum expression in roots tissue. This gene was found to be induced by heat (42oC), desiccation, NaCl (250 mM), mannitol (200 mM) and oxidative stress; however maximum upregulation was observed under heat stress up to 40 folds. Expression analysis of this gene in 16 finger millet genotypes under heat stress revealed its maximum expression after 2h which remained upregulated up to 4h showing it as early responsive gene under heat stress. As this gene is well characterized for stress tolerance in other crops, the alleles of HSP17.8 from crop like finger millet can be used as better candidate for generating heat tolerant crop plants.Not Availabl

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Not AvailableAn extreme temperature regime beyond desired level imposes significant stress in crop plants. The low and high temperature stresses are one of the primary constraints for plant development and yield. Finger millet, being a climate resilient crop, is a potential source of novel stress tolerant genes. In this study, functional characterization of finger millet DREB2A gene in different abiotic stress conditions was done. This novel EcDREB2A transcription factor isolated from finger millet is a truncated version of DREB2A gene compared to previously reported DREB genes from other plant species. The overexpression of EcDREB2A in transgenic tobacco exhibits improved tolerance against heat stress 42 ◦C for up to 7 days, by altering physiology and biochemical means. However, same transgenic lines were unable to provide tolerance to 200 mM NaCl and 200 mM Mannitol stress. Under heat stress conditions, increased seed germination with improved lateral roots, fresh and dry weight relative to wild type (WT) was observed. The EcDREB2A transgenics exposed to heat stress showed improved rate of stomatal conductance, chlorophyll and carotenoids contents, and other photosynthesis parameters compared to WT plants. EcDREB2A overexpression also resulted in increased antioxidant enzyme activity (SOD, CAT, GR, POD and, APX) with decreased electrolyte leakage (EL), H2O2, and malondialdehyde (MDA) content than WT plants under heat stress. Quantitative real time expression analysis demonstrated that all eight downstream genes were significantly upregulated in transgenic plants only after heat stress. Our data provide a clear demonstration of the positive impact of overexpression of EcDREB2A providing heat stress tolerance to plants.NICR