Browsing by Author "Misra, Gopal"

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An advanced draft genome assembly of a desi type chickpea (Cicer arietinum L.)
(Nature Publishing Group, 2015) Parween, Sabiha; Nawaz, Kashif; Roy, Riti; Pole, Anil K.; Venkata Suresh, B.; Misra, Gopal; Jain, Mukesh; Yadav, Gitanjali; Parida, Swarup K.; Tyagi, Akhilesh K.; Bhatia, Sabhyata; Chattopadhyay, Debasis
Chickpea (Cicer arietinum L.) is an important pulse legume crop. We previously reported a draft genome assembly of the desi chickpea cultivar ICC 4958. Here we report an advanced version of the ICC 4958 genome assembly (version 2.0) generated using additional sequence data and an improved genetic map. This resulted in 2.7-fold increase in the length of the pseudomolecules and substantial reduction of sequence gaps. The genome assembly covered more than 94% of the estimated gene space and predicted the presence of 30,257 protein-coding genes including 2230 and 133 genes encoding potential transcription factors (TF) and resistance gene homologs, respectively. Gene expression analysis identified several TF and chickpea-specific genes with tissue-specific expression and displayed functional diversification of the paralogous genes. Pairwise comparison of pseudomolecules in the desi (ICC 4958) and the earlier reported kabuli (CDC Frontier) chickpea assemblies showed an extensive local collinearity with incongruity in the placement of large sequence blocks along the linkage groups, apparently due to use of different genetic maps. Single nucleotide polymorphism (SNP)-based mining of intra-specific polymorphism identified more than four thousand SNPs differentiating a desi group and a kabuli group of chickpea genotypes.
The chickpea genomic web resource: visualization and analysis of the desi-type Cicer arietinum nuclear genome for comparative exploration of legumes
(BioMed Central Ltd, 2014) Misra, Gopal; Priya, Piyush; Bandhiwal, Nitesh; Bareja, Neha; Jain, Mukesh; Bhatia, Sabhyata; Chattopadhyay, Debasis; Tyagi, Akhilesh K.; Yadav, Gitanjali
Background: Availability of the draft nuclear genome sequences of small-seeded desi-type legume crop Cicer arietinum has provided an opportunity for investigating unique chickpea genomic features and evaluation of their biological significance. The increasing number of legume genome sequences also presents a challenge for developing reliable and information-driven bioinformatics applications suitable for comparative exploration of this important class of crop plants. Results: The Chickpea Genomic Web Resource (CGWR) is an implementation of a suite of web-based applications dedicated to chickpea genome visualization and comparative analysis, based on next generation sequencing and assembly of Cicer arietinum desi-type genotype ICC4958. CGWR has been designed and configured for mapping, scanning and browsing the significant chickpea genomic features in view of the important existing and potential roles played by the various legume genome projects in mutant mapping and cloning. It also enables comparative informatics of ICC4958 DNA sequence analysis with other wild and cultivated genotypes of chickpea, various other leguminous species as well as several non-leguminous model plants, to enable investigations into evolutionary processes that shape legume genomes. Conclusions: CGWR is an online database offering a comprehensive visual and functional genomic analysis of the chickpea genome, along with customized maps and gene-clustering options. It is also the only plant based web resource supporting display and analysis of nucleosome positioning patterns in the genome. The usefulness of CGWR has been demonstrated with discoveries of biological significance made using this server. The CGWR is compatible with all available operating systems and browsers, and is available freely under the open source license at http://www.nipgr.res.in/CGWR/home.php.
Development of eSSR-markers in Setaria italica and their applicability in studying genetic diversity, cross-transferability and comparative mapping in millet and non-millet species
(PLOS, 2013) Kumari, Kajal; Muthamilarasan, Mehanathan; Misra, Gopal; Gupta, Sarika; Subramanian, Alagesan; Parida, Swarup K.; Chattopadhyay, Debasis; Prasad, Manoj
Foxtail millet (Setariaitalica L.) is a tractable experimental model crop for studying functional genomics of millets and bioenergy grasses. But the limited availability of genomic resources, particularly expressed sequence-based genic markers is significantly impeding its genetic improvement. Considering this, we attempted to develop EST-derived-SSR (eSSR) markers and utilize them in germplasm characterization, cross-genera transferability and in silico comparative mapping. From 66,027 foxtail millet EST sequences 24,828 non-redundant ESTs were deduced, representing ~16 Mb, which revealed 534 (~2%) eSSRs in 495 SSR containing ESTs at a frequency of 1/30 kb. A total of 447 pp were successfully designed, of which 327 were mapped physically onto nine chromosomes. About 106 selected primer pairs representing the foxtail millet genome showed high-level of cross-genera amplification at an average of ~88% in eight millets and four non-millet species. Broad range of genetic diversity (0.02-0.65) obtained in constructed phylogenetic tree using 40 eSSR markers demonstrated its utility in germplasm characterizations and phylogenetics. Comparative mapping of physically mapped eSSR markers showed considerable proportion of sequence-based orthology and syntenic relationship between foxtail millet chromosomes and sorghum (~68%), maize (~61%) and rice (~42%) chromosomes. Synteny analysis of eSSRs of foxtail millet, rice, maize and sorghum suggested the nested chromosome fusion frequently observed in grass genomes. Thus, for the first time we had generated large-scale eSSR markers in foxtail millet and demonstrated their utility in germplasm characterization, transferability, phylogenetics and comparative mapping studies in millets and bioenergy grass species.
A draft genome sequence of the pulse crop chickpea (Cicer arietinum L.)
(John Wiley & Sons, 2013) Jain, Mukesh; Misra, Gopal; Patel, Ravi K.; Priya, Pushp; Jhanwar, Shalu; Khan, Aamir W.; Shah, Niraj; Singh, Vikas K.; Garg, Rohini; Jeena, Ganga; Yadav, Manju; Kant, Chandra; Sharma, Priyanka; Yadav, Gitanjali; Bhatia, Sabhyata; Tyagi, Akhilesh K.; Chattopadhyay, Debasis
Cicer arietinum L. (chickpea) is the third most important food legume crop. We have generated the draft sequence of a desi-type chickpea genome using next-generation sequencing platforms, bacterial artificial chromosome end sequences and a genetic map. The 520-Mb assembly covers 70% of the predicted 740-Mb genome length, and more than 80% of the gene space. Genome analysis predicts the presence of 27,571 genes and 210 Mb as repeat elements. The gene expression analysis performed using 274 million RNA-Seq reads identified several tissue-specific and stress-responsive genes. Although segmental duplicated blocks are observed, the chickpea genome does not exhibit any indication of recent whole-genome duplication. Nucleotide diversity analysis provides an assessment of a narrow genetic base within the chickpea cultivars. We have developed a resource for genetic markers by comparing the genome sequences of one wild and three cultivated chickpea genotypes. The draft genome sequence is expected to facilitate genetic enhancement and breeding to develop improved chickpea varieties.
FmMDb: A versatile database of foxtail millet markers for millets and bioenergy grasses research
(PLOS, 2013) Suresh B, Venkata; Muthamilarasan, Mehanathan; Misra, Gopal; Prasad, Manoj
The prominent attributes of foxtail millet (Setaria italica L.) including its small genome size, short life cycle, inbreeding nature, and phylogenetic proximity to various biofuel crops have made this crop an excellent model system to investigate various aspects of architectural, evolutionary and physiological significances in Panicoid bioenergy grasses. After release of its whole genome sequence, large-scale genomic resources in terms of molecular markers were generated for the improvement of both foxtail millet and its related species. Hence it is now essential to congregate, curate and make available these genomic resources for the benefit of researchers and breeders working towards crop improvement. In view of this, we have constructed the Foxtail millet Marker Database (FmMDb; http://www.nipgr.res.in/foxtail.html), a comprehensive online database for information retrieval, visualization and management of large-scale marker datasets with unrestricted public access. FmMDb is the first database which provides complete marker information to the plant science community attempting to produce elite cultivars of millet and bioenergy grass species, thus addressing global food insecurity.
Genome-wide development and use of microsatellite markers for large-scale genotyping applications in foxtail millet [Setaria italica (L.)]
(Oxford University Press, 2013) Pandey, Garima; Misra, Gopal; Kumari, Kajal; Gupta, Sarika; Parida, Swarup K.; Chattopadhyay, Debasis; Prasad, Manoj
The availability of well-validated informative co-dominant microsatellite markers and saturated genetic linkage map has been limited in foxtail millet (Setaria italica L.). In view of this, we conducted a genome-wide analysis and identified 28 342 microsatellite repeat-motifs spanning 405.3 Mb of foxtail millet genome. The trinucleotide repeats (∼48%) was prevalent when compared with dinucleotide repeats (∼46%). Of the 28 342 microsatellites, 21 294 (∼75%) primer pairs were successfully designed, and a total of 15 573 markers were physically mapped on 9 chromosomes of foxtail millet. About 159 markers were validated successfully in 8 accessions of Setaria sp. with ∼67% polymorphic potential. The high percentage (89.3%) of cross-genera transferability across millet and non-millet species with higher transferability percentage in bioenergy grasses (∼79%, Switchgrass and ∼93%, Pearl millet) signifies their importance in studying the bioenergy grasses. In silico comparative mapping of 15 573 foxtail millet microsatellite markers against the mapping data of sorghum (16.9%), maize (14.5%) and rice (6.4%) indicated syntenic relationships among the chromosomes of foxtail millet and target species. The results, thus, demonstrate the immense applicability of developed microsatellite markers in germplasm characterization, phylogenetics, construction of genetic linkage map for gene/quantitative trait loci discovery, comparative mapping in foxtail millet, including other millets and bioenergy grass species.
IGMAP: An interactive mapping and clustering platform for plants
(Cell Press, 2015) Priya, Piyush; Bandhiwal, Nitesh; Misra, Gopal; Mondal, Subhashish; Yadav, Gitanjali
Next-generation sequencing (NGS) technologies have resulted in a massive surge of high-throughput genomic data, particularly for the plant kingdom, boosting the development of methods for gene family discovery and identification of clustering patterns at genomic scales. Plants are well known for the occurrence of both genic and chromosomal duplications that have resulted in the widespread existence of gene families in this kingdom, apart from being associated with subsequent evolutionary divergence via sub-functionalization or neo- functionalization (Flagel and Wendel, 2009). These divergent mechanisms eventually lead to the formation of gene clusters, which in turn, have been shown to confer selective advantages to the genome including co-inheritance and co- regulation (Fischbach et al., 2008). Although gene clusters have been conventionally understood to be the genetic building blocks of prokaryotic genomes, comparative genomic studies have revealed the presence of functionally related genes that are clustered in lower nematodes, fungi, and several higher eukaryotes (Zorio et al., 1994; Blumenthal, 1998; Lee and Sonnhammer, 2003; Hurst et al., 2004; Thomas, 2006). However, very few data are available on the modularity or clustered linkage of genes in plants, despite the widespread occurrence of duplication events in the kingdom. In this regard, recent reports of biosynthetic modules and clustered organization of genes spotted in several major classes of plant-derived secondary metabolites arising through neo-functionalization and relocation of duplicated or existing genes, have offered an exciting niche (Osbourn, 2010). The presently available approaches for the identification and analysis of plant gene clusters include map-based cloning, forward and reverse genetics, and genome mining. As the wealth of plant genome sequence data continues to increase exponentially, newer methods are going to be required to carry out genome mining in a rational and useful manner.
Structural and biochemical perspectives in plant isoprenoid biosynthesis
(Springer, 2013) Kumari, Sangita; Priya, Piyush; Misra, Gopal; Yadav, Gitanjali
The isoprenoid family represents one of the most ancient and widespread classes of structurally and functionally rich biomolecules known to man. Although these natural products are synthesized in all organisms, the plant kingdom exhibits tremendous variation in their chemistry and roles, ranging from primary metabolism to secondary metabolism and specialized ecological interactions with the environment. Despite enormous diversity in structure and function, all isoprenoids are derived from the universal C5 precursor isoprene. The isoprenoid biosynthetic pathway has three major stages, viz., (1) synthesis of the isoprene building blocks, followed by their (2) assembly into flexible linear and branched hydrocarbon substrates, which then undergo (3) multistep reaction cascades to generate the vast assortment of isoprenoid end products. One of the most interesting aspects of isoprenoid biosynthesis is its being finely tuned by a multilayered and complex regulatory network, which excellently controls the machinery producing one of the most heterogeneous groups of molecules in plants. Terpene synthases, enzymes of the final stage, are key players in the generation of isoprenoid diversity, catalyzing one of the most complex reactions known to chemistry and biology. Unraveling the mechanism by which a minimal pool of substrates is thus converted into tens of thousands of regiospecific and stereospecific products, is a promising research avenue: This knowledge may be practically used for rational design of novel compounds by metabolic engineering, in order to yield plants with improved nutritional efficacy, stress resistance, bio-pharmaceutical properties etc. This review is an attempt to summarize the biochemical, molecular, physiological, structural, genomic and evolutionary aspects of isoprenoid biosynthesis, providing new insights into how these enzymes utilize various innovative strategies for creation of the so-called final terpenome.
Tomato genomic resources database: an integrated repository of useful tomato genomic information for basic and applied research
(PLOS, 2014) Suresh, B. Venkata; Roy, Riti; Sahu, Kamlesh; Misra, Gopal; Chattopadhyay, Debasis
Tomato Genomic Resources Database (TGRD) allows interactive browsing of tomato genes, micro RNAs, simple sequence repeats (SSRs), important quantitative trait loci and Tomato-EXPEN 2000 genetic map altogether or separately along twelve chromosomes of tomato in a single window. The database is created using sequence of the cultivar Heinz 1706. High quality single nucleotide polymorphic (SNP) sites between the genes of Heinz 1706 and the wild tomato S. pimpinellifolium LA1589 are also included. Genes are classified into different families. 5'-upstream sequences (5'-US) of all the genes and their tissue-specific expression profiles are provided. Sequences of the microRNA loci and their putative target genes are catalogued. Genes and 5'-US show presence of SSRs and SNPs. SSRs located in the genomic, genic and 5'-US can be analysed separately for the presence of any particular motif. Primer sequences for all the SSRs and flanking sequences for all the genic SNPs have been provided. TGRD is a user-friendly web-accessible relational database and uses CMAP viewer for graphical scanning of all the features. Integration and graphical presentation of important genomic information will facilitate better and easier use of tomato genome. TGRD can be accessed as an open source repository at http://59.163.192.91/tomato2/.