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  1. Article ; Online: Comparative Metagenomics Reveal Phylum Level Temporal and Spatial Changes in Mycobiome of Belowground Parts of Crocus sativus.

    Ambardar, Sheetal / Singh, Heikham Russiachand / Gowda, Malali / Vakhlu, Jyoti

    PloS one

    2016  Volume 11, Issue 9, Page(s) e0163300

    Abstract: Plant-fungal associations have been explored by routine cultivation based approaches and cultivation based approaches cannot catalogue more than 5% of fungal diversity associated with any niche. In the present study, an attempt has been made to catalogue ...

    Abstract Plant-fungal associations have been explored by routine cultivation based approaches and cultivation based approaches cannot catalogue more than 5% of fungal diversity associated with any niche. In the present study, an attempt has been made to catalogue fungal diversity associated with belowground parts i.e. rhizosphere and cormosphere, of Crocus sativus (an economically important herb) during two growth stages, using cultivation independent ITS gene targeted approach, taking bulk soil as reference. The 454 pyrosequencing sequence data analysis suggests that the fungal diversity was niche and growth stage specific. Fungi diversity, in the present case, was not only different between the two organs (roots and corm) but the dominance pattern varies between the cormosphere during two growth stages. Zygomycota was dominant fungal phylum in the rhizosphere whereas Basidiomycota was dominant in cormosphere during flowering stage. However in cormosphere though Basidiomycota was dominant phylum during flowering stage but Zygomycota was dominant during dormant stage. Interestingly, in cormosphere, the phyla which was dominant at dormant stage was rare at flowering stage and vice-versa (Basidiomycota: Flowering = 93.2% Dormant = 0.05% and Zygomycota: Flowering = 0.8% Dormant = 99.7%). At genus level, Rhizopus was dominant in dormant stage but was rare in flowering stage (Rhizopus: Dormant = 99.7% Flowering = 0.55%). This dynamics is not followed by the bulk soil fungi which was dominated by Ascomycota during both stages under study. The genus Fusarium, whose species F. oxysporum causes corm rot in C. sativus, was present during both stages with slightly higher abundance in roots. Interestingly, the abundance of Rhizopus varied a great deal in two stages in cormosphere but the abundance of Fusarium was comparable in two growth stages (Bulk soil Flowering = 0.05%, Rhizosphere Flowering = 1.4%, Cormosphere Flowering = 0.06%, Bulk soil Dormant = 2.47% and cormosphere dormant = 0.05%). This is the first report on the fungal diversity associated with the root of Crocus sativus and first report on the fungi associated with corm of any plant with the temporal and spatial variation in the fungal community structure.
    Language English
    Publishing date 2016
    Publishing country United States
    Document type Journal Article
    ISSN 1932-6203
    ISSN (online) 1932-6203
    DOI 10.1371/journal.pone.0163300
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Transcriptome sequencing of rhizome tissue of Sinopodophyllum hexandrum at two temperatures.

    Kumari, Anita / Singh, Heikham Russiachand / Jha, Ashwani / Swarnkar, Mohit Kumar / Shankar, Ravi / Kumar, Sanjay

    BMC genomics

    2014  Volume 15, Page(s) 871

    Abstract: Background: Sinopodophyllum hexandrum is an endangered medicinal herb, which is commonly present in elevations ranging between 2,400-4,500 m and is sensitive to temperature. Medicinal property of the species is attributed to the presence of ... ...

    Abstract Background: Sinopodophyllum hexandrum is an endangered medicinal herb, which is commonly present in elevations ranging between 2,400-4,500 m and is sensitive to temperature. Medicinal property of the species is attributed to the presence of podophyllotoxin in the rhizome tissue. The present work analyzed transcriptome of rhizome tissue of S. hexandrum exposed to 15°C and 25°C to understand the temperature mediated molecular responses including those associated with podophyllotoxin biosynthesis.
    Results: Deep sequencing of transcriptome with an average coverage of 88.34X yielded 60,089 assembled transcript sequences representing 20,387 unique genes having homology to known genes. Fragments per kilobase of exon per million fragments mapped (FPKM) based expression analysis revealed genes related to growth and development were over-expressed at 15°C, whereas genes involved in stress response were over-expressed at 25°C. There was a decreasing trend of podophyllotoxin accumulation at 25°C; data was well supported by the expression of corresponding genes of the pathway. FPKM data was validated by quantitative real-time polymerase chain reaction data using a total of thirty four genes and a positive correlation between the two platforms of gene expression was obtained. Also, detailed analyses yielded cytochrome P450s, methyltransferases and glycosyltransferases which could be the potential candidate hitherto unidentified genes of podophyllotoxin biosynthesis pathway.
    Conclusions: The present work revealed temperature responsive transcriptome of S. hexandrum on Illumina platform. Data suggested expression of genes for growth and development and podophyllotoxin biosynthesis at 15°C, and prevalence of those associated with stress response at 25°C.
    MeSH term(s) Berberidaceae/cytology ; Berberidaceae/enzymology ; Berberidaceae/genetics ; Berberidaceae/metabolism ; Gene Expression Profiling ; Gibberellins/metabolism ; Indoleacetic Acids/metabolism ; Molecular Sequence Annotation ; Podophyllotoxin/biosynthesis ; Rhizome/cytology ; Rhizome/enzymology ; Rhizome/genetics ; Rhizome/metabolism ; Sequence Analysis ; Signal Transduction/genetics ; Starch/metabolism ; Temperature ; Transcription Factors/metabolism
    Chemical Substances Gibberellins ; Indoleacetic Acids ; Transcription Factors ; Starch (9005-25-8) ; Podophyllotoxin (L36H50F353)
    Language English
    Publishing date 2014-10-07
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ISSN 1471-2164
    ISSN (online) 1471-2164
    DOI 10.1186/1471-2164-15-871
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: miR-BAG: bagging based identification of microRNA precursors.

    Jha, Ashwani / Chauhan, Rohit / Mehra, Mrigaya / Singh, Heikham Russiachand / Shankar, Ravi

    PloS one

    2012  Volume 7, Issue 9, Page(s) e45782

    Abstract: Non-coding elements such as miRNAs play key regulatory roles in living systems. These ultra-short, ∼21 bp long, RNA molecules are derived from their hairpin precursors and usually participate in negative gene regulation by binding the target mRNAs. ... ...

    Abstract Non-coding elements such as miRNAs play key regulatory roles in living systems. These ultra-short, ∼21 bp long, RNA molecules are derived from their hairpin precursors and usually participate in negative gene regulation by binding the target mRNAs. Discovering miRNA candidate regions across the genome has been a challenging problem. Most of the existing tools work reliably only for limited datasets. Here, we have presented a novel reliable approach, miR-BAG, developed to identify miRNA candidate regions in genomes by scanning sequences as well as by using next generation sequencing (NGS) data. miR-BAG utilizes a bootstrap aggregation based machine learning approach, successfully creating an ensemble of complementary learners to attain high accuracy while balancing sensitivity and specificity. miR-BAG was developed for wide range of species and tested extensively for performance over a wide range of experimentally validated data. Consideration of position-specific variation of triplet structural profiles and mature miRNA anchored structural profiles had a positive impact on performance. miR-BAG's performance was found consistent and the accuracy level was observed to be >90% for most of the species considered in the present study. In a detailed comparative analysis, miR-BAG performed better than six existing tools. Using miR-BAG NGS module, we identified a total of 22 novel miRNA candidate regions in cow genome in addition to a total of 42 cow specific miRNA regions. In practice, discovery of miRNA regions in a genome demands high-throughput data analysis, requiring large amount of processing. Considering this, miR-BAG has been developed in multi-threaded parallel architecture as a web server as well as a user friendly GUI standalone version.
    MeSH term(s) Animals ; Artificial Intelligence ; Cattle ; Computational Biology/methods ; Dogs ; Drosophila ; Humans ; Mice ; MicroRNAs/metabolism ; Models, Statistical ; Nematoda ; Predictive Value of Tests ; Probability ; Programming Languages ; Rats ; Reproducibility of Results ; Sequence Analysis, DNA/methods ; Species Specificity
    Chemical Substances MicroRNAs
    Language English
    Publishing date 2012-09-25
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ISSN 1932-6203
    ISSN (online) 1932-6203
    DOI 10.1371/journal.pone.0045782
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: Transcriptomic resources for the medicinal legume Mucuna pruriens: de novo transcriptome assembly, annotation, identification and validation of EST-SSR markers.

    Sathyanarayana, N / Pittala, Ranjith Kumar / Tripathi, Pankaj Kumar / Chopra, Ratan / Singh, Heikham Russiachand / Belamkar, Vikas / Bhardwaj, Pardeep Kumar / Doyle, Jeff J / Egan, Ashley N

    BMC genomics

    2017  Volume 18, Issue 1, Page(s) 409

    Abstract: Background: The medicinal legume Mucuna pruriens (L.) DC. has attracted attention worldwide as a source of the anti-Parkinson's drug L-Dopa. It is also a popular green manure cover crop that offers many agronomic benefits including high protein content, ...

    Abstract Background: The medicinal legume Mucuna pruriens (L.) DC. has attracted attention worldwide as a source of the anti-Parkinson's drug L-Dopa. It is also a popular green manure cover crop that offers many agronomic benefits including high protein content, nitrogen fixation and soil nutrients. The plant currently lacks genomic resources and there is limited knowledge on gene expression, metabolic pathways, and genetics of secondary metabolite production. Here, we present transcriptomic resources for M. pruriens, including a de novo transcriptome assembly and annotation, as well as differential transcript expression analyses between root, leaf, and pod tissues. We also develop microsatellite markers and analyze genetic diversity and population structure within a set of Indian germplasm accessions.
    Results: One-hundred ninety-one million two hundred thirty-three thousand two hundred forty-two bp cleaned reads were assembled into 67,561 transcripts with mean length of 626 bp and N50 of 987 bp. Assembled sequences were annotated using BLASTX against public databases with over 80% of transcripts annotated. We identified 7,493 simple sequence repeat (SSR) motifs, including 787 polymorphic repeats between the parents of a mapping population. 134 SSRs from expressed sequenced tags (ESTs) were screened against 23 M. pruriens accessions from India, with 52 EST-SSRs retained after quality control. Population structure analysis using a Bayesian framework implemented in fastSTRUCTURE showed nearly similar groupings as with distance-based (neighbor-joining) and principal component analyses, with most of the accessions clustering per geographical origins. Pair-wise comparison of transcript expression in leaves, roots and pods identified 4,387 differentially expressed transcripts with the highest number occurring between roots and leaves. Differentially expressed transcripts were enriched with transcription factors and transcripts annotated as belonging to secondary metabolite pathways.
    Conclusions: The M. pruriens transcriptomic resources generated in this study provide foundational resources for gene discovery and development of molecular markers. Polymorphic SSRs identified can be used for genetic diversity, marker-trait analyses, and development of functional markers for crop improvement. The results of differential expression studies can be used to investigate genes involved in L-Dopa synthesis and other key metabolic pathways in M. pruriens.
    Language English
    Publishing date 2017-05-25
    Publishing country England
    Document type Journal Article
    ISSN 1471-2164
    ISSN (online) 1471-2164
    DOI 10.1186/s12864-017-3780-9
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article ; Online: De novo sequencing and characterization of Picrorhiza kurrooa transcriptome at two temperatures showed major transcriptome adjustments.

    Gahlan, Parul / Singh, Heikham Russiachand / Shankar, Ravi / Sharma, Niharika / Kumari, Anita / Chawla, Vandna / Ahuja, Paramvir Singh / Kumar, Sanjay

    BMC genomics

    2012  Volume 13, Page(s) 126

    Abstract: Background: Picrorhiza kurrooa Royle ex Benth. is an endangered plant species of medicinal importance. The medicinal property is attributed to monoterpenoids picroside I and II, which are modulated by temperature. The transcriptome information of this ... ...

    Abstract Background: Picrorhiza kurrooa Royle ex Benth. is an endangered plant species of medicinal importance. The medicinal property is attributed to monoterpenoids picroside I and II, which are modulated by temperature. The transcriptome information of this species is limited with the availability of few hundreds of expressed sequence tags (ESTs) in the public databases. In order to gain insight into temperature mediated molecular changes, high throughput de novo transcriptome sequencing and analyses were carried out at 15 °C and 25 °C, the temperatures known to modulate picrosides content.
    Results: Using paired-end (PE) Illumina sequencing technology, a total of 20,593,412 and 44,229,272 PE reads were obtained after quality filtering for 15 °C and 25 °C, respectively. Available (e.g., De-Bruijn/Eulerian graph) and in-house developed bioinformatics tools were used for assembly and annotation of transcriptome. A total of 74,336 assembled transcript sequences were obtained, with an average coverage of 76.6 and average length of 439.5. Guanine-cytosine (GC) content was observed to be 44.6%, while the transcriptome exhibited abundance of trinucleotide simple sequence repeat (SSR; 45.63%) markers.Large scale expression profiling through "read per exon kilobase per million (RPKM)", showed changes in several biological processes and metabolic pathways including cytochrome P450s (CYPs), UDP-glycosyltransferases (UGTs) and those associated with picrosides biosynthesis. RPKM data were validated by reverse transcriptase-polymerase chain reaction using a set of 19 genes, wherein 11 genes behaved in accordance with the two expression methods.
    Conclusions: Study generated transcriptome of P. kurrooa at two different temperatures. Large scale expression profiling through RPKM showed major transcriptome changes in response to temperature reflecting alterations in major biological processes and metabolic pathways, and provided insight of GC content and SSR markers. Analysis also identified putative CYPs and UGTs that could help in discovering the hitherto unknown genes associated with picrosides biosynthesis.
    MeSH term(s) Base Composition ; Cinnamates/metabolism ; Cluster Analysis ; Computational Biology ; Cytochrome P-450 Enzyme System/genetics ; Exons/genetics ; Expressed Sequence Tags/metabolism ; Gene Expression Profiling/methods ; Glycosyltransferases/genetics ; Iridoid Glucosides/metabolism ; Microsatellite Repeats/genetics ; Molecular Sequence Annotation ; Picrorhiza/enzymology ; Picrorhiza/genetics ; Picrorhiza/metabolism ; RNA, Messenger/genetics ; RNA, Plant/genetics ; Reproducibility of Results ; Sequence Analysis, RNA/methods ; Sequence Homology, Nucleic Acid ; Temperature
    Chemical Substances Cinnamates ; Iridoid Glucosides ; RNA, Messenger ; RNA, Plant ; picroside I (27409-30-9) ; picroside II (39012-20-9) ; Cytochrome P-450 Enzyme System (9035-51-2) ; Glycosyltransferases (EC 2.4.-)
    Language English
    Publishing date 2012-03-31
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2041499-7
    ISSN 1471-2164 ; 1471-2164
    ISSN (online) 1471-2164
    ISSN 1471-2164
    DOI 10.1186/1471-2164-13-126
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  6. Article ; Online ; Research data: (with research data) Genome sequencing of herb Tulsi (Ocimum tenuiflorum) unravels key genes behind its strong medicinal properties.

    Upadhyay, Atul K / Chacko, Anita R / Gandhimathi, A / Ghosh, Pritha / Harini, K / Joseph, Agnel P / Joshi, Adwait G / Karpe, Snehal D / Kaushik, Swati / Kuravadi, Nagesh / Lingu, Chandana S / Mahita, J / Malarini, Ramya / Malhotra, Sony / Malini, Manoharan / Mathew, Oommen K / Mutt, Eshita / Naika, Mahantesha / Nitish, Sathyanarayanan /
    Pasha, Shaik Naseer / Raghavender, Upadhyayula S / Rajamani, Anantharamanan / Shilpa, S / Shingate, Prashant N / Singh, Heikham Russiachand / Sukhwal, Anshul / Sunitha, Margaret S / Sumathi, Manojkumar / Ramaswamy, S / Gowda, Malali / Sowdhamini, Ramanathan

    BMC plant biology

    2015  Volume 15, Page(s) 212

    Abstract: Background: Krishna Tulsi, a member of Lamiaceae family, is a herb well known for its spiritual, religious and medicinal importance in India. The common name of this plant is 'Tulsi' (or 'Tulasi' or 'Thulasi') and is considered sacred by Hindus. We ... ...

    Abstract Background: Krishna Tulsi, a member of Lamiaceae family, is a herb well known for its spiritual, religious and medicinal importance in India. The common name of this plant is 'Tulsi' (or 'Tulasi' or 'Thulasi') and is considered sacred by Hindus. We present the draft genome of Ocimum tenuiflurum L (subtype Krishna Tulsi) in this report. The paired-end and mate-pair sequence libraries were generated for the whole genome sequenced with the Illumina Hiseq 1000, resulting in an assembled genome of 374 Mb, with a genome coverage of 61 % (612 Mb estimated genome size). We have also studied transcriptomes (RNA-Seq) of two subtypes of O. tenuiflorum, Krishna and Rama Tulsi and report the relative expression of genes in both the varieties.
    Results: The pathways leading to the production of medicinally-important specialized metabolites have been studied in detail, in relation to similar pathways in Arabidopsis thaliana and other plants. Expression levels of anthocyanin biosynthesis-related genes in leaf samples of Krishna Tulsi were observed to be relatively high, explaining the purple colouration of Krishna Tulsi leaves. The expression of six important genes identified from genome data were validated by performing q-RT-PCR in different tissues of five different species, which shows the high extent of urosolic acid-producing genes in young leaves of the Rama subtype. In addition, the presence of eugenol and ursolic acid, implied as potential drugs in the cure of many diseases including cancer was confirmed using mass spectrometry.
    Conclusions: The availability of the whole genome of O.tenuiflorum and our sequence analysis suggests that small amino acid changes at the functional sites of genes involved in metabolite synthesis pathways confer special medicinal properties to this herb.
    MeSH term(s) Gene Expression Regulation, Plant ; Genome, Plant ; India ; Ocimum/genetics ; Ocimum/metabolism ; Plant Leaves/metabolism ; Plants, Medicinal/genetics ; Plants, Medicinal/metabolism
    Language English
    Publishing date 2015-08-28
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ISSN 1471-2229
    ISSN (online) 1471-2229
    DOI 10.1186/s12870-015-0562-x
    Database MEDical Literature Analysis and Retrieval System OnLINE

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