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<document id="FD7117C9A8E19107F3400324BA571F41" ID-DOI="10.1016/j.phytochem.2018.09.013" ID-ISSN="1873-3700" ID-Zenodo-Dep="10484692" IM.bibliography_approvedBy="karina" IM.illustrations_approvedBy="karina" IM.materialsCitations_approvedBy="felipe" IM.metadata_approvedBy="felipe" IM.tables_requiresApprovalFor="GgImagineBatch,operationResults" IM.taxonomicNames_approvedBy="karina" IM.treatments_approvedBy="karina" checkinTime="1704941816124" checkinUser="felipe" docAuthor="Deshmukh, Aaditi B., Datir, Sagar S., Bhonde, Yogesh, Kelkar, Natasha, Samdani, Pawan &amp; Tamhane, Vaijayanti A." docDate="2018" docId="03881B76835D7F05FCCDE87484A2F934" docLanguage="en" docName="Phytochemistry.156.201-213.pdf" docOrigin="Phytochemistry 156" docSource="http://dx.doi.org/10.1016/j.phytochem.2018.09.013" docStyle="DocumentStyle:9E596C34F4E94307D29315B03ACE1007.6:Phytochemistry.2014-2019.journal_article" docStyleId="9E596C34F4E94307D29315B03ACE1007" docStyleName="Phytochemistry.2014-2019.journal_article" docStyleVersion="6" docTitle="Oroxylum indicum subsp. root" docType="treatment" docVersion="4" lastPageNumber="202" masterDocId="FFB1630E835C7F04FFFFEC71806BFFAE" masterDocTitle="De novo root transcriptome of a medicinally important rare tree Oroxylum indicum for characterization of the flavonoid biosynthesis pathway" masterLastPageNumber="213" masterPageNumber="201" pageNumber="202" updateTime="1706220466884" updateUser="karina">
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<mods:title id="2ABD82447155953E4E9DDA6E7EA4615F">De novo root transcriptome of a medicinally important rare tree Oroxylum indicum for characterization of the flavonoid biosynthesis pathway</mods:title>
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<mods:namePart id="3E155B014D389C4FAC0D548EEAF3F7A1">Deshmukh, Aaditi B.</mods:namePart>
<mods:affiliation id="5DE6CE3A6128DC7D1BE3B772B6126610"> &amp; Institute of Bioinformatics and Biotechnology (IBB), Savitribai Phule Pune University, Ganeshkhind Road, Pune, 411007, Maharashtra, India</mods:affiliation>
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<mods:namePart id="902C4E7274F279DD2C6EAAA0072937C9">Datir, Sagar S.</mods:namePart>
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<mods:namePart id="8A8BC43BAA7333463ACFA2A135C7C3BB">Bhonde, Yogesh</mods:namePart>
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<mods:namePart id="E50F4F7CBD8CEEBE213936A7A52EFD55">Kelkar, Natasha</mods:namePart>
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<heading id="D0D61D0C835D7F05FCCDE87485C9FBB6" bold="true" box="[818,1442,1029,1048]" fontSize="36" level="1" pageId="1" pageNumber="202" reason="1">
<emphasis id="B9557672835D7F05FCCDE87485C9FBB6" bold="true" box="[818,1442,1029,1048]" italics="true" pageId="1" pageNumber="202">
2.1. Sequencing and De novo assembly of
<taxonomicName id="4C21D1E3835D7F05FB4EE874857FFBB6" authority="(L.) Kurz" box="[1201,1300,1029,1048]" class="Magnoliopsida" family="Bignoniaceae" genus="Oroxylum" kingdom="Plantae" order="Lamiales" pageId="1" pageNumber="202" phylum="Tracheophyta" rank="subSpecies" species="indicum" subSpecies="root">O. indicum</taxonomicName>
root transcripts
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<emphasis id="B9557672835D7F05FCACE84C83F6FBFE" bold="true" box="[851,925,1085,1104]" italics="true" pageId="1" pageNumber="202">De novo</emphasis>
transcriptome analysis is an excellent platform for generation of comprehensive information to deduce the basic biological, molecular and cellular processes for non-model organisms lacking whole genome sequences. To generate a transcriptome database for
<taxonomicName id="4C21D1E3835D7F05FA45E8E18312FB11" authority="(L.) Kurz" class="Magnoliopsida" family="Bignoniaceae" genus="Oroxylum" kingdom="Plantae" order="Lamiales" pageId="1" pageNumber="202" phylum="Tracheophyta" rank="species" species="indicum">
<emphasis id="B9557672835D7F05FA45E8E18312FB11" bold="true" italics="true" pageId="1" pageNumber="202">O. indicum</emphasis>
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, cDNA libraries of root tissue were sequenced using Illumina NextSeq 500. A total of 24,625,398 raw reads were generated comprising 3,718,435,098 nucleotides with an average size of 151 bp and 44% GC content (
<tableCitation id="C6A39FDB835D7F05FC21E971844EFABD" box="[990,1061,1280,1299]" captionStart="Table 1" captionStartId="1.[886,936,1728,1745]" captionTargetPageId="1" captionText="Table 1 Throughput and quality of Illumina sequencing of O. indicum root transcriptome." pageId="1" pageNumber="202">Table 1</tableCitation>
). Reads with Q &lt;
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20 were removed. Reads were mapped to human reference transcriptome which showed absence of contamination of human transcripts. These high quality reads were used for
<emphasis id="B9557672835D7F05FC7AE92583A0FAC9" bold="true" box="[901,971,1364,1383]" italics="true" pageId="1" pageNumber="202">de novo</emphasis>
assembly using Trinity (
<bibRefCitation id="EFB0D791835D7F05FB4AE9258538FAC9" author="Hass, B. J. &amp; Papanicolaou, A. &amp; Yassour, M. &amp; Grabherr, M. &amp; Blood, P. D. &amp; Bowden, J. &amp; Couger, M. B. &amp; Eccles, D. &amp; Li, B. &amp; Lieber, M. &amp; MacManes, M. D. &amp; Ott, M. &amp; Orvis, J. &amp; Pochet, N. &amp; Strozzi, F. &amp; Weeks, N. &amp; Westerman, R. &amp; William, T. &amp; Dewey, C. N. &amp; Henschel, R. &amp; LeDuc, R. D. &amp; Friedman, N. &amp; Regev, A." box="[1205,1363,1364,1383]" pageId="1" pageNumber="202" pagination="1494 - 1512" refId="ref11622" refString="Hass, B. J., Papanicolaou, A., Yassour, M., Grabherr, M., Blood, P. D., Bowden, J., Couger, M. B., Eccles, D., Li, B., Lieber, M., MacManes, M. D., Ott, M., Orvis, J., Pochet, N., Strozzi, F., Weeks, N., Westerman, R., William, T., Dewey, C. N., Henschel, R., LeDuc, R. D., Friedman, N., Regev, A., 2013. De novo transcript sequence reconstruction from RNA-seq using the trinity platform for reference generation and analysis. Nat. Protoc. 8, 1494 - 1512." type="journal article" year="2013">Hass et al., 2013</bibRefCitation>
). It has been reported that the quality of a
<emphasis id="B9557672835D7F05FBAEE90184F3FA2D" bold="true" box="[1105,1176,1392,1411]" italics="true" pageId="1" pageNumber="202">de novo</emphasis>
assembly is dependent on many factors, such as the
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of assembler used, followed by parameters like
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value and coverage (
<bibRefCitation id="EFB0D791835D7F05FBD5E9D684B0FA14" author="Reddy, N. &amp; Mehta, R. H. &amp; Soni, P. H. &amp; Makasana, J. &amp; Gajbhiye, N. A. &amp; Ponnuchamy, M. &amp; Kumar, J." box="[1066,1243,1447,1466]" pageId="1" pageNumber="202" pagination="0129422" refId="ref13148" refString="Reddy, N., Mehta, R. H., Soni, P. H., Makasana, J., Gajbhiye, N. A., Ponnuchamy, M., Kumar, J., 2015. Next generation sequencing and transcriptome analysis predicts biosynthetic pathway of Sennosides from Senna (Cassia angustifolia Vahl.), a non-model plant with potent laxative properties. PloS One 10 (6), e 0129422. https: // doi. org / 10.1371 / journal. pone. 0129422." type="journal article" year="2015">Reddy et al., 2015</bibRefCitation>
). The data has been submitted to NCBI,
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Sequence Read Archive (
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) submission:
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.
</paragraph>
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The assembly resulted in a total of 121,286 transcripts with N50 value of 1783 bp with an average contig length of 1080 bp (
<tableCitation id="C6A39FDB835D7F05FA85EA6685AAF984" box="[1402,1473,1559,1578]" captionStart="Table 2" captionStartId="2.[100,150,161,178]" captionTargetPageId="2" captionText="Table 2 Assembly and CDS statistics of Illumina sequencing of O. indicum in root transcriptome." pageId="1" pageNumber="202">Table 2</tableCitation>
). Furthermore, transcript clustering (CD-HIT-EST) resulted into 81,002 transcripts with an average contig length of 1071 bp and 1788 N50 value (
<tableCitation id="C6A39FDB835D7F05FC88EA1A83A9F9D0" box="[887,962,1643,1662]" captionStart="Table 2" captionStartId="2.[100,150,161,178]" captionTargetPageId="2" captionText="Table 2 Assembly and CDS statistics of Illumina sequencing of O. indicum in root transcriptome." pageId="1" pageNumber="202">Table 2</tableCitation>
). These 81,002 transcripts were further targeted for BLASTx analysis for functional annotation.
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