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<document id="3297130F9781D9632BF4CFEC6AA3F46E" 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="03881B7683597F01FCCDECEE8315F98B" 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="1" lastPageNumber="206" 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="206" updateTime="1706220466884" updateUser="karina">
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<mods:title id="2BC994FACDBBEC1FA46706B720E8FD72">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="8864293D6672E1E8A1A83C2DDD792A47">Deshmukh, Aaditi B.</mods:namePart>
<mods:affiliation id="B10D34E485BA2B7DD0D3E1D4C598564A"> &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="1F0BAC0FB60A9060346A057DAB5D15AE">Datir, Sagar S.</mods:namePart>
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<mods:namePart id="F59E19684A8E2F30726BD164D8D5E82B">Kelkar, Natasha</mods:namePart>
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<mods:namePart id="AC4DF6E0D6EB013AA16627517E27372E">Samdani, Pawan</mods:namePart>
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<mods:title id="84DDACFB8ADA4712141FC5791BFA5458">Phytochemistry</mods:title>
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<treatment id="03881B7683597F01FCCDECEE8315F98B" LSID="urn:lsid:plazi:treatment:03881B7683597F01FCCDECEE8315F98B" httpUri="http://treatment.plazi.org/id/03881B7683597F01FCCDECEE8315F98B" lastPageNumber="206" pageId="5" pageNumber="206">
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<paragraph id="8B9EAA6083597F01FCCDECEE8445FF44" blockId="5.[818,1412,159,178]" lastBlockId="5.[818,1487,187,234]" pageId="5" pageNumber="206">
<emphasis id="B955767283597F01FCCDECEE85EFFF1C" bold="true" box="[818,1412,159,178]" italics="true" pageId="5" pageNumber="206">
2.8. Identification and phylogenetic analysis of cytochrome
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450s
</emphasis>
<emphasis id="B955767283597F01FCCDECCA8445FF44" bold="true" italics="true" pageId="5" pageNumber="206">
(CYP450s) putatively involved in biosynthesis of specialized flavonoids like baicalein in
<taxonomicName id="4C21D1E383597F01FC5FECA68468FF44" authority="(L.) Kurz" box="[928,1027,215,234]" class="Magnoliopsida" family="Bignoniaceae" genus="Oroxylum" kingdom="Plantae" order="Lamiales" pageId="5" pageNumber="206" phylum="Tracheophyta" rank="subSpecies" species="indicum" subSpecies="root">O. indicum</taxonomicName>
root
</emphasis>
</paragraph>
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<paragraph id="8B9EAA6083597F01FCACED7E8304FCB6" blockId="5.[818,1488,271,1573]" pageId="5" pageNumber="206">
Command line
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was performed on the
<taxonomicName id="4C21D1E383597F01FAD3ED7E83E5FE90" class="Magnoliopsida" family="Bignoniaceae" genus="Oroxylum" kingdom="Plantae" order="Lamiales" pageId="5" pageNumber="208" phylum="Tracheophyta" rank="subSpecies" species="indicum" subSpecies="transcriptome">
<emphasis id="B955767283597F01FAD3ED7E85FEFE8C" bold="true" box="[1324,1429,271,290]" italics="true" pageId="5" pageNumber="206">O. indicum</emphasis>
transcriptome
</taxonomicName>
using flavone synthase II, flavone-6-hydroxylase, glucuronidase and transferase genes reported from
<taxonomicName id="4C21D1E383597F01FB1FED3685CEFEF4" box="[1248,1445,327,346]" class="Magnoliopsida" family="Lamiaceae" genus="Scutellaria" kingdom="Plantae" order="Lamiales" pageId="5" pageNumber="206" phylum="Tracheophyta" rank="species" species="baicalensis">
<emphasis id="B955767283597F01FB1FED3685CEFEF4" bold="true" box="[1248,1445,327,346]" italics="true" pageId="5" pageNumber="206">Scutellaria baicalensis</emphasis>
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and shown to be involved in the biosynthesis of specialized flavonoids. This led to the identification of the
<taxonomicName id="4C21D1E383597F01FBA2ED0E84AAFE3C" box="[1117,1217,383,402]" class="Magnoliopsida" family="Bignoniaceae" genus="Oroxylum" kingdom="Plantae" order="Lamiales" pageId="5" pageNumber="206" phylum="Tracheophyta" rank="species" species="indicum">
<emphasis id="B955767283597F01FBA2ED0E84AAFE3C" bold="true" box="[1117,1217,383,402]" italics="true" pageId="5" pageNumber="206">O. indicum</emphasis>
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homologs of those genes as listed (Table 7). Additionally, 164 transcripts annotated as
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CYP
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50s from
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<emphasis id="B955767283597F01FC9BEDC783ADFE67" bold="true" box="[868,966,438,457]" italics="true" pageId="5" pageNumber="206">O. indicum</emphasis>
transcriptome
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were analyzed (Supplementary
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.) Of these, the ones with high
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value (more than 5.5), were screened for identification of full length CYP450 ORFs from the
<taxonomicName id="4C21D1E383597F01FA64ED9F83FDFDB3" class="Magnoliopsida" family="Bignoniaceae" genus="Oroxylum" kingdom="Plantae" order="Lamiales" pageId="5" pageNumber="206" phylum="Tracheophyta" rank="subSpecies" species="indicum" subSpecies="root">
<emphasis id="B955767283597F01FA64ED9F8303FDB3" bold="true" italics="true" pageId="5" pageNumber="206">O. indicum</emphasis>
root
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transcriptome.
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total of 31 genes (
<emphasis id="B955767283597F01FB0BEE7B8530FDB3" bold="true" box="[1268,1371,522,541]" italics="true" pageId="5" pageNumber="206">OinCyp450</emphasis>
) were identified (
<tableCitation id="C6A39FDB83597F01FC8DEE5783D3FD97" box="[882,952,550,569]" captionStart="Table 8" captionStartId="8.[100,150,383,400]" captionTargetPageId="8" captionText="Table 8 O. indicum Cytochrome P450 (OinCyp) full length genes identified from the transcriptome. No. New name Transcript FPKM Protein name and References for reported functions Predicted First hit of BLAST [organism] (Accession Activities reported (KEGG enzyme data base) protein (Amino number) acids) 1 OinCyp34 TR39327| c0_g2_i1 5.58 Abscisic acid 8-hydroxylase 2 (ABA 8-hydroxylase 2) (EC 415 hypothetical protein CDL12_11573 Catalyses the first step in the oxidative degradation of 1.14.13.93) (Cytochrome P450 707A6) (OsABA8ox2) [Handroanthus impetiginosus] PIN15764.1 abscisic acid. PMID:9808729 2 OinCyp33 TR37750| c0_g1_i8 5.93 Geraniol 8-hydroxylase (EC 1.14.13.152) (Cytochrome P450 500 Cytochrome P450 CYP2 subfamily [H. 3-hydroxylation of naringenin (cf. EC 1.14.13.21, flavonoid 76B6) (Geraniol 10-hydroxylase) (CrG10H) impetiginosus] PIM97815.1 3-monooxygenase) 1.14.13.152 transferred to 1.14.14.83 PMID:11718718 PMID:21504162 PMID:20699579 3 OinCyp32 TR28501| c0_g1_i2 6.05 Cytochrome P450 704C1 (EC 1.14.-.-) 500 Cytochrome P450 CYP4/CYP19/CYP26 Oxidoreductase activity, acting on paired donors, with subfamily [H. impetiginosus] (PIN26646.1) incorporation or reduction of molecular oxygen. 4 OinCyp21 TR44876|c0_g2_i1 12.43 Probable inactive linolenatehydroperoxidelyase (Cytochrome 143 Cytochrome P450 [H. impetiginosus] P450 74B2) (Hydroperoxidelyase 1) (PIN22161.1) 5 OinCyp19 TR31362| c0_g1_i4 12.96 Cytokinin hydroxylase (EC 1.14.13.-) (Cytochrome P450 517 35A1) 6 OinCyp16 TR31653| c0_g2_i1 16.39 Protein CYPRO4 641 PREDICTED: protein CYPRO4 [Erythranthe guttata] (XP_012836945.1) 7 OinCyp15 TR21276| c0_g1_i1 16.42 Cytochrome P450 81E8 (EC 1.14.13.-) 496 Cytochrome P450 81D11-like [Sesamum indicum] (XP_011078365.1) 8 OinCyp9 TR45834| c0_g1_i1 48.09 Cytochrome P450 704C1 (EC 1.14.-.-) (Cytochrome P450 497 Cytochrome P450 CYP4/CYP19/CYP26 CYPD) subfamily [H. impetiginosus] (PIN00373.1) 9 OinCyp31 TR10190|c0_g1_i12 6.06 Cytochrome P450 4V2 (EC 1.14.13.-) (Docosahexaenoic acid 460 Cytochrome P450 CYP4/CYP19/CYP26 Defects in the enzyme are associated with Bietti crystalline omega-hydroxylase CYP4V2) (EC 1.14.13.199) subfamily [H. impetiginosus] (PIN06076.1) corneoretinal dystrophy. The enzyme also produces some 21- 1.14.13.199 Transferred to 1.14.14.79 hydroxydocosahexaenoate. Acts in a similar way on eicosapentaenoic acid. 10 OinCyp30 TR5995|c0_g2_i3 6.34 Psoralen synthase (EC 1.14.13.102) (Cytochrome P450 510 Cytochrome P450 71A21-like [S. indicum] This microsomal cytochrome P-450-dependent enzyme is CYP71AJ2) (Fragment) (XP_011085047.1) specific for (+)-marmesin, and to a much lesser extent 5- PMID:17068340 hydroxymarmesin, as substrate. 11 OinCyp29 TR29646| c0_g1_i1 6.43 Cytochrome P450 704C1 (EC 1.14.-.-) (Cytochrome P450 504 Cytochrome P450 CYP4/CYP19/CYP26 Acts on a number of flavonoids, including naringenin and CYPD) subfamily [H. impetiginosus] (PIN06076.1) dihydrokaempferol. 12 OinCyp28 TR40383| c0_g1_i1 6.99 Cytochrome P450 704C1 (EC 1.14.-.-) (Cytochrome P450 293 Cytochrome P450 704C1 [S. indicum] (XP_ CYPD) 011076303.1) 13 OinCyp22 TR25900| c0_g1_i1 11.77 Cytochrome P450, putative (EC 1.14.13.21) 526 Cytochrome P450 734A1 [S. indicum] (XP_01109186) 14 OinCyp18 TR3312| c0_g1_i1 15.51 Cytochrome P450, putative (EC 1.14.13.88) 516 Cytochrome P450 CYP2 subfamily [H. impetiginosus] (PIN04379.1) 15 OinCyp14 TR39103| c0_g2_i1 30.21 Cytochrome P450, putative (EC 1.14.13.21) 536 Cytochrome P450 CYP2 subfamily [H. \ impetiginosus] (PIN06001.1) The EC no 1.14.13.21 has been changed to 1.14.14.82 (flavonoid 3-monooxygenase; CYP75B1 (gene name)) PMID:10504566 PMID:11030432 16 OinCyp4 TR45085| c0_g1_i1 152.57 Cytochrome P450, putative (EC 1.14.13.21) 511 Cytochrome P450 CYP2 subfamily [H. impetiginosus] (PIN10800.1) 17 OinCyp12 TR492| c0_g1_i1 33.03 Cytochrome P450, putative (EC 1.14.13.21) 528 Cytochrome P450 89A2 [S. indicum] (XP_ 011085115.1) 18 OinCyp11 TR34154| c0_g1_i1 44.58 Cytochrome P450, putative (EC 1.14.13.21) 199 Cytochrome P450 CYP2 subfamily [H. impetiginosus] (PIN04070.1) 19 OinCyp8 TR35703| c1_g1_i2 53.85 Peptidyl-prolylcis-trans isomerase CYP23 (PPIase CYP23) (EC 229 peptidylprolylisomerase [H. impetiginosus] 5.2.1.8) (PIN24972.1) 20 OinCyp27 TR25360| c0_g1_i1 7.12 305" pageId="5" pageNumber="206">Table 8</tableCitation>
). The phylogenetic tree of these
<emphasis id="B955767283597F01FB0EEE57852BFD97" bold="true" box="[1265,1344,550,569]" italics="true" pageId="5" pageNumber="206">
OinCyp
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</emphasis>
50s indicated a high diversity of CYP450s in the
<taxonomicName id="4C21D1E383597F01FB8FEE338569FDFB" box="[1136,1282,578,597]" class="Magnoliopsida" family="Bignoniaceae" genus="Oroxylum" kingdom="Plantae" order="Lamiales" pageId="5" pageNumber="206" phylum="Tracheophyta" rank="subSpecies" species="indicum" subSpecies="root">
<emphasis id="B955767283597F01FB8FEE3384BFFDFB" bold="true" box="[1136,1236,578,597]" italics="true" pageId="5" pageNumber="206">O. indicum</emphasis>
root
</taxonomicName>
(
<figureCitation id="131AB6E583597F01FAEEEE33852CFDFB" box="[1297,1351,578,597]" captionStart="Fig" captionStartId="10.[100,130,1231,1248]" captionTargetBox="[264,1322,154,1207]" captionTargetId="figure-160@10.[263,1325,152,1208]" captionTargetPageId="10" captionText="Fig. 5. Phylogenetic analysis of full length O. indicum Cytochrome P450 using maximum likelihood method. Phylogenetic tree of the newly identified, full length OinCyps (Tables 7 and 8) was performed using MEGA7 by maximum likelihood method." figureDoi="http://doi.org/10.5281/zenodo.10484702" httpUri="https://zenodo.org/record/10484702/files/figure.png" pageId="5" pageNumber="206">Fig. 5</figureCitation>
). At least five
<emphasis id="B955767283597F01FCCDEE2F8317FDDF" bold="true" box="[818,892,606,625]" italics="true" pageId="5" pageNumber="206">OinCyps</emphasis>
showed a high degree of similarity to biochemically well characterized enzymes from
<taxonomicName id="4C21D1E383597F01FBBEEE0B84D3FD23" box="[1089,1208,634,653]" class="Magnoliopsida" family="Lamiaceae" genus="Scutellaria" kingdom="Plantae" order="Lamiales" pageId="5" pageNumber="206" phylum="Tracheophyta" rank="species" species="baicalensis">
<emphasis id="B955767283597F01FBBEEE0B84D3FD23" bold="true" box="[1089,1208,634,653]" italics="true" pageId="5" pageNumber="206">S. baicalensis</emphasis>
</taxonomicName>
namely, flavones synthase II, baicalin-ss-D-glucuronidase, flavone-6-hydroxylase, baicalin-7-O-glucuronosyltransferase (
<bibRefCitation id="EFB0D79183597F01FBFCEEC084FEFD6A" author="Liu, J. &amp; Hou, J. &amp; Jiang, C. &amp; Li, G. &amp; Lu, H. &amp; Meng, F." box="[1027,1173,689,708]" pageId="5" pageNumber="206" pagination="0136397" refId="ref12494" refString="Liu, J., Hou, J., Jiang, C., Li, G., Lu, H., Meng, F., et al., 2015. Deep sequencing of the Scutellaria baicalensis Georgi transcriptome reveals flavonoid biosynthetic profiling and organ-specific gene expression. PloS One 10 (8), e 0136397. https: // doi. org / 10. 1371 / journal. pone. 0136397." type="journal article" year="2015">Liu et al., 2015</bibRefCitation>
; Zhao et al., 2016, 2018). These enzymes catalyze the biosynthesis of specialized flavonoids like baicalin and wogonin through a root specific alternative pathway (Zhao et al., 2016).
</paragraph>
<paragraph id="8B9EAA6083597F01FCACEF5085C6FB2D" blockId="5.[818,1488,271,1573]" pageId="5" pageNumber="206">
The
<emphasis id="B955767283597F01FC83EF5083A0FC9A" bold="true" box="[892,971,801,820]" italics="true" pageId="5" pageNumber="206">OinCyp3</emphasis>
and
<emphasis id="B955767283597F01FC06EF508423FC9A" bold="true" box="[1017,1096,801,820]" italics="true" pageId="5" pageNumber="206">OinCyp4</emphasis>
were abundant in roots of
<taxonomicName id="4C21D1E383597F01FABAEF5085CCFC9A" box="[1349,1447,801,820]" class="Magnoliopsida" family="Bignoniaceae" genus="Oroxylum" kingdom="Plantae" order="Lamiales" pageId="5" pageNumber="206" phylum="Tracheophyta" rank="species" species="indicum">
<emphasis id="B955767283597F01FABAEF5085CCFC9A" bold="true" box="[1349,1447,801,820]" italics="true" pageId="5" pageNumber="206">O. indicum</emphasis>
</taxonomicName>
and showed a high degree of similarity with flavone synthase II (CYP93B; Zhao et al., 2016) along with five more
<emphasis id="B955767283597F01FB3CEF288566FCC2" bold="true" box="[1219,1293,857,876]" italics="true" pageId="5" pageNumber="206">OinCyps</emphasis>
(
<figureCitation id="131AB6E583597F01FAE0EF288533FCC2" box="[1311,1368,857,876]" captionStart="Fig" captionStartId="10.[100,130,1231,1248]" captionTargetBox="[264,1322,154,1207]" captionTargetId="figure-160@10.[263,1325,152,1208]" captionTargetPageId="10" captionText="Fig. 5. Phylogenetic analysis of full length O. indicum Cytochrome P450 using maximum likelihood method. Phylogenetic tree of the newly identified, full length OinCyps (Tables 7 and 8) was performed using MEGA7 by maximum likelihood method." figureDoi="http://doi.org/10.5281/zenodo.10484702" httpUri="https://zenodo.org/record/10484702/files/figure.png" pageId="5" pageNumber="206">Fig. 5</figureCitation>
). Moreover, OinCyp2 (with similarity to
<taxonomicName id="4C21D1E383597F01FBB8EF0484ABFC26" box="[1095,1216,885,904]" class="Magnoliopsida" family="Lamiaceae" genus="Scutellaria" kingdom="Plantae" order="Lamiales" pageId="5" pageNumber="206" phylum="Tracheophyta" rank="species" species="baicalensis">
<emphasis id="B955767283597F01FBB8EF0484ABFC26" bold="true" box="[1095,1216,885,904]" italics="true" pageId="5" pageNumber="206">S. baicalensis</emphasis>
</taxonomicName>
baicalin-ss-D-glucuronidase) clustered close to OinCyp27. OinCyp2 and OinCyp35 (both flavone-6- hydroxylase) clustered away from each other forming two different OinCyp clades (
<figureCitation id="131AB6E583597F01FC34EFB98469FC75" box="[971,1026,968,987]" captionStart="Fig" captionStartId="10.[100,130,1231,1248]" captionTargetBox="[264,1322,154,1207]" captionTargetId="figure-160@10.[263,1325,152,1208]" captionTargetPageId="10" captionText="Fig. 5. Phylogenetic analysis of full length O. indicum Cytochrome P450 using maximum likelihood method. Phylogenetic tree of the newly identified, full length OinCyps (Tables 7 and 8) was performed using MEGA7 by maximum likelihood method." figureDoi="http://doi.org/10.5281/zenodo.10484702" httpUri="https://zenodo.org/record/10484702/files/figure.png" pageId="5" pageNumber="206">Fig. 5</figureCitation>
). OinCyp5 (with similarity to
<taxonomicName id="4C21D1E383597F01FAD4EFB985C8FC75" box="[1323,1443,968,987]" class="Magnoliopsida" family="Lamiaceae" genus="Scutellaria" kingdom="Plantae" order="Lamiales" pageId="5" pageNumber="206" phylum="Tracheophyta" rank="species" species="baicalensis">
<emphasis id="B955767283597F01FAD4EFB985C8FC75" bold="true" box="[1323,1443,968,987]" italics="true" pageId="5" pageNumber="206">S. baicalensis</emphasis>
</taxonomicName>
baicalin-7-O-glucuronosyl transferase) was highly abundant in
<taxonomicName id="4C21D1E383597F01FA93EF9585A4FC59" box="[1388,1487,996,1015]" class="Magnoliopsida" family="Bignoniaceae" genus="Oroxylum" kingdom="Plantae" order="Lamiales" pageId="5" pageNumber="206" phylum="Tracheophyta" rank="species" species="indicum">
<emphasis id="B955767283597F01FA93EF9585A4FC59" bold="true" box="[1388,1487,996,1015]" italics="true" pageId="5" pageNumber="206">O. indicum</emphasis>
</taxonomicName>
roots and formed a cluster with at least five more OinCyps. Based on the phylogenetic analysis of the OinCyps, it can be hypothesized that they are involved in the specialized flavonoid biosynthesis in
<taxonomicName id="4C21D1E383597F01FA96E84985A4FBE5" box="[1385,1487,1080,1099]" class="Magnoliopsida" family="Bignoniaceae" genus="Oroxylum" kingdom="Plantae" order="Lamiales" pageId="5" pageNumber="206" phylum="Tracheophyta" rank="species" species="indicum">
<emphasis id="B955767283597F01FA96E84985A4FBE5" bold="true" box="[1385,1487,1080,1099]" italics="true" pageId="5" pageNumber="206">O. indicum</emphasis>
</taxonomicName>
roots. Further, the functional validation of OinCyps may highlight their specific biochemical roles in metabolite biosynthesis in
<taxonomicName id="4C21D1E383597F01FABAE80185C3FB2D" box="[1349,1448,1136,1155]" class="Magnoliopsida" family="Bignoniaceae" genus="Oroxylum" kingdom="Plantae" order="Lamiales" pageId="5" pageNumber="206" phylum="Tracheophyta" rank="species" species="indicum">
<emphasis id="B955767283597F01FABAE80185C3FB2D" bold="true" box="[1349,1448,1136,1155]" italics="true" pageId="5" pageNumber="206">O. indicum</emphasis>
</taxonomicName>
.
</paragraph>
<paragraph id="8B9EAA6083597F01FCACE8FD8315F98B" blockId="5.[818,1488,271,1573]" pageId="5" pageNumber="206">
Phylogenetic variations in
<emphasis id="B955767283597F01FB9FE8FD84CCFB31" bold="true" box="[1120,1191,1164,1183]" italics="true" pageId="5" pageNumber="206">Cyp450</emphasis>
genes play a central role in evolution of metabolic complexity (
<bibRefCitation id="EFB0D79183597F01FB74E8D9854DFB15" author="Bak, S. &amp; Beisson, F. &amp; Bishop, G. &amp; Hamberger, B. &amp; Hofer, R. &amp; Paquette, S. &amp; Werck-Reichhart, D." box="[1163,1318,1192,1211]" pageId="5" pageNumber="206" pagination="0144" refId="ref10413" refString="Bak, S., Beisson, F., Bishop, G., Hamberger, B., Hofer, R., Paquette, S., Werck-Reichhart, D., 2011. Cytochromes P 450, vol. 9 The Arabidopsis Book / American Society of Plant Biologists, e 0144. http: // doi. org / 10.1199 / tab. 0144." type="book chapter" year="2011">Bak et al., 2011</bibRefCitation>
).
<emphasis id="B955767283597F01FAC2E8D98500FB15" bold="true" box="[1341,1387,1192,1211]" italics="true" pageId="5" pageNumber="206">Cyp9</emphasis>
8
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and its duplicated sister genes evolved functional diversification in phenolic and flavonoid biosynthesis pathways (
<bibRefCitation id="EFB0D79183597F01FB5EE8AE855EFB5C" author="Liu, Z. &amp; Tavares, R. &amp; Forsythe, E. S. &amp; Andre, F. &amp; Lugan, R. &amp; Jonasson, G. &amp; Mercey, S. B. &amp; Tohge, T. &amp; Beilstein, M. A. &amp; Daniele Werck-Reichhart, D. W. &amp; Renault, H." box="[1185,1333,1247,1266]" pageId="5" pageNumber="206" pagination="13026" refId="ref12394" refString="Liu, Z., Tavares, R., Forsythe, E. S., Andre, F., Lugan, R., Jonasson, G., Mercey, S. B., Tohge, T., Beilstein, M. A., Daniele Werck-Reichhart, D. W., Renault, H., 2016. Evolutionary interplay between sister cytochrome P 450 genes shapes plasticity in plant metabolism. Nat. Commun. 7, 13026. https: // doi. org / 10.1038 / ncomms 13026." type="journal article" year="2016">Liu et al., 2016</bibRefCitation>
). The flavonoid biosynthesis pathway is known to involve the formation of metabolons or protein complexes (
<bibRefCitation id="EFB0D79183597F01FBF5E96684A3FA84" author="Bassard, J. E. &amp; Moller, L. B. &amp; Laursen, T." box="[1034,1224,1303,1322]" pageId="5" pageNumber="206" pagination="37 - 51" refId="ref10483" refString="Bassard, J. E., Moller, L. B., Laursen, T., 2017. Assembly of dynamic P 450 - mediated metabolons - order versus chaos. Curr. Mol. Biol. Rep. 3, 37 - 51." type="journal article" year="2017">Bassard et al., 2017</bibRefCitation>
). It has been demonstrated that Cyp450s like flavone synthase and cinnamate 4-hydroxylase are ER membrane anchored enzymes and provide a nucleation platform for the assembly of other soluble enzymes like chalcone synthase, chalcone isomerase and reductase. Together, these complexes drive the flavonoid metabolism (
<bibRefCitation id="EFB0D79183597F01FC50E9D284E4FA18" author="Dastmalchi, M. &amp; Mark, A. &amp; Bernards, M. A. &amp; Dhaubhadel, S." box="[943,1167,1443,1462]" pageId="5" pageNumber="206" pagination="689 - 706" refId="ref10860" refString="Dastmalchi, M., Mark, A., Bernards, M. A., Dhaubhadel, S., 2016. Twin anchors of the soybean isoflavonoid metabolon: evidence for tethering of the complex to the endoplasmic reticulum by IFS and C 4 H. Plant J. 85, 689 - 706." type="journal article" year="2016">Dastmalchi et al., 2016</bibRefCitation>
) and plant specific diversity in a flavonoid metabolon organization (
<bibRefCitation id="EFB0D79183597F01FB7CE9CE855FFA7C" author="Fujino, N. &amp; Tenma, N. &amp; Waki, T. &amp; Ito, K. &amp; Komatsuzaki, Y. &amp; Sugiyama, K. &amp; Yamazaki, T. &amp; Yoshida, S. &amp; Hatayama, M. &amp; Yamashita, S. &amp; Tanaka, Y. &amp; Motohashi, R. &amp; Denessiouk, K. &amp; Takahashi, S. &amp; Nakayama, T." box="[1155,1332,1471,1490]" pageId="5" pageNumber="206" pagination="372 - 392" refId="ref11291" refString="Fujino, N., Tenma, N., Waki, T., Ito, K., Komatsuzaki, Y., Sugiyama, K., Yamazaki, T., Yoshida, S., Hatayama, M., Yamashita, S., Tanaka, Y., Motohashi, R., Denessiouk, K., Takahashi, S., Nakayama, T., 2018. Physical interactions among flavonoid enzymes in snapdragon and torenia reveal the diversity in the flavonoid metabolon organization of different plant species. Plant J. 94, 372 - 392." type="journal article" year="2018">Fujino et al., 2018</bibRefCitation>
). Detailed functional validation of OinCyp diversity will open up their precise biochemical roles and novelty in specialized flavonoid metabolism in
<taxonomicName id="4C21D1E383597F01FA45E9878312F98B" class="Magnoliopsida" family="Bignoniaceae" genus="Oroxylum" kingdom="Plantae" order="Lamiales" pageId="5" pageNumber="206" phylum="Tracheophyta" rank="species" species="indicum">
<emphasis id="B955767283597F01FA45E9878312F98B" bold="true" italics="true" pageId="5" pageNumber="206">O. indicum</emphasis>
</taxonomicName>
.
</paragraph>
</subSubSection>
</treatment>
</document>