02549nas a2200265   4500008004100000022001400041245016000055210006900215260000800284300001400292490000700306520161000313653003601923653001201959653001001971653003101981653001602012100002002028700001802048700002602066700002402092700001802116700001902134856013002153       2020                        eng d  a0971-589400aTIAs pathway genes and associated miRNA identification in Vinca minor: supporting aspidosperma and eburnamine alkaloids linkage via transcriptomic analysis0 aTIAs pathway genes and associated miRNA identification in Vinca   cAUG  a1695-17110 v263 a<p>V. minorcontains monomeric eburnamine-type of indole alkaloids having utilization as a neuro-medicinal plant. The biosynthetic pathway studies using miRNAs has been the focal point for plant genomic research in recent years and this technique is utilized to get an insight into a possible pathway level study inV. minoras understanding of genes in this prized medicinal plant is meagrely understood. The de novo transcriptomic analysis using Illumina Next gen sequencing has been performed in glasshouse shifted plant and transformed roots to elucidate the possible non confirmed steps of terpenoid indole alkaloids (TIAs) pathway inV. minor. A putative TIA pathway is elucidated in the study including twelve possible TIAs biosynthetic genes. The specific miRNA associated with TIAs pathway were identified and their roles were discussed for the first time inV. minor. The comparative analysis of transcriptomic data of glasshouse shifted plant and transformed roots showed that the raw reads of transformed roots were higher (83,740,316) compared to glasshouse shifted plant (67,733,538). The EST-SSR prediction showed the maximum common repeats among glasshouse shifted plant and transformed roots, although small variation was found in trinucleotide repeats restricted to glasshouse shifted plant. The study reveals overall 37 miRNAs which were observed to be true and can have a role in pathway as they can regulate the growth and alkaloid production. The identification of putative pathway genes plays an important role in establishing linkage between Aspidosperma and Eburnamine alkaloids.</p>
10ade novo transcriptomic analysis10aEST-SSR10amiRNA10aTerpenoid indole alkaloids10aVinca minor1 aVerma, Priyanka1 aSingh, Noopur1 aKhan, Shamshad, Ahmad1 aMathur, Ajay, Kumar1 aSharma, Ashok1 aJamal, Farrukh  uhttps://library.ncl.res.in/content/tias-pathway-genes-and-associated-mirna-identification-vinca-minor-supporting-aspidosperma02729nas a2200253   4500008004100000022001400041245018000055210006900235260000800304300001400312490000700326520183900333653001802172653001002190653001502200653000902215653001602224653001402240100002002254700002602274700002402300700002402324856012702348       2021                        eng d  a0971-589400aZCTs knockdown using antisense LNA GapmeR in specialized photomixotrophic cell suspensions of Catharanthus roseus: Rerouting the flux towards mono and dimeric indole alkaloids0 aZCTs knockdown using antisense LNA GapmeR in specialized photomi  cJUL  a1437-14530 v273 aThe present study was carried out to silence the transcription factor genes ZCT1, ZCT2 and ZCT3 via lipofectamine based antisense LNA GapmeRs transfection into the protoplasts of established photomixotrophic cell suspensions. The photomixotrophic cell suspensions with a threshold of 0.5% sucrose were raised and established using two-tiered CO2 providing flasks kept under high light intensity. The photomixotrophic cell suspensions showed morphologically different thick-walled cells under scanning electron microscopic analysis in comparison to the simple thin-walled parenchymatous control cell suspensions. The LC-MS analysis registered the vindoline production (0.0004 +/- 0.0001 mg/g dry wt.) in photomixotrophic cell suspensions which was found to be absent in control cell suspensions. The protoplasts were isolated from the photomixotrophic cell suspensions and subjected to antisense LNA GapmeRs silencing. Three lines, viz. Z1A, Z2C and Z3G were obtained where complete silencing of ZCT1, ZCT2 and ZCT3 genes, respectively, was observed. The Z3G line was found to show maximum production of vindoline (0.038 +/- 0.001 mg/g dry wt.), catharanthine (0.165 +/- 0.008 mg/g dry wt.) and vinblastine (0.0036 +/- 0.0003 mg/g dry wt.). This was supported by the multifold increment in the gene expression of TDC, SLS, STR, SGD, d4h, dat, CrT16H and Crprx. The present work indicates the master regulation of ZCT3 knockdown among all three ZCTs transcription factors in C. roseus to enhance the terpenoid indole alkaloids production. The successful silencing of transcription repressor genes has been achieved in C. roseus plant system by using photomixotrophic cell cultures through GapmeR based silencing. The present study is a step towards metabolic engineering of the TIAs pathway using protoplast transformation in C. roseus.10aCatharanthine10aLC-MS10aProtoplast10aTIAs10aVinblastine10aVindoline1 aVerma, Priyanka1 aKhan, Shamshad, Ahmad1 aParasharami, Varsha1 aMathur, Ajay, Kumar  uhttps://library.ncl.res.in/content/zcts-knockdown-using-antisense-lna-gapmer-specialized-photomixotrophic-cell-suspensions