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Transcriptome analysis reveals in vitro cultured Withania somnifera leaf and root tissues as a promising source for targeted withanolide biosynthesis

BACKGROUND: The production of metabolites via in vitro culture is promoted by the availability of fully defined metabolic pathways. Withanolides, the major bioactive phytochemicals of Withania somnifera, have been well studied for their pharmacological activities. However, only a few attempts have b...

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Autores principales: Senthil, Kalaiselvi, Jayakodi, Murukarthick, Thirugnanasambantham, Pankajavalli, Lee, Sang Choon, Duraisamy, Pradeepa, Purushotham, Preethi M, Rajasekaran, Kalaiselvi, Nancy Charles, Shobana, Mariam Roy, Irene, Nagappan, Arul Kumar, Kim, Gon Sup, Lee, Yun Sun, Natesan, Senthil, Min, Tae-Sun, Yang, Tae Jin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4310147/
https://www.ncbi.nlm.nih.gov/pubmed/25608483
http://dx.doi.org/10.1186/s12864-015-1214-0
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author Senthil, Kalaiselvi
Jayakodi, Murukarthick
Thirugnanasambantham, Pankajavalli
Lee, Sang Choon
Duraisamy, Pradeepa
Purushotham, Preethi M
Rajasekaran, Kalaiselvi
Nancy Charles, Shobana
Mariam Roy, Irene
Nagappan, Arul Kumar
Kim, Gon Sup
Lee, Yun Sun
Natesan, Senthil
Min, Tae-Sun
Yang, Tae Jin
author_facet Senthil, Kalaiselvi
Jayakodi, Murukarthick
Thirugnanasambantham, Pankajavalli
Lee, Sang Choon
Duraisamy, Pradeepa
Purushotham, Preethi M
Rajasekaran, Kalaiselvi
Nancy Charles, Shobana
Mariam Roy, Irene
Nagappan, Arul Kumar
Kim, Gon Sup
Lee, Yun Sun
Natesan, Senthil
Min, Tae-Sun
Yang, Tae Jin
author_sort Senthil, Kalaiselvi
collection PubMed
description BACKGROUND: The production of metabolites via in vitro culture is promoted by the availability of fully defined metabolic pathways. Withanolides, the major bioactive phytochemicals of Withania somnifera, have been well studied for their pharmacological activities. However, only a few attempts have been made to identify key candidate genes involved in withanolide biosynthesis. Understanding the steps involved in withanolide biosynthesis is essential for metabolic engineering of this plant to increase withanolide production. RESULTS: Transcriptome sequencing was performed on in vitro adventitious root and leaf tissues using the Illumina platform. We obtained a total of 177,156 assembled transcripts with an average unigene length of 1,033 bp. About 13% of the transcripts were unique to in vitro adventitious roots but no unique transcripts were observed in in vitro-grown leaves. A putative withanolide biosynthetic pathway was deduced by mapping the assembled transcripts to the KEGG database, and the expression of candidate withanolide biosynthesis genes -were validated by qRT PCR. The accumulation pattern of withaferin A and withanolide A varied according to the type of tissue and the culture period. Further, we demonstrated that in vitro leaf extracts exhibit anticancer activity against human gastric adenocarcinoma cell lines at sub G1 phase. CONCLUSIONS: We report here a validated large-scale transcriptome data set and the potential biological activity of in vitro cultures of W. somnifera. This study provides important information to enhance tissue-specific expression and accumulation of secondary metabolites, paving the way for industrialization of in vitro cultures of W. somnifera. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12864-015-1214-0) contains supplementary material, which is available to authorized users.
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spelling pubmed-43101472015-01-30 Transcriptome analysis reveals in vitro cultured Withania somnifera leaf and root tissues as a promising source for targeted withanolide biosynthesis Senthil, Kalaiselvi Jayakodi, Murukarthick Thirugnanasambantham, Pankajavalli Lee, Sang Choon Duraisamy, Pradeepa Purushotham, Preethi M Rajasekaran, Kalaiselvi Nancy Charles, Shobana Mariam Roy, Irene Nagappan, Arul Kumar Kim, Gon Sup Lee, Yun Sun Natesan, Senthil Min, Tae-Sun Yang, Tae Jin BMC Genomics Research Article BACKGROUND: The production of metabolites via in vitro culture is promoted by the availability of fully defined metabolic pathways. Withanolides, the major bioactive phytochemicals of Withania somnifera, have been well studied for their pharmacological activities. However, only a few attempts have been made to identify key candidate genes involved in withanolide biosynthesis. Understanding the steps involved in withanolide biosynthesis is essential for metabolic engineering of this plant to increase withanolide production. RESULTS: Transcriptome sequencing was performed on in vitro adventitious root and leaf tissues using the Illumina platform. We obtained a total of 177,156 assembled transcripts with an average unigene length of 1,033 bp. About 13% of the transcripts were unique to in vitro adventitious roots but no unique transcripts were observed in in vitro-grown leaves. A putative withanolide biosynthetic pathway was deduced by mapping the assembled transcripts to the KEGG database, and the expression of candidate withanolide biosynthesis genes -were validated by qRT PCR. The accumulation pattern of withaferin A and withanolide A varied according to the type of tissue and the culture period. Further, we demonstrated that in vitro leaf extracts exhibit anticancer activity against human gastric adenocarcinoma cell lines at sub G1 phase. CONCLUSIONS: We report here a validated large-scale transcriptome data set and the potential biological activity of in vitro cultures of W. somnifera. This study provides important information to enhance tissue-specific expression and accumulation of secondary metabolites, paving the way for industrialization of in vitro cultures of W. somnifera. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12864-015-1214-0) contains supplementary material, which is available to authorized users. BioMed Central 2015-01-22 /pmc/articles/PMC4310147/ /pubmed/25608483 http://dx.doi.org/10.1186/s12864-015-1214-0 Text en © Senthil et al.; licensee Biomed Central. 2015 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Senthil, Kalaiselvi
Jayakodi, Murukarthick
Thirugnanasambantham, Pankajavalli
Lee, Sang Choon
Duraisamy, Pradeepa
Purushotham, Preethi M
Rajasekaran, Kalaiselvi
Nancy Charles, Shobana
Mariam Roy, Irene
Nagappan, Arul Kumar
Kim, Gon Sup
Lee, Yun Sun
Natesan, Senthil
Min, Tae-Sun
Yang, Tae Jin
Transcriptome analysis reveals in vitro cultured Withania somnifera leaf and root tissues as a promising source for targeted withanolide biosynthesis
title Transcriptome analysis reveals in vitro cultured Withania somnifera leaf and root tissues as a promising source for targeted withanolide biosynthesis
title_full Transcriptome analysis reveals in vitro cultured Withania somnifera leaf and root tissues as a promising source for targeted withanolide biosynthesis
title_fullStr Transcriptome analysis reveals in vitro cultured Withania somnifera leaf and root tissues as a promising source for targeted withanolide biosynthesis
title_full_unstemmed Transcriptome analysis reveals in vitro cultured Withania somnifera leaf and root tissues as a promising source for targeted withanolide biosynthesis
title_short Transcriptome analysis reveals in vitro cultured Withania somnifera leaf and root tissues as a promising source for targeted withanolide biosynthesis
title_sort transcriptome analysis reveals in vitro cultured withania somnifera leaf and root tissues as a promising source for targeted withanolide biosynthesis
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4310147/
https://www.ncbi.nlm.nih.gov/pubmed/25608483
http://dx.doi.org/10.1186/s12864-015-1214-0
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