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

Abstract

BackgroundThe 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.ResultsTranscriptome 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.ConclusionsWe 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 materialThe online version of this article (doi:10.1186/s12864-015-1214-0) contains supplementary material, which is available to authorized users.

Withaferin A inhibits the proliferation of gastric cancer cells by inducing G2/M cell cycle arrest and apoptosis

“…Transcription factors, including nuclear factor-κB (NF-κB) and mitogen-activated protein kinases, contribute to WA-mediated induction of apoptosis of leukemia (4,24), glioblastoma (25) and breast cancer (26). The antitumor effects of WA on human AGS were not previously studied, although W. somnifera leaf and root extract have been assessed for their effect on AGS cells (10). In the present study, it was identified that apoptosis was significantly increased in AGS cells treated with WA.…”

“…Indeed, WA has been demonstrated to produce antitumorigenic effects in multiple cancer cell lines, including breast, prostate and lung cancer cells (7)(8)(9). Senthil et al (10) recently reported that an extract from W. somnifera inhibits the proliferation of AGS cells by inducing apoptosis and cell cycle arrest. Furthermore, we have recently reported that WA efficiently decreased pro-inflammatory processes in gastric epithelial cells, as well as immune cells, in response to Helicobacter pylori (11,12), which serves a pivotal role in the high incidence of gastric cancer (2).…”

Comparative Characterization of the Leaf Tissue of Physalis alkekengi and Physalis peruviana Using RNA-seq and Metabolite Profiling

“…RNA sequencing (RNA-seq) on NGSs can be used for both model and non-model plants (Gongora-Castillo and Buell, 2013; Fukushima and Kusano, 2014; Weber, 2015). Successful transcriptome studies on non-model plants, e.g., crop plants such as the eggplant (Ramesh et al, 2016), pepper (Gordo et al, 2012) and tobacco (Lei et al, 2014) and medicinal plants, e.g., Withania somnifera (Gupta et al, 2013; Senthil et al, 2015) and P. peruviana (Garzon-Martinez et al, 2012) from the Solanaceae family have been published. Resources for transcriptome data from medicinal plants, e.g., the Medicinal Plant Genomics Resource (MPGR 1 ) and the Medicinal Plants Transcriptomics (Medplants 2 ; Gongora-Castillo et al, 2012a,b; Yeo et al, 2013) are available.…”

Ethnopharmacological Review of Natural Products in Cancer Prevention and Therapy

“…A shrub of Solanacea family regarded as one of the important members of generally regarded as safe plants found all over in India [148], having anticarcinogenic properties [149]. It has been suggested that this plant is used as an alternative long-term remedy to avoid the spread of cancer cells, after the analysis of root extracts against vimentin pro-metastatic protein, and thus, this plant is used to create motility inhibition of cells of breast tumors in different formulations [150].…”