Abstract

Rheum australe is an endangered medicinal herb of high altitude alpine region of Himalayas and is known to possess anti-cancerous properties. Unlike many herbs of the region, R. australe has broad leaves. The species thrives well under the environmental extremes in its niche habitat, therefore an understanding of transcriptome of R. australe to environmental cues was of significance. Since, temperature is one of the major environmental variables in the niche of R. australe, transcriptome was studied in the species growing in natural habitat and those grown in growth chambers maintained at 4 °C and 25 °C to understand genes associated with different temperatures. A total of 39,136 primarily assembled transcripts were obtained from 10,17,74,336 clean read, and 21,303 unigenes could match to public databases. An analysis of transcriptome by fragments per kilobase of transcript per million, followed by validation through qRT-PCR showed 22.4% up- and 22.5% down-regulated common differentially expressed genes in the species growing under natural habitat and at 4 °C as compared to those at 25 °C. These genes largely belonged to signaling pathway, transporters, secondary metabolites, phytohormones, and those associated with cellular protection, suggesting their importance in imparting adaptive advantage to R. australe in its niche.

Highlights

  • Of human breast carcinoma (MDA-MB-435S), and liver carcinoma (Hep3B) cell l­ines[10]

  • Three cDNA libraries were generated from the plant growing naturally in the niche location and the plants grown in plant growth chambers maintained at 4 °C and 25 °C (Supplementary Fig. S1a-f)

  • Leaf tissues collected from natural habitat at High altitude (HA), 4 °C, and 25 °C were termed as LHA, L4, and L25, respectively

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Summary

Introduction

Of human breast carcinoma (MDA-MB-435S), and liver carcinoma (Hep3B) cell l­ines[10]. Since temperature is one of the most dominant perceivable parameters at HA, the objective of the present work was to (i) compare the transcriptome profile of plant samples collected in-situ from its natural habitat with those grown at 4 °C and 25 °C in plant growth chambers and (ii) identify the common genes which show a similar trend of expression in the niche location and those exposed to 4 °C as compared to those at 25 °C. The present work identified a wide range of genes through RNA-Seq data generated on Illumina platform (Genome Analyzer IIx) which showed overexpression at the niche location and at 4 °C when compared to those at 25 °C and 28 genes were validated by quantitative real-time PCR (qRT-PCR). Analysis showed positive correlation between qRT-PCR and fragments per kilobase of transcript per million (FPKM) data

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