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Exploring drought stress-regulated genes in senna (Cassia angustifolia Vahl.): a transcriptomic approach

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Abstract

De novo assembly of reads produced by next-generation sequencing (NGS) technologies offers a rapid approach to obtain expressed gene sequences for non-model organisms. Senna (Cassia angustifolia Vahl.) is a drought-tolerant annual undershrub of Caesalpiniaceae, a subfamily of Fabaceae. There are insufficient transcriptomic and genomic data in public databases for understanding the molecular mechanism underlying the drought tolerance of senna. Therefore, the main purpose of this study was to know the transcriptome profile of senna, with special reference to drought stress. RNA from two different stages of leaf development was extracted and sequenced separately using the Illumina technology. A total of 200 million reads were generated, and a de novo assembly of processed reads in the pooled transcriptome using Trinity yielded 43,413 transcripts which were further annotated using NCBI BLAST with “green plant database (txid 33090),” Swiss Prot, Kyoto Encyclopedia of Genes and Genomes (KEGG), Clusters of Orthologous Groups (COG), and Gene Ontology (GO). Out of the total transcripts, 42,280 (95.0 %) were annotated by BLASTX against the green plant database of NCBI. Senna transcriptome showed the highest similarity to Glycine max (41 %), followed by Phaseolus vulgaris (16 %), Cicer arietinum (15 %), and Medicago trancatula (5 %). The highest number of GO terms were enriched for the molecular functions category; of these “catalytic activity” (GO: 0003824) (25.10 %) and “binding activity” (GO: 0005488) (20.10 %) were most abundantly represented. We used InterProscan to see protein similarity at domain level; a total of 33,256 transcripts were annotated against the Pfam domains. The transcripts were assigned with various KEGG pathways. Coding DNA sequences (CDS) encoding various drought stress-regulated pathways such as signaling factors, protein-modifying/degrading enzymes, biosynthesis of phytohormone, phytohormone signaling, osmotically active compounds, free radical scavengers, chlorophyll metabolism, leaf cuticular wax, polyamines, and protective proteins were identified through BLASTX search. The lucine-rich repeat kinase family was the most abundantly found group of protein kinases. Orphan, bHLH, and bZIP family TFs were the most abundantly found in senna. Six genes encoding MYC2 transcription factor, 9-cis-epoxycarotenoid dioxygenase (NCED), l -ascorbate peroxidase (APX), aminocyclopropane carboxylate oxidase (ACO), abscisic acid 8′-hydroxylase (ABA), and WRKY transcription factor were confirmed through reverse transcriptase-PCR (RT-PCR) and Sanger sequencing for the first time in senna. The potential drought stress-related transcripts identified in this study provide a good start for further investigation into the drought adaptation in senna. Additionally, our transcriptome sequences are the valuable resource for accelerated genomics-assisted genetic improvement programs and facilitate manipulation of biochemical pathways for developing drought-tolerant genotypes of crop plants.

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Acknowledgments

The authors gratefully acknowledge the Department of Science and Technology (DST), Government of India (GOI), for supporting this work under the Fast Track Scheme for Young Scientist under grant number SB/FT/LS-329/2012 and the project entitled “Transcriptome analysis of senna (Cassia angustifolia Vahl.) to identify potential genes involved in the biosynthesis of sennosides” and the Director, ICAR-Directorate of Medicinal and Aromatic Plants Research, Boriavi, Anand, Gujarat, India, and Indian Council of Agricultural research (ICAR), New Delhi, India for the facilities to undertake the study.

Authors’ contributions

Conceived and designed the experiments: NRRR. Performed the experiments: NRRR RHM. Analyzed the data: NRRR MP JK. Contributed reagents/materials/analysis tools: NRRR MP JK. Wrote the paper: RHM NRRR MP JK

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Authors and Affiliations

  1. ICAR-Directorate of Medicinal and Aromatic Plants Research (ICAR-DMAPR), Anand, 387310, Gujarat, India

    Rucha Harishbhai Mehta, Manivel Ponnuchamy, Jitendra Kumar & Nagaraja Reddy Rama Reddy

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  1. Rucha Harishbhai Mehta
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  2. Manivel Ponnuchamy
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  3. Jitendra Kumar
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  4. Nagaraja Reddy Rama Reddy
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Correspondence to Nagaraja Reddy Rama Reddy.

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The authors declare that they have no conflicts of interest.

Availability of supporting data

The datasets supporting the results of this article are available at the National Center for Biotechnology Information (NCBI) BioProject database (Short Read Archive) under accession number PRJNA273534 and the Transcriptome Shotgun Assembly (TSA) at DDBJ/EMBL/GenBank under the accession GEEB00000000. The version described in this paper is the first version, GEEB01000000.

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Figure S1

Transcript size (bp) distribution (DOC 61 kb)

Figure S2

Size distribution of CDS (DOC 67 kb)

Figure S3

Sequence alignment to show conformity between transcriptome assembly (trans) and Sanger sequencing (seq) of the in vitro amplified PCR product of drought stress regulated genes in senna (DOCX 30 kb)

Table S1

KEGG categories of CDS in the transcriptome of leaf of C. angustifolia (PDF 3957 kb)

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Mehta, R.H., Ponnuchamy, M., Kumar, J. et al. Exploring drought stress-regulated genes in senna (Cassia angustifolia Vahl.): a transcriptomic approach. Funct Integr Genomics 17, 1–25 (2017). https://doi.org/10.1007/s10142-016-0523-y

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  • DOI: https://doi.org/10.1007/s10142-016-0523-y

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