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JXB Advance Access published online on October 18, 2006
Journal of Experimental Botany, doi:10.1093/jxb/erl146
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© 2006 The Author(s). This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Received May 25, 2006
Accepted August 16, 2006

RESEARCH PAPER

Transcriptome analysis in Catharanthus roseus leaves and roots for comparative terpenoid indole alkaloid profiles

Ashutosh K. Shukla 1, Ajit K. Shasany 1, Madan M. Gupta 2, and Suman P. S. Khanuja 1 *

1 Genetic Resources and Biotechnology Division, Central Institute of Medicinal and Aromatic Plants, Lucknow 226015, India
2 Analytical Chemistry Division, Central Institute of Medicinal and Aromatic Plants, Lucknow 226015, India

* To whom correspondence should be addressed.
Suman P. S. Khanuja, E-mail: khanujazy{at}yahoo.com


  Abstract

In Catharanthus roseus (L.) G. Don each tissue is known to produce a distinct spectrum of terpenoid indole alkaloids. Since the invaluable antineoplastic bisindole alkaloids are restricted to the aerial parts of the plant and do not occur in its underground tissues, identification of the structural and regulatory factors operating distinctly in the shoot/leaf of the plant will be a necessity for modulation of bisindole alkaloid biosynthesis. This study aimed at elucidating the differential gene expression in the two main tissues (leaf and root) of the plant, well known for their distinct terpenoid indole alkaloid profiles. The leaf and root transcriptomes of C. roseus were comparatively analysed using two different approaches: (i) indirectly through construction and characterization of separate cDNA libraries; and (ii) directly through a strategically designed suppression subtractive hybridization, using the leaf and root cDNA populations as tester and driver, respectively. A total of 155 ESTs (55 and 45 from the separate leaf and root cDNA libraries, respectively, and 55 from the subtracted leaf-specific cDNA library) were subjected to homology-based classification and submitted to dbEST. The direct approach yielded an EST for sgd (strictosidine {beta}-D-glucosidase) and 16 novel ESTs. Dat (acetyl-CoA: 4-O-deacetylvindoline 4-O-acetyl-transferase) and sgd transcripts could not be detected in the root system of the plant (cv. ‘Dhawal’) at any developmental stage (6 d, 6 weeks, or 6 months). The growth-related decrease in shoot/leaf dat and sgd transcript levels was paralleled by a concomitant decrease in shoot/leaf vindoline content.

Keywords: Catharanthine; dat; ESTs; semi-quantitative RT-PCR; sgd; suppression subtractive hybridization; TIAs; vinblastine; vincristine; vindoline.
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