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

Ashutosh K Shukla, Ajit K Shasany, Madan M Gupta, Suman P S Khanuja
Journal of Experimental Botany 2006, 57 (14): 3921-32
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.

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