Overexpression of an apoplastic peroxidase gene CrPrx in transgenic hairy root lines of Catharanthus roseus
Peroxidases are a family of isoenzymes found in all higher plants and are known to be involved in a broad range of physiological processes. However, very little information is available concerning their role in Catharanthus roseus. The present study describes the impact of both overexpression and suppression of a peroxidase gene, CrPrx in C. roseus transgenic hairy root lines. Real-time PCR analysis in 35S-CrPrx and CrPrx-RNAi transgenic lines indicated differential transcript profile for peroxidases as well as for genes and regulators involved in MIA (monoterpenoid indole alkaloid) pathway of C. roseus. Comparative analysis revealed that MIA pathway genes showing elevated levels of expression in 35S-CrPrx transgenic lines showed a significant reduction in their transcript level in CrPrx-RNAi transgenic lines. Metabolite analysis detected higher levels of ajmalicine and serpentine accumulation in overexpressed lines. It was observed that all overexpressed transgenic lines produced more amount of H2O2. These results indicate a role of CrPrx gene in the regulation of MIA pathway genes and regulators, thus affecting the production of specific alkaloids.
KeywordsAgrobacterium rhizogenes Catharanthus roseus Monoterpenoid indole alkaloids Peroxidases Transgenic hairy roots
MJ and SK thank Council of Scientific and Industrial Research (CSIR), India for the award of senior research fellowships. The authors are grateful to Dr. Peter Waterhouse of CSIRO Plant Industry, Canberra, Australia, for kindly providing vectors pKANNIBAL and pART27. The work is supported by financial assistance from the core grant of National Institute of Plant Genome Research, New Delhi, India.
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