Growth and terpenoid indole alkaloid production in Catharanthus roseus hairy root clones in relation to left- and right-termini-linked Ri T-DNA gene integration
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Hairy root cultures of Catharanthus roseus var. Prabal were established by infecting the leaves with Agrobacterium rhizogenes agropine-type A4 strain. Two hundred and fifty independent root clones were evaluated for growth, morphology, number of integration of Ri T-DNA genes and alkaloid contents. On the basis of growth pattern, type of branching and number of lateral roots we were able to separate the hairy root clones into four categories. However based on the integration of the Ri TL-DNA and TR-DNA genes, there were only three different categories of independent hairy root clones—C1 (rolA&B+/ags+), C2 (rolA&B-/ags+) and C3 (rolA&B+/ags−). Southern hybridization analysis revealed both single and multiple copies of T-DNA integration in the root clones. The accumulation of considerable amounts of the root-specific alkaloids ajmalicine and serpentine was observed in the presence of both the TL-DNA and TR-DNA genes (C1) and the TL-DNA gene (C3) alone. Two rolA&B− but ags+ clones (C2) accumulated much less or only very negligible amounts of ajmalicine. The possible role of the TL-DNA and TR-DNA genes on growth and alkaloid accumulation in these root clones is discussed.
KeywordsHairy root culture Catharanthus roseus Ri T-DNA genes Morphological evaluation Alkaloid production
Terpenoid indole alkaloids
We sincerely thank the Department of Biotechnology, Government of India, for financial support. We gratefully acknowledge Dr. S. Dwivedi, Central Institute of Medicinal and Aromatic Plants, Lucknow, India for kindly providing the seeds of C. roseus var. Prabal.
Junior Research Fellowship to JB by CSSR, Govt. of India, is thankfully acknowledged.
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