Induction and in vitro culture of Wedelia trilobata hairy roots / 生物工程学报

ABSTRACT

To study the possibilities for improvement of the ornamental character and production of secondary metabolites by using Wedelia trilobata hairy roots, we investigated the induction of W. trilobata L. hairy roots and its consumption changes of carbon resource, nitrogen resource, phosphate and calcium in the medium during liquid culture. The results showed that hairy roots could be incited from the cut edges of leaf explants 7 days after inoculation with Agrobacterium rhizogenes ATCC15834 and could have an autonomous growth on the medium without phytohormones. The PCR amplification showed that rol genes of Ri plasmid of A. rhizogenes was integrated and expressed into the genome of transformed hairy roots. The hairy root line grew very slowly in 0-7 days, very fast from 7 to 21 days. During the liquid culture of hairy roots, sucrose, NO3(-)-N, PO4(3-) and Ca2+ in the medium could be gradually absorbed and utilized with time. The content of NO3(-)-N in the medium was 5.8% of the initial amount at day 7, while sucrose content was about 50% of the initial amount. At day 35, the NO3(-)-N and sucrose content in the medium was 1.82% and 3.39% of the initial amount, respectively. In combination with Ca2+ consumption, PO4(3-) of the medium was rapidly absorbed and utilized. At day 7, the content of PO4(3-) in the spent medium was only 1.76% of the initial amount; but even at day 35, the content of Ca2+ in the spent medium was still 61.3% of the initial amount. The results presented here had provided the possibilities on improvement the ornamental character and how to prepare optimum medium for large scale cultivation and production of secondary metabolites from W. trilobata L. hairy roots.