[Induction and in vitro culture of hairy roots of Solanum nigrum L. var. pauciflorum Liou and its solasodine production].

Abstract

By using genetic transformation of Agrobacterium rhizogenes and liquid culture, induction and culture conditions for Solanum nigrum L. var. pauciflorum Liou hairy roots and its solasodine production and consumption changes of N resource and calcium in the medium during liquid culture were investigated. The results showed that hairy roots could be initiated from the cut edges of leaf explants 5 days after inoculation with the strain of A. rhizogenes ATCC15834. The percentage of rooted leaf explants 25 days after infection was more than 90%. Hairy roots could grow rapidly on solid or liquid growth regulator-free MS medium. The PCR amplification of rol genes and virC gene showed that rol genes of Ri plasmid of A. rhizogenes were integrated in the genome of transformed hairy roots of S. nigrum L var pauciflorum. The hairy roots could produce medicinal secondary metabolites solasodine and the amount of solasodine in the hairy root cultures reached a level of 582.05 microg/g dry weight and was 1.31 times as much as those in the untransformed roots. The hairy roots grew very slowly in 0-5 days in the liquid medium, then, very fast from 5 to 15 days. During liquid culture, NO3(-) and NH4(+) in the medium were gradually absorbed and utilized by hairy roots. NH4(+) -N of the medium was used up at day 15 of the culture, while NO3(-) in the medium was not used up, the content of which was 44.7% of the initial amount. Ca2+ of the medium was gradually absorbed and utilized during liquid culture and it was not used up at day 25, the content of which was still 43.5% of the initial amount. The results presented here had provided the possibilities on how to prepare optimum medium for large scale cultivation and production of solasodine from S. nigrum L. var. pauciflorum hairy roots.