Establishment of Stevia rebaudiana Bertoni Hairy Root Culture System and Induction of Chlorogenic Acids
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Abstract:
Stevia rebaudiana hairy roots were induced using Agrobacterium rhizogenes A4. A culture system to produce chlorogenic acids in hairy roots was then established, and the effects of methyl jasmonate and salicylic acid on the accumulation of chlorogenic acids in hairy roots were studied. The results showed that Stevia rebaudiana hairy roots were successfully reproduced after 14 days’ culture of Stevia rebaudiana leaf explants infected with Agrobacterium rhizogenes A4. Polymerase chain reaction (PCR) results indicated that genes rolB and rolC from Agrobacterium rhizogenes were integrated into the genome of Stevia rebaudiana. Hairy roots were cultured in Murashige and Skoog (MS) liquid medium for 14 d, and various concentrations (50, 100, and 200 μmol/L) of methyl jasmonate and salicylic acid were added to the cultures, respectively. After 1 d, 3d, and 6 d treatments, both plant elicitors inhibited the growth of hairy roots. However, methyl jasmonate promoted the accumulation of chlorogenic acids in hairy roots, and salicylic acid inhibited the accumulation of chlorogenic acids. Thus, Stevia rebaudiana hairy root was induced by Agrobacterium rhizogenes A4 and could be used to produce chlorogenic acids; this could be promoted by methyl jasmonate elicitor.
