Effect of elicitors on chromene and total isoflavonoid accumulation in p. candollei var. mirifica hairy root culture
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Abstract
Introduction: Pueraria candollei Wall, ex Benth. (White Kwao Krua) is belong to the family Leguminosae and commonly known as a Thai herbal medicine having been used for rejuvenation. In Thailand there are two plants varieties, p. candollei var. candollei (PC) and p. candollei var. mirifica (PM) which have similar botanical characteristic but both vary in chemical component containing. Their tuberous roots contain chemical compounds, known as phytoestrogen, such as isoflavonoids, chromenes, etc. In the present study, we established a hairy root culture of p. candollei var. mirifica and optimal conditions for growth, and investigated the effects of plants elicitors (methyl jasmonate and yeast extract) on enhancing isoflavonoid and chromene accumulation. We also comparative studied on isoflavonoid and chromene production between hairy root and native root of both varieties of p. candollei. Materials and methods: Chromene analysis was performed using HPLC method (mobile phase consisted of 20% acetonitrile containing 1.5% acetic acid at 1.0 mL/min flow rate and 254 nm detection wavelengths). Total isoflavonid analysis was investigated using competitive ELISA (anti-puerarin and anti-daidzin polyclonal antibodies). The data were analyzed using one-way analysis of variance (ANOVA). Results: The results found 200 µM methyl jasmonate and 0.5 mg/L yeast extract significantly increased chromene production after 3 days and 6 days of elicitation, respectively (17.03 and 17.47 µg/g dry wt, 2-fold higher than control, respectively). They also increased total isoflavonoid production after 3 days of elicitation. (16.06 and 14.65 mg/g dry wt, 2-fold higher than control, respectively) For the comparative analysis of each phytoestrogen between hairy root and native root, chromene content in PM hairy root, PC hairy root, PM root and PC root were 5.51, 4.62, 18.86 and 17.10 µg/g dry wt., respectively. Total isoflavonoid content in PM hairy root, PC hairy root, PM root and PC root were 7.72, 9.10, 6.64 and 7.05 mg/g dry wt., respectively. Conclusion: Methyl jasmonate and yeast extract can enhance both total chromene and isoflavonoid accumulation in p. candollei var. mirifica hairy root culture. The comparative analysis of hairy root and native root of both varieties of p. candollei found that both native roots produced chromenes higher than hairy roots about 4-fold. เท contrast total isoflavonoid content in both hairy roots were higher than native roots.
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