Effect Of Elicitors On The Growth And Total Saponin Content Of Gynostemma Pentaphyllum Hairy Roots

Gynostemma pentaphyllum(Thunb.)Makino is a perennial herbaceous vine of the genus Gynostemma in Cucurbitaceae.It can be used as a herb and has extensive pharmacological effects.It has the functions of enhancing immunity,delaying senility,improving mental power,lowering blood lipid,blood sugar and regulating nervous system.it is called “Southern ginseng”.Therefore,the market demand is big,but with the medicine and the health care product increasing rapidly,the natural disease and pest control is disadvantageous and so on aspect reason,causes the Gynostemma pentaphyllum medicinal ingredient in short supply.In the field of medicinal plants,the regulation of secondary metabolites synthesis from hairy roots of medicinal plants by elicitors has become a new research hotspot.Studies on the chemical constituents,pharmacological effects and germplasm resources of Gynostemma Pentaphyllum are in the majority,but the effects of elicitors on the growth of hairy roots and the accumulation of total saponins of Gynostemma pentaphyllum have not been reported.The hairy roots of Gynostemma pentaphyllum were induced by inoculation of Agrobacterium rhizogenes C58C1 into Gynostemma pentaphyllum leaves and expanded in 1/2 MS liquid medium,the effects of elicitors MJ,SA and YE on the growth and total saponin accumulation of Gynostemma Pentaphyllum hairy roots were further studied.It laid a foundation for the expansion of Gynostemma Pentaphyllum hairy roots production,the increase of total saponin content and the innovation of Gynostemma Pentaphyllum Germplasm.The main findings are as follows:1.After the co-cultivation is over,the leaves were transferred into the sterile medium to continue dark culture,and after 2 weeks,the hairy roots continued to grow,and then transferred into 1/2 MS liquid culture to expand the culture,by drawing the growth curve of the Gynostemma pentaphyllum hairy roots,roughly determine that the elicitors addition time is the 10 th day and the harvest time is 25 d to reduce the error.2.The hairy roots of Gynostemma pentaphyllum were cultured in 1/2MS liquid medium with dark shaking.The effects of different concentrations of MJ(0,50,100,150,200 μmol/L),SA(0,50,100,150,200 μmol/L),YE(0,50,100,150,200 mg/L)on the growth and total saponin accumulation of hairy roots of Gynostemma pentaphyllum were studied.The best concentration of MJ was 50 μmol/L,the best time of MJ was 10 th day,the best time of action was 7 days.The growth of hairy roots was 2.05 times of that of control group,the content of total saponin was 20.82%,which was 1.58 times higher than that of control group.The best addition concentration of SA was 100 μmol/L,the best addition time was 10 th day,the best action time was 7 days.The growth of hairy roots was 2.11 times as that of control group,the content of total saponin was19.01%,which was 1.45 times higher than that of control group.The best concentration of YE was 150 mg/L,the best addition time was 14 th day,the best time of action was 7 days.The growth of hairy roots was3.08 times as that of control group,the content of total saponin was17.49%,which was 1.34 times higher than that of control group.3.The results of orthogonal test showed that the best combination for the hairy root of Gynostemma pentaphyllum was 40 μmol/L MJ,100μmol/L SA and 150 mg/L YE.The growth amount and the content of total saponin were 2.73 and 1.47 times of the control respectively.The effect of the combination of the inducer on the growth of hairy roots was stronger than that of adding MJ and SA alone,but weaker than that of adding YE alone,and the accumulation effect of total saponins was stronger than that of adding SA and YE alone,weaker than that of adding MJ alone.The results of range analysis and variance analysis were consistent: the effect of three elicitors on the total saponin content of Gynostemma pentaphyllum hairy roots was MJ > SA > YE.