Establishment Of Jatropha Curcas L.Hairy Root Culture System And The Effect Of Elicitors On Content Of Active Ingredient

The seeds of Jatropha curcas L. contain munch more oil, so J. curcas L. was used to produce biological diesel oil. Also, it is a kind of medicinal plant, with antitumor and antivirus activities and has been used in industry and medicine broadly.In this study, J. curcas L. hairy root induction and culture system were set up with Agrobacterium rhizogenes, and the hairy root induction conditions were optimized, and also he best induction conditions were established. Analysis the effect of several elecitors on the growth of J.curcas hairy root and the contention of Jatropholone B.The major conclusions of this experiment are as follow:1. Establishment of J.curcas L.hairy root induction and culture system:Single factor experiments for several influential factors were carried out to find condition for inducement of J.curcas hairy roots.The influential factors contain explant type, concentration of bacteria and infection time, pre-culture time, Agrobacterium type, different antibiotic medium on the hairy induction. In the article, we found that the explant was used the hypocotyl of J. curcas, pre-culture time was2days, the most appropriate OD600of A. rhizogenes suspension was about0.5, the induced time was8minutes, and the most appropriate concentration of AS was50μmol/L, the best co-culture time was2to3days.In conclusion, A. rhizogenes R1000reached the highest induction rate,75.862%.2.J. curcas hairy root liquid culture conditions optimization:Different media had different effects on J.curcas hairy root growth. It was obtained that1/2MS culture medium was the most favorable one growth of hairy roots of J. curcas, among MS,1/2MS,B5,1/2B5, White and1/2White basic culture medium, in which the average growth fold of fresh weight was1.59.3. According to experiments, three biotic elicitors (yeast extract, the bacteria suspension of KLBMP1111and KLBMP4804) and one abiotic elicitor (methyl jasmonate) all had effect on the hairy roots growth and the content of Jatropholone B. Research found that different concentrations and elicitors showed different level effects on the hairy roots growth and the content of Jatropholone B. Three biotic elicitors could promote the growth percentage of hairy roots, and the abiotic elicitor--ethyl jasmonate inhibited the hairy root growth.The induced effect was the most obvious, when the elicitor KLBMP1111concentration was100μg/ml, the result was the control group1.56fold.4. Analysis of the content of and the diterpenoids synthesis key enzyme gene—PS had correlation with synthesis of Jatropholone B, when the elicitor KLBMP1111was used to treat the hairy roots, FPS gene expression was significantly higher than control. This result indicated that elicitor promoted Jatropholone B accumulation.