The Research On High-frequency Plant Regeneration And Nano-scale Genic Carriers In Transformation Of Jatropha Curcas L

Jatropha curcas(Jatropha curcas L.) is a very important tree species of biodiesel. Jatropha curcas is characterized by rapid growth, yoshimitsu and drought tolerance, which is also has high value in Economic and Medicinal. This article studies on the plant regeneration of Jatropha curcas which is based on taking aseptic seedling stem section of planlet as explant. Meanwhile, establishing suspension cell lines of Jatropha curcas,exploring the effects of nanometer gene vector on genetic transformation of Jatropha curcas,so as to establish a new way of genetic transformation.The basic results as follows:(1)Taking aseptic seedling stem section of planlet as explant, preliminary establishes the plant regeneration of Jatropha curcas.The result shows that:The most suitable hormone combination of adventitious bud induction by stem section of Jatropha curcas is: BA1.0mg/L+TDZ1.5mg/L, the average induction rate of adventitious bud has reached 69.25%.:In the culture of root induction, using auxin NAA, The most suitable hormone combination is:MS+NAA1.0mg/L+Sucrose 30g/L+Agar7.2g/L, and the rooting rate has reached 66.67%, thick roots and well developed. The explants has adventitious bud both in the MS solid medium and liquid medium,and in the solid medium,it has a higher induction rate of adventitious bud and a lower browning rate.(2)In the culture of suspension cell of Jatropha curcas, the effects on the growth of suspension cell in different scheme of loose callus induction and the different culture condition. The result shows that:The most suitable hormone combination of loose callus induction is:MS+2,4-D0.6mg/L+KT0.4mg/L,in which the callus is humid, loose and colorful. The most suitable hormone combination of liquid culture callus is NAA0.2mg/L+2,4-D1.0mg/L+BA0.5mg/L.When using the callus of first generation, callus of subculture two times and callus of subculture four times, the first generation is the best one, which has the highest cell dispersion in suspension cell lines.Adding 500mg/L casein hydrolysate in medium can promote the growth of suspension cell and increase the biomass of suspension cell lines. In the culture of oscillation,the rotation speed should lower than 120rpm,and 110rpm is the most suitable.(3)Studies on the preparation of starch nanometer gene vector and the modification in the surface of the vector, the result shows that:the optimal experimental scheme of the preparation of starch nanometer gene vector(SNGC) is:starch concentration 15%, stirring speed 2000rpm, volume ratio of oil and water 15:1,phosphorus oxychloride 0.05%.Taking modification in the surface of the starch nanometer gene vector (SNGC) by using polylysine (PLL).When the mass ratio between the PLL and the SNGC are different,the electric charge on the vector are different. When SNGC:PLL(M:M)=2:1, the electric charge on the vector are saturated. The modified PLL-SNGC are spherical particle and the size are almost uniform and well dispersive. The PLL-SNGC and Cs-PLL-SNGC vector which modified by polylysine (PLL) and water-soluble quantum dots(CdSe) both can combined with DNA molecular.(4)Studies on nanometer gene vector on callus and suspension cell transformation of Jatropha curcas show that:The callus treated by PLL-SNGC and Cs-PLL-SNGC grow well, the vectors has no cytotoxicity or little. There is no effect on normal cell growth an division. So they can both used as vector in genetic transformation of Jatropha curcas.Eithei of two kind of vector has no significantly effect on callus transformation. But both of them has significantly effect on suspension cell transformation and can be self-degraded in cells, which have good biocompatibility. In the suspension cell transformation of Jatropha curcas,the best ultrasonic time is 10min.