Research On The Technique Of Tissue Culture In Different Strains Of Pinellia Ternata In JingZhou

Pinellia ternate(Thunb.)Breit is a important kind of Chinese herbal medicine.In this paper the tissue culture of three steains of P.ternate,which were XY,TY and KY,were mainly studies,the variety of the three strains P.ternate were studied in the culture process in the same time.In this thesis,several experiments about tissue culture were conducted including different explants,different hormone concentration and ingredient of culture medium.Because its low tradition multiplication and quality degradation,the advanced tissue culture method was researched to multiple in order to gain more plant in the short time to fulfill the market demand.In addition,UV and HPLC were employed to analyze the extract of guanosine and total alkaloid from tissue culture tubers.The results as follows.(1)Different explant had different competence of.The best explant is tender pedicel;(2)This experiment inspected the effect of three different phytohormones on callus induction with the orthogonal design.The result showed that the respective best initiation medium of TY,XY,KY were NAA 0.2mg/L+2,4-D 1.0mg/L+KT 0.5mg/L(the rate of initiation was 96.0%),NAA0.5mg/L+2,4-D1.0mg/L+KT0.2mg/L (the rate of initiation was 100.0%) and NAA 0.2mg/L+2,4-D 0.5mg/L+KT 1.0mg/L(the rate of initiation was 90.0%).In addition 2,4-D was essential to induction of callus.(3)MS medium supplemented with different concentration NAA,2,4-D and KT could induce multiple shoots in three strains.MS medium supplemented with 0.5mg/L2,4-D and 0.5mg/LKT was fit for TY and XY,while 0.5mg/LNAA and 0.5mg/LKT was fit for KY.XY and TY showed much more properties than KY.The callus which was off-white or light green and pultaceous have good proliferation of adventitious buds.(4)Stage of rooting:The rooting effect which induced by 1/2MS+NAN0.1mg/L was fit for three strains P.ternate.But the TY P.ternate could get the best rooting effect on medium of MS+NAN0.1mg/L.So there was some different between different strains P.ternate.1/2MS+NAA0.1mg/L+0.8%AC and 1/2MS+NAA0.5mg/L +0.8%AC were fit for the induction of tuber.AC would play a important role in inducting tuber from callus.(5)The survival rate of transplantation were 92%in the medium with 40%soil in garden,40%vermiculite and 20%perlite.If the plantlet have little tuber,it can improve the rate of survive.(6) The content of total alkaloid and guanosine which were extract from three strains tissue culture tubers were all lower than field cultivation.In total alkaloid content,the TY was the best,it reached 0.0125%;in guanosine content,the XY was the best,it reached 0.0122%.