Establishment Of Tissue Culture System And Genetic Transformation System Of Pinellia Ternata

Pinellia ternata(Thunb.)Breit is a perennial herbaceous plant of Araceae,origin only in China and Japan.P.ternata in the use of Chinese herbal medicine is ranked No.22,as a traditional Chinese herbal medicine,itself and its processed products with a variety of pharmacological effects,such as dampness phlegm,and stomach vomiting,Vomiting,cough and so on.Under natural ecological conditions,P.ternata mainly relies on tubers and buds for asexual reproduction,and the reproductive coefficient is low.In order to meet the demand of medicine,human beings have long been dependent on the collection of wild resources of P.ternata,but the environmental pollution and the reform of farming system have led to the shortage of raw materials in the market.In order to meet the market demand,artificial long-term asexual reproduction and high density cultivation,not only harm the P.ternata plant viruses,endophytic bacteria and pathogenic microorganisms rapid horizontal diffusion,but also cause the virus to spread in the plant a large number of vertical accumulation,Seriously affecting the yield and quality.At the same time,it was found that weed in the field also threatened the yield of P.ternata.Cultivation of P.ternata seedlings can be obtained in large quantities using plant tissue culture.the use of Meristem-tip detoxification technology for P.ternata virus detoxification,can effectively guarantee the quality of P.ternata.The use of genetic engineering technology to get glyphosate Pinellia varieties,reduce the cost of cultivation and management of P.ternata.In this study,P.ternata was chosen as the research material to explore the effects of different culture media on the induction of dedifferentiation,callus proliferation,differentiation and rooting.The results showed that the bestmedium for inducing dedifferentiation of P.ternata was MS+1.0mg/L2,4-D+0.2mg/L 6-BA.The optimum medium for callus proliferation was MS+1.0mg/L 2,4-D+0.2 mg/L 6-BA.The optimum differentiation mediumwas MS+1.0mg/L 6-BA+ 0.2 mg/L NAA.The optimum rooting medium was1/2 MS+0.2 IBA+0.01 mg/L mg/L(6-BA).The best medium for the formation of P.ternata was MS + 1.5 mg / L 6-BA + 0.2 mg / L NAA.In this study,meristem-tip detoxification technology was used to obtain virus-free P.ternata.Virus removal was detected by RT-PCR of cucumber mosaic virus and soybean mosaic virus,Among the five plants detected,one was successfully removed from cucumber mosaic virus,and four were successfully removed from soybean mosaic virus,Only one plant succeeded in removing two viruses.In this study,the genetic transformation of Pinellia ternata was carried out by Agrobacterium tumefaciens,and the exogenous gene vector was pM3301UbiSpCP4.The transformed plant was detected by PCR.and 50 strains in 2 strains of transgenic plants showed positive.