Fast Propagation And Polyploidy Induction Of Gynostemma Pentaphyllum (Thunb.) Makino

The plant Gynostemma pentaphyllum (Thunb.) Makino is a kind of perennial herb liane, which belongs to Cucurbitaceae. All of it can be made into medicine. It can clear heat and expell miasma, dispell phlegm, arrest cough, strengthen health, activate cell, tranquilize, assuage pain, reduce blood fat, enhance immunizing ability etc. Our country is the distribution and differentiation center of the Gynostemma, and has abundant resource. Now most of the cultivated varieties are wild, new varieties are few found in the breeding field.The Gynostemma wild resource is limited to satisfy our need. The medicine raw material is mainly from crop. And the crop is asexual reproduct. For a long time, no variety has been selected, and no variety is purified or rejuvenated, so its property is degenerating, its yield and quality are declining, and its chemistry components are unsteady. To improve Gynostemma yield quality and resistance, firstly we have established fast propagation system through tissue culture, and then induced autopolyploid with colchicines.To establish fast propagation system, through complete design and blank control, with MS culture medium, 6-BA, NAA, IBA, we got the results. In the condition of temperature 25 °C, intensity of illumination 1300-15001x and illumination time 14 h/d, the best inducement medium is MS +NAA 0.2 mg/L+ 6-BA 2.0 mg/L, the best medium for propagation is MS+6-BA1.0mg/L+IAA0.5mg/L, and the best medium for root is MS+IBA 2.0 mg/L+ NAA 1.0 mg/L. After that the Gynostemma can transplant and survive in field through exercise.To induce autopolyploid with colchicines, we took two ways marination and filter paper with colchicines. In this two ways, we selected three colchicines concentration, three processing time to process the same target-shoots. We got the result that the best way can get 72.73% autopolyploid and 87.50% homozygote by marination with 0.25% colchicines for two days. The proceeding shoots were fast propagated, and then were karyotype analyzed. We took the pressed disc method to get the chromosomes number. Because the karyotype analysis is a part of our experiment, we selected three preproceeding time, three immobilization time and three decomposed time to finish, and then sheet formate, colorate, observe, took pictures, and then karyotype analyzed. The result of karyotype analysis is as follow: the diploid karyotype formulae 2n=2x=22=1M+3m+7sm, the tetraploid karyotype formulae 2n=4x=44.