Studies On Genetic Diversity Of Pinellia Ternata Resources From Sichuan Province

Pinellia ternate (Thunb.)Breit. is a common used Chinese medicine which is from rhizome of P. ternata, family Araceae. During the past years, wild resources in P. ternata are close to exhaustion, because of non-limited herborization and environmental transition disruption. Artificial cultivation of P. ternatat is benefit to providing more tuber resources for marketable demands. However, there are many problems including the confusion of various germplasm materials, the decline of tuber quality caused by virus and the lowing self-propagation coefficient. Sichuan Province is the one of main producing regions for P.ternata, and there are abundant wild resource but also has the special variety that own premium quality and yield. This study used 27 materials of P. ternata which mainly came from Sichuan Province.The soluble protein fingerprinting pattern and the chromosome number analysis were used to evaluate the genetic diversity of P. ternata. Soluble protein from leaves and tubers were analyzed by SDS-PAGE. Then we observed the microspore development of different meisis stage and determinated pollen viability and stigma receptivity. And 12 different origin of P. ternata researched growth rhythm, yield and quality. With a view to lay the foundation for P. ternata on identification, protection, further development and utilization. The main results were displayed here:1. P. ternata exhibits highly complex variation in chromosome number with the number from 26 to 117, of which 101 are reported for the first time.The basic number of this species might be considered to be 13. Polyploidy complex (2n=26,78,91,104,117) and minority (2n=80,84,88,94,96,98,101) commonly exist. This phenomenon may be related to geographical distribution and ecological environment.2.We could find the corresponding relationship between anthotaxy morpholgics and different meiosis stage and observed laggard chromosomes,chromosome bright and micronuclei at every phase of meisis. The pollen viability and stigma receptivity are 1.9% and 85.9%. Efficient asexual reproduction may be the most important reason to the high polyploidy complex and minority level in P. ternata.3. Ice water extraction method, Tris-HCl buffer solution method, Na3P03 solution extraction and PVP solution containing were used to extract soluble protein in P. ternata, Then the key steps affecting the SDS-PAGE were discussed for establishing a set of applicable soluble protein extraction and electrophoresis method. That is directly electrophoresis after extraction of the sample by ice water extraction method, discontinuous separation system,10% separating gel,4% stacking gel, Coomassie brilliant blue staining, distilled water washed and destaining. The ice water extraction method was simple and rapid, SDS-PAGE showed the clear bands and high resolution.4. The SDS-PAGE results suggested that there was rich genetic polymorphism among the tested resources of P. ternata. The electrophoresis pattern of soluble protein in leaves was different from soluble protein in tubers. There were 19 bands in leaves,3 bands were same in all materials. Every material may have 11-18 bands. The average bands were 14.5. There were 12 bands in tubers,3 bands were same in all materials. Every material may have 4-12 bands. The average bands were 9.4. The clustering was not completely associated with their geographic origin but chromosome number. The electrophoresis result of soluble protein in leaves can offer a better identification method between Pinellia ternata (Thunb.) Breit and Pinellia pedatisecta Scotte.5. Twelve populations of P. ternata. were collected and cultivated in expetimental farm of Sichuan Agricultural University with same cultural conditions, whose differences in growth rhythm, yield and quality have been observed and compared. There are obvious differences in sprouting, bolting, bulbils produced. There are two times of "tumber" from late May to mid-July and mid-August to October. The fast-growing period for bolting, bulbils produced is May. The weight gain and growth rate were large difference. Through the determination of soluble protein,β-sitosterol, alkaloids by uv spectrophotometry, we found large difference in different area. By comparision from Banqiao,Quxian Sichuan S07-28 have high levels of three material chemical composition relatively, may be an excellent materials in Sichuan Province.