Study On Tissue Culture And Rapid Propagation Optimization Of Pinellia Ternate And Its Quality Analysis

Pinelliae tuber,the tuber of Pinellia ternata (Thunb.) breit., is a commonly used traditionally Chinese materia medica with long history and notable curative effect. In recent years, along with the market demand continuously increasing and the wild resources continuously reducing of Pinelliae tuber, the resources deposit and production of Pinelliae tuber in China descended greatly, very difficult to satisfy the need of medicial use and export currently. Techniques of artificial cultivation for Pinellia ternata, existing several problems as following:germplasm sources mixing, evaluation criterion backward, serious occurrence of pest and disease cultivation technique developed slowly, Species degradation。Tissue culture technology of establishing an asexual rapid propagation of the system to improve its reproduction, and to prevent a variety of problems. the organization of training and study on rapid propagation of the report is induction differentiation, but Little research on Culture Technique, especially quality control research is a blank.The work in this thesis is part of the National 11th Five-Year scientific and technological support projects, in the basis of prophase preliminary assessment on Pinelliae tuber germplasm resource。This research take quality and purity and grow strong, the disease of the summer as explants. has carried on one Step by Tissue Culture and Culture Technique, and Growth and Development, and the quality of tissue culture Pinellia ternata。This topic is not only ensure the quality of Pinellia ternata, but also give an impetus the industrialization of the production of Pinellia ternate.1. Study on optimization for Pinellia ternata in one Step by tissue cultureThis research take the pinellia tuber, the bead bud, the leaf blade, the petiole as explants, designing different disinfection method, through explant spectrums of the different growth regulating substances and additives (compound additives, activated carbon), different sugar concentration, light cultivation and dark to step into seedlings cultivation impact study. Sure different explant appropriate disinfects processing methods; Meanwhile in inducing differentiation into seedlings, was set up for the first time in four explant in the same medium (MS+6-BA B2+A1NAA+H1olumes) step into seedlings rapid propagation system, four explant sprout rate are in the into above 85%, especially in tuber and bead buds as the explant into sprout rate of over 95%, another study also found that by petiole explant medium for when joining C1～C3 compound additives, obviously reduced growth cycle 25 to 30 days. This system can be simplified medium and vaccination steps prepared, easy operation and reduce the cost.In vitro rule induction, found little tuber sprouting from the body of light can promote the formation, light nduced generated little tubers, green, growth-well; The same lighting conditions and hormone concentration conditions, when sugar concentration is Z1～Z3. single tubers fresh weight and average is not big, but few differences sprout> Z3of sucrose, each sprout significantly lower average number of tuber. Further combining sucrose, activated carbon three factors three levels orthogonal experimental analysis, and the results indicate that Z3sucrose+B16-BA+H2 activated carbon, inducing rules from the body small tubers effect is better.2. Study on culture technique of tissue culture Pinellia ternateThis research is on cultivation of sprouting, cultivation of tissue matrix methods of management, transplanting period and harvest time and other multiple research investigation, preliminarily determined the best cultivation plan of tissue sprouting. Seedling height in 3～8cm,0.20 cm< diameter< 0.16 cm tuber tissue seedlings transplant.problems the survival rate are more than 90%; Through the five kinds of cultivation of the matrix of the investigation, were retention of osteoporosis in fertilizer capacity good cultivation 3 and cultivation 4 tissue transplant seedlings transplant. and the best survival rate was 90%; Cultivation mode and transplanting time aspects with field cultivation and seedling dish+field is better, the survival rate cultivation with more than 90%, and the further growth of tissue seedling yield and quality of comprehensive evaluation, sowing 2 is the best; Through the study on harvest time, it is determined primarily harvest timeⅢ～harvest timeⅣ, as the most suitable tissue quantitative sprouting。3. Study on growth and development of tissue culturing seedlingsFrom the plant height, leaf type, bead buds, fast-growing, emergence rate, pour seedlings status on research of tissue growth situation sprouting dynamic variation, and compared with the traditional cultivation sprouting observation, preliminaris master tissue in different seedling sprouting growth phase of the growth situation. The results found that the first generation of tissue, plant is small, more rare bead and flowering in the whole county; the second generation of tissue in the whole county, the foliage is compound leaves, stable. And their parents were compared, second generation of tissue in bead sprouting shoots is not big, but bead buds differences of its parents. Meanwhile subject tracking different first tissue growth cycle of stem character and sprouting microstructure characteristics. Found that as the exten- sion of tissue growth, sprouting in the original plant tissue sprouting tuber morphologic, and microscopic characters (leaf surface, tuber) compared with traditional cultivation Pinellia ternate without obvious difference. It gave a basis seedling field management in future.4. Study on quality of tissue culture Pinellia ternateScreening tissue culture tuber with above the average diameter of 1.0 cm, this paper studies morphologi-cal and histological, extracting substances, total alkaloids, total acid content with traditional cultivation, and commercial samples quality comparison. The results found tissue culture is similar in morphological and histological. extracting substances, microscopic characters, moisture, ash and moisture content traditi--onal cultivation sprouting. In chem ical composition, the study found, among the total acid (similar to extract tissue with butanedioicacid plan) sprouting slightly lower than traditional cultivation sprouting and commodit ies, but spro uting sprouting total alkaloid content of tissue significant higher, respectively, between traditional cultivation of sprouting, commodity sprouting 1.2 times and 1.6 times. Initially established a HPLC fingerprint, tissue sprouting sprouting tissue with traditional cultivation were mode maps of sprouting similarity is higher, correlation coefficient in 0.90 above.The results of the study can realize the scale, the industry manufacture; Also ensure the stable quality of tissue medicinal sprouting, effective and controllable.It provided the basic data for Clincal Application of tissue culture tuber。...