Studies On Tissue Culture And Virus Elimination For Lycium Barbarum L.

Lycium barbarum L. belonged to solanaceae, was perennial deciduous shrub. In recent years, with the development of Chinese wolfberry medicinal function, the nutrition and health role, In northwest area Chinese wolfberry had became one of the main economic species. In many places,Chinese wolfberry had been largly cultivated, and the demand of seedlings also increased,in the process of cultivation also had a lot of problems, especially the increase of pests had great effecton to wolfberry grow and fruit quality and output etc. In the production of Chinese wolfberry seedling mainly reproduced by cuttings, long-term asexual reproduction is accumulated inside of body, make the virus caused virus disease,had seriously influenced of fruit production and quality as well as the growth of branches. This experiment chosed germination branches of Chinese wolfberry as explant, systematically studied for stem cultivated in Vitro invovled of various factors, including the explant clumps of disinfection, induction of cluster buds,cluster buds proliferation culture medium choosing,the best root culture medium and hormone concentration,ratio of type selection. At the same time,in order to obtain non-toxic plant,also studied different handle methods of lycium virus removal effection.The specific study was as follows:1.The best explant disinfection method was:used 0.1%HgCl2 for seven minutes had the best sterilization effection. While processing time was too long or short were all adverse to explant disinfection, short time the sterilization was not thoroughly; But for a long time, HgCl2 may had poison effection to explant, and also could make glomerating stage become slow growth stage.2.The best culture medium for inducing cluster buds was 6-BA0.5 mg/L+NAAO.5 mg/L,and the germination rate of lateral bud was up to90%.And germination, growth rate was also faster,cluster buds number were more, the bud seedling growth were more robust.3.For cluster buds proliferation,the best culture medium was put 6-BA1.5 mg/L +NAA 0.6 mg/L+activated carbon 0.5 g/L in MS,the growth coefficient could reach 4.68. Proliferation culture medium was not only had high differentiation rate,but also the focus formation of adventitious buds growed well,and the buds were well and strong,and aslo had no invalid buds form. 4.Put different concentrations of GA3 in the culture medium with 6-BA0.5 mg/L +IBA0.6 mg/L, With different concentrations change of GA3 the stems and buds'elongation and proliferation also had changed,when the concentration of GA3 was 2.0mg/L,the stems and buds'elongation were well,buds growed higher and buds proliferation number also increased, and when reached a certain concentration, the stems and buds were no longer elongation,and with the concentration continued to increase,the buds number of proliferation were decreased and plants growed slowly,and the leaves small and not flatten.5.For rooting,1/2MS+IBA 0.5 mg/L had a better effect.The rate of rooting was 95.0%and the number of growth was 11.6,and the root was brawny,callus was less and the growth rate was faster.6.Long-term higher temperature meristem-tip training and combined with continuous subculture virus-free treatment for the stem tip in different lengths, which had a good effection to tobacco Mosaic removal., when taking stem length< 0.3 mm,young stem tip achieved the highest rate 92.0%of virus-free,But the survival rate was lowest,and was only 20.0%.7.Through the different processing methods for removal of virus treatment comparision, high temperature combined with subculture virus-free treatment of virus-free removal, was superior to other dispell virus treatment method. When taking stem length for< 0.3,0.3-.0.5,0.5-0.8and0.8-1.0mm,the virus-free rate was 92.0%,86.7%,76.5%,83.3%respectively, because at high temperatures, the virus were easily passivation, and its fecundity weakened or lost fertility,so as to achieve the objective of virus removal. Therefore,high temperature combined with meristem-tip treatment method were more more successful removal than other treatment to remove medlar tobacco Mosaic virus.