| Pinus massoniana is a important native species of fast-growing conifers which grows quickly and has higher comprehensive utilization value.To investigate the culture in vitro technology and build-up fleetness probing that seeds of trees breed system can establish the fine theory and technology basis for the job ameliorating the engineering study studying and carrying out a gene to the inheritance being seeds of trees's turn.Two types of initial explants from Pinus massoniana,including immature embryos and stems of seedlings,were taken to study its organogenesis and somaticembryogenesis in this thesis.Factors have been tested for their effects on organogenesis and somatic embryogenesis. The main findings are as follows:First,The difference studying originally had adopt the masson pine immature embryo and the young stems to explant, in the process of sterilisatio,we had carried out different treatment on them and had analysed their respective sterilisatio effect comparatively,The results indicated:In the process of sterilisatio of the two explants, the effect of alcohol was all comparatively notable,If the time of sterilisatio was too short,sterilisatio was not thorough and the expants easy to be communicated; Sterilisatio long time, although sterilisatio effect was good. However, the explants were comparatively young and were lossed easily,even led the explants to death.In the process of the sterilisatio to immature embryo of Pinus massoniana,the best way to disinfect:75% alcohol surface sterilisatioed 30 seconds; afterwards,to be soaked in 0.1% mercuric chloride 8 mins and washed 5-6 times with sterile water finally;To soak the explants for 20 second with75% alcohol and soak them for 8 mins with 0.1%mercuric chloride and washed 5-6 times with sterile water was the best sterilisatio method to young seedling of Pinus massoniana,the pollution and mortality rates being a minimum without exception,was 3.71% only.Second,During the direct organogenesis of Pinus massoniana culturing, to add 0.15g/LVc in medium and connect once every two days was the effective method of preventing brown and can reduce that the time of explants settled in medium and the probability of being browned. At the same time,by the lots of analysis indicated:During the period of initiation culture,the germination rate in WPM was higher than in GD and DCR.The proper concentration of BA,KT and NAA was 1mg/L,0.5mg/L and 0.15mg/L,if the concentration of NAA was too high, the degree of callus will be large. Specifically speaking,the proper combination of medium to induce adventitious buds was WPM+1mg/LBA+0.5mg/L KT+0.15mg/L NAA+0.15g/L Vc;During the multiplication and elongation of adventitious buds,the most feasible combination of medium was WPM+0.15mg/L NAA+0.15g/L Vc+0.5g/L Ac;During the rootage culture, the best effect combination of medium was WPM+2mg/L IBA+0.15mg/L NAA+0.5g/L Ac,and the proper concentration of sugar was 20g/L.Third,During the callus culturing, the ratio of auxin and cytokinin was playing an important regulate and control role to explants dedifferenting to callus. So it is therefore necessary to constantly adjust the rate of hormones in favor of dedifferenting to callus of Pinus massoniana.This study reached the best medium combination of inducing callus of Pinus massoniana was GD+2.5mg/LNAA. Simultaneous, a great of quantity testing and studying analysis found that NAA was playing an regulate and control role,if short of NAA,the callus will be difficult to form.Finally, it can be seen that NAA is the callus induction essential factor.Forth,By the lots of analysis, we found that the immature embryo of Pinus massoniana had larger and easier regeneration ability than young stems Pinus massoniana.
|
PDF Full Text Request