茶树类黄酮基因功能验证相关体系的筛选-生理学专业论文.docxVIP

PAGE PAGE IV Abstract The enzymes involved in the biosynthetic pathways of tea polyphenols, such as catechin, flavonoid and proanthocyanidin, have been basically clear, but the functions of the structural gene and the regulator gene have not been proved effectively. The main difficulty is to establish a stable transgenic system of tea plant, and the cycle of current regeneration system is long with low efficiency. Thus, it is still a very urgent task that how to improve the efficiency of regeneration system and achieve an instantaneous genetic transformation system. In addition, transgenic system of model plant could be used to verify the functions of related genes in tea. The main results obtained were as follows: A regeneration system of tea plant In this paper, the regeneration system of callus in tea including five steps (callus induction, embryogenic callus proliferation, adventitious bud induction, adventitious proliferation and rooting in vitro) was established. It consists of two proliferation processes, callus proliferation and adventitious bud proliferation. Main steps are as follows: seeds of tea plant (Camellia sinensis) in early September were picked as explants, then, they were disinfected and cultured in vitro on B5 medium supplemented with 0.5mg/L 2,4-D and 0.1mg/L KT. The callus was induced in two weeks and subcultured for four years steadily. The callus was subcultured on MS medium supplemented with 0.1mg/L IBA and 2.0mg/L 6-BA. The shoot induction rate was 65.71% after being cultured for four weeks. The shoots were cultured on MS medium supplemented with 0.1mg/L IBA and 0.01mg/L TDZ. The large number of cluster buds was induced in four weeks, which were then cultivated to strong plantlets on MS medium supplemented with 0.2mg/L IBA. The seedlings with height above 5cm were selected and transplanted in soil, and the survival rate of it was above 80.8%. The whole culture period was about twenty-five weeks. The study of transient expression system