Studies On Male Gametophytes Observation With Microscope Andculture In Vitro Of Endangered Plant-Kolkwitzia Amabilils Graebn

kolkwitzia amabilils graebn. is the third class protected plant of china .Now, it is rare and endangered, but only little parts of ovules develop to seeds and the quality of seeds are worse. In this paper, the whole process of it microsporogenesis and male gametophytes development was observed with microscope to sure weather stamen development is normal. At the same time, in order to provide techniques on biotechnology conservation and the foundation of its resources gene pool in cell engineering, its techniques on culture in vitro was studied.The results were as follow: in the process of its male organ development, these are many abnormalities, such as: premature tapetal degradation,giant tapetal cells ,the great vocalization of the tapetal cells and inadaptable function of callose enzyme, which lead abnormal and premature degradation microspores and male gametophytes. The highest rate of pollen germination is 15.4%. Rare and sterile pollen because of these is the weak links in the sexual reproduction of kolkwitzia amabilils graebn.The results of its culture in vitro were as follow: young leaves were the best explants in callus induction, the suitable culture medium was MS+BA2.0mg/L+NAA0.1mg/L.the results of organ differentiation were that BA was better than ZT in the induction of shoots, but ZT was better than BA in the induction of root and callus quantity. The results of analysis of variance for orthogonal experiment have shown that the changes of BA concentration have significant effects on the numbers of shoots, the weight of callus . The changes of NAA concentration have the most significant effects on the numbers of shoots and significant effects on numbers of roots. The suitable culture medium of bud induction was BA4.0 mg/L+ NAA0.05mg/L, which had the suitable callus quantity and the rate of shoots differentiation was 21.2%.The results of stem culture were as follow: B5 medium had faster bud germination and low browning. When subculture medium was B5+BA2mg/L+NAA0.2mg/L, the numbers of propagation were 3.2 times of explants, what's more, the seedlings were robust.