【作者】 李凤霞；

【导师】 杨鹏； 王玉国；

【作者基本信息】 山西农业大学 ， 植物学， 2003， 硕士

【摘要】 猬实为我国国家三级保护植物，珍稀濒危。其由胚珠转化成种子困难，种子质量低劣。本论文通过对猬实小孢子发生和雄配子体发育全过程进行细胞观察，探寻猬实的雄性器官的发育是否是猬实有性生殖的薄弱环节，并对猬实的离体培养进行了初步的研究，为猬实生物技术保存、建立猬实种质资源基因库提供细胞工程方面的途径和技术。实验结果表明： 1．猬实的雄性生殖器官在发育过程中，出现绒毡层细胞的液泡化、肥大、提前降解及不适时释放胼胝质酶等异常现象，造成猬实花粉发育过程中出现畸形、解体。经猬实花粉萌发实验发现猬实花粉萌发率最高只有15.4％。由于上述原因造成猬实花粉稀少、败育，成为限制猬实有性生殖的一个原因。 2．猬实愈伤组织诱导以幼嫩叶片为外植体较好，培养MS+BA2.0mg／L+NAA0.1mg／L为好。进一步诱导器官分化，结果发现BA对芽的诱导效应强于ZT，而ZT在对根的诱导、愈伤组织生长量上优于BA。方差分析结果表明，在BA、ZT、NAA各七水平的49种组合中，BA浓度变化对芽的诱导、愈伤组织的生长量有显著的影响，NAA浓度变化对芽的诱导有极显著的影响，对根的诱导有显著影响。猬实愈伤组织不定芽诱导的最优激素组合为BA4.0mg／L+NAA0.05mg／L，其愈伤组织生长量适中，芽的分化率达21.2％。 猬实茎段培养以B₅为基本培养基时，腋芽萌发速度快、褐变率低。适宜猬实腋芽萌发的培养基为B₅+BA1.0mg／L。猬实继代培养基B₅+BA2mg／L+NAA0.2mg／L最为有利于猬实芽的增殖，其芽的增殖数为接种数的3.2倍，小苗生长健壮。更多还原

【Abstract】 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.更多还原