【作者】 王为；

【导师】 王万军；

【作者基本信息】 西南交通大学 ， 生物化学与分子生物学， 2008， 硕士

【摘要】 光是植物生长发育最重要的环境因子之一,现已广泛应用于植物组织培养过程中的形态发生、生长控制、光形态建成及其各种反应机理的研究中,大量研究结果表明,各种光质对植物体细胞胚胎发育中的形态发生、器官分化、生化指标等都有一定程度的影响。本实验以铁皮石斛原球茎诱导得到的胚性愈伤组织为原始材料,经过红光NO.1、红光NO.2、绿光、蓝光、白光、黑暗6种光质处理,在形态学水平上观察体胚发育过程中一些形态特征的变化,并对过氧化物酶（POD）、酯酶（EST）、细胞色素氧化酶（COD）的同工酶谱进行了分析。对各种光质下铁皮石斛体胚的形态学研究发现,在42d的培养期中,光质对铁皮石斛体胚生长量有不同程度的促进作用:蓝光＞白光＞红光NO.1＞红光NO.2＞绿光＞黑暗;而不同光质下每个生长时期内每克鲜重的增长率顺序为:白光＞蓝光＞红光NO.1＞绿光＞红光NO.2＞黑暗。对铁皮石斛体胚颜色变化的观察发现,绿光和白光能够使组织块更快地变绿,蓝光次之。本实验对6种光质条件下POD、EST、COD的同工酶谱的酶带数、酶带强度、酶带Rf值进行了分析,并且找到了每种光质都共有的酶带:POD同工酶带Rf_P16、Rf_P20;EST同工酶带Rf_E7、Rf_E11;COD同工酶带Rf_C13、Rf_C15。此外也发现了某种光质特有的酶带14条:红光NO.1下3条;绿光和蓝光下各1条:白光下3条;黑暗下6条。将酶带的特征与形态学特征结合,我们可以通过同工酶的指示初步的了解铁皮石斛体胚在不同光质下的发育过程。更多还原

【Abstract】 Light is one of the most important environmental factors for plant growth and development.As a physical factor,it has been widely used in plant tissue cultures for morphogenesis,growth control,photomorphogenesis and its reaction mechanism.Results of many researches have shown that the light has a great effect on morphogenesis,organ differentiation and biochemical indicators in plant callus growth.In this study,the embryonic calli was used as the original material,which were induced by protocorm-like bodies（PLBs）of Dendrobium candidum Wall ex Lind.6 kinds of light quality:red No.1,red No.2,green,blue, white and dark were applied in the culture.During somatic embryogenesis, morphogenesis and isozymes of peroxidase（POD）,esterase（EST）and cytochrome oxidase（COD）were analyzed.The somatic embryos of Dendrobium candidum were observed in morphology and it was found that different lights could take different extents effects on somatic embryos during 42 days of culture:the Blue＞White＞red No.1＞Red No.2＞Green＞dark;and in each growth period the order of the growth rate of fresh weight per gram was:White＞Blue＞red No.1＞Green＞red No.2＞Dark.The color of somatic embryos in Dendrobium candidum changed from yellow to green.Under white light and green light,the speed of greening was faster than under blue.In this study,through analysis of POD,EST,COD isozyme zymogram,it was found that the light was related with gene expression of the isozymes.And the common isozyme band was found for each kind of the light:POD isozyme with Rf_P1Rf_P2,Rf_P3,Rf_P1and Rf_P2were in each of the eight periods;EST isozyme with Rf_E1,Rf_E2,and both of them were in each of the eight periods;COD isozyme with Rf_C1,Rf_C2,Rf_C3,Rf_C4,Rf_C5,they were in great difference in each period.In addition,we also found 14 specific bands:red No.1 light with three; green light and blue light each with one;white light with three;darkness with six. To combine with the morphological characteristics and the isozyme zymogram characteristics,we could preliminary understand the development process of the somatic embryos in Dendrobium candidum through the indication of isozyme zymogram.更多还原