黄瓜单性花决定基因M的功能分析-作物遗传育种专业毕业论文.docxVIP

上海交通大学硕士学位论文摘要 上海交通大学硕士学位论文 摘要 II II 转化技术将CsACS2 基因置于两种启动子（黄瓜CsACS2 基因原始启 动子，PM；和花椰菜花叶病毒 35S启动子，P35S）的控制之下，利用 植物叶盘法转化烟草，并对转基因T0代部分植株进行Southern，斑点 印记和RT-PCR检测。结果表明PM-CsACS2 转化烟草中CsACS2 基因没 有表达，植株与野生型表型相同。但经外源乙烯处理后，PM-CsACS2 转化烟草的花芽和嫩叶中可检测到CsACS2 基因的诱导表达，且花芽 中的诱导效果更强。另一方面，已检测的P35S-CsACS2 转化烟草叶片 中均有CsACS2 基因的表达。P35S-CsACS2 转化烟草在生长的不同阶 段出现幼苗白化，成株节间距缩短，叶片变小，开花延迟和部分植株 花丝缩短等现象，与植物激素乙烯的作用密切相关。 关键词：黄瓜（Cucumis sativus L.）；单性花决定基因 M；CsACS2； 原核功能分析；遗传转化； 上海交通大学硕士学位论文Ab 上海交通大学硕士学位论文 Abstract III III Function analysis of the unisexual-determination gene M in cucumber plant Abstract Due to its diversity in sex types and to the extensive physiological and genetics studies, cucumber (Cucumis sativus L.) is becoming a model plant for sex-determination research. Recently, the gynoecious-influence gene F and unisexual-determination gene M have been cloned, and showed that they both encode homologies of 1-aminocyclopropane-1-carboxylic acid synthase (ACS), which is related with phytohormone ethylene. However, the function and regulation pattern of the M gene (CsACS2) haven’t been elucidated very well. In this study, alignment analysis between CsACS2-M (encoded by wild M gene) and CsACS2-m (encoded by mutant m gene) reveals that the mutant amino residue (Gly33Cys) may reside in a putative conserved domain. With the E. coli expression system, we study the original enzymology activity of different isoforms of CsACS-M mutant. The results indicated that CsACS2-M has ACS activity, and it can synthesize ACC in E. coli strain JAde6, which can use ACC as its C source. Nevertheless, CsACS2-m has lost its original activity. Meanwhile, the mutagenic isoforms, CsACS2-A (Gly33Ala) and CsACS2-S (Gly33Ser) PAGE PAGE IV also show the disordered activity. Combined with some previous studies, we think there is a new putative conserved domain (S-YF-GW) at the 28~34 position (in CsACS2), which may influence the final enzymology activity by folding an α helix. To study the CsACS2 gene function in plant, w