棉花抗逆相关基因的克隆与功能研究及低温胁迫下棉花蛋白质差异表达分析-植物学专业毕业论文.docx

3．采用TcA一丙酮沉淀法，改良优化了棉花蛋白质组研究中的双向电泳技术。通过对根和子叶 3．采用TcA一丙酮沉淀法，改良优化了棉花蛋白质组研究中的双向电泳技术。通过对根和子叶 全细胞蛋白的提取、蛋白的溶解、胶条的选择及电泳等环节的优化，得到了重复性很高、 分辨率很好的蛋白质图谱。进一步对5天苗龄的棉花进行4℃低温处理12小时后，提取子 叶的蛋白质，利用双向电泳技术分离全细胞蛋白，研究棉花在低温处理下子叶中蛋白表达 谱的变化。并用PDQuest软件分析比较棉花子叶在低温处理下的蛋白表达谱，得到了50个 有显著差异的蛋白点。其中，低温处理上调表达的蛋白点有28个。减弱表达的蛋白点有22 个，推测这些差异蛋白点可能与低温胁迫相关，为研究棉花蛋白质组学奠定了基础。 关键词：棉花；水孔蛋白：Dil9蛋白；逆境胁迫；植物基因工程；转基因；蛋白质组学 双向电泳； AbstractDrought,high Abstract Drought,high salt and low temperature are major environmental factors that influence the growth and output of plant．These faotom damago plant irreversibly and load to pinto death．It is 扯j玎】portam field of modern agriculture to study the mechanisml of stress-tolerance，obtain related genes in plant and cultivate tress-toleran[t crops through molecular biology,molecular genetics methods and∞011．In this study,3捌_潮-related genes wore isolated fiom cotton．Then the expression and the preliminary function of the genes we∞analyze吐Meanwhile the different expression proteins ofcottonu出cold stresswore studiedby2-DE．The resultsobtained8re asfollows： 1．One aquaperin gene was isolated fiom more 30 putative stress-related genes of cotton and was designated GhTIPI．It contains 251amino aci凼and the／dip family 8ignatllre sequence HVNPAVTFG At the amino acid level,the劬n7,1 shares high identity with other plant TIPs． For example,90％identity to bobTIP26-1(Brassica oleracea var．botry出册and 88％idenC畸 toAtT-TlParabidopsisthaliana—∽．TIPbelongstotonoplastmfmbrsnointrinsicprotein and plays key role in water transportation．RT-PCR analysis showed that accumulations of GhTIPl mRNAwereabundantin rootsandlowerlevelsinothertisSUes，whichwasdevelopment-regulated in roots．]1lrthoITnoro,GhTIPl Was regulated in cotton under drought，salt and cold翻r‰Funher experiments showed that the pattern of GhTIPl extneasion Was different under different stresses and in different tissues．Especially,GhTIPl transcript Was increased in cotyledon,however, decreased in roots trader cold treated,angges=ting that the GhTIPl