黄瓜CsAFB基因影响拟南芥盐胁迫耐受性的研究
时间:2020-06-26 09:18 来源:毕业论文 作者:毕业论文 点击:次
| 摘要拟南芥（Arabidopsis thaliana）生长素受体参与盐胁迫下的转录调控，影响了植株的盐胁迫耐受性。本实验室前期实验已克隆获得黄瓜（Cucumis sativus L.）生长素受体的同源基因CsAFB(生长素信号F-box蛋白基因，auxin signaling F box protein)，但其功能尚未有明确的报道。实验室已成功获取黄瓜CsAFB转基因高表达拟南芥，因此本研究以纯合转基因拟南芥为材料，探讨了高盐胁迫下黄瓜CsAFB基因对拟南芥抗盐性的影响，并且分析了其影响机理。本实验用300 mM NaCl处理转基因拟南芥和野生型拟南芥的种子、幼苗与成年植株，统计了转基因种子相较野生型在高盐培养基上的萌发率，观测比较了幼苗生长情况以及测量成年植株抗逆境生理指标。获得的主要实验结果如下：300 mM NaCl处理下，转基因拟南芥种子萌发率和萌发速度相较野生型有一定程度的提高，转基因幼苗侧根数量增加、对叶绿素合成的抑制作用有所减弱。检测发现，在黄瓜CsAFB基因影响下，拟南芥成年植株叶片SOD活性有一定程度的提高，丙二醛含量下降。本研究说明，黄瓜生长素受体同源基因CsAFB提高了拟南芥盐胁迫耐受性，其功能类似拟南芥TIR1基因，为进一步研究耐盐机制和黄瓜生长素受体提供新的数据和参考。51642 毕业论文关键词：盐胁迫；转基因拟南芥；CsAFB；耐盐机制 Abstract Auxin receptors participate in transcriptional regulation under salt stress, which influence the salt stress tolerance in Arabidopsis. We had cloned the auxin receptor homologous genes CsAFB (auxin signaling F box protein) in cucumber (Cucumis sativus L.) in our previous work, whose function had not been reported yet. The transgenic CsAFB over-expressed Arabidopsis were obtained successfully before，so we used homozygous transgenic plants as materials to explore the potential influence of CsAFB in salt stress tolerance and study its mechanism in Arabidopsis. In this work, both transgenic CsAFB and wild-type plants were treated with 300 mM NaCl, germination rate of seeds were counted, and the growth of seedings were observed and measured. The main results are as follows: under 300 mM NaCl, transgenic plants had an enhanced seed germination percentage and rate, increased numbers of lateral roots, and decreased inhibition of chlorophyll synthesis. The SOD activity increased as the content of MDA decreased in adult plant leaves in transgenic CsAFB plants. This work illustrated that transgene of CsAFB enhanced salt stress tolerance in arabidopsis thaliana. which functioned similarly as AtTIR1. This work also contributes to the further study of their functions and mechanisms in salt stress tolerance in cucumber. Keywords：Salt stress; Transgenic Arabidopsis; CsAFB; Salt tolerance mechanism 目 录 引言 4 1 黄瓜CsAFB基因影响拟南芥盐胁迫耐受性 6 1.1 材料与方法 6 1.1.1 实验材料 6 1.1.2 实验主要试剂和仪器 6 1.1.3 盐胁迫下种子萌发率统计 6 1.1.4 盐胁迫下幼苗根的生长情况统计 7 1.1.5 盐胁迫下土培苗生长状态统计 8 1.2 实验结果 8 1.2.1 黄瓜CsAFB基因影响种子在处理下的萌发率 8 1.2.2 黄瓜CsAFB基因影响盐处理下根的生长 9 1.2.3 黄瓜CsAFB基因影响植株在盐处理下的生长状态 10 1.3 讨论 11 2 黄瓜CsAFB基因影响拟南芥耐受性的机理研究 12 2.1 材料与方法 12 2.1.1 材料 12 2.1.2 主要试剂和主要使用仪器 12 2.1.3 SOD酶活性测定 12 2.1.4 丙二醛含量测定 13 (责任编辑：qin) |