高粱低磷胁迫响应相关WRKY转录因子克隆和表达分析
时间:2019-08-04 12:10 来源:毕业论文 作者:毕业论文 点击:次
| 摘要：高粱（Sorghum bicolor L.）是重要的粮食、饲料和能源作物。磷参与植物大部分重要的生理过程，是作物生长发育所必需的营养元素，是蛋白质、核酸、氨基酸和叶绿素等的组成成分。然而，土壤中磷含量十分有限，远不能满足植物对磷含量的需求。近年来WRKY转录因子在植物中被发现为特有的新型转录调控因子。本实验克隆了高粱中与低磷胁迫响应相关的WRKY转录因子，命名为SbWRKY2，共编码246个氨基酸。氨基酸序列分析和同源比对发现，SbWRKY2属于第Ⅱ类WRKY转录因子。分析SbWRKY2在低磷处理不同时间下的表达模式，结果表明：SbWRKY2在高粱根系和地上部中都有表达，根系中的表达量均比地上部中的表达量高；低磷处理后，SbWRKY2的表达受低磷诱导，在低磷处理的第8天表达量较对照高达10倍以上。38001 毕业论文关键词：高粱；WRKY；磷胁迫；克隆；表达分析 Isolation and Expression Analysis of WRKY transcription factor Respond to low-phosphorous stress in Sorghum Abstract: Sorghum (Sorghum bicolor L.) is an important food, feed and energy crops. Phosphorous join in most of important physiological process in plants, which is a necessary element for crop growth. It is proteins, nucleic acids, amino acids and chlorophyll composition. However, the content of phosphate in the soil is not enough to satisfy the demand of plant. WRKY transcription factors are a new type transcription regulatory factors found in plants in recent years. In this study, WRKY transcription factors respond to low-phosphorous stress in sorghum was cloned, named SbWRKY2. SbWRKY2 encodes 246 amino acids. Amino acid sequence analysis and homology comparison analysis showed that SbWRKY2 belongs to group Ⅱ of the WRKY transcription factor family. Expression pattern of SbWRKY2 is analyzed under different time in low phosphorus treatment, the results showed that SbWRKY2 were expressed in both roots and shoots. The relative express quantity of SbWRKY2 in the roots is higher than in the shoot’s. Expression of SbWRKY2 induced by low-phosphorus stress and it reached the highest level after treated by low phosphorus for 8 days. Keywords: sorghum; WRKY; low-phosphate stress; cloning; expression analysis 目 录 摘要：.1 关键词：….1 Abstract1 Keywords.…1 引言… 2 1 材料与方法…..5 1.1 实验材料及处理…..…...5 1.2 主要试剂与仪器…..…...5 1.2.1 主要试剂5 1.2.2 主要仪器...….5 1.3 植物组织总RNA的提取及检测.….….5 1.3.1 植物组织总RNA的提取5 1.3.2 总RNA质量的检测…..6 1.4 cDNA 的合成及检测.….6 1.4.1 cDNA 的合成.6 1.4.2 cDNA 的检测.6 1.5 高粱低磷相关WRKY基因cDNA全长的分离.....7 1.5.1 cDNA全长的分离..7 1.5.2 DNA片段琼脂糖凝胶回收…7 1.6 克隆载体的构建..….8 1.6.1 目的基因与克隆载体的连接…8 1.6.2 大肠杆菌的转化与培养…8 1.6.3 阳性单克隆筛选及测序..8 1.7 生物信息学分析….….….8 1.8 表达特性研究.....9 2 结果与分析...10 2.1 植物组织总RNA与cDNA的检测10 2.2 高粱低磷相关WRKY基因全长的克隆..10 2.3 WRKY基因序列分析及蛋白质特性分析…...11 2.3.1 序列分析及氨基酸预测...11 2.3.2 氨基酸组成及理化性质….…12 2.3.3 氨基酸的同源性分析.…12 2.3.4 蛋白二级结构预测….…13 2.4 SbWRKY2基因的表达分析….14 3 讨论.....15 3.1 SbWRKY2序列特征…...15 (责任编辑：qin) |