| 刘婷婷1，殷幼平1，林亚玉1，贤家旭2，陈世伟2，王中康1*.柑橘黄龙病侵染相关抗病基因同源序列的克隆鉴定与表达分析[J].植物保护,2012,38(2):23-28. |
| 柑橘黄龙病侵染相关抗病基因同源序列的克隆鉴定与表达分析 |
| Isolation and expression analysis of resistance gene analogs related to infection of Citrus Huanglongbing |
| |
| DOI： |
| 中文关键词: 柚cDNA 抗病基因相似序列 定量PCR 表达分析 |
| 英文关键词:cDNA of Citrus maxima resistance gene analogs(RGAs) quantitative Real-time PCR expression analysis |
| 基金项目: |
|
| 摘要点击次数: 1120 |
| 全文下载次数: 791 |
| 中文摘要: |
| ［目的］ 从黄龙病耐病寄主植物cDNA中筛选抗病基因相关序列并对其进行表达分析研究。［方法］ 根据已克隆的植物抗性基因表达产物NBS-LRR保守区域设计简并引物，以耐HLB的柑橘属柚cDNA为模板扩增RGAs，并进行实时荧光定量PCR。［结果］ 通过RFLP分析及克隆测序共得到5个NBS类抗病基因相似序列(RGAs)片段，在GenBank上登录号为HM777043~HM777047。通过Clustalx、DNAMAN等软件分析5个RGAs及其推导的氨基酸的相似性，结果显示它们均含有典型NBS-LRR类抗性基因所具有的保守区域：P-loop、Kinase-2a、GLPLAL，其与已克隆的烟草N、亚麻L6、拟南芥RPS2、RPS5、RPP8、RPM1等抗病基因在保守区域氨基酸水平上的相似性为19.71%~42.86%。根据得到的序列设计特异性引物，对5个RGAs在HLB侵染过程中的表达进行定量PCR，结果显示嫁接病芽接穗后的8次连续采样中5个RGA的表达受到不同程度的调控。［结论］ 表明5个RGAs可能与黄龙病的侵染有关。 |
| 英文摘要: |
| Resistance gene analogs were isolated from cDNA of plant tolerant to HLB and analyzed by expression. The resistance gene analogs were amplified from cDNA of Citrus maxima using degenerated primers based on the conserved regions of NBS (Nucleotide Binding Site) LRR (Leucine Rich Repeat) domain from plant resistance genes cloned, and the quantitative Real time PCR was used to check the expression of 5 RGAs.［Result］ A total of 5 RGA fragments were successfully obtained, with GenBank accession numbers of HM777043 HM777047. The deduced amino acid of these RGAs were analyzed by Clustalx and DNAMAN softwares. Sequence analysis showed that these RGAs contained the conserved domains P loop, kinase 2a and GLPL, which were conserved in NBS-LRR type disease resistance genes. The 5 RGAs shared 19.71%-42.86% identity with the resistance genes of tobacco N, flax L6, and〖WTBX〗 Arabidopsis thaliana〖WTBZ〗 〖WTBX〗RPS2, RPS5, RPP8 〖WTBZ〗and〖WTBX〗 RPM1〖WTBZ〗. The quantitative Real time PCR results showed that expression of the 5 RGAs was different. ［Conclusion］ Therefore, it suggested that the 5 RGAs might be related with pathogen infection. |
| 查看全文 查看/发表评论 下载PDF阅读器 |
| 关闭 |
