摘要
本研究利用同源克隆方法,根据已知植物抗病(R)基因保守结构域设计一对简并引物,采用RTPCR方法对花生抗旱品种J11进行扩增,获得了一个通读的NBS-LRR类基因片段,命名为PDR1。PDR1长度为474 bp,核苷酸比较分析表明,PDR1与已克隆的大豆抗性基因(XM006573278)相似性为75%,与豌豆(AF123699)、苜蓿(XM003604518)抗性基因的相似性为74%;生物信息学预测其全长CDS编码框为5 838bp,编码的蛋白质含1 945个氨基酸,含有NB-ARC等保守结构域。荧光定量PCR分析表明,PDR1基因在花生幼苗根、茎、叶中均有表达,相对表达量为叶>根>茎;干旱胁迫后PDR1的相对表达量在12 h达到顶峰,随后下降,趋于平稳。本研究从花生中成功获得了NBS-LRR类基因的同源序列,为筛选新的抗旱花生种质提供了理论依据。
In this study, one resistance gene homology fragment was isolated from peanut by homology cloning method. A pair of degenerate primers was designed according to the conserved domains of the known plant disease resistance genes, and then, the fragment with continuous open reading frame was obtained from the peanut variety Jll with drought tolerance by RT - PCR. The sequence was named PDR1. The length of PDR1 was 474 bp. The homology research results showed that the nueleotide of PDR1 was 75% identical to a cloned Glycine max resistance gene （XM006573278）, and 74% identical to the sequences from Pisum sati- rum （AF123699） and Medicago truncatula （ XM003604518 ）, respectively. By bioinformatics method, it was predicted that the CDS of PDR1 was 5 838 bp, encoding 1 945 amino acids and containing the conserved do- main of NB - ARC. The results of real - time fluorescence quantitative PCR showed that PDRI expressed in peanut root, stem and leaves. The relative expression in different organs was leaves 〉 root 〉 stem. After drought stress, the relative expression of PDR1 in Jll reached the peak on the 12th hour, then decreased and moved towards stabilization. The obtain of this resistance homology sequence provided theoretical basis for the screening of drought resistant ~ermDlasms form peanut.
出处
《山东农业科学》
2015年第5期1-5,14,共6页
Shandong Agricultural Sciences
基金
国家科技支撑计划项目(2014BAD11B00)
国家国际科技合作专项(2015DFA31190)
山东省现代农业产业技术体系项目(SDAIT-05-022-02)
山东省农业良种工程项目"花生抗逆
广适种质资源创新与利用"
山东省农业科学院青年基金项目(2015YQN07)
