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【目的】在花生野生种、栽培种及栽野杂交后代中,克隆抗病基因类似物(Resistance gene analog,RGA)相关基因片段,为抗病基因的进一步分离、鉴定及利用奠定基础。【方法】以14个花生栽野杂交种和2个抗病花生亲本为材料,应用PCR方法扩增来自基因组DNA的抗病基因类似物序列,并分析其同源性。【结果】扩增出15条来自基因组DNA的抗病基因类似物序列,序列长度在500bp左右,由此推导出的15条氨基酸序列含有典型的NBS保守区的N端糖基化位点、蛋白激酶C磷酸化位点、酪蛋白激酶Ⅱ磷酸化位点、N-豆蔻酰化位点。15条花生RGA氨基酸序列间的同源性在73.2%~100.0%,与已发表的花生抗病基因类似物的相似性较高,达72.1%~100.0%;与已知的其他作物的抗病基因编码的氨基酸序列有34.1%~54.6%的同源性。【结论】分离到的15个花生抗病基因同源片段为抗病基因的部分序列。
【Objective】 The aim of this study was to clone cloned gene fragments of resistance gene analog (RGA) in the wild, cultivated and hybridized offspring of peanut, which laid the foundation for the further isolation, identification and utilization of disease-resistant genes. 【Method】 The 14 resistant peanut hybrids and 2 resistant peanut parents were used as materials to amplify the sequences of the resistance gene analogues from genomic DNA by PCR and their homologies were analyzed. 【Result】 Fifteen DNA sequences of resistance genes were amplified from genomic DNA. The length of the sequence was about 500 bp. The deduced 15 amino acid sequences contained the N-terminal glycosylation sites of typical NBS conserved regions, Kinase C phosphorylation site, casein kinase II phosphorylation site, N-myristoylation site. The homologies of 15 amino acid sequences of peanut RGA were between 73.2% and 100.0%, and their similarity with published peanut resistance gene analogues was high (72.1% -100.0%). Compared with other known crops, Genes encoded amino acid sequences 34.1% ~ 54.6% homology. 【Conclusion】 The 15 homologous fragments of the peanut resistance genes isolated were partial sequences of disease resistance genes.