菠菜NBS-LRR类抗病基因同源序列的克隆及分析
发布时间：2018-11-02 08:06
【摘要】：依据已知NBS-LRR类抗病基因的P-loop和GLPL两个保守结构域设计简并引物,对抗菠菜霜霉病商业杂交种RZ51-147、自交系12S3和12S4的基因组DNA进行PCR扩增、克隆和序列测定,共获得23条具有完整开放阅读框且含有NBS结构的抗病基因同源序列(resistancegeneanalogs,RGAs),编号为SP1~SP17、SP19~SP24,其在NCBI中的登录号为KX914865~KX914887。核苷酸比较分析结果表明,这23条RGAs大多与甜菜中推测的一些抗病蛋白基因或其他相关蛋白基因具有较高的同源性,其中SP1、SP2、SP5、SP6、SP10、SP11、SP12、SP13等8条序列均与甜菜RF45类抗病蛋白有着86%以上的同源性;SP3、SP4、SP8、SP9、SP24等5条序列与甜菜RPP13类抗病蛋白的同源性在85%以上,与向日葵抗霜霉病基因PI8相应区域的氨基酸序列相似性为32.20%~33.52%,且在进化树上聚为一类,推测其可能与抗霜霉病相关。同源进化分析结果表明,除SP7外,其余序列均为non-TIR-NBS-LRR类抗病基因,并与推导的氨基酸序列多重比较结果一致。
[Abstract]:Based on the two conserved domains of P-loop and GLPL of known NBS-LRR resistance genes, degenerate primers were designed to amplify, clone and sequence the genomic DNA of RZ51-147, inbred lines 12S3 and 12S4 against downy mildew of spinach. A total of 23 disease-resistance gene homologous sequences (resistancegeneanalogs,RGAs) with complete open reading frame and NBS structure were obtained, numbered SP1~SP17,SP19~SP24, and its accession number in NCBI was KX914865~KX914887.. The results of nucleotide comparison showed that most of the 23 RGAs had high homology with some disease-resistant protein genes or other related protein genes in sugarbeet, among which SP1,SP2,SP5,SP6,SP10,SP11,SP12, was highly homologous. SP13 and other 8 sequences were more than 86% homology with beet RF45 protein. The homology of five SP3,SP4,SP8,SP9,SP24 sequences with beet RPP13 resistance proteins was more than 85%, and the amino acid sequence similarity between SP3,SP4,SP8,SP9,SP24 and sunflower resistance gene PI8 was 32.20% and 33.52%, respectively. The phylogenetic tree may be associated with resistance to downy mildew. The results of homologous evolution analysis showed that all the sequences except SP7 were non-TIR-NBS-LRR resistance genes, and the results were consistent with the deduced amino acid sequences.
【作者单位】： 中国农业科学院蔬菜花卉研究所;中蔬种业科技(北京)有限公司;
【基金】：国家“十二五”科技支撑计划项目(2012BAD02B04,2014BAD01B08) 国家自然科学基金项目(31401872) 中国农业科学院科技创新工程项目(CAAS-ASTIP-IVFCAAS) 农业部园艺作物生物学与种质创制重点实验室项目
【分类号】：S436.36
本文编号：2305427
[Abstract]:Based on the two conserved domains of P-loop and GLPL of known NBS-LRR resistance genes, degenerate primers were designed to amplify, clone and sequence the genomic DNA of RZ51-147, inbred lines 12S3 and 12S4 against downy mildew of spinach. A total of 23 disease-resistance gene homologous sequences (resistancegeneanalogs,RGAs) with complete open reading frame and NBS structure were obtained, numbered SP1~SP17,SP19~SP24, and its accession number in NCBI was KX914865~KX914887.. The results of nucleotide comparison showed that most of the 23 RGAs had high homology with some disease-resistant protein genes or other related protein genes in sugarbeet, among which SP1,SP2,SP5,SP6,SP10,SP11,SP12, was highly homologous. SP13 and other 8 sequences were more than 86% homology with beet RF45 protein. The homology of five SP3,SP4,SP8,SP9,SP24 sequences with beet RPP13 resistance proteins was more than 85%, and the amino acid sequence similarity between SP3,SP4,SP8,SP9,SP24 and sunflower resistance gene PI8 was 32.20% and 33.52%, respectively. The phylogenetic tree may be associated with resistance to downy mildew. The results of homologous evolution analysis showed that all the sequences except SP7 were non-TIR-NBS-LRR resistance genes, and the results were consistent with the deduced amino acid sequences.
【作者单位】： 中国农业科学院蔬菜花卉研究所;中蔬种业科技(北京)有限公司;
【基金】：国家“十二五”科技支撑计划项目(2012BAD02B04,2014BAD01B08) 国家自然科学基金项目(31401872) 中国农业科学院科技创新工程项目(CAAS-ASTIP-IVFCAAS) 农业部园艺作物生物学与种质创制重点实验室项目
【分类号】：S436.36
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