【摘要】：依據已知NBS-LRR類抗病基因的P-loop和GLPL兩個保守結構域設計簡并引物,對抗菠菜霜霉病商業雜交種RZ51-147、自交系12S3和12S4的基因組DNA進行PCR擴增、克隆和序列測定,共獲得23條具有完整開放閱讀框且含有NBS結構的抗病基因同源序列(resistancegeneanalogs,RGAs),編號為SP1~SP17、SP19~SP24,其在NCBI中的登錄號為KX914865~KX914887。核苷酸比較分析結果表明,這23條RGAs大多與甜菜中推測的一些抗病蛋白基因或其他相關蛋白基因具有較高的同源性,其中SP1、SP2、SP5、SP6、SP10、SP11、SP12、SP13等8條序列均與甜菜RF45類抗病蛋白有著86%以上的同源性;SP3、SP4、SP8、SP9、SP24等5條序列與甜菜RPP13類抗病蛋白的同源性在85%以上,與向日葵抗霜霉病基因PI8相應區域的氨基酸序列相似性為32.20%~33.52%,且在進化樹上聚為一類,推測其可能與抗霜霉病相關。同源進化分析結果表明,除SP7外,其余序列均為non-TIR-NBS-LRR類抗病基因,并與推導的氨基酸序列多重比較結果一致。
[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.
【作者單位】： 中國農業科學院蔬菜花卉研究所;中蔬種業科技(北京)有限公司;
【基金】：國家“十二五”科技支撐計劃項目(2012BAD02B04,2014BAD01B08) 國家自然科學基金項目(31401872) 中國農業科學院科技創新工程項目(CAAS-ASTIP-IVFCAAS) 農業部園藝作物生物學與種質創制重點實驗室項目
【分類號】：S436.36

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