【摘要】：依據(jù)已知NBS-LRR類抗病基因的P-loop和GLPL兩個保守結(jié)構(gòu)域設(shè)計簡并引物,對抗菠菜霜霉病商業(yè)雜交種RZ51-147、自交系12S3和12S4的基因組DNA進(jìn)行PCR擴(kuò)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(jié)構(gòu)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(jié)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(guān)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(yīng)區(qū)域的氨基酸序列相似性為32.20%~33.52%,且在進(jìn)化樹上聚為一類,推測其可能與抗霜霉病相關(guān)。同源進(jìn)化分析結(jié)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(dǎo)的氨基酸序列多重比較結(jié)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(nóng)業(yè)科學(xué)院蔬菜花卉研究所;中蔬種業(yè)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(xué)基金項目(31401872) 中國農(nóng)業(yè)科學(xué)院科技創(chuàng)新工程項目(CAAS-ASTIP-IVFCAAS) 農(nóng)業(yè)部園藝作物生物學(xué)與種質(zhì)創(chuàng)制重點(diǎn)實驗室項目
【分類號】：S436.36

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