Plant disease resistance gene (R gene); defense response gene encode some conserved motifs. In the present work, a PCR strategy was used to clone resistance gene analogs (RGAs); defense gene analogs (DGAs) from Sea-is...Plant disease resistance gene (R gene); defense response gene encode some conserved motifs. In the present work, a PCR strategy was used to clone resistance gene analogs (RGAs); defense gene analogs (DGAs) from Sea-island cotton variety Hai7124 using oligonucleotide primers based on the nucleotide-binding site (NBS); serine/threonine kinase (STK) in the R-gene; pathogenesis-related proteins of class 2 (PR2) of defense response gene. 79 NBS sequences, 21 STK sequences; 11 DGAs were cloned from disease-resistance cotton. Phylogenic analysis of 79 NBS-RGAs; NBS-RGAs nucleotide sequences of cotton already deposited in GenBank identified one new sub-cluster. The deduced amino acid sequences of NBS-RGAs; STK-RGAs were divided into two distinct groups respectively: Toll/Interleukin-1 receptor (TIR) group; non-TIR group, A group; B group. The expression of RGAs; DGAs having consecutive open reading frame (ORF) was also investigated; it was found that 6 NBS-RGAs; 1 STK-RGA were induced,; 1 DGA was up-regulated by infection of Verticillium dahliae strain VD8. 4 TIR-NBS-RGAs; 4 non-TIR-NBS-RGAs were arbitrarily used as probes for Southern-blotting. There existed 2–10 blotted bands. In addition, since three non-TIR-NBS-RGAs have the same hybridization pattern, we conjecture that these three RGAs form a cluster distribution in the genome.展开更多
One resistance gene analog fragment named RGA-CIN14 was isolated from TcLr19 wheat,which contains kinase-2,kinase-3a,and the GLPL motif of the NBS-spanning region,using degenerated primers according to the nucleotide ...One resistance gene analog fragment named RGA-CIN14 was isolated from TcLr19 wheat,which contains kinase-2,kinase-3a,and the GLPL motif of the NBS-spanning region,using degenerated primers according to the nucleotide binding site （NBS） conserved domain.Based on the RGA-CIN14,a full-length cDNA,CIN14,which was 2 987 bp encoding 880 amino acids,was obtained by using the method of the rapid amplification cDNA ends （RACE）.Bioinformatics analysis showed that the deduced amino acids of CIN14 protein consisted of a NB-ARC conserved domain and many leucine-rich repeats （LRR） domains.The phylogenetic tree analysis indicated a considerable identity of the protein encoded by CIN14 with that of wheat leaf rust resistance gene Lr1,but a lower similarity with Lr21.The expression profile of the CIN14 gene detected by semi-quantitative RT-PCR showed that the CIN14 gene was not induced by Puccinia triticina and it was a constitutive gene with low abundance in the wheat leaf tissue.The resistance homology sequence was successfully obtained,which provides the shortcut for cloning of the resistance gene in TcLr19 wheat.展开更多
为了加快小麦抗性亲本的选配,培育具有持久抗性的小麦新品种,根据大多数已克隆的植物抗病基因所编码蛋白产物的NBS-LRR(核苷酸结合位点和富含亮氨酸区域,N ucleotide b ind ing s ite-leuc ine richrepeat)和STK(丝氨酸/苏氨酸蛋白激酶,...为了加快小麦抗性亲本的选配,培育具有持久抗性的小麦新品种,根据大多数已克隆的植物抗病基因所编码蛋白产物的NBS-LRR(核苷酸结合位点和富含亮氨酸区域,N ucleotide b ind ing s ite-leuc ine richrepeat)和STK(丝氨酸/苏氨酸蛋白激酶,Ser/T hr K inase)保守结构域,合成24条简并引物,对四川省主要利用的82份小麦育种抗性亲本材料进行PCR扩增和聚丙烯酰胺凝胶电泳检测,筛选出多态性高的三对引物组合:P tok in1N/P tok in1N、C er3 l/XLRR INV 2、A S3/S2,进一步扩增,共获得123条带,其中54条带具有多态性,多态率达43.9%。U PGM A分析把82份材料分为两大类,其中LB 0485、C 866-1、R 116、R 185为I类,其余78份材料为Ⅱ类;Ⅱ类材料在遗传距离0.397处又分为Ⅱ-1和Ⅱ-2两个亚类。分析结果表明:(1)82份小麦供试材料间存在丰富的抗病基因同源序列的多样性,聚类分析结果与相应的抗性鉴定结果基本吻合;(2)国外引进的抗性材料与四川省广泛使用的小麦育种亲本材料的抗性背景不同,两者之间能够很好地区分开来;(3)R系材料具有广泛的抗性遗传基础,在大多数类群中均有分布。展开更多
基金supported by the National Natural Science Foundation of China(Grant Nos.30270806 and 30370899)Program for Changjiang Scholars and Innovative Research Team in University in Ministry of Education in China.
文摘Plant disease resistance gene (R gene); defense response gene encode some conserved motifs. In the present work, a PCR strategy was used to clone resistance gene analogs (RGAs); defense gene analogs (DGAs) from Sea-island cotton variety Hai7124 using oligonucleotide primers based on the nucleotide-binding site (NBS); serine/threonine kinase (STK) in the R-gene; pathogenesis-related proteins of class 2 (PR2) of defense response gene. 79 NBS sequences, 21 STK sequences; 11 DGAs were cloned from disease-resistance cotton. Phylogenic analysis of 79 NBS-RGAs; NBS-RGAs nucleotide sequences of cotton already deposited in GenBank identified one new sub-cluster. The deduced amino acid sequences of NBS-RGAs; STK-RGAs were divided into two distinct groups respectively: Toll/Interleukin-1 receptor (TIR) group; non-TIR group, A group; B group. The expression of RGAs; DGAs having consecutive open reading frame (ORF) was also investigated; it was found that 6 NBS-RGAs; 1 STK-RGA were induced,; 1 DGA was up-regulated by infection of Verticillium dahliae strain VD8. 4 TIR-NBS-RGAs; 4 non-TIR-NBS-RGAs were arbitrarily used as probes for Southern-blotting. There existed 2–10 blotted bands. In addition, since three non-TIR-NBS-RGAs have the same hybridization pattern, we conjecture that these three RGAs form a cluster distribution in the genome.
基金funded by the National Natural Science Foundation of China (30771391,30700505)the Natural Science Foundation of Hebei Province,China (C2008000281)
文摘One resistance gene analog fragment named RGA-CIN14 was isolated from TcLr19 wheat,which contains kinase-2,kinase-3a,and the GLPL motif of the NBS-spanning region,using degenerated primers according to the nucleotide binding site （NBS） conserved domain.Based on the RGA-CIN14,a full-length cDNA,CIN14,which was 2 987 bp encoding 880 amino acids,was obtained by using the method of the rapid amplification cDNA ends （RACE）.Bioinformatics analysis showed that the deduced amino acids of CIN14 protein consisted of a NB-ARC conserved domain and many leucine-rich repeats （LRR） domains.The phylogenetic tree analysis indicated a considerable identity of the protein encoded by CIN14 with that of wheat leaf rust resistance gene Lr1,but a lower similarity with Lr21.The expression profile of the CIN14 gene detected by semi-quantitative RT-PCR showed that the CIN14 gene was not induced by Puccinia triticina and it was a constitutive gene with low abundance in the wheat leaf tissue.The resistance homology sequence was successfully obtained,which provides the shortcut for cloning of the resistance gene in TcLr19 wheat.
文摘为了加快小麦抗性亲本的选配,培育具有持久抗性的小麦新品种,根据大多数已克隆的植物抗病基因所编码蛋白产物的NBS-LRR(核苷酸结合位点和富含亮氨酸区域,N ucleotide b ind ing s ite-leuc ine richrepeat)和STK(丝氨酸/苏氨酸蛋白激酶,Ser/T hr K inase)保守结构域,合成24条简并引物,对四川省主要利用的82份小麦育种抗性亲本材料进行PCR扩增和聚丙烯酰胺凝胶电泳检测,筛选出多态性高的三对引物组合:P tok in1N/P tok in1N、C er3 l/XLRR INV 2、A S3/S2,进一步扩增,共获得123条带,其中54条带具有多态性,多态率达43.9%。U PGM A分析把82份材料分为两大类,其中LB 0485、C 866-1、R 116、R 185为I类,其余78份材料为Ⅱ类;Ⅱ类材料在遗传距离0.397处又分为Ⅱ-1和Ⅱ-2两个亚类。分析结果表明:(1)82份小麦供试材料间存在丰富的抗病基因同源序列的多样性,聚类分析结果与相应的抗性鉴定结果基本吻合;(2)国外引进的抗性材料与四川省广泛使用的小麦育种亲本材料的抗性背景不同,两者之间能够很好地区分开来;(3)R系材料具有广泛的抗性遗传基础,在大多数类群中均有分布。
基金This study was financially supported by the Fujian Natural Science Foundation (2009J05050)the National High Technology Research and Develop-ment Program of China (863 Program) (2007AA100701)+2 种基金the Program of Introducing International Super Agricultural Science and Technology (948 Program) (2006-G37)National Science and Technology Project File of Science and Technology Commission, Fujian Science Department (F2007AA100701)the earmarked fund for Modern Agro-industry Technology Research System
