Isolation and Characterization of NBS-LRR Class Resistance Homologous Gene from Wheat

• Published in: Agricultural sciences in China Vol. 10; no. 8; pp. 1151 - 1158
• Main Authors: ZHANG, Nan, WANG, Shen, WANG, Hai-yan, LIU, Da-qun
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 01.08.2011
• Subjects: NBS; NBS-LRR; rapid amplification cDNA end (RACE); resistance gene analogs (RGAs); RT-PCR; wheat leaf rust resistance gene; 同源基因; 国家统计局; 富含亮氨酸重复序列; 小麦叶锈病; 快速扩增cDNA末端; 抗性基因; 核苷酸结合位点; rapid amplification cDNA end (RACE); NBS-LRR; wheat leaf rust resistance gene; resistance gene analogs (RGAs); RT-PCR
• ISSN: 1671-2927
• DOI: 10.1016/S1671-2927(11)60105-3

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.