Isolation and Characterization of NBS-LRR Class Resistance Homologous Gene from Wheat
2011
ZHANG Nan, WANG Shen, WANG Hai-yan, LIU Da-qun (School of Agriculture and Biology, Shanghai Jiao Tong University)
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 nucleotidebinding site (NBS) conserved domain. Based on the RGA-CIN14, a full-length cDNA, CIN14, which was 2 987 bpencoding 880 amino acids, was obtained by using the method of the rapid amplification cDNA ends (RACE). Bioinformaticsanalysis showed that the deduced amino acids of CIN14 protein consisted of a NB-ARC conserved domain and manyleucine-rich repeats (LRR) domains. The phylogenetic tree analysis indicated a considerable identity of the proteinencoded by CIN14 with that of wheat leaf rust resistance gene Lr1, but a lower similarity with Lr21. The expression profileof the CIN14 gene detected by semi-quantitative RT-PCR showed that the CIN14 gene was not induced by Pucciniatriticina and it was a constitutive gene with low abundance in the wheat leaf tissue. The resistance homology sequencewas successfully obtained, which provides the shortcut for cloning of the resistance gene in TcLr19 wheat.
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