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

doi:10.1016/S1671-2927(11)60105-3

摘要/Abstract

摘要： 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.

关键词: wheat leaf rust resistance gene, NBS-LRR, resistance gene analogs (RGAs), rapid amplification cDNA end (RACE), RT-PCR

Abstract: 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.

Key words: wheat leaf rust resistance gene, NBS-LRR, resistance gene analogs (RGAs), rapid amplification cDNA end (RACE), RT-PCR

ZHANG Nan, WANG Shen, WANG Hai-yan, LIU Da-qun. Isolation and Characterization of NBS-LRR Class Resistance Homologous Gene from Wheat[J]. Journal of Integrative Agriculture, 2011, 10(8): 1151-1158.

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