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Frontiers of Agriculture in China

ISSN 1673-7334

ISSN 1673-744X(Online)

CN 11-5729/S

Front Agric Chin    2009, Vol. 3 Issue (3) : 240-246     DOI: 10.1007/s11703-009-0035-9
RESEARCH ARTICLE |
Cloning and structure analysis of an NBS-LRR disease-resistant gene from Setaria italica Beauv
Qiaoyun WENG1, Zhiyong LI2, Jihong XING1, Zhiping DONG2(), Jingao DONG1()
1. Molecular Plant Pathology Lab, College of Life Science, Agricultural University of Hebei, Baoding 071001, China; 2. Institute of Millet Crops, Hebei Academy of Agriculture and Forestry Sciences, Shijiazhuang 050000, China
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Abstract  

Degenerate PCR primers targeting conserved motifs of most NBS-LRR disease-resistant genes in plants were tested in Setaria italica Beauv. cultivar Shilixiang, which is resistant to Uromyces setariae-italicae. A sequence with a length of 2673 bp has been obtained by using Genomic Walking technology. The nucleotide sequence contained an open reading frame that encoded 891 amino acid residues with a calculated molecular mass of 101.44 kDa. It was named RUS1 (Resistance against Uromyces setariae-italicae, GenBank No. FJ467296). It contained an NB-ARC domain and three conserved motifs P-loop, kinase 2, and kinase 3, which had the characteristics of NBS-LRR type resistant gene of plant. Phylogenetic analysis indicated that it was similar to RPM1 and might belong to LZ-NBS-LRR type disease resistance gene. Southern blotting result displayed that there were at least three copies of RUS1 in the foxtail millet genome.

Keywords Setaria italica      RUS1      NB-ARC domain     
Corresponding Authors: DONG Zhiping,Email:dzping001@163.com; DONG Jingao,Email:shmdjg@hebau.edu.cn   
Issue Date: 05 September 2009
URL:  
http://academic.hep.com.cn/fag/EN/10.1007/s11703-009-0035-9     OR     http://academic.hep.com.cn/fag/EN/Y2009/V3/I3/240
Fig.1  Symptom of cultivar inoculated with virulent isolate 93-5
Note: Left and right represent Shilixiang and Yugu No. 1, respectively.
Fig.2  PCR amplification result of disease resistant gene of
Note: Arrowhead indicates the specific band presented in Shilixiang; M represents pBR322.
Fig.3  Amplification result of Genome Walking
Note: M represents DL2000, Lanes 1-5 represent PCR product of 3' end, Lanes 6-9 represent PCR product of 5' end, and arrowhead indicates the specific band.
Fig.4  Alignment of deduced amino acid sequences of with NB-ARC domains of five known -gene
Fig.5  Conserved domain of gene of
Fig.6  Phylogenetic tree based on alignment of the deduced amino-acid sequences of and the NBS domains of known -genes
Fig.7  Copy number analysis of in genome of
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