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doi:10.1534/genetics.107.080648
A more recent version of this article appeared on November 1, 2007.
REGULAR RESEARCH PAPERS |
The Blast Resistance Gene Pi37 Encodes an NBS-LRR Protein and is a Member of a Resistance Gene Cluster on Rice Chromosome 1
Fei Lin 1, Shen Chen 1, Zhiqun Que 1, Ling Wang 1, Xinqiong Liu 1 and Qinghua Pan 1*
1 College of Resources and Environmental Sciences, South China Agricultural University
* To whom correspondence should be addressed. E-mail: panqh{at}scau.edu.cn.
Submitted on August 15, 2007
Revised on September 4, 2007
Accepted on 4 September 2007
The resistance (R) gene Pi37, present in the rice cultivar St. No. 1, was isolated by an in silico map-based cloning procedure. The equivalent genetic region in Nipponbare contains four NBS-LRR type loci. These four candidates for Pi37 (Pi37-1, -2, -3 and -4) were amplified separately from St. No. 1 via long-range PCR, and cloned into a binary vector. Each construct was individually transformed into the highly blast susceptible cultivar Q1063. The subsequent complementation analysis revealed Pi37-3 to be the functional gene, while -1, -2 and -4 are probably pseudogenes. Pi37 encodes a 1290 peptide NBS-LRR product, and the presence of substitutions at two sites in the NBS region (V239A and I247M) is associated with the resistance phenotype. Semi-quantitative expression analysis showed that in St. No. 1, Pi37 was constitutively expressed and only slightly induced by blast infection. Transient expression experiments indicated that the Pi37 product is restricted to the cytoplasm. Pi37-3 is thought to have evolved recently from -2, which in turn was derived from an ancestral -1 sequence. Pi37-4 is likely the most recently evolved member of the cluster, and probably represents a duplication of -3. The four Pi37 paralogs are more closely related to maize rp1 than to any of the currently isolated rice blast R genes Pita, Pib, Pi9, Pi2, Piz-t and Pi36.
Key Words: gene evolution, gene isolation, in silico map-based cloning, resistance gene cluster
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