Genetics. Published Articles Ahead of Print: December 30, 2005, Copyright © 2005
doi:10.1534/genetics.105.044891


A more recent version of this article appeared on March 1, 2006.

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The Broad-Spectrum Blast Resistance Gene Pi9 Encodes an NBS-LRR Protein and is a Member of a Multigene Family in Rice

Shaohong Qu 1, Guifu Liu 2, Bo Zhou 1, Maria Bellizzi 1, Lirong Zeng 1, Liangying Dai 3, Bin Han 4 and Guo-Liang Wang 1*

1 Ohio State University
2 Ohio State University and Chinese Academy of Sciences
3 Hunan Agricultural University
4 National Center for Gene Research, Chinese Academy of Sciences

* To whom correspondence should be addressed. E-mail: wang.620{at}osu.edu.

Submitted on April 26, 2005
Revised on July 29, 2005
Accepted on 17 December 2005


Abstract

The broad-spectrum rice blast resistance gene Pi9 was cloned using a map-based cloning strategy. Sequencing of a 76-kb bacterial artificial chromosome (BAC) contig spanning the Pi9 locus led to identification of six tandemly-arranged resistance-like genes with a nucleotide-binding site (NBS) and leucine-rich repeats (LRRs) (named Nbs1-Pi9 to Nbs6-Pi9). Analysis of selected Pi9 deletion mutants and transformation of a 45-kb fragment from the BAC contig into the susceptible rice cultivar TP309 narrowed down Pi9 to the candidate genes Nbs2-Pi9 and Nbs3-Pi9. Disease evaluation of the transgenic lines carrying the individual candidate genes confirmed that Nbs2-Pi9 is the Pi9 gene. Sequence comparison analysis revealed that the six paralogs at the Pi9 locus belong to four classes and gene duplication might be one of the major evolutionary forces contributing to the formation of the NBS-LRR gene cluster. Semi-quantitative reverse transcriptase (RT)-PCR analysis showed that Pi9 was constitutively expressed in the Pi9 resistant plants and was not induced by blast infection. The cloned Pi9 gene provides a starting point to elucidate the molecular basis of the broad-spectrum disease resistance and the evolutionary mechanisms of blast resistance gene clusters in rice.

Key Words: Magnaporthe grisea, NBS-LRR gene family, broad-spectrum resistance gene




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