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Published on February 27, 2009; 10.1104/pp.108.134551

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Received December 17, 2008
Accepted February 24, 2009

Identification and Analyses of Candidate Genes for Rpp4-mediated Resistance to Asian Soybean Rust in Soybean (Glycine max (L.) Merr.)

Jenelle D.F. Meyer , Danielle C.G. Silva , Chunling Yang , Kerry F. Pedley , Chunquan Zhang , Martijn van de Mortel , John H. Hill , Randy C. Shoemaker , Ricardo V. Abdelnoor , Steven A. Whitham , and Michelle A. Graham *

USDA-ARS Corn Insects and Crop Genetics Research Unit, Ames, IA, U.S.A; Embrapa Soja, Londrina, PR, Brazil, FFALM Bandeirantes, PR Brazil and UNESP, Jaboticabal, SP Brazil; Department of Plant Pathology, Iowa State University, Ames, IA, U.S.A; USDA-ARS Foreign Disease-Weed Science Research Unit, Ft. Detrick, MD, U.S.A; Department of Agronomy, Iowa State University, Ames IA, U.S.A; Embrapa Soja, Londrina, PR, Brazil

* Corresponding author; email: Michelle.Graham{at}ars.usda.gov.

Asian Soybean Rust (ASR) is a formidable threat to soybean production in many areas of the world including the United States. Only five sources of resistance have been identified (Rpp1, Rpp2, Rpp3, Rpp4, and Rpp5). Rpp4 was previously identified in the resistant genotype PI459025B and mapped within 2 cM of Satt288 on soybean chromosome 18 (linkage group G). Using simple sequence repeat markers, we developed a bacterial artificial chromosome contig for the Rpp4 locus in the susceptible cultivar Williams82 (Wm82). Sequencing within this region identified three Rpp4candidate disease resistance genes (Rpp4C1 - Rpp4C3 (Wm82)) with greatest similarity to the lettuce RGC2 family of coiled coil-nucleotide binding site-leucine rich repeat (CC-NBS-LRR) disease resistance genes. Constructs containing regions of the Wm82 Rpp4 candidate genes were used for virus-induced gene silencing experiments to silence resistance in PI459025B, confirming that orthologous genes confer resistance. Using primers developed from conserved sequences in the Wm82 Rpp4 candidate genes, we identified five Rpp4 candidate genes (Rpp4C1 - Rpp4C5 (PI459025B)) from the resistant genotype. Additional markers developed from the Wm82 Rpp4 BAC contig further defined the region containing Rpp4 and eliminated Rpp4C1 (PI459025B) and Rpp4C3 (PI459025B) as candidate genes. Sequencing of RT-PCR products revealed that Rpp4C4 (PI459025B) was highly expressed in the resistant genotype while expression of the other candidate genes was nearly undetectable. These data support Rpp4C4 (PI459025B) as the single candidate gene for Rpp4-mediated resistance to ASR.






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