title: Soybean cyst nematode resistance in soybean is independent of the Rhg4 locus LRR-RLK gene.
abstract: To test the function of candidate genes in soybean for resistance to the soybean cyst nematode (SCN), a large collection of EMS-mutants from the SCN-resistant soybean cultivar "Forrest" was developed for Targeting Induced Local Lesions IN Genomes (TILLING). Additionally, due to the complexity of the soybean genome, an integrated set of genomic and genetic analysis tools was employed to complement the TILLING approach. The efficiency of this integrated set of tools was tested using a candidate soybean gene for resistance to SCN, encoding a leucine-rich repeat receptor-like kinase (LRR-RLK) that was identified by map-based cloning at the Rhg4 locus. The Rhg4 locus is one of the major quantitative trait loci controlling soybean resistance against SCN race 3 (HG type 0) in cv. Forrest, but the gene(s) sequence for resistance remains to be determined. Using TILLING, a Forrest mutant containing a nonsense mutation in the LRR domain of the candidate resistance protein was identified and confirmed; however, the SCN-resistant phenotype of the mutant was not altered. Haplotyping and EcoTILLING of recombinant inbred lines along with complementation analysis corroborated the TILLING result and ruled out the possibility of functional redundancy by a second copy of the LRR-RLK gene identified in the soybean genome. This study validates the use of TILLING, in combination with an integrated set of genomic tools, as an efficient means of testing candidate genes for SCN resistance in soybean.
authors.affiliation: Division of Plant Sciences and Bond Life Sciences Center, University of Missouri, Columbia, MO 65211, USA.
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Liu
authors.forename: Shiming
authors.initials: S
authors.lastname: Liu
authors.forename: Aziz
authors.initials: A
authors.lastname: Jamai
authors.forename: Abdelhafid
authors.initials: A
authors.lastname: Bendahmane
authors.forename: David A
authors.initials: DA
authors.lastname: Lightfoot
authors.forename: Melissa G
authors.initials: MG
authors.lastname: Mitchum
authors.forename: Khalid
authors.initials: K
authors.lastname: Meksem
mesh_terms: D000818:Animals; D001483:Base Sequence; D060467:Disease Resistance; D056726:Genetic Association Studies; D005816:Genetic Complementation Test; D056426:Genetic Loci; D006239:Haplotypes; D007178:Inbreeding; D008969:Molecular Sequence Data; D009154:Mutation; D010641:Phenotype; D010935:Plant Diseases / Q000235:genetics / Q000276:immunology / Q000469:parasitology; D010940:Plant Proteins / Q000235:genetics*; D020641:Polymorphism, Single Nucleotide; D011494:Protein Kinases / Q000235:genetics*; D017422:Sequence Analysis, DNA; D013025:Glycine max / Q000235:genetics / Q000276:immunology*; D014431:Tylenchoidea
chemical_list: D010940:Plant Proteins; D011494:Protein Kinases