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- Title
Isolation, characterization, and development of WRKY genes as useful genetic markers in Theobroma cacao.
- Authors
Borrone, James W; Kuhn, David N; Schnell, Raymond J
- Abstract
There is currently an international effort in improving disease resistance and crop yield in Theobroma cacao L., an economically important crop of the tropics, using marker-assisted selection for breeding. We are developing molecular genetic markers focusing upon gene families involved with disease resistance. One such family is the WRKY proteins, which are plant-specific transcriptional factors associated with regulating defense responses to both abiotic and biotic stresses. Degenerate PCR primers were designed to the highly conserved DNA-binding domain and other conserved motifs of group I and group II, subgroups a-c, WRKY genes. Sixteen individual WRKY fragments were isolated from a mixture of T. cacao DNA using one pair of primers. Of the 16 WRKY loci investigated, seven contained single nucleotide polymorphisms within the intron as detected by sequence comparison of the PCR products. Four of these were successfully converted into molecular markers and mapped in an F2 population by capillary electrophoresis-single strand conformation polymorphism analysis. This is the first report of a pair of degenerate primers amplifying WRKY loci directly from genomic DNA and demonstrates a simple method for developing useful genetic markers from members of a large gene family.
- Publication
TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik, 2004, Vol 109, Issue 3, p495
- ISSN
0040-5752
- Publication type
Journal Article
- DOI
10.1007/s00122-004-1662-4