The identification of candidate genes associated with Pch2 eyespot resistance in wheat using cDNA-AFLP |
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Authors: | Natalie H Chapman Christopher Burt Paul Nicholson |
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Institution: | (1) John Innes Centre, Norwich Research Park, Colney, Norwich, NR4 7UH, UK;(2) Present address: Division of Plant Sciences, School of Biosciences, University of Nottingham, Sutton Bonington Campus, Loughborough, LE12 5RD, UK |
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Abstract: | Eyespot is a fungal disease of the stem base of cereal crops and causes lodging and the premature ripening of grain. Wheat
cultivar Cappelle Desprez contains a highly durable eyespot resistance gene, Pch2 on the long arm of chromosome 7A. A cDNA-amplified fragment length polymorphism (AFLP) platform was used to identify genes
differentially expressed between the eyespot susceptible variety Chinese Spring (CS) and the CS chromosome substitution line
Cappelle Desprez 7A (CS/CD7A) which contains Pch2. Induced and constitutive gene expression was examined to compare differences between non-infected and plants infected with
Oculimacula acuformis. Only 34 of approximately 4,700 cDNA-AFLP fragments were differentially expressed between CS and CS/CD7A. Clones were obtained
for 29 fragments, of which four had homology to proteins involved with plant defence responses. Fourteen clones mapped to
chromosome 7A and three of these mapped in the region of Pch2 making them putative candidates for involvement in eyespot resistance. Of particular importance are two fragments; 4CD7A8
and 19CD7A4, which have homology to an Oryza sativa putative callose synthase protein and a putative cereal cyst nematode NBS-LRR disease resistance protein (RCCN) respectively.
Differential expression associated with Pch2 was examined by semi-quantitative RT-PCR. Of those genes tested, only four were differentially expressed at 14 days post
inoculation. We therefore suggest that a majority of the differences in the cDNA-AFLP profiles are due to allelic polymorphisms
between CS and CD alleles rather than differences in expression.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. |
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