Comparing two microarray platforms for identifying mutated genes in barley (Hordeum vulgare L.).

Area of Science:

• Plant molecular biology
• Genetics and genomics
• Biochemistry

Background:

• Previous work established a cDNA microarray method for identifying mutated barley genes.
• Competitive hybridization was effective in highlighting specific mutated gene clones.

Purpose of the Study:

• To evaluate the suitability of the Affymetrix microarray platform for identifying mutated barley genes.
• To test the previously developed competitive hybridization approach on a new platform.

Main Methods:

• Utilized the Affymetrix barley microarray with 22,792 probe sets.
• Employed two barley mutant strains (xantha-h.57 and xantha-f.27) with known mutations in magnesium chelatase genes.
• Applied competitive hybridization between phenotypically similar mutants.

Main Results:

• The Affymetrix platform successfully identified the arrayed Xantha-h clone, corresponding to a mutation in a magnesium chelatase subunit gene.
• Xantha-f was identified as one of the top fourteen candidate clones, indicating successful detection of another mutation.
• The Affymetrix platform met the requirements for this gene identification approach.

Conclusions:

• The Affymetrix microarray platform is suitable for identifying mutated genes in barley using competitive hybridization.
• This approach effectively highlights specific mutated gene clones, aiding in the discovery of genetic mutations.
• The study confirms the utility of microarrays in dissecting genetic mutations in barley (Hordeum vulgare L.).