Figure 4.

Methylation and Transcription Analysis of Genes Associated with the Barley Mla Region.

Primers, amplification conditions, and a summary of results are provided in Table 4.

(A) Different patterns of DNA methylation of genes adjacent to and within the Mla locus. Positive amplification occurs if a particular 5′-C/CGG-3′ site is methylated and cannot be cut by the restriction enzyme. Likewise, amplification cannot occur if the particular 5′-C/CGG-3′ site is not methylated and is digested to completion by the restriction enzyme.

(B) RT-PCR analysis of predicted genes adjacent to and within the Mla locus. The templates in each one-step RT-PCR analysis are shown at top. RNA or DNA indicates the use of RNA or genomic DNA as a template for the reaction. In the analysis of each gene, the first lane shows the result of the RT-PCR. The second and the third lanes represent a DNA-free RNA negative control without RT and a positive DNA control, respectively.