Development of specific and degenerate primers for CesA genes encoding cellulose synthase in flax (Linum usitatissimum L.)

Representatives of the CesA multigene family that control the synthesis of the catalytic subunits of the cellulose synthase complex were described for a number of higher plants. It has been established that the HVR2 region of these genes is class-specific and determines the involvement of the gene product in the synthesis of either the primary or secondary cell wall. The purpose of the current research was to develop degenerate and specific primers for parts of the CesA genes to allow the construction of molecular markers for the class-specific HVR2 region. Two pairs of specific primers for the CesA-1 and CesA-6 genes as well as a pair of degenerate primers for the HVR2 region of all flax CesA genes were developed, based on analysis of the CesA ESTs as well as the full-length cDNA sequences of the CesA genes in Arabidopsis, poplar, maize, and cotton that are available in the GenBank. Fragments of the expected size were amplified using flax cDNA as a template (201 bp for CesA-1, 300 bp for CesA-6, and 600 bp for HVR2). The markers developed in this research can be used for CesA gene cloning and sequencing, analysis of gene copy numbers as well as characterization of tissue- and development specific gene expression.