Screening Genes Expressed by Rhizobium vitis Inoculation and Salicylic Acid Treatment in Grapevines Using GeneFishing

Abstract

To search for specifically expressed genes during resistant responses against Rhizobium vitis infection from grapevine leaves, the project employed a new and accurate reverse transcription-polymerase chain reaction (RT-PCR) method using annealing control primers (ACPs) to minimize the incidence of false-positive clones. Twenty ACPs were used to identify and sequence 31 differentially expressed genes (DEGs). Basic Local Alignment Search Tool (BLAST) searches revealed that 27 of these genes are known. Of the known genes, six were selected and further characterized using semi-quantitative RT-PCR analysis to assess their specific expression in plants infected with R. vitis. Results of the analysis suggest that the ACP system is a very useful tool for identifying disease resistance-specific genes in grapevines. Further analysis of the cloned differentially expressed genes will provide insights into the molecular basis of disease resistance expression.