Detection of Pectobacterium carotovorum subsp. actinidiae, a Bacterial Pathogen in Yellow Kiwifruit Actinidia chinensis, by PCR

摘要

A bacterial disease characterized by die back or blight symptoms on young cane and often red-rust exudation on cane or trunk during summer and autumn seasons has been occurred in yellow kiwifruit, Actinidia chinensis. The pathogen was identified as Pectobacterium carotovorum subsp. actinidiae. A PCR-based method that utilizes primers to target the carA, carbamoylphosphate synthetase gene, was developed for the rapid and reliable detection of the pathogen. The PCR product amplified with primers specific to the carA gene of enterobacteria and genomic DNA of P. carotovorum subsp. actinidiae was cloned and the nucleotide sequence was determined. On the basis of the sequence of the amplified carA region, a PCR primer pair, PcaF/R, specific to P. carotovorum subsp. actinidiae was designed. This primer pair yielded a 343-bp product from DNA of P. carotovorum subsp. actinidiae, but no products from five other Pectobacterium spp. and eight plant-related bacteria. The PCR assay using PcaF/R primer pair was successfully applied to identify the pathogen isolated from typical disease symptoms.