RESULTS: A polymerase chain reaction restriction fragment length polymorphism capillary electrophoresis (PCR-RFLP-CE) technology-based quarantine approach was developed for 16 Phytophthora species associated with the imported fruits in China. The Ypt1 gene, exhibiting abundant interspecific variations, was selected as the marker gene for PCR. The restriction endonuclease AluI was proven to be capable and compatible in simultaneously separating different Phytophthora species during CE. By combining with a fast and efficient DNA extraction kit, the developed PCR-RFLP-CE technique was successfully applied to identify Phytophthora species in artificially infested fruits.
CONCLUSIONS: We provide a quick, practical, and high-throughput detection approach for hazardous and invasive Phytophthora species associated with imported fruits in China. This strategy can give good convenience and technological support for carrying out massive quarantine activities at Chinese ports. © 2024 Society of Chemical Industry.
结果:开发了一种基于聚合酶链反应限制性片段长度多态性毛细管电泳(PCR-RFLP-CE)技术的检疫方法,对与中国进口水果相关的16种疫霉菌进行了检疫。Ypt1基因,表现出丰富的种间变异,选择作为PCR的标记基因。限制性核酸内切酶AluI被证明能够并且兼容于CE期间同时分离不同的疫霉物种。通过与快速高效的DNA提取试剂盒相结合,开发的PCR-RFLP-CE技术已成功用于鉴定人工侵染水果中的疫霉。
结论:我们提供了一个快速,实用,与中国进口水果相关的有害和入侵疫霉的高通量检测方法。该策略可以为在中国港口开展大规模检疫活动提供良好的便利和技术支持。©2024化学工业学会。