植物保护学报2025，Vol.52Issue(4)：852-858,7.DOI:10.13802/j.cnki.zwbhxb.2025.2025034
枣疯病植原体巢式PCR检测方法的建立及应用
Establishment and application of a nested PCR method for the dection of Candidatus Phytoplasma ziziphi causing jujube witches'broom
摘要
Abstract
To establish an efficient and sensitive molecular method for the detection of Candidatus Phy-toplasma ziziphi causing jujube witches'broom,specific primers were designed and screened using phytoplasma secA gene as the target sequence.A nested PCR method with high specificity and sensitiv-ity for the detection of Ca.P.ziziphi was developed,and a total of 34 jujube samples were tested.Results showed that,the nested PCR method based on two pairs of specific primers could accurately detect Ca.P.ziziphi from jujube samples.The sensitivity of this nested PCR detection method for the phy-toplasma was 3×10-4 ng/μL,which was 1 000 times more sensitive than that of conventional PCR.The detection rate of this method for 16 symptomatic and 18 asymptomatic jujube samples was 100.0%and 66.7%,respectively,which was significantly higher than that of conventional PCR.These findings indi-cated that the established nested PCR method for Ca.P.ziziphi has strong specificity,high sensitivity and stability,making it suitable for monitoring,early warning,and scientific prevention of jujube witches'broom.关键词
枣疯病/枣疯病植原体/secA基因/巢式PCR/病原检测/灵敏度/特异性Key words
jujube witches'broom/Candidatus Phytoplasma ziziphi/secA gene/nested PCR/patho-gen detection/sensitivity/specificity引用本文复制引用
赵娟,程艳丽,刘亚勇,孟盼盼,王甦,秦文韬..枣疯病植原体巢式PCR检测方法的建立及应用[J].植物保护学报,2025,52(4):852-858,7.基金项目
国家桃产业技术体系枣植保技术岗位科学家(CARS-30),北京市农林科学院青年科研基金(QNJJ202313),北京市农林科学院创新能力建设专项(KJCX20240405) （CARS-30）
