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农学学报

农学学报 ›› 2024, Vol. 14 ›› Issue (1): 39-43.doi: 10.11923/j.issn.2095-4050.cjas2023-0040

• 植物保护 • 上一篇    下一篇

柑橘黄龙病菌亚洲种微滴数字PCR检测方法的建立

宋晓兵1(), 黄峰1, 崔一平1, 彭埃天1, 岳茂峰2   

  1. 1 广东省农业科学院植物保护研究所/农业农村部华南果蔬绿色防控重点实验室/广东省植物保护新技术重点实验室，广州 510640
    2 广东石油化工学院，广东茂名 525000
  • 收稿日期:2023-02-21 修回日期:2023-08-22 出版日期:2024-01-16 发布日期:2024-01-16
  • 作者简介:

    宋晓兵，男，1980年出生，山东胶南人，副研究员，博士，主要从事柑橘病害防控技术研究。通信地址：510640 广州市天河区五山金颖街7号 广东省农业科学院植物保护研究所，Tel：020-87597446，E-mail：。

  • 基金资助:
    “十四五”广东省农业科技创新十大主攻方向“柑橘黄龙病综合防控技术”(2023SDZG06); 广州市重点研发计划项目“柑橘黄龙病早期诊断及监测预警技术研发应用”(2024B03J1317); 茂名市科技计划项目“柑橘黄龙病新型绿色防控技术研发与应用示范”(2023S017084); 广东省现代农业产业技术体系创新团队建设项目“柑橘病害综合防控技术岗位”(2023KJ108)

Establishment of Droplet Digital PCR Detection Method of Candidatus Liberibacter asiaticus

SONG Xiaobing1(), HUANG Feng1, CUI Yiping1, PENG Aitian1, YUE Maofeng2   

  1. 1 Institute of Plant Protection, Guangdong Academy of Agricultural Sciences/Key Laboratory of Green Prevention and Control on Fruits and Vegetables in South China, Ministry of Agriculture and Rural Affairs/Guangdong Provincial Key Laboratory of High Technology for Plant Protection, Guangzhou 510640, Guangdong, China
    2 Guangdong University of Petrochemical Technology, Maoming 525000, Guangdong, China
  • Received:2023-02-21 Revised:2023-08-22 Online:2024-01-16 Published:2024-01-16

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摘要：

柑橘黄龙病是世界柑橘产业的毁灭性病害，带病苗木、带菌接穗和田间病株是病害的初侵染源，建立一套病原菌超灵敏检测方法对柑橘黄龙病的早期预警及防控具有重要意义。根据柑橘黄龙病亚洲种16S rDNA基因序列设计特异引物，建立基于微滴数字PCR技术的检测方法，并评价该方法的灵敏度和检测准确性。结果表明，研究建立的柑橘黄龙病亚洲种微滴数字PCR检测方法特异性强、灵敏度高，其检测灵敏度是荧光定量PCR的10倍。建立的微滴数字PCR检测技术为柑橘种质资源的早期黄龙病检测提供了一种新方法。

关键词: 柑橘, 黄龙病菌亚洲种, ddPCR, 早期检测, 柑橘种苗

Abstract:

Huanglongbing disease is a devastating disease of citrus industry in the world. Diseased seedlings, scions and field diseased plants are the primary infection sources of the disease. The establishment of a set of ultra-sensitive detection method for pathogen is important for the early warning and prevention of Huanglongbing disease. In this study, specific primers were designed according to the 16S rDNA gene sequence of Candidatus Liberibacter asiaticus, and a detection method based on droplet digital PCR technology was explored and established, and the sensitivity and detection accuracy of the method was evaluated. The test results showed that the microdroplet digital PCR detection method for Huanglongbing disease established in this study had strong specificity and high sensitivity, and its detection sensitivity was 10 times that of fluorescence quantitative PCR. The microdroplet digital PCR detection technology established in this study provides a new method for the early detection of Huanglongbing disease in citrus seedlings.

Key words: citrus, Candidatus Liberibacter asiaticus, droplet digital PCR, early detection, citrus seedlings