园艺学报 ›› 2022, Vol. 49 ›› Issue (3): 590-596.doi: 10.16420/j.issn.0513-353x.2021-0114

• 研究报告 • 上一篇    下一篇


贾瑾1, 徐云龙1, 周佳乐1, 杨小芬1, 钟永辉2, 曾继吾3, 洪霓1, 王国平1, 彭抒昂4, 丁芳1,4,*()   

  1. 1华中农业大学植物科学技术学院,湖北省作物病虫害检测与安全控制重点实验室,武汉 430070
    2广东省梅州市梅县区农业科学研究所,广东梅州 514021
    3广东农业科学院果树研究所,广州 510640
    4华中农业大学园艺植物生物学教育部重点实验室,武汉 430070
  • 收稿日期:2021-06-22 修回日期:2021-12-23 出版日期:2022-03-25 发布日期:2022-03-25
  • 通讯作者: 丁芳
  • 基金资助:

Detection and Identification of Huanglongbing Bacteria in‘Cocktail’Grapefruit

JIA Jin1, XU Yunlong1, ZHOU Jiale1, YANG Xiaofen1, ZHONG Yonghui2, ZENG Jiwu3, HONG Ni1, WANG Guoping1, PENG Shu’ang4, DING Fang1,4,*()   

  1. 1Hubei Key Laboratory of Plant Pathology,College of Plant Science and Technology,Huazhong Agricultural University,Wuhan 430070,China
    2Institute of Agricultural Sciences,Meixian District,Meizhou,Guangdong 514021,China
    3Institute of Fruit Trees,Guangdong Academy of Agricultural Sciences,Guangzhou 510640,China
    4Key Laboratory of Horticultural Plant Biology,Ministry of Education,Huazhong Agricultural University,Wuhan 430070,China
  • Received:2021-06-22 Revised:2021-12-23 Online:2022-03-25 Published:2022-03-25
  • Contact: DING Fang


采用直接组织免疫印迹(Direct tissue blot immunoassay,DTBIA)和常规PCR技术,对采自广东梅州新建‘Cocktail’葡萄柚园内表现疑似黄龙病症状的样品进行检测。DTBIA检测发现疑似样品的韧皮部存在特征性紫红色反应,与黄龙病阳性对照反应一致。PCR检测发现枝条、叶片和果实样品中均检测出了黄龙病菌特异性条带,其中果蒂、中柱和种皮内病菌含量较高。进一步测序分析发现,来自‘Cocktail’葡萄柚病样的16S rDNA和efTu基因与NCBI中的柑橘黄龙病菌亚洲种(Candidatus Liberibacter asiaticus,CLas)对应基因的序列相似性均为100%,证实‘Cocktail’葡萄柚感染了柑橘黄龙病菌亚洲菌株。

关键词: 葡萄柚, 柑橘黄龙病菌, 检测, 鉴定, DTBIA, 16S rDNA, efTu, 序列分析


DTBIA and regular PCR were both adopted to identify Huanglongbing(HLB)bacteria in the samples showing symptoms similar to HLB in orchards in Meizhou,Guangdong. Typical purple spots were observed in the phloem of leaf petioles and branches of cocktail grapefruit by DTBIA assay,similar as those produced in HLB positive control. Further analysis with branch,leaf,fruit peduncle,columella,seed coat,and ovary by PCR revealed positive target bands were got from all the above samples. The titer of Candidatus Liberibacter asiaticus in fruit peduncle,columella,and seed coat was relatively higher as compared to other parts. 16S rDNA and efTu gene were both cloned and sequenced. The sequence identity between isolates of CLas deposited in GenBank and the ones from‘Cocktail’grapefruit was 100%. In conclusion,the pathogen infecting‘Cocktail’grapefruit was identified as CLas.

Key words: grapefruit, Candidatus Liberibacter asiaticus, detection, identification, DTBIA, 16S rDNA, efTu, sequence analysis