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

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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


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

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