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ACTA HORTICULTURAE SINICA ›› 2018, Vol. 45 ›› Issue (8): 1605-1612.doi: 10.16420/j.issn.0513-353x.2018-0093

• New Technologies and New Methods • Previous Articles     Next Articles

Research on the Fast Detection of Candidatus Liberibacter Asiaticus with DTBIA

DING Fang1,2,6,WANG Qinglian2,XU Guoliang1,ZHOU Jiale1,ZENG Jiwu3,YAN Xiang4,XIA Changxiu4,GUO Jun5,LIU Yongzhong2,HONG Ni1,6,WANG Guoping1,6,PENG Shu’ang2,*,and John S HARTUNG7,*   

  1. 1Hubei Key Laboratory of Plant Pathology,Huazhong Agricultural University,Wuhan 430070,China;2Key Laboratory of Horticultural Plant Biology(Huazhong Agricultural University),Ministry of Education,Wuhan 430070,China;3Institute of Fruit Trees,Guangdong Academy of Agricultural Sciences,Guangzhou 510640,China;4Ganzhou Citrus Research Institute,Ganzhou,Jiangxi 341000,China;5Institute of Tropical and Subtropical Cash Crops,Yunnan Academy of Agricultural Sciences,Ruili,Yunnan 678600,China;6State Key Laboratory of Agricultural Microbiology,Huazhong Agricultural University,Wuhan 430070,China;7USDA-ARS-MPPL,MD,20705,USA
  • Online:2018-08-25 Published:2018-08-25

Abstract: Citrus Huanglongbing(HLB)is caused by phloem-limited bacteria named Candidatus Liberibacters. Up to date,despite of all kinds of efforts to get pure culture,HLB bacteria still resist in vitro growth. Fast detection especially the high-throughput methods based on the specificity of serological methods has not yet been developed. In our previous work,we constructed fusion OmpCLas protein and heterogonous expressed to get antigen to produce corresponding polyclonal antibody(Pab). In the present study,we carried out experiment on the assessment of the detection efficiency of Anti-OmpCLas-Pab antibody with direct tissue blot immunoassay(DTIBA). Citrus samples collected from different geographical regions were tested. Different organs of petiole,midrib,stem,root,peduncale and seeds were all analyzed. Detection efficiency of both DTBIA and PCR were compared. Our result indicated:Anti- OmpCLas-Pab could specifically recognize HLB bacteria in different isolates originated from different geographical areas in China,including samples from Jiangxi,Fujian,Yunnan,Guangdong and Hainan provinces. Tests with different tissues of petiole,leaf mid rib,stem,root,peduncale and seeds revealed the presence of HLB bacteria. However,less HLB bacteria were found in roots and seeds;higher titer was found in leaf mid-ribs and stems. Our results indicated that Anti-OmpCLas-Pab is a promising antibody for the fast detection of HLB bacteria which laid a basic foundation for the product development on the fast detection kits and dipsticks.

Key words: Citrus, HLB bacteria, fast detection, DTBIA, polyclonal antibody, specificity

CLC Number: