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ACTA HORTICULTURAE SINICA ›› 2017, Vol. 44 ›› Issue (3): 487-494.doi: 10.16420/j.issn.0513-353x.2016-0733

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Serological Detection and Molecular Identification of Tomato spotted wilt virus in Pepper in Chongqing

SUN Miao1,JING Chenchen1,CHU Chengru1,WU Gentu1,SUN Xianchao1,XIE Yan2,LIU Yong3,and QING Ling1,*   

  1. 1College of Plant ProtectionSouthwest UniversityChongqing 400716,China2College of Agriculture and BiotechnologyZhejiang UniversityHangzhou310058,China3Institute of Plant ProtectionHunan ProvinceChangsha 410125,China
  • Online:2017-03-25 Published:2017-03-25

Abstract:

To identify the occurrence and variation of the coat protein gene(cp)of Tomato spotted wilt virus(TSWV)in pepper in Chongqing,a total of 298 diseased pepper samples were collected and subjected to the detection of Dot-ELISA with specific antiserum of TSWV and RT-PCR with specific primers. Results of Dot-ELISA detection revealed that 40 samples were positive with a detection ratio of 13.42%. Typical enveloped spherical particles were observed in the TSWV-positive samples through electron microscopy. Six positive samples were randomly selected and amplified by RT-PCR with primers specific for the cp gene of TSWV,and results revealed that an approximate 750 bp fragment was obtained in each sample. Sequencing showed that cp gene of the isolate TSWV-CQ(KX611497)shared 96.1% to 99.4% identities with those of the other 22 previously reported TSWV isolates. Phylogenetic analysis showed that the isolate TSWV-CQ was most closely related to TSWV isolates of China and suggested that geographical selection force played a role in virus evolution.

Key words: pepper, Tomato spotted wilt virus, Dot-ELISA, RT-PCR