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Acta Horticulturae Sinica ›› 2021, Vol. 48 ›› Issue (8): 1531-1540.doi: 10.16420/j.issn.0513-353x.2020-0914

• Research Papers • Previous Articles     Next Articles

Characterization of the Subcellular Localization and Pathogenicity of P6 Encoded by Cucurbit Chlorotic Yellows Virus

TU Liqin1,2, GAN Shexiang1,3, WU Shuhua1, REN Chunmei1, CHENG Zhaobang1, ZHANG Songbai3, ZHU Yuelin2, ZHOU Yijun1, JI Yinghua1,**()   

  1. 1Institute of Plant Protection,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China
    2College of Horticulture,Nanjing Agricultural University,Nanjing 210095,China
    3College of Agriculture,Yangtze University,Jingzhou,Hubei 434025,China
  • Received:2020-12-17 Revised:2021-05-06 Online:2021-08-25 Published:2021-09-06
  • Contact: JI Yinghua E-mail:jiyinghua@jaas.ac.cn

Abstract:

Cucurbit chlorotic yellows virus(CCYV)is an important whitefly-transmitted Crinivirus,which has become an emerging infectious agent of cucurbits leading to severe disease and significant economic losses. P6 is a small protein encoded by ORF2 of RNA1,one segment of CCYV,and its functions still remains unknown. In this study,P6 was cloned from an isolate of CCYV from Shouguang,Shandong Provience using specific primers by RT-PCR. The results of sequence analysis revealed that P6 gene was 159 bp in length,encoding a protein of approximately 6 kD with a transmembrane domain. To clarify the subcellular localization of P6,a YFP-tagged P6(P6-YFP)was constructed using a 35S:YFP vector and transiently expressed in the Nicotiana benthamiana by agrobacterium infiltration. Fluorescence signal was observed in the cytoplasm and nucleus of the epidermal cells of infiltrated N. benthamiana leaves,which indicated that P6-YFP was distributed in the cytoplasm and nucleus. To test the pathogenicity of P6,a potato virus X(PVX)-based vector was introduced and PVX-P6 was constructed. Chlorotic,crumple and necrosis symptoms were observed in the emerging leaves of N. benthamiana plants inoculated by PVX-P6 and only slight mosaic symptoms was observed in the plant inoculated by PVX, which showed that heterologous expression of P6 in N. benthamiana enhanced the pathogenicity of PVX,indicating that P6 might be an important protein involved in the pathology of CCYV.

Key words: cucurbit chlorotic yellows virus, P6 protein, subcellular localization, pathogenicity

CLC Number: