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Journal of Virology, May 2004, p. 5382-5389, Vol. 78, No. 10
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.10.5382-5389.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Rice Dwarf Phytoreovirus Segment S6-Encoded Nonstructural Protein Has a Cell-to-Cell Movement Function

Yi Li,1* Yi M. Bao,2,3 Chun H. Wei,1 Zhen S. Kang,4 Yong W. Zhong,1 Peng Mao,1 Gang Wu,1 Zhang L. Chen,1 Joachim Schiemann,5 and Richard S. Nelson2*

Peking-Yale Joint Center for Plant Molecular Genetics and Agrobiotechnology, The National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing 100871,1 Northwestern Agriculture and Forestry University, Department of Plant Protection, Shaanxi, Yangling, 712100, China,4 Plant Biology Division, Samuel Roberts Noble Foundation, Inc., Ardmore, Oklahoma 73401,2 National Center for Biotechnology Information, National Institutes of Health, Bethesda, Maryland 20892-6510,3 Federal Biological Research Centre for Agriculture and Forestry, Institute for Biochemistry and Plant Virology, D-38104 Braunschweig, Germany5

Received 8 August 2003/ Accepted 21 November 2003

Rice dwarf virus (RDV) is a member of the genus Phytoreovirus, which is composed of viruses with segmented double-stranded RNA genomes. Proteins that support the intercellular movement of these viruses in the host have not been identified. Microprojectile bombardment was used to determine which open reading frames (ORFs) support intercellular movement of a heterologous virus. A plasmid containing an infectious clone of Potato virus X (PVX) defective in cell-to-cell movement and expressing either ß-glucuronidase or green fluorescent protein (GFP) was used for cobombardment with plasmids containing ORFs from RDV gene segments S1 through S12 onto leaves of Nicotiana benthamiana. Cell-to-cell movement of the movement-defective PVX was restored by cobombardment with a plasmid containing S6. In the absence of S6, no other gene segment supported movement. Identical results were obtained with Nicotiana tabacum, a host that allows fewer viruses to infect and spread within its tissue. S6 supported the cell-to-cell movement of the movement-defective PVX in sink and source leaves of N. benthamiana. A mutant S6 lacking the translation start codon did not complement the cell-to-cell movement of the movement-defective PVX. An S6 protein product (Pns6)-enhanced GFP fusion was observed near or within cell walls of epidermal cells from N. tabacum. By immunocytochemistry, unfused Pns6 was localized to plasmodesmata in rice leaves infected with RDV. S6 thus encodes a protein with characteristics identical to those of other viral proteins required for the cell-to-cell movement of their genome and therefore is likely required for the cell-to-cell movement of RDV.

* Corresponding author. Mailing address for Yi Li: Peking-Yale Joint Center for Plant Molecular Genetics and Agrobiotechnology, The National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing 100871, China. Phone: 86-10-62759690. Fax: 86-10-62754427. E-mail: liyi{at}pku.edu.cn. Mailing address for Richard S. Nelson: Plant Biology Division, The Samuel Roberts Noble Foundation, Inc., 2510 Sam Noble Parkway, Ardmore, OK 73401. Phone: (580) 224-6600. Fax: (580) 224-6692. E-mail: rsnelson{at}noble.org.

Journal of Virology, May 2004, p. 5382-5389, Vol. 78, No. 10
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.10.5382-5389.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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