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Journal of Virology, December 2004, p. 13966-13974, Vol. 78, No. 24
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.24.13966-13974.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

A DNAß Associated with Tomato Yellow Leaf Curl China Virus Is Required for Symptom Induction

Xiaofeng Cui,1,{dagger} Xiaorong Tao,1,{dagger} Yan Xie,1 Claude M. Fauquet,2 and Xueping Zhou1*

Institute of Biotechnology, Zhejiang University, Hangzhou, People's Republic of China,1 International Laboratory for Tropical Agricultural Biotechnology, Danforth Plant Science Center, St. Louis, Missouri2

Received 24 February 2004/ Accepted 8 August 2004

We report here that all 25 isolates of Tomato yellow leaf curl China virus (TYLCCNV) collected from tobacco, tomato, or Siegesbeckia orientalis plants in different regions of Yunnan Province, China, were associated with DNAß molecules. To investigate the biological role of DNAß, full-length infectious clones of viral DNA and DNAß of TYLCCNV isolate Y10 (TYLCCNV-Y10) were agroinoculated into Nicotiana benthamiana, Nicotiana glutinosa, Nicotiana. tabacum Samsun (NN or nn), tomato, and petunia plants. We found that TYLCCNV-Y10 alone could systemically infect these plants, but no symptoms were induced. TYLCCNV-Y10 DNAß was required, in addition to TYLCCNV-Y10, for induction of leaf curl disease in these hosts. Similar to TYLCCNV-Y10, DNAß of TYLCCNV isolate Y64 was also found to be required for induction of typical leaf curl diseases in the hosts tested. When the ßC1 gene of TYLCCNV-Y10 DNAß was mutated, the mutants failed to induce leaf curl symptoms in N. benthamiana when coinoculated with TYLCCNV-Y10. However, Southern blot hybridization analyses showed that the mutated DNAß molecules were replicated. When N. benthamiana and N. tabacum plants were transformed with a construct containing the ßC1 gene under the control of the Cauliflower mosaic virus 35S promoter, many transgenic plants developed leaf curl symptoms similar to those caused by a virus, the severity of which paralleled the level of ßC1 transcripts, while transgenic plants transformed with the ßC1 gene containing a stop codon after the start codon remained symptomless. Thus, expression of a ßC1 gene is adequate for induction of symptoms of viral infection in the absence of virus.

* Corresponding author. Mailing address: Institute of Biotechnology, Zhejiang University, Hangzhou 310029, People’s Republic of China. Phone: 0086 571 86971680. Fax: 0086 571 86971498. E-mail: zzhou{at}zju.edu.cn.

{dagger} These two people contributed equally to this paper.

Journal of Virology, December 2004, p. 13966-13974, Vol. 78, No. 24
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.24.13966-13974.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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