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Journal of Virology, June 2003, p. 6785-6798, Vol. 77, No. 12
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.12.6785-6798.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Tomato Yellow Leaf Curl Sardinia Virus Rep-Derived Resistance to Homologous and Heterologous Geminiviruses Occurs by Different Mechanisms and Is Overcome if Virus-Mediated Transgene Silencing Is Activated

Alessandra Lucioli,1 Emanuela Noris,2 Angela Brunetti,1 Raffaela Tavazza,1 Valentino Ruzza,1 Araceli G. Castillo,3 Eduardo R. Bejarano,3 Gian Paolo Accotto,2 and Mario Tavazza1*

ENEA C.R. Casaccia, Settore Biotec, 00060 Rome,1 Istituto di Virologia Vegetale, CNR, 10135 Torino, Italy,2 Departamento de Biologia Celular, Genética y Fisiología, Universidad de Malaga, Campus de Teatinos, Malaga 29071, Spain3

Received 10 January 2003/ Accepted 19 March 2003

The replication-associated protein (Rep) of geminiviruses is involved in several biological processes brought about by the presence of distinct functional domains. Recently, we have exploited the multifunctional character of the Tomato yellow leaf curl Sardinia virus (TYLCSV) Rep to develop a molecular interference strategy to impair TYLCSV infection. We showed that transgenic expression of its N-terminal 210 amino acids (Rep-210) confers resistance to the homologous virus by inhibiting viral transcription and replication. We have now used biochemical and transgenic approaches to carry out a fuller investigation of the molecular resistance mechanisms in transgenic plants expressing Rep-210. We show that Rep-210 confers resistance through two distinct molecular mechanisms, depending on the challenging virus. Resistance to the homologous virus is achieved by the ability of Rep-210 to tightly inhibit C1 gene transcription, while that to heterologous virus is due to the interacting property of the Rep-210 oligomerization domain. Furthermore, we present evidence that in Rep-210-expressing plants, the duration of resistance is related to the ability of the challenging virus to shut off transgene expression by a posttranscriptional homology-dependent gene silencing mechanism. A model of Rep-210-mediated geminivirus resistance that takes transgene- and virus-mediated mechanisms into account is proposed.

* Corresponding author. Mailing address: ENEA C.R. Casaccia, Settore Biotec, Via Anguillarese 301, Rome CP2400, Italy. Phone: 39-06-30486373. Fax: 39-06-30484448. E-mail: tavazza_m{at}casaccia.enea.it.

Journal of Virology, June 2003, p. 6785-6798, Vol. 77, No. 12
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.12.6785-6798.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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