Plant Biotechnology
EJB Electronic Journal of Biotechnology ISSN: 0717-3458 Vol.2 No.1, Issue of April 15, 1999.
© 1999 by Universidad Católica de Valparaíso -- Chile Received 17 November, 1998 / Accepted 23 December, 1998
RESEARCH ARTICLE

Sequence variability in p27 gene of Citrus Tristeza Virus (CTV) revealed by SSCP analysis

Selma Gago-Zachert
Instituto de Bioquímica y Biología Molecular, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, La Plata, Argentina.
E-mail: selma_gz@biol.unlp.edu.ar

Norma Costa
EEA INTA-Concordia, Concordia, Argentina
E-mail: ncosta@concordia.com.ar

Liliana Semorile*
Departamento de Ciencia y Tecnología, Universidad Nacional de Quilmes, Roque Saenz Pena 180, 1876 Bernal, Argentina.
Tel: (54-1) 365-7161, Fax: (54-1) 365-7132
E-mail:
lsemorile@unq.edu.ar

Oscar Grau
Instituto de Bioquímica y Biología Molecular, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, La Plata, Argentina.
Email: grau@biol.unlp.edu.ar

*Corresponding author

Keywords: Citrus tristeza virus (CTV), p27 gene, SSCP

Abstract Full Text

Citrus tristeza closterovirus (CTV), is a phloem-limited virus transmitted by aphids in a semipersistent manner. The genome of CTV is composed of a ssRNA with two capsid proteins: CP, covering about 95% of the particle length, and a diverged coat protein (dCP), present only in one end of the particle, forming a rattlesnake structure. dCP is the product of p27 gene for which it is also postulated a function in the transmissibility by aphid vectors.

Hybridization analysis showed a p27 gene region, which exhibits different patterns with two probes derived from two biological distinct CTV isolates. In an attempt to screen whether that gene region differs in mild and severe strains, six CTV isolates belonging to different biogroups were compared for variations in their p27 gene by analysis of single-strand conformation polymorphism (SSCP). The p27 gene was reverse transcribed and amplified by PCR and thirty clones of each isolate were obtained. From each clone, two fragments of the gene were amplified by PCR: fragment (a), 459 bp long, and fragment (b), 281 bp long. Sequence variations in both gene fragments were studied by SSCP analysis. A variety of SSCP patterns was obtained from each isolate, being isolates belonging to the groups II-IV and III those with the higher and lower number of them. Moreover, SSCP analysis provided a rapid procedure to screen the genetic heterogeneity of the viral isolates reducing considerably the amount of nucleic acid sequenciation necessary to gain that knowledge.

Supported by UNESCO / MIRCEN network
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