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Applied and Environmental Microbiology, February 2009, p. 1030-1035, Vol. 75, No. 4
0099-2240/09/$08.00+0     doi:10.1128/AEM.01572-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Pseudomonas savastanoi pv. savastanoi Contains Two iaaL Paralogs, One of Which Exhibits a Variable Number of a Trinucleotide (TAC) Tandem Repeat{triangledown} ,{dagger}

Isabel M. Matas,1 Isabel Pérez-Martínez,1,§ José M. Quesada,2 José J. Rodríguez-Herva,3,{ddagger} Ramón Penyalver,2 and Cayo Ramos1*

Área de Genética, Facultad de Ciencias, Universidad de Málaga, Campus de Teatinos s/n, E-29071 Málaga, Spain,1 Centro de Protección Vegetal y Biotecnología, Instituto Valenciano de Investigaciones Agrarias (IVIA), E-46113 Moncada, Valencia, Spain,2 Departamento de Protección Ambiental, Estación Experimental del Zaidín, Consejo Superior de Investigaciones Científicas, C/Albareda, 1, E-18008 Granada, Spain3

Received 10 July 2008/ Accepted 12 December 2008

In this study, Pseudomonas savastanoi pv. savastanoi isolates were demonstrated to contain two iaaL paralogs, which are both chromosomally located in most strains. Comparative analysis of iaaL nucleotide sequences amplified from these two paralogs revealed that one paralog, iaaLPsn, is 100% identical to iaaL from P. savastanoi pv. nerii, while the other paralog, iaaLPsv, exhibited 93% identity to iaaL from Pseudomonas syringae pv. tomato (iaaLPto). A 3-nucleotide motif (TAC) comprised of 3 to 15 repeats, which remained stable after propagation of the strains in olive plants, was found in iaaLPsv. Based on the observed nucleotide sequence variations, a restriction fragment length polymorphism assay was developed that allowed differentiation among iaaLPsn, iaaLPsv, and iaaLPto. In addition, reverse transcriptase PCR on total RNA from P. savastanoi pv. savastanoi strains demonstrated that both iaaLPsv and iaaLPsn containing 14 or fewer TAC repeats are transcribed. Capillary electrophoresis analysis of PCR-amplified DNA fragments containing the TAC repeats from iaaLPsv allowed the differentiation of P. savastanoi pv. savastanoi isolates.

* Corresponding author. Mailing address: Área de Genética, Facultad de Ciencias, Universidad de Málaga, Campus de Teatinos s/n, E-29071 Málaga, Spain. Phone: 34-952131955. Fax: 34-952132001. E-mail: crr{at}uma.es

{triangledown} Published ahead of print on 19 December 2008.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

§ Present address: Département de Microbiologie Fondamentale (DMF), Quartier UNIL-Sorge, Bâtiment Biophore, CH-1015 Lausanne, Switzerland.

{ddagger} Present address: Centro de Biotecnología y Genómica de Plantas, Universidad Politécnica de Madrid, Campus Montegancedo, E-28223 Pozuelo de Alarcón, Madrid, Spain.

Applied and Environmental Microbiology, February 2009, p. 1030-1035, Vol. 75, No. 4
0099-2240/09/$08.00+0     doi:10.1128/AEM.01572-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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