Protein pattern changes induced by culture filtrate of Pseudomonas syringae pv. phaseolicola in bean (Phaseolus vulgaris L.) explants cultured in vitro

Issue		Agron. Sustain. Dev. Volume 20, Number 6, September-October 2000


Page(s)		673 - 682
DOI		10.1051/agro:2000159

DOI: 10.1051/agro:2000159

Agronomie 20 (2000) 673-682

Protein pattern changes induced by culture filtrate of Pseudomonas syringae pv. phaseolicola in bean (Phaseolus vulgaris L.) explants cultured in vitro

Ana Isabel González - Carlos Polanco - María Luisa Ruiz

Area de Genética, Facultad de Biología, Universidad de León, 24071 León, Spain

(Received 29 February 2000; accepted 15 May 2000)

Abstract:

Cotiledonary node explants of two bean cultivars were cultured in vitro in a medium supplemented with filtrate from liquid cultures of race 1 of the bean pathogen Pseudomonas syringae pv. phaseolicola in order to study changes in gene expression induced by elicitors present in the filtrate. The protein patterns of explants from cultivars Palmeña (susceptible to the pathogen race 1) and Red Mexican (resistant to race 1) cultured for 3 and 5 days in filtrate-free and filtrate-supplemented media have been obtained by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and comparison made between them. On the basis of their reproducibility, 550 polypeptides were scored and 58 of them showed differences among the treatments. These differences revealed possible changes in gene expression associated to stress and/or presence of filtrate, and a differential response between the cultivars susceptible and resistant to the pathogen.

Keywords: Phaseolus vulgaris / Pseudomonas syringae pv. phaseolicola / 2-D PAGE / in vitro culture / gene expression

Résumé:

Modifications du patron protéique induites par le filtrat de culture de Pseudomonas syringae pv. phaseolicola chez des explants de haricot (Phaseolus vulgaris L.) cultivés in vitro. Des explants de n $\oe$ uds cotylédonnaires de 2 variétés de haricot ont été cultivés " in vitro '' dans un milieu supplémenté avec un filtrat liquide provenant de la race 1 du pathogène Pseudomonas syringae pv. phaseolicola pour étudier les altérations d'expression des gènes induites par l'éliciteur présent dans le filtrat. Les diagrammes protéiques des explants issus de : Palmeña (sensible à la race 1 du pathogène) et de Red Mexican (résistante à la race 1) cultivés durant 3 et 5 jours dans un milieu sans et avec filtrat bactérien, obtenus par électrophorèse bidimensionnelle (2-D PAGE), ont été comparés entre eux. On a observé 550 polypeptides reproductibles, 58 d'entre eux ont montré des différences entre les différents traitements qui révèlent des possibles changements dans l'expression des gènes associés au stress et/ou à la présence du filtrat et ce entre les cultures résistantes et sensibles au pathogène.

Mots clé : Phaseolus vulgaris / Pseudomonas syringae pv. phaseolicola / 2-D PAGE / culture in vitro / expression génétique

Correspondence and reprints: María Luisa Ruiz
e-mail: degmrs@unileon.es

Communicated by Nicolás Jouve (Alcalá de Henares, Spain)

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