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<i>Rhizobium</i> cellulase CelC2 is essential for primary symbiotic infection of legume host roots

M. Robledo*Departamento de Microbiología y Genética and Centro Hispano Luso de Investigaciones Agrarias, Universidad de Salamanca, 37007 Salamanca, Spain;José I. Jiménez‐ZurdoGrupo de Ecología Genética de la Rizosfera. Estación Experimental del Zaidín, Consejo Superior de Investigaciones Científicas, 18008 Granada, Spain; andEncarna Velázquez*Departamento de Microbiología y Genética and Centro Hispano Luso de Investigaciones Agrarias, Universidad de Salamanca, 37007 Salamanca, Spain;Martha E. Trujillo*Departamento de Microbiología y Genética and Centro Hispano Luso de Investigaciones Agrarias, Universidad de Salamanca, 37007 Salamanca, Spain;José Luis Zurdo‐Piñeiro*Departamento de Microbiología y Genética and Centro Hispano Luso de Investigaciones Agrarias, Universidad de Salamanca, 37007 Salamanca, Spain;Martha-Helena Ramírez-Bahena*Departamento de Microbiología y Genética and Centro Hispano Luso de Investigaciones Agrarias, Universidad de Salamanca, 37007 Salamanca, Spain;Brisa Ramos*Departamento de Microbiología y Genética and Centro Hispano Luso de Investigaciones Agrarias, Universidad de Salamanca, 37007 Salamanca, Spain;J. M. Díaz‐Mínguez*Departamento de Microbiología y Genética and Centro Hispano Luso de Investigaciones Agrarias, Universidad de Salamanca, 37007 Salamanca, Spain;Frank B. DazzoDepartment of Microbiology and Molecular Genetics, Michigan State University, East Lansing, MI 48824Eustoquio Martı́nez-Molina*Departamento de Microbiología y Genética and Centro Hispano Luso de Investigaciones Agrarias, Universidad de Salamanca, 37007 Salamanca, Spain;Pedro F. Mateos*Departamento de Microbiología y Genética and Centro Hispano Luso de Investigaciones Agrarias, Universidad de Salamanca, 37007 Salamanca, Spain;
2008en
ABI

Аннотация

The rhizobia-legume, root-nodule symbiosis provides the most efficient source of biologically fixed ammonia fertilizer for agricultural crops. Its development involves pathways of specificity, infectivity, and effectivity resulting from expressed traits of the bacterium and host plant. A key event of the infection process required for development of this root-nodule symbiosis is a highly localized, complete erosion of the plant cell wall through which the bacterial symbiont penetrates to establish a nitrogen-fixing, intracellular endosymbiotic state within the host. This process of wall degradation must be delicately balanced to avoid lysis and destruction of the host cell. Here, we describe the purification, biochemical characterization, molecular genetic analysis, biological activity, and symbiotic function of a cell-bound bacterial cellulase (CelC2) enzyme from Rhizobium leguminosarum bv. trifolii, the clover-nodulating endosymbiont. The purified enzyme can erode the noncrystalline tip of the white clover host root hair wall, making a localized hole of sufficient size to allow wild-type microsymbiont penetration. This CelC2 enzyme is not active on root hairs of the nonhost legume alfalfa. Microscopy analysis of the symbiotic phenotypes of the ANU843 wild type and CelC2 knockout mutant derivative revealed that this enzyme fulfils an essential role in the primary infection process required for development of the canonical nitrogen-fixing R. leguminosarum bv. trifolii-white clover symbiosis.

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