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Development of Functional Symbiotic White Clover Root Hairs and Nodules Requires Tightly Regulated Production of Rhizobial Cellulase CelC2

July 2011 , Volume 24 , Number  7
Pages  798 - 807

Marta Robledo,1 José I. Jiménez-Zurdo,2 M. José Soto,2 Encarnación Velázquez,1 Frank Dazzo,3 Eustoquio Martínez-Molina,1 and Pedro F. Mateos1

1Departamento de Microbiología y Genética and Centro Hispano Luso de Investigaciones Agrarias, Universidad de Salamanca, Salamanca, 37185. Spain; 2Estación Experimental del Zaidín, CSIC, Granada, 18008, Spain; 3Department of Microbiology and Molecular Genetics, Michigan State University, East Lansing 48824. U.S.A.


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Accepted 3 March 2011.

The establishment of rhizobia as nitrogen-fixing endosymbionts within legume root nodules requires the disruption of the plant cell wall to breach the host barrier at strategic infection sites in the root hair tip and at points of bacterial release from infection threads (IT) within the root cortex. We previously found that Rhizobium leguminosarum bv. trifolii uses its chromosomally encoded CelC2 cellulase to erode the noncrystalline wall at the apex of root hairs, thereby creating the primary portal of its entry into white clover roots. Here, we show that a recombinant derivative of R. leguminosarum bv. trifolii ANU843 that constitutively overproduces the CelC2 enzyme has increased competitiveness in occupying aberrant nodule-like root structures on clover that are inefficient in nitrogen fixation. This aberrant symbiotic phenotype involves an extensive uncontrolled degradation of the host cell walls restricted to the expected infection sites at tips of deformed root hairs and significantly enlarged infection droplets at termini of wider IT within the nodule infection zone. Furthermore, signs of elevated plant host defense as indicated by reactive oxygen species production in root tissues were more evident during infection by the recombinant strain than its wild-type parent. Our data further support the role of the rhizobial CelC2 cell wall–degrading enzyme in primary infection, and show evidence of its importance in secondary symbiotic infection and tight regulation of its production to establish an effective nitrogen-fixing root nodule symbiosis.



© 2011 The American Phytopathological Society