Abstract
Primary infection of legumes by rhizobia involves the controlled localized enzymatic breakdown of cell walls at root hair tips. Previous studies determined the role of rhizobial CelC2 cellulase in different steps of the symbiotic interaction Rhizobium leguminosarum-Trifolium repens. Recent findings also showed that CelC2 influences early signalling events in the Ensifer meliloti-Medicago truncatula interaction. Here, we have monitored the root hair phenotypes of two legume plants, T. repens and M. sativa, upon inoculation with strains of their cognate and non-cognate rhizobial species, R. leguminosarum bv trifolii and E. meliloti, (over)expressing the CelC2 coding gene, celC. Regardless of the host, CelC2 specifically elicited ‘hole-on-the-tip’ events (Hot phenotype) in the root hair apex, consistent with the role of this endoglucanase in eroding the noncrystalline cellulose found in polarly growing cell walls. Overproduction of CelC2 also increased root hair tip redirections (RaT phenotype) events in both cognate and non-cognate hosts. Interestingly, heterologous celC expression also induced non-canonical alterations in ROS (Reactive Oxygen Species) homeostasis at root hair tips of Trifolium and Medicago. These results suggest the concurrence of shared unspecific and host-related plant responses to CelC2 during early steps of symbiotic rhizobial infection. Our data thus identify CelC2 cellulase as an important determinant of events underlying early infection of the legume host by rhizobia.
Highlights
The symbiotic relationship between a legume and its compatible rhizobia is a complex and highly regulated process involving early signalling steps for specific recognition between both partners
Previous studies showed that the purified cellulase CelC2 isozyme from wild-type ANU843 degrades the cell wall at the apex of the root hair tip (“Hole on the Tip (HoT)” phenotype) when incubated with intact seedling roots of its compatible host, white clover[10]
CelC2 has two described functions: (i) it modulates the length of cellulose microfibrils, affecting biofilm formation, and (ii) it catalyses the localized hydrolysis of the root cell-wall, thereby promoting primary and secondary infection into the legume host tissues[9,10,12,16,44,45]
Summary
The symbiotic relationship between a legume and its compatible rhizobia is a complex and highly regulated process involving early signalling steps for specific recognition between both partners. We have monitored early responses of Trifolium and Medicago root hairs to inoculation with R. leguminosarum bv trifolii and E. meliloti wild-type and celC-expressing strains by microscopic and molecular techniques.
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