NAD1 Controls Defense-Like Responses in Medicago truncatula Symbiotic Nitrogen Fixing Nodules Following Rhizobial Colonization in a BacA-Independent Manner.

Ágota Domonkos,Gabriella Endre,Ferhan Ayaydin,Mónika Tóth,Beatrix Horváth,Szilárd Kovács,Attila Farkas,Lili Fodor,Péter Kaló,Károly Bóka,Pascal Ratet,Ernő Kiss,Attila Kereszt,Gyöngyi Kováts,Anikó Gombár

doi:10.3390/genes8120387

Abstract

Legumes form endosymbiotic interaction with host compatible rhizobia, resulting in the development of nitrogen-fixing root nodules. Within symbiotic nodules, rhizobia are intracellularly accommodated in plant-derived membrane compartments, termed symbiosomes. In mature nodule, the massively colonized cells tolerate the existence of rhizobia without manifestation of visible defense responses, indicating the suppression of plant immunity in the nodule in the favur of the symbiotic partner. Medicago truncatula DNF2 (defective in nitrogen fixation 2) and NAD1 (nodules with activated defense 1) genes are essential for the control of plant defense during the colonization of the nitrogen-fixing nodule and are required for bacteroid persistence. The previously identified nodule-specific NAD1 gene encodes a protein of unknown function. Herein, we present the analysis of novel NAD1 mutant alleles to better understand the function of NAD1 in the repression of immune responses in symbiotic nodules. By exploiting the advantage of plant double and rhizobial mutants defective in establishing nitrogen-fixing symbiotic interaction, we show that NAD1 functions following the release of rhizobia from the infection threads and colonization of nodule cells. The suppression of plant defense is self-dependent of the differentiation status of the rhizobia. The corresponding phenotype of nad1 and dnf2 mutants and the similarity in the induction of defense-associated genes in both mutants suggest that NAD1 and DNF2 operate close together in the same pathway controlling defense responses in symbiotic nodules.

Highlights

Medicago truncatula and other leguminous plants are able to establish nitrogen-fixing symbiotic associations with soil bacteria that are belonging to the genus rhizobia
The brown pigmentation associated with strong autofluorescence that appeared after inoculation suggested the induction of defense responses in 7Y mutant nodules
Nodules of nad1-3 (B,G), nad1-4 (C,H), nad1-3/dnf2 (D,I), and dnf2 (E,J) displayed extensive brown pigmentation that corresponded to the area showing autofluorescence which is pseudocolored in red

Summary

Introduction

Medicago truncatula and other leguminous plants are able to establish nitrogen-fixing symbiotic associations with soil bacteria that are belonging to the genus rhizobia. The symbiotic interaction between the rhizobia and the host plant is initiated by exchange of chemical signals between the two partners resulting in the formation the root nodules, wherein nitrogen fixation takes place [1,2]. Plant flavonoids secreted into the rhizosphere induce the production of the bacterial signal molecule, the nodulation factor (NF) that triggers morphological changes, root hair curling and cortical cell division, and physiological responses, such as ion fluxes, calcium oscillations, and transcriptional activation of symbiosis associated genes [1], in the host plant. Rhizobia are required for the initiation and growth of infection threads (ITs) that develop towards the newly formed nodule primordia by penetrating the outer cortical cell layers [3]. The bacteria are accommodated in a cytoplasmic structure referred to as the symbiosome that is delimited by a plant derived peribacteroid membrane

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