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Sinorhizobium meliloti-induced chitinase gene expression in Medicago truncatula ecotype R108-1: a comparison between symbiosis-specific class V and defence-related class IV chitinases

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Abstract

The Medicago truncatula (Gaertn.) ecotypes Jemalong A17 and R108-1 differ in Sinorhizobium meliloti-induced chitinase gene expression. The pathogen-inducible class IV chitinase gene, Mtchit 4, was strongly induced during nodule formation of the ecotype Jemalong A17 with the S. meliloti wild-type strain 1021. In the ecotype R108-1, the S. meliloti wild types Sm1021 and Sm41 did not induce Mtchit 4 expression. On the other hand, expression of the putative class V chitinase gene, Mtchit 5, was found in roots of M. truncatula cv. R108-1 nodulated with either of the rhizobial strains. Mtchit 5 expression was specific for interactions with rhizobia. It was not induced in response to fungal pathogen attack, and not induced in roots colonized with arbuscular mycorrhizal (AM) fungi. Elevated Mtchit 5 gene expression was first detectable in roots forming nodule primordia. In contrast to Mtchit 4, expression of Mtchit 5 was stimulated by purified Nod factors. Conversely, Mtchit 4 expression was strongly elevated in nodules formed with the K-antigen-deficient mutant PP699. Expression levels of Mtchit 5 were similarly increased in nodules formed with PP699 and its parental wild-type strain Sm41. Phylogenetic analysis of the deduced amino acid sequences of Mtchit 5 (calculated molecular weight = 41,810 Da, isoelectric point pH 7.7) and Mtchit 4 (calculated molecular weight 30,527 Da, isoelectric point pH 4.9) revealed that the putative Mtchit 5 chitinase forms a separate clade within class V chitinases of plants, whereas the Mtchit 4 chitinase clusters with pathogen-induced class IV chitinases from other plants. These findings demonstrate that: (i) Rhizobium-induced chitinase gene expression in M. truncatula occurs in a plant ecotype-specific manner, (ii) Mtchit 5 is a putative chitinase gene that is specifically induced by rhizobia, and (iii) rhizobia-specific and defence-related chitinase genes are differentially influenced by rhizobial Nod factors and K antigens.

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Abbreviations

AM :

Arbuscular mycorrhiza(l)

BAC :

Bacterial artificial chromosome

bp :

Base pairs

RT–PCR :

Polymerase chain reaction after reverse transcription

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Acknowledgements

We are grateful to Peter Putnoky (University of Pécs, Pécs, Hungary) and Eva Kondorosi (Institut des Sciences du Végétal, CNRS, Gif-sur-Yvette, France) for providing bacterial strains. We also thank our colleagues from the Botanical Institute in Basel, Dirk Redecker, Philipp Raab, and Virgine Galati, for their help in phylogenetic analysis and for technical assistance.

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  1. Christian Staehelin

    Present address: Sciences III, Université de Genève, 30 Quai Ernest-Ansermet, 1211, Genève 4, Switzerland

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  1. Botanisches Institut der Universität Basel, Hebelstrasse 1, 4056, Basel, Switzerland

    Peter Salzer, Nadja Feddermann, Andres Wiemken, Thomas Boller & Christian Staehelin

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  1. Peter Salzer
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Salzer, P., Feddermann, N., Wiemken, A. et al. Sinorhizobium meliloti-induced chitinase gene expression in Medicago truncatula ecotype R108-1: a comparison between symbiosis-specific class V and defence-related class IV chitinases. Planta 219, 626–638 (2004). https://doi.org/10.1007/s00425-004-1268-8

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