Leguminous plants establish endosymbiotic associations with both rhizobia (nitrogen fixation) and arbuscular mycorrhizal fungi (phosphate uptake). These associations involve controlled entry of the soil microsymbiont into the root and the coordinated differentiation of the respective partners to generate the appropriate exchange interfaces. As part of a study to evaluate analogies at the molecular level between these two plant-microbe interactions, we focused on genes from Medicago truncatula encoding putative cell wall repetitive proline-rich proteins (RPRPs) expressed during the early stages of root nodulation. Here we report that a novel RPRP-encoding gene, MtENOD11, is transcribed during preinfection and infection stages of nodulation in root and nodule tissues. By means of reverse transcription-polymerase chain reaction and a promoter-reporter gene strategy, we demonstrate that this gene is also expressed during root colonization by endomycorrhizal fungi in inner cortical cells containing recently formed arbuscules. In contrast, no activation of MtENOD11 is observed during root colonization by a nonsymbiotic, biotrophic Rhizoctonia fungal species. Analysis of transgenic Medicago spp. plants expressing pMtENOD11-gusA also revealed that this gene is transcribed in a variety of nonsymbiotic specialized cell types in the root, shoot, and developing seed, either sharing high secretion/metabolite exchange activity or subject to regulated modifications in cell shape. The potential role of early nodulins with atypical RPRP structures such as ENOD11 and ENOD12 in symbiotic and nonsymbiotic cellular contexts is discussed.

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This paper will review work mainly done during the last twenty years on the involvement of phytoalexin and phenolic compounds in mycorrhizal interactions. It has been observed that phytoalexins and associated molecules accumulate in roots after mycorrhizal infection, but less intensively and more slowly than in pathogenic interactions. Following mycorrhizal infection, enzymes of phenylpropanoid metabolism have been shown to be activated differentially. Some flavonoids and isoflavonoids have been reported to stimulate in vitro germination of mycorrhizal fungi or in vitro mycorrhizal infection, but their biological significance in signalling between the two symbiotic partners, and in biocontrol of plant disease by arbuscular mycorrhizal fungi, have not yet been elucidated.

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BackgroundThe purpose of this project was to identify metabolites, proteins, genes, and promoters associated with water stress responses in soybean. A number of these may serve as new targets for the biotechnological improvement of drought responses in soybean (Glycine max).ResultsWe identified metabolites, proteins, and genes that are strongly up or down regulated during rapid water stress following removal from a hydroponics system. 163 metabolites showed significant changes during water stress in roots and 93 in leaves. The largest change was a root-specific 160-fold increase in the coumestan coumestrol making it a potential biomarker for drought and a promising target for improving drought responses. Previous reports suggest that coumestrol stimulates mycorrhizal colonization and under certain conditions mycorrhizal plants have improved drought tolerance. This suggests that coumestrol may be part of a call for help to the rhizobiome during stress. About 3,000 genes were strongly up-regulated by drought and we identified regulators such as ERF, MYB, NAC, bHLH, and WRKY transcription factors, receptor-like kinases, and calcium signaling components as potential targets for soybean improvement as well as the jasmonate and abscisic acid biosynthetic genes JMT, LOX1, and ABA1. Drought stressed soybean leaves show reduced mRNA levels of stomatal development genes including FAMA-like, MUTE-like and SPEECHLESS-like bHLH transcription factors and leaves formed after drought stress had a reduction in stomatal density of 22.34 % and stomatal index of 17.56 %. This suggests that reducing stomatal density may improve drought tolerance. MEME analyses suggest that ABRE (CACGT/CG), CRT/DRE (CCGAC) and a novel GTGCnTGC/G element play roles in transcriptional activation and these could form components of synthetic promoters to drive expression of transgenes. Using transformed hairy roots, we validated the increase in promoter activity of GmWRKY17 and GmWRKY67 during dehydration and after 20 μM ABA treatment.ConclusionsOur toolbox provides new targets and strategies for improving soybean drought tolerance and includes the coumestan coumestrol, transcription factors that regulate stomatal density, water stress-responsive WRKY gene promoters and a novel DNA element that appears to be enriched in water stress responsive promoters.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-016-2420-0) contains supplementary material, which is available to authorized users.

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From a pool of Medicago truncatula mutants--obtained by gamma-irradiation or ethyl methanesulfonate mutagenesis--impaired in symbiosis with the N-fixing bacterium Sinorhizobium meliloti, new mutants are described and genetically analysed, and for already reported mutants, complementary data are given on their phenotypic and genetic analysis. Phenotypic data relate to nodulation and mycorrhizal phenotypes. Among the five new mutants, three were classified as [Nod+ Fix- Myc+] and the mutations were ascribed to two loci, Mtsym20 (TRV43, TRV54) and Mtsym21 (TRV49). For the two other new mutants, one was classified as [Nod-/+ Myc+] with a mutation ascribed to gene Mtsym15 (TRV48), and the other as [Nod- Myc-/+] with a mutation ascribed to gene Mtsym16 (TRV58). Genetic analysis of three previously described mutants has shown that [Nod-/+ Myc+] TR74 mutant can be ascribed to gene Mtsym14, and that [Nod-/+ Myc-/+] TR89 and TRV9 mutants are ascribed to gene Mtsym2 (dmi2). Using a detailed analysis of mycorrhizal phenotype, we have observed a delayed typical arbuscular mycorrhizal formation on some mutants that present thick lens-shaped appressoria. This phenotype was called [Myc-/+] and mutants TR25, TR26, TR89, TRV9, P1 and Y6 were reclassified as [Myc-/+]. Mutant P1 was reclassified as [Nod-/+] because of a late nodulation observed on roots of this mutant.

SUMMARYThe ultrastructural organization and some cytochemical features (protein and polysaccharide distribution) of the mycorrhiza formed by Glomus tenuis in raspberry roots have been investigated. Certain aspects of the fine mycorrhizal endophyte (smaller hyphae, thinner walls, distinct two-layered wall structure following the PATAg test for polysaccharides, complete absence of septa) distinguish it from the coarse vesicular-arbuscular mycorrhizal fungi. The modifications occurring in the host-fungus interface during Glomus tenuis mycorrhiza development are however very similar to those that have been described in several mycorrhizae formed by coarse vesicular-arbuscular endophytes.