Piriformospora indica, an endophytic fungus of the Sebacinaceae family, colonizes the roots of a wide variety of plant species and promotes their growth, in a manner similar to arbuscular mycorrhizal fungi. The results of the present study demonstrate that the fungus interacts also with the non‐mycorrhizal host Arabidopsis thaliana and promotes its growth. The interaction is detectable by the appearance of a strong autofluorescence in the roots, followed by the colonization of root cells by fungal hyphae and the generation of chlamydospores. Promotion of root growth was detectable even before noticeable root colonization. Membrane‐associated proteins from control roots and roots after cultivation with P. indica were separated by two‐dimensional gel‐electrophoresis and identified by electrospray ionization mass spectrometry and tandem mass spectrometry. Differences were found in the expression of glucosidase II, beta‐glucosidase PYK10, two glutathione‐S‐transferases and several so‐far uncharacterized proteins. Based on conserved domains present in the latter proteins their possible roles in plant–microbe interaction are predicted. Taken together, the present results suggest that the interaction of Arabidopsis thaliana with P. indica is a powerful model system to study beneficial plant–microbe interaction at the molecular level. Furthermore, the successful accommodation of the fungus in the root cells is preceded by protein modifications in the endoplasmatic reticulum as well as at the plasma membrane of the host.
In mammals, signalling components at the cell surface are clustered in Triton X‐100 insoluble plasma membrane microdomains. We isolated plasma membrane microdomains from Arabidopsis and mustard cotyledons and determined their protein composition by mass spectrometry. Although the protein composition of the plant vesicles differ from the composition of the animal vesicles, they are also enriched in signalling components. We identified at least seven receptor kinases with leucine‐rich repeats, 10 other kinases, the β subunit of heterotrimeric G‐proteins and five small GTP‐binding proteins. Thus, specific signalling components are highly enriched in plant plasma membrane microdomains while others are excluded.
SummaryRoot colonization by the beneficial fungus Piriformospora indica is controlled by plant innate immunity, but factors that channel this interaction into a mutualistic relationship are not known. We have explored the impact of abscisic acid (ABA) and osmotic stress on the P. indica interaction with Arabidopsis thaliana.The activation of plant innate immunity in roots was determined by measuring the concentration of the phytoalexin camalexin and expression of transcription factors regulating the biosynthesis of tryptophan-related defence metabolites. Furthermore, the impact of the fungus on the content of ABA, salicylic acid, jasmonic acid (JA) and JA-related metabolites was examined.We demonstrated that treatment with exogenous ABA or the ABA analogue pyrabactin increased fungal colonization efficiency without impairment of plant fitness. Concomitantly, ABA-deficient mutants of A. thaliana (aba1-6 and aba2-1) were less colonized, while plants exposed to moderate stress were more colonized than corresponding controls. Sustained exposure to ABA attenuated expression of transcription factors MYB51, MYB122 and WRKY33 in roots upon P. indica challenge or chitin treatment, and prevented an increase in camalexin content.The results indicate that ABA can strengthen the interaction with P. indica as a consequence of its impact on plant innate immunity. Consequently, ABA will be relevant for the establishment and outcome of the symbiosis under stress conditions.
The basidiomycete Piriformospora indica interacts with Arabidopsis roots and mimics an arbuscular mycorrhiza. A MATH [meprin and TRAF (tumour necrosis factor receptor‐associated factor) homology] domain‐containing (MATH) protein at the plasma membrane of Arabidopsis roots is one of the first components to respond to the presence of this fungus. MATH proteins are involved in nodule formation in Medicago and protein degradation in the Arabidopsis cytosol. They exhibit sequence similarities to meprins, extracellular peptidases which cleave (signal) peptides, and to TRAFs, intracellular proteins which interact with receptor kinases at the plasma membrane. Fifty‐nine genes for MATH proteins are present in the Arabidopsis genome. Members of this protein family are predicted to be found in the ER–plasma membrane–extracellular space continuum, in the nucleus–cytosol compartment and in organelles. In this article, we describe this novel class of plant genes. We also use MS‐MS analyses to identify the subcellular localization of individual members of the MATH protein family in Arabidopsis thaliana.
The role of the heterotrimeric G-protein beta-subunit in plant development was studied in transgenic tobacco (Nicotiana tabacum L.) plants with reduced beta-subunit levels due to the antisense expression of the beta-subunit mRNA. The antisense plants had aberrant anther shape and produced non-germinating pollen. The anthers were sporadically transformed to petals, whereas other floral organs were not affected. The pollen grains were smaller than the wild-type pollen and had abnormal cell walls. The architecture of mature antisense plants was altered. The plants had long branched panicles and short stems. These data suggest that the beta-subunit of the plant heterotrimeric G-proteins is involved in the regulation of the reproductive phase of the tobacco life cycle, particularly in stamen development and pollen maturation.
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