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Arbuscular mycorrhizal (AM) fungi are soil-borne fungi belonging to the ancient phylum Glomeromycota and are important symbionts of the arbuscular mycorrhiza, enhancing plant nutrient acquisition and resistance to various abiotic stresses. In contrast to their significant physiological implications, the molecular basis involved is poorly understood, largely due to their obligate biotrophism and complicated genetics. Here, we identify and characterize three genes termed  Fm201 ,  Ri14-3-3  and  RiBMH2  that encode 14-3-3-like proteins in the AM fungi  Funneliformis mosseae  and  Rhizophagus irregularis , respectively. The transcriptional levels of  Fm201 ,  Ri14-3-3  and  RiBMH2  are strongly induced in the pre-symbiotic and symbiotic phases, including germinating spores, intraradical hyphae- and arbuscules-enriched roots. To functionally characterize the  Fm201 ,  Ri14-3-3  and  RiBMH2  genes, we took advantage of a yeast heterologous system owing to the lack of AM fungal transformation systems. Our data suggest that all three genes can restore the lethal  Saccharomyces cerevisiae bmh1 bmh2  double mutant on galactose-containing media. Importantly, yeast one-hybrid analysis suggests that the transcription factor RiMsn2 is able to recognize the STRE (CCCCT/AGGGG) element present in the promoter region of  Fm201  gene. More importantly, Host-Induced Gene Silencing of both  Ri14-3-3  and  RiBMH2  in  Rhizophagus irregularis  impairs the arbuscule formation in AM symbiosis and inhibits the expression of symbiotic  PT4  and  MST2  genes from plant and fungal partners, respectively. We further subjected the AM fungus- Medicago truncatula  association system to drought or salinity stress. Accordingly, the expression profiles in both mycorrhizal roots and extraradical hyphae reveal that these three 14-3-3-like genes are involved in response to drought or salinity stress. Collectively, our results provide new insights into molecular functions of the AM fungal 14-3-3 proteins in abiotic stress responses and arbuscule formation during AM symbiosis.

Arbuscular Mycorrhizal Fungal 14-3-3 Proteins Are Involved in Arbuscule Formation and Responses to Abiotic Stresses During AM Symbiosis