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The formation of arbuscular mycorrhizal (AM) symbiosis requires plant root host cells to undergo major structural and functional reprogramming to house the highly branched AM fungal structure for the reciprocal exchange of nutrients. These morphological modifications are associated with cytoskeleton remodelling. However, molecular bases and the role of microtubules (MTs) and actin filament dynamics during AM formation are largely unknown. In this study, the tomato tsb (tomato similar to SB401) gene, belonging to a Solanaceae group of genes encoding MT-associated proteins (MAPs) for pollen development, was found to be highly expressed in root cells containing arbuscules. At earlier stages of mycorrhizal development, tsb overexpression enhanced the formation of highly developed and transcriptionally active arbuscules, while tsb silencing hampers the formation of mature arbuscules and represses arbuscule functionality. However, at later stages of mycorrhizal colonization, tsb overexpressing (OE) roots accumulate fully developed transcriptionally inactive arbuscules, suggesting that the collapse and turnover of arbuscules might be impaired by TSB accumulation. Imaging analysis of the MT cytoskeleton in cortex root cells OE tsb revealed that TSB is involved in MT bundling. Taken together, our results provide unprecedented insights into the role of novel MAP in MT rearrangements throughout the different stages of the arbuscule life cycle.

A Novel Putative Microtubule-Associated Protein Is Involved in Arbuscule Development during Arbuscular Mycorrhiza Formation