Search bioRxiv⌕ Search

Biology subjects

Akamatsu, A.

Publications and source records attributed to Akamatsu, A..

2 recordsLinked to original sources

Rhizobial infection-specific accumulation ofphosphatidylinositol 4,5-bisphosphate inhibits the excessive infection of rhizobia in Lotus japonicus

O_LIDuring the symbiosis of legume with nitrogen-fixing bacteria, collectively called rhizobia, suppression of excessive rhizobial infection by host plants is important to maximize the benefits of symbiotic nitrogen fixation. However, it remains relatively poorly understood the molecular mechanism involved in the suppression. C_LIO_LIWe performed LC-MS and RNA-seq analysis using rhizobia-infected Lotus japonicus roots and investigated the role of phosphatidylinositol (PI) and phosphatidylinositol phosphates (PIPs) in the symbiosis. Phosphatidylinositol transfer protein (PITP)-like proteins 4 (PLP4), phosphatidylinositol 3-phosphate 5-kinase 4 (PIP5K4) and PIP5K6 mutants, which are involved in the vesicular transport of lipids and phosphorylation of PIPs, were used to show the involvement of the signaling of PI and PIPs. Accumulation of phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] during rhizobial infection were examined by a fluorescent marker 1xTUBBY-C (TUBBY). C_LIO_LIWe found that PI signaling-related genes were upregulated, and the amount of PIP2 increased in L. japonicus roots during rhizobial infection. In the PLP4, PIP5K4 and PIP5K6 mutants, rhizobial infection increased and PIP2 accumulation was failed. Furthermore, the observation of PI(4,5)P2 in rhizobia-infected roots revealed that the ectopic accumulation was closely related to suppression of rhizobial infection. C_LIO_LIOur findings indicate that the accumulation of PI(4,5)P2, which is mediated by PLP and PIP5Ks, suppresses excessive rhizobial infection in the root epidermis and cortex, leading to the optimal number of nodules. C_LI

plant biology↗

In vivo monitoring of plant small GTPase activation using a Förster resonance energy transfer biosensor

Small GTPases act as molecular switches that regulate various plant responses such as disease resistance, pollen tube growth, root hair development, cell wall patterning and hormone responses. Thus, to monitor their activation status within plant cells is believed to be the key step in understanding their roles. We have established a plant version of a Forster resonance energy transfer (FRET) probe called Ras and interacting protein chimeric unit (Raichu) that can successfully monitor activation of the rice small GTPase OsRac1 during various defence responses in rice cells. Here, we describe the protocol for visualizing spatiotemporal activity of plant Rac/ROP GTPase in living plant cells, transfection of rice protoplasts with Raichu-OsRac1 and acquisition of FRET images. Our protocol should be widely adaptable for monitoring activation for other plant small GTPases and for other FRET sensors in various plant cells.

plant biology↗