LORELEI-LIKE GPI-ANCHORED PROTEINS 2/3 Regulate Pollen Tube Growth as Chaperones and Coreceptors for ANXUR/BUPS Receptor Kinases in Arabidopsis

Abstract

Pollen tube growth is crucial for successful fertilization. In Arabidopsis thaliana, the ANXUR (ANX)/BUPS receptor kinase complex controls and maintains pollen tube growth in response to autocrine rapid alkalinization factor 4/19 (RALF4/19) signaling; however, the molecular and cellular mechanisms underlying the ANX/BUPS-mediated regulation of pollen tube growth remain unclear. In this study, we found that pollen-specific LORELEI-like GPI-anchored proteins 2 and 3 (LLG2/3) promote pollen tube growth in vitro and in vivo. LLG2/3 interacte with ANX/BUPS in a RALF4-concentration-dependent manner, suggesting that ANX/BUPS and LLG2/3 might form a receptor-coreceptor complex for perceiving RALF peptide signals. Disruption of the ANX/BUPS-LLG2/3 interaction led to the cytoplasmic retention of ANX1/2, in either llg2/3 knockdown mutants or in anx1/2 mutants lacking the J region, which mediates the ANX/BUPS-LLG2/3 interaction. Moreover, we found that RALF4 induced the production of reactive oxygen species (ROS), which stimulate pollen tube growth and reduce pollen burst rate. ROS levels are reduced in the pollen tubes of LLG2/3 RNAi lines, and application of exogenous H₂O₂ could partially rescue the defective pollen tube growth of LLG2/3 RNAi lines. Taken together, our study identifies LLG2/3 as novel regulatory components of pollen tube growth, and shows that they chaperone ANX/BUPS for secretion to the apical plasma membrane of pollen tube and act as coreceptors of ANX/BUPS in the activation of ROS production for promoting pollen tube growth.

Keywords: cell wall integrity; reactive oxygen species; reception complex; secretion.