“花期施钙”是梨生产的一项重要农艺措施，因为花粉胞质钙离子是其萌发和生长的决定性因子。外源的钙离子主要通过质膜钙通道进入胞内。近年来，申请人已在梨花粉质膜上检测到多种钙通道，但其编码基因尚不清楚。谷氨酸类受体通道是一类重要的钙通道，在植物中广泛存在。基于梨全基因组序列分析，申请人发现梨谷氨酸类受体通道基因家族共有34个成员，其中PbrGLR3.3表达量随着花粉管的生长，呈现先上升后下降的现象。本研究中，申请人拟综合利用膜片钳、激光共聚焦显微镜、反义寡核苷酸基因抑制表达等前沿技术方法，开展PbrGLR3.3的克隆与功能分析，PbrGLR3.3配体的鉴定及其介导的钙电流特性解析，以及PbrGLR3.3的花粉胞内互作蛋白鉴定及其调控分析等方面的研究，最终阐明梨花粉管质膜谷氨酸类受体通道PbrGLR3.3介导钙离子内流、调控花粉管生长的信号转导机制，从而为梨“花期施钙”的农艺措施提供理论依据。

Fertilization with calcium ion (Ca2+) in flowering period is an important agronomic measures in pear production. Main reason of this is that the Ca2+ is one of the main factors for controlling pollen germination and pollen tube growth. Exogenous Ca2+ enter in the cytosolic of pollen is mainly through the Ca2+ channel that localized in the plasma membrane. Recent years, we have detected several types of Ca2+ channels in the pollen of pear, however, the encoding genes of these Ca2+ channels are still unknown. The glutamate receptor-like (GLR) channel extensively exists in the plants. Based on analysis of pear whole genome sequence, we found that the pear GLR gene family have 34 members. The expression of PbrGLR3.3 in pollen tube showed after rising to decline phenomenon during pollen tube growth, but the specific function of PbrGLR3.3 need to be further exploration. In this study, we intends to comprehensive utilization of multiple techniques, such as patch clamp, laser confocal microscope, antisense oligonucleotide gene expression suppression, to clone and functional analysis the PbrGLR3.3, identification the ligand of PbrGLR3.3, characterization of calcium current mediated by PbrGLR3.3, and identification the PbrGLR3.3 interaction protein in the intracellular of pollen. Finally, we want to clarify the signal transduction mechanisms of PbrGLR3.3 mediation Ca2+ influx in the pear pollen tube and controlling their growth, and provide the theoretical basis for the agronomic measures of fertilization with Ca2+ in flowering period.