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花粉外壁蛋白参与多种植物生理进程,在花粉发育过程中起着重要的作用。本试验以青花菜保持系‘WN12-95B’为材料,利用RT-PCR技术克隆得到青花菜花粉外壁蛋白(PCP)基因。结果显示,该基因全长578 bp其中开放阅读框长度为561 bp共编码186个氨基酸。该蛋白为疏水性蛋白,在第1~24位有一个信号肽序列,预测相对分子量为18.82 kD等电点为10.91。青花菜PCP蛋白的二级结构主要由a-螺旋和不规则卷曲构成,不含β-转角。系统进化分析表明青花菜花粉外壁蛋白与同属植物的进化关系相近,其中与甘蓝进化关系最近。采用qRT-PCR技术分析青花菜PCP基因的表达特性,结果显示该基因仅在花蕾中表达,具有明显的组织特异性。青花菜Ogu不育系及其保持系不同发育阶段花蕾中PCP基因差异明显,表明其在花粉发育中具有重要的作用。本研究结果为进一步研究PCP基因在青花菜花粉发育过程中的作用奠定基础。
Pollen outer wall protein is involved in many plant physiological processes and plays an important role in pollen development. In this study, the broccoli maintainer line ’WN12-95B’ was used as material to clone the bromelain pollen wall protein (PCP) gene by RT-PCR. The results showed that the full length of the gene was 578 bp, of which the open reading frame length of 561 bp encoding a total of 186 amino acids. The protein is a hydrophobic protein with a signal peptide sequence at positions 1 ~ 24 and a predicted relative molecular weight of 18.82 kD with an isoelectric point of 10.91. The secondary structure of PCP protein in broccoli is mainly composed of a-helix and irregular curl, without β-turn. Phylogenetic analysis showed that the evolutionary relationship between broccoli exosomes and the same species was similar, and the relationship between them was closely related to the evolution of cabbage. QRT-PCR analysis of PCP gene expression characteristics of broccoli, the results show that the gene is only expressed in the bud, with significant tissue specificity. The differences of PCP gene in flower buds between Ogu CMS and their maintainer lines at different developmental stages were significant, indicating that they play an important role in pollen development. The results of this study lay the foundation for further study on the role of PCP gene in the development of pollen in broccoli.