论文部分内容阅读
脂质转移蛋白是植物生命活动中重要的活性蛋白质,在花粉发育中起着重要作用。本研究以青花菜保持系‘WN12-95B’为试验材料,采用RT-PCR技术获得脂质转移蛋白(LTP)基因c DNA序列。通过生物信息学分析,结果表明:该基因含有一个360 bp的开放阅读框,推导编码119个氨基酸,预测蛋白分子量12.47 k D,p I值7.48,属疏水性蛋白。青花菜LTP蛋白氨基酸序列含有24个氨基酸长度的信号肽,α-螺旋和不规则卷曲是其二级结构的主要成分,三级结构则具有典型的LTP蛋白结构特征。分子进化分析表明该基因与油菜亲缘关系最近,相似性最高,其次为芥菜和芜菁。时空表达特性分析显示,青花菜LTP基因特异表达于花蕾中,且表达丰度较高。保持系和Ogu不育系不同发育阶段花蕾LTP基因的表达丰度差异明显,表达量随花蕾发育呈下降趋势。本研究可为深入研究LTP基因在青花菜绒毡层和小孢子发育过程中的作用提供一定的理论基础。
Lipid transfer protein is an important active protein in plant life activities and plays an important role in pollen development. In this study, the broccoli maintainer line ’WN12-95B’ was used as the experimental material, and the cDNA sequence of the lipotransfer protein (LTP) gene was obtained by RT-PCR. The bioinformatics analysis showed that the gene contained a 360 bp open reading frame and deduced a protein of 119 amino acids. The predicted molecular weight was 12.47 kD, and the p I value was 7.48, which was a hydrophobic protein. The amino acid sequence of LTP protein in broccoli contains a signal peptide of 24 amino acids in length. Alpha-helix and irregular curl are the main components of its secondary structure. The tertiary structure has the typical LTP protein structure. Molecular phylogenetic analysis showed that this gene has the closest genetic relationship with rape, the highest similarity, followed by mustard and turnip. Analysis of spatiotemporal expression showed that LTP was expressed specifically in flower buds and its expression abundance was higher. There were significant differences in the expression abundance of LTP gene in flower bud between maintainer line and Ogu CMS at different developmental stages. The expression level of LTP decreased with the bud development. This study may provide some theoretical basis for further study on the role of LTP gene in the development of tapetum and microspore in broccoli.