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糖转运蛋白SWEET作为一种新发现的蛋白在动植物中广泛存在,其对植物花蜜的生产和种子、花粉的发育有重要作用。本研究通过对本课题组获得的新基因MdSWEET1利用生物信息学软件进行分析,并利用实时荧光定量PCR对经低温预处理不同时间段后的苹果花药中该基因的表达量进行分析。生物信息学分析结果表明:MdSWEET1的cDNA中包含342 bp的开放阅读框(ORF),编码113个氨基酸,相对分子质量12.985 kD,等电点8.93,该基因编码蛋白的二级结构主要由无规则卷曲和α-螺旋构成,中间夹杂分布β-折叠;具有三个跨膜结构域,N端位于膜外;序列分析表明MdSWEET1与其他物种中发现的该蛋白氨基酸序列同源性很低。实时荧光定量分析表明,经低温预处理的苹果花药中MdSWEET1基因表达量随着冷处理时间的增加呈下调趋势,由此预测低温预处理阻碍了MdSWEET1基因的表达而影响其功能。该研究为进一步了解MdSWEET1对苹果花粉育性的影响提供理论依据。
As a newly discovered protein, SWEET is widely found in animals and plants and plays an important role in plant nectar production and seed and pollen development. In this study, the new gene MdSWEET1 obtained by our group was analyzed by bioinformatics software, and the expression level of the gene in apple anthers was analyzed by real-time fluorescence quantitative PCR after different time pretreatment. The results of bioinformatics analysis showed that the cDNA of MdSWEET1 contained a 342 bp open reading frame (ORF) encoding 113 amino acids with a relative molecular mass of 12.985 kD and an isoelectric point of 8.93. The secondary structure of this gene was mainly composed of random Curl and α-helix. The three transmembrane domains were located in the N-terminal region of the membrane. The sequence analysis showed that MdSWEET1 shared low homology with the amino acid sequences of other proteins. Real-time fluorescence quantitative analysis showed that MdSWEET1 gene expression in apple anthers pretreated with low temperature showed a downward trend with the increase of cold treatment time, and thus predicted that hypothermic preconditioning hinder the expression of MdSWEET1 gene and affect its function. This study provides a theoretical basis for further understanding of the effect of MdSWEET1 on apple pollen fertility.