芹菜韧皮部蛋白基因的分离与表达分析

Isolation of Phloem Protein Gene and Expression Analysis in Celery

韧皮部蛋白在维持植物形态、物质转运以及植物伤口保护等方面起着重要作用。本研究以地域来源和性状特性差异均较大的2个芹菜品种六合黄心芹和美国西芹为试验材料,利用RT-PCR技术获得这2种芹菜韧皮部蛋白基因的cDNA序列。结果显示:这2种芹菜来源的韧皮部蛋白基因全长均为546 bp,编码181个氨基酸。两者核苷酸序列有3个位点的不同,分别为:88G/A、399T/C和489T/C;在氨基酸序列上有1个位点的不同,为30T/A。预测其蛋白质分子量为19 kD,pI值为9.18。六合黄心芹和美国西芹的韧皮部蛋白与忽地笑等植物的韧皮部蛋白相似度较高,在保守位置分别具有5个亮氨酸残基和4个色氨酸残基。实时定量PCR表达分析表明,该基因主要在芹菜的茎和根等部位表达,具有明显的组织特异性。

The phloem proteins play important role in the maintenance of morphology, material transportation and wound protection in higher plant. Here,eDNA sequences of phloem protein were obtained using RT-PCR technology from two celery varieties Liu He Huang Xin Qin and Mei Guo Xi Qin. The full-length of the two phloem-protein genes were 546 bp, and encoded 181 amino acid residues, respectively. The nueleotide sequences alignment showed that there were 3 loci differences： 88G/A,399T/C and 489T/C. The deduced amino acid sequences showed that there was 1 residue difference,30T/A. Prediction of the proteins molecular weight were 19 kDa, and pI were 9.18, respectively. The P-proteins from Liu He Huang Xin Qin and Mei Guo Xi Qin have highly identity with Lycoris aurea. There were 5 leueine and 4 tryptophan amino acid residues in the conservative position, respectively. Real time RT-PCR analysis showed that the genes expression were tissue specificity,which mainly expressed in the stem and root.