云南野生黄牡丹谷胱甘肽转移酶(GST)基因的分离及表达分析

doi:10.11707/j.1001-7488.20141209

摘要/Abstract

摘要：

从已构建的云南野生黄牡丹花瓣转录组数据库中得到10个与花色素转运相关的谷胱甘肽转移酶(GST)蛋白同源性高的Unigene序列,分别命名为 PlGST1-10.利用RT-PCR和RACE技术对PlGST 1-10 最大阅读框(ORF)序列扩增并进行氨基酸序列比较和系统进化树分析,结果显示: PlGST5 可能与花色素转运相关,包含1个 675 bp的开放阅读框,编码1个224个氨基酸的蛋白,含有2个内含子和3个外显子,属于phi型GST; 其氨基酸序列与葡萄VvGST4、矮牵牛PhAn9、仙客来 CkmGST3、拟南芥AtTT19、瓜叶菊ScGST3和香石竹DcGSTF2等花色素转运GST具有较高的相似性. PlGST5 在不同花色的花发育时期及盛开期的不同组织的qRT-PCR分析表明 PlGST5 在花色素积累较多的组织中高丰度表达,在黄牡丹和紫牡丹的盛开期表达量最高,而在牡丹品种‘赵粉’和‘玉板白’的花发育早期表达量最高.推测 PlGST5 参与黄牡丹花色素转运.

关键词: 黄牡丹, 谷胱甘肽转移酶, 花色素, 基因克隆, 基因表达

Abstract:

In this study, 10 unigene sequences that share high homology with glutathione S-transferase(GST) protein involved in plant anthocyanin transfering were obtained from previously-constructed tree peony (Paeonia delavayi var. lutea) petal transcriptome database and named PlGST 1-10. Sequences of Open Reading Frame (ORF) in PlGST 1-10 were amplified with designed specific primers using RT-PCR and RACE techniques, and the amino acid sequences were compared and the phylogenetic tree was analy zed. Results showed that PlGST5 was considered to be an alternative gene which is related to anthocyanin transferring, contained a 675 bp ORF encoding 224 amino acid residues with 2 introns and 3 excons and belonged to phi type GST. The predicted protein sequence of PlGST5 also shared high similarity with other GSTs that are related to anthocyanin transferring, such as VvGST4 (Vitis vinifera), PhAn9 (Petunia hybrida), CkmGST3 (Cyclamen persicum), AtTT19 (Arabidopsis thaliana), ScGST3 (Senecio cruentus ) and DcGSTF2 (Dianthus caryophyllus). The expression patterns of PlGST 5 in different tissues and petals with different flower colors during flower development were investigated using qRT-PCR. The results indicated that PlGST5 had a higher expression level in the tissues containing more anthocyanin. The expression level was the highest when flowers fully opened in P. delavayi var. lutea and P. delavayi, while the expression reached the highest abundance at early developmental stage of P. suffruticosa ‘Zhao Fen' and P. suffruticosa ‘Yu Banbai’. In conclusion, we inferred that PlGST5 might associate with the transfer of anthocyanin in P. delavayi var. lutea and this would provide the basis for insight into the molecular mechanisms underlying tree peony yellow flower pigmentation.

Key words: Paeonia delavayi var. lutea, glutathione S-transferase, anthocyanin, gene clone, gene expression

中图分类号:

S718.46

史倩倩, 周琳, 王雁. 云南野生黄牡丹谷胱甘肽转移酶(GST)基因的分离及表达分析[J]. 林业科学, 2014, 50(12): 63-72.

Shi Qianqian, Zhou Lin, Wang Yan. Isolation and Expression Analysis of GST Gene Encoding Glutathione S-Transferase of Paeonia delavayi var. lutea Wild Population in Yunnan[J]. Scientia Silvae Sinicae, 2014, 50(12): 63-72.

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