滇牡丹环阿屯醇合成酶基因的克隆及表达分析

李云琴<sup>1; 2</sup>; 原晓龙<sup>1; 2</sup>; 陈中华<sup>1</sup>; 王毅<sup>1; 2*</sup>

引用本文:	李云琴,原晓龙,陈中华,王毅.滇牡丹环阿屯醇合成酶基因的克隆及表达分析[J].广西植物,2021,41(4):567-572.[点击复制]
	LI Yunqin, YUAN Xiaolong, CHEN Zhonghua, WANG Yi.Cloning and expression analysis of cycloartenol synthase gene from Paeonia delavayi[J].Guihaia,2021,41(4):567-572.[点击复制]

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滇牡丹环阿屯醇合成酶基因的克隆及表达分析
李云琴^1,2,原晓龙^1,2,陈中华¹,王毅^1,2*
1.云南省林业和草原科学院,昆明650201;2.云南省森林植物培育与开发利用重点实验室,

摘要:

为研究环阿屯醇合成酶(cycloartenolsynthase)基因在滇牡丹生物合成中的作用机制,该文采用RT-PCR技术首次从滇牡丹(Paeoniadelavayi)中克隆得到一个具完整开放阅读框的环阿屯醇合成酶基因(PdCAS),该基因全长2274bp,共编码757个氨基酸,PdCAS蛋白序列与桃(Prunuspersica)、日本裸樱(Prunusyedoensisvar.nudiflora)、葡萄(Vitisvinifera)蛋白序列相似性在86%以上。序列比对分析显示PdCAS具有氧化鲨烯环化酶超家族典型的DCTAE结构域和三萜合成酶的标志性序列DGSWYGSWGVCFTYG。滇牡丹PdCAS蛋白与蔷薇科植物苹果(MalusdomesticaXP008391430.1)、白梨(PyrusbretschneideriXP009370034.1)、月季(RosachinensisXP024193310.1)、日本裸樱(Prunusyedoensisvar.nudifloraPQQ11009.1)、桃(PrunuspersicaXP007225240.1)的CAS蛋白聚为一类。PdCAS基因在滇牡丹各组织中均有表达,但是在花瓣中表达量较高。该研究克隆的PdCAS基因与滇牡丹甾醇类化合物的合成密切相关。

关键词: 滇牡丹,环阿屯醇合成酶,cDNA克隆,基因功能分析

DOI：10.11931/guihaia.gxzw201904021

分类号:Q943

文章编号:1000-3142(2021)04-0567-06

基金项目:国家自然科学基金项目(31860177);云南省应用基础研究青年项目(2016FD100)[SupportedbytheNationalNaturalScienceFoundationofChina(31860177);YunnanAppliedBasicResearchYouthProgram(2016FD100)]。

Cloning and expression analysis of cycloartenol synthase gene from Paeonia delavayi

LI Yunqin^1,2, YUAN Xiaolong^1,2, CHEN Zhonghua¹, WANG Yi^1,2*

1. Yunnan Academy of Forestry and Grassland, Kunming 650201, China;2. Yunnan Key Laboratory of Forestry Plant Cultivation and Utilization, State Forestry and Grassland Administration Key Laboratory of Rare and Endangered Species Conservation and Propagation, Kunming 650201, China

Abstract:

To study the effect of cycloartenol synthase gene(CAS)in the sterol biosynthesis of Paeonia delavayi, using the RT-PCR technology, a complete open reading frame of PdCAS gene was cloned from Paeonia delavayi for the first time. The full-length cDNA of PdCAS gene was 2 274 bp, encoded 757 amino acids. The PdCAS protein sequence of P. delavayi had more than 86% similarty with the Prunus persica, Prunus yedoensis var. nudiflora and Vitis vinifera. Sequence alignment analysis showed that PdCAS had a typical conserved DCTAE motif of oxidosaualene cyclase and the marker sequence DGSWYGSWGVCFTYG of triterpenoid synthase. The phylogenetic analysis revealed that PdCAS clustered together with CAS proteins of Malus domestica (XP 008391430.1), Pyrus bretschneideri(XP 009370034.1), Rosa chinensis(XP 024193310.1), Prunus yedoensis var. nudiflora(PQQ11009.1)and Prunus persica (XP 007225240.1). PdCAS expressed significantly in different tissues and had the highest transcript profile in flowers. PdCAS was closely related to the synthesis of sterols from Paeonia delavayi.

Key words: Paeonia delavayi, cycloartenol synthase(CAS), cDNA clone, gene function analysis