牡丹蔗糖转运蛋白基因PsSUT2的克隆及表达分析  

Cloning and Expression Analysis of Sucrose Transporter Gene PsSUT2 from Tree Peony (Paeonia suffruticosa)

作　　者：栗燕[1] 杨旭升[1] 杨秋生[1] 崔洋 常毅洪 高梦媛 宋佳[1] 

机构地区：[1]河南农业大学林学院,郑州450002 [2]鹤壁市淇滨区园林局,鹤壁458030

出　　处：《农业生物技术学报》2017年第4期567-578,共12页Journal of Agricultural Biotechnology

基　　金：河南省科技攻关项目(No.102102110033);郑州市重点科技攻关计划项目(No.30800472)

摘　　要：蔗糖是植物体中重要的碳源物质,参与植物的生长、发育乃至信号转导等多个生理过程。植物体在长距离运输蔗糖的过程中,蔗糖转运蛋白(sucrose transporters,SUT)基因起关键作用。为了解该基因在牡丹(Paeonia suffruticosa)植株体内的转运机制,以洛阳红牡丹(P.suffruticosa'Luoyanghong')为材料,利用反转录PCR(reverse transcription-PCR,RT-PCR)和RACE(rapid-amplification of cDNA ends)方法从叶片中克隆了蔗糖转运蛋白基因,命名为PsSUT2。该基因全长2 347 bp,包含1 803 bp的开放阅读框,编码601个氨基酸,Gen Bank登录号为KC542396,相对分子量为64.6 k D,等电点为6.14,与葡萄(Vitis vinifera)中的蔗糖转运蛋白基因同源性最高,为75.27%。该蛋白属于易化子超家族(major facilitator superfamily,MFS)成员,包含12个跨膜螺旋。采用qRT-PCR和离子色谱法研究牡丹不同生育期与不同组织部位中PsSUT2基因的表达量及可溶性糖的含量变化,结果表明,在牡丹的年生长周期中,PsSUT2在牡丹的根、老茎、叶和鳞芽(花瓣)中均有表达,但主要在盛花期的花瓣中大量表达,相对表达量为11.18,是同时期根的4.26倍、小风铃期根的4.40倍、圆桃期花瓣的10.48倍、鳞芽分化期鳞芽的30.55倍,与同时期的其他部位乃至其他时期的所有部位相比差异显著,因此推测该基因可能参与了"库"端蔗糖的卸载。蔗糖、葡萄糖和果糖的含量在不同生育期、不同部位间存在较大差异,盛花期到来之前,鳞芽(花瓣)中蔗糖含量几乎为0,而葡萄糖和果糖水平较高,说明洛阳红牡丹花瓣属于己糖(葡萄糖和果糖)积累型器官,己糖的不断积累有利于花朵的开放。相关性分析表明,PsSUT2的表达量与与叶片中蔗糖的含量及根中葡萄糖含量成显著负相关,说明该基因的表达可能受叶片中蔗糖及根中葡萄糖的反馈抑制。本研究结果为进一步了解PsSUT2基因的功能、利用该基因进行牡丹分子育种和�Sucrose which involved in the plant growth and development processes, the signal transduction pathway and other physiological activities is an important carbon source in plants. Sucrose transporter genes （SUT） play a key role in the long distance transport activities of sucrose in plants. In order to understand the transport mechanism of the SUT in Paeonia suffruticosa, P. suffruticosa ＇Luoyanghong＇ was used as experimental materials in this study. The sucrose transporter gene named PsSUT2 was separated and cloned from the leaf by reverse transcription-PCR （RT-PCR） and rapid-amplification of cDNA ends （RACE）. The full length cDNA of PsSUT2 is 2 347 bp, and the ORF encoding a protein of 601 amino acids, was 1 803 bp. The accession number is KC542396 in GenBank. Its molecular mass was 64.6 kD and pI was 6.14, The homologous amino acid sequence of PsSUT2 compared with that of SUT gene from Vitis vinifera was 75.27%. PsSUT2 protein contained 12 transmembrane helical structures, which belongs to the major facilitator superfamily （MFS）. The relative expression of PsSUT2 gene and the soluble content in different organs were determined with qRT-PCR and chromatography technology at the different developmental stage of tree peony. The main results showed that the PsSUT2 gene expressed abundantly in the petals at the flowering stage, and the relative expression was 11.18, which was significantly different from other organs of the same stage even every organ of other stages. Meanwhile, the PsSUT2 gene expressed in every organ such as the root, the perennial stem, the leaf, the bulbil, the petal at different developmental stage, but the relative expression were low, for example, the relative expression of petals in the flowering stage was 4.26 times as much as that of roots in the same stage, and 4.40 times as much as that of roots in the little bell stage, and 10.48 times as much as that of petals in the peach stage, and 30.55 times as much as that of bulbil in the bulbil differentiation stage. So, it ind

关 键 词：牡丹 蔗糖 牡丹蔗糖转运蛋白基因(PsSUT2) 表达分析 

分 类 号：S685.11[农业科学—观赏园艺]