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https://www.ahs.ac.cn/images/0513-353X/images/top-banner12.jpg|#|菊花

园艺学报 ›› 2017, Vol. 44 ›› Issue (7): 1235-1243.doi: 10.16420/j.issn.0513-353x.2016-0726

• 研究论文 • 上一篇    下一篇

苹果液泡膜蔗糖转运蛋白基因MdSUT4 的表达分析与功能鉴定

许海峰,曲常志,刘静轩,王意程,王得云,左卫芳,姜生辉,王 楠,张宗营,陈学森*   

  1. 山东农业大学园艺科学与工程学院/作物生物学国家重点实验室,山东泰安 271018
  • 出版日期:2017-07-25 发布日期:2017-07-25

Expression Analysis and Functional Identification of a Vacuolar Sucrose Transporter Gene MdSUT4 in Apple

XU Haifeng,QU Changzhi,LIU Jingxuan,WANG Yicheng,WANG Deyun,ZUO Weifang,JIANG Shenghui,WANG Nan,ZHANG Zongying,and CHEN Xuesen*   

  1. College of Horticulture Science and Engineering,Shandong Agricultural University,State Key Laboratory of Crop
    Biology,Tai’an,Shandong 271018,China
  • Online:2017-07-25 Published:2017-07-25

摘要:

以新疆红肉苹果杂种一代优系‘红脆1 号’为试材,克隆MdSUT4 并原核诱导获得其重组
蛋白,对其进行生物信息学分析,测定其在不同组织及不同发育时期果实中的表达,通过亚细胞定位及
转基因鉴定其功能。结果发现,MdSUT4 编码的蛋白大小为55 kD 左右,定位于8 号染色体,由5 个外显
子和4 个内含子组成;糖代谢相关基因系统进化树分析发现,MdSUT4 与AtSUC4、PpSUT4 在同一个进
化支上;定量表达分析发现,MdSUT4 在苹果的花、叶和幼果中均有较高的表达,且在果实发育中的表达
水平与蔗糖含量显著负相关;MdSUT4 启动子含有与糖信号及激素信号相关的顺式作用元件;在‘王林’
苹果愈伤组织中过表达MdSUT4,能降低蔗糖含量,但却提高类黄酮含量;MdSUT4 定位于‘王林’愈伤
组织原生质体的液泡膜上。以上结果表明,MdSUT4 可能参与了蔗糖从液泡膜中的外排,并可能促进类黄
酮的合成。

关键词: 苹果, 液泡膜蔗糖转运蛋白, SUT4, 表达分析, 功能鉴定, 亚细胞定位

Abstract:

MdSUT4 was cloned from‘Hongcui 1’apple and the recombinant protein was obtained by
the prokaryotic induction technology,then,it was analyzed using bio-information. The expression level of
MdSUT4 in different tissues and different development stages were studied by the qRT-PCR,and we also
identified the function of MdSUT4 by the subcellular localization and transgenosis. The results showed that
the molecular mass of MdSUT4 was about 55 kD,it was located in the chromosome 8 of the apple
genome,included 5 exons and 4 introns. A phylogenetic tree indicated MdSUT4,AtSUC4 and PpSUT4
were located in the same evolutionary branch. The qRT-PCR showed that the MdSUT4 has the higher expression level in flowers,leaves and young fruits of apples,and its expression had a significant negative
correlation with the content of sucrose during the development stage of fruits. The promoter of MdSUT4
contained several typical cis-acting elements , including sugar signaling responsive elements and
phytohormone responsive elements. We overexpressed MdSUT4 in‘Orin’callus,then found that it could
increase the content of flavonoid and decrease the content of sucrose. MdSUT4 was located in the
tonoplast of‘Orin’callus. These results indicated that MdSUT4 might participate in the efflux of sucrose
from the vacuole and promote the synthesis of flavonoid.

Key words: apple, vacuolar sucrose transporter, SUT4, expression analysis, functional identification, subcellular localization