[1]邹晶晶,陆玲,曾祥玲,等.桂花DAD-1基因克隆与表达分析[J].北方园艺,2017,41(08):96-102.[doi:10.11937/bfyy.201708022]
　ZOU Jingjing,LU Ling,ZENG Xiangling,et al.Cloning and Expression Analysis of DAD-1 Gene in Osmanthus fragrans[J].Northern Horticulture,2017,41(08):96-102.[doi:10.11937/bfyy.201708022]
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桂花DAD-1基因克隆与表达分析

《北方园艺》[ISSN:1001-0009/CN:23-1247/S] 卷: 第41卷 期数: 2017年08 页码: 96-102 栏目: 出版日期: 2017-04-30
Title:
Cloning and Expression Analysis of DAD-1 Gene in Osmanthus fragrans
作者:
邹晶晶1; 陆玲2; 曾祥玲1; 王彩云3
（1.湖北科技学院 核技术与化学生物学院，湖北 咸宁 437100；2.孝感市农业科学院 蔬菜研究所，湖北 孝感 432000； 3.华中农业大学 园艺植物生物学教育部重点实验室，湖北 武汉 430070）
Author(s):
ZOU Jingjing1; LU Ling2; ZENG Xiangling1; WANG Caiyun3
(1.School of Nuclear Technology and Chemistry & Biology，Hubei University of Science and Technology，Xianning，Hubei 437100;2.Institute of Vegetables,Xiaogan Academy of Agricultural Sciences，Xiaogan,Hubei 432000;3.Key Laboratory for Biology of Horticultural Plants，Ministry of Education，Huazhong Agricultural University,Wuhan,Hubei 430070)
关键词:
桂花; DAD-1基因; 细胞程序性死亡; 花瓣衰老
Keywords:
Osmanthus fragrans; DAD-1 gene; program cell death; flower senescence
DOI:
10.11937/bfyy.201708022
文献标志码:
A
摘要:
以“柳叶金桂”桂花(Osmanthus fragrans ‘Liuye Jingui’)为试材，采用TAIL-PCR技术，克隆了“柳叶金桂”花瓣中DAD-1基因的全长序列，并分析了其在桂花不同组织部位及开花阶段的时空表达模式，以期为研究该基因在桂花花瓣衰老过程中的功能提供参考依据。结果表明：DAD-1基因由560个碱基组成，包含一个351 bp的完整开放阅读框和一个3′-Poly A尾巴；DAD-1基因编码的蛋白包含116个氨基酸，其结构预测表明DAD-1蛋白属于疏水性膜整合蛋白，通过3个α-螺旋构成的跨膜区行使功能。半定量PCR结果表明，该基因在“柳叶金桂”的根、茎、叶、花等各组织器官中均有表达，且在花苞、新叶等幼嫩组织器官中相对表达量较高，在老叶等成熟组织器官中相对表达量较低。实时荧光定量PCR结果表明，在“柳叶金桂”花瓣衰老过程中，DAD-1基因在铃梗期开始显著下调表达。
Abstract:
Osmanthus fragrans ‘Liuye Jingui’ was used as tested material.The complete coding sequence (CDS) of DAD-1 gene was cloned by the method of thermal asymmetric interlaced PCR (TAIL-PCR)，and its expression patterns in different organs and different flowering stages in O.fragrans were analyzed，to provide reference for studying the function of this gene during the flower senescence.The results showed that the length of DAD-1 gene was constituted of 560 bases with an open reading frame of 351 bp and a 3′-Poly A.The DAD-1 protein encoded 116 amino acids，and it was a hydrophobic integration membrane protein with three alpha-helixs in the structure.The results of semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) showed that DAD-1 expressed in most tissues of sweet Osmanthus，such as the stem，root，flower and leaf，and it was higher in the young organs such as young flowers and new leaves，but lower in old organs such as the old leaves.The results of quantitative real-time polymerase chain reaction (real-time PCR) showed that DAD-1 abundance rapidly decreased at the initial flowering stage，much earlier than the visible senescence symptoms.

参考文献/References:

 

[1]KELLEHER D J，GILMORE R．DAD1，the defender against apoptotic cell death，is a subunit of the mammalian oligosaccharyltransferase[J]．P Natl Acad Sci USA，1997，94(10)：4994-4999.

[2]ROBOTI P,HIGH S.The oligosaccharyltransferase subunits OST48，DAD1 and KCP2 function as ubiquitous and selective modulators of mammalian N-glycosylation[J]．J Cell Sci,2012,125(14):3474-3484.

[3]NAKASHIMA T,SEKIGUCHI T,KURAOKA A,et al.Molecular cloning of a human cDNA encoding novel protein，DAD1，whose defect causes apoptotic cell death in hamster BHK21 cells[J].Molecular and Cellular Biology,1993,13(10):6367-6374.

[4]GALLOIS P,MAKISHIMA T,HECHT V,et al.An Arabidopsis thaliana cDNA complementing a hamster apoptosis suppressor mutant[J].Plant J,1997,11(6):1325-1331.

[5]TANAKA Y,MAKISHIMA T,SASABE M,et al.DAD-1,a putative programmed cell death suppressor gene in rice[J].Plant Cell Physiol,1997,38(3):379-383.

[6]WANG X J,TANG C L,ZHANG H C,et al.TaDAD2，a negative regulator of programmed cell death，is important for the interaction between wheat and the stripe rust fungus[J].Mol Plant Microbe In,2011,24(1):79-90.

[7]贾志蓉,李丹,吕应堂.拟南芥AtDAD1超量表达植株对H2O2抗性的研究[J].武汉植物学研究,2004(22):373-379.

[8]蔡璇,苏蘩,金荷仙,等.四季桂花瓣色素的初步鉴定与提取方法[J].浙江林学院学报,2010,27(4):559-564.

[9]邹晶晶,袁斌,高微,等.桂花未成熟合子胚诱导体细胞胚再生[C]//中国观赏园艺研究进展.2015年中国观赏园艺学术研讨会，2015:238-242.

[10]阳韶昆,蔡璇,邹晶晶,等．桂花开放与衰老过程中花瓣蛋白酶活性与种类的变化[J]．园艺学报,2012,39(10):1967-1974.

[11]周媛.桂花花瓣衰老过程中细胞程序性死亡特征与机制研究[D].武汉：华中农业大学，2008.

[12]ZOU J J，ZHOU Y，CAI X，et al．Increase in DNA fragmentation and the role of ethylene and reactive oxygen species in petal senescence of Osmanthus fragrans[J].Postharvest Biol Tec,2014,93:97-105.

[13]陆铃.桂花DAD-1同源基因片段的克隆及表达分析[D].武汉:华中农业大学,2009.

[14]龚文芳.棉花抗细胞凋亡基因GhDADl的克隆、定位及表达分析[D].武汉:华中农业大学,2010.

[15]孙建富,陈尚珂,王爱英,等.新疆雪莲DAD1基因提高烟草的抗旱性与抗盐性[J].江苏农业科学,2011(39):84-86.

[16]杨舒雅,史娟,马斌芳,等.抗细胞凋亡因子DAD1的研究进展[J].生理科学进展,2012(43):315-318.

[17]国栋,和四梅,张丽莉,等.杂色鲍DAD1的克隆及在发育、弧菌感染、高温和缺氧下的表达分析[J].水产学报,2016,40(7):1018-1027.

[18]潘永娟.烟草ZX4D7基因功能初探[D].武汉:华中农业大学,2014.

[19]LINDHOLM P,KUITTINEN T,SORRI O,et al.Glycosylation of phytepsin and expression of dad1，dad2 and ost1 during onset of cell death in germinating barley scutella[J].Mech Dev,2000,93(12):169-173.

[20]WAGSTAFF C,MALCOLM P,RAFIQ A,et al．Programmed cell death (PCD) processes begin extremely early in Alstroemeria petal senescence[J].New Phytol,2003,160(1):49-59.

[22]YAMADA T,TAKATSU Y,KASUMI M,et al.A homolog of the defender against apoptotic death gene (DAD1) in senescing gladiolus petals is down-regulated prior to the onset of programmed cell death[J].J Plant Physiol,2004,161(11):1281-1283.

[23]van der KOP D A M，RUYS G，DEES D，et al．Expression of defender against apoptotic death (DAD-1) in Iris and Dianthus petals[J].Physiol Plantarum,2003,117(2):256-263.

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备注/Memo

第一作者简介：邹晶晶（1985-），女，博士，讲师，研究方向为观赏植物采后生理与分子生物学。E-mail：xingxingzou@163.com.责任作者：王彩云（1963-），女，博士，教授，研究方向为观赏植物栽培及采后生理。E-mail：248361509@qq.com.基金项目：湖北省教育厅科学技术研究资助项目（Q20162805）；湖北科技学院博士启动基金资助项目（BK1429）。

更新日期/Last Update: 2017-05-05