姜花脱落酸生物合成关键酶基因HcNCED的克隆及表达分析
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- 英文论文题名：Molecular Cloning and Expression Analysis of HcNCED,an Abscisic Acid Biosynthesis Key Enzyme Gene in Hedychium coronarium
- 论文作者：曾燕燕 ; 岳跃冲 ; 范燕萍
- 英文论文作者：ZENG Yan-yan ; YUE Yue-chong ; FAN Yan-ping ; College of Forestry and Landscape Architecture ; Flower Research Center ; South China Agricultural University
- 年：2016
- 作者机构：华南农业大学林学与风景园林学院华南农业大学花卉研究中心;
- 论文关键词：姜花 ; 脱落酸 ; 9-顺式-环氧类胡萝卜素双氧合酶 ; 表达分析
- 英文论文关键词：Hedychium coronarium ; ABA ; 9-cis-epoxycarotenoid dioxygenase ; Expression analysis
- 会议召开时间：2016-07-19
- 会议录名称：中国观赏园艺研究进展2016
- 语种：中文
- 分类号：S682.19
- 学会代码：EGYP
- 会议名称：2016年中国观赏园艺学术研讨会
- 会议地点：中国湖南长沙
- 主办单位：中国园艺学会观赏园艺专业委员会、国家花卉工程技术研究中心
- 学会名称：中国园艺学会
- 页数：7
- 文件大小：1902k
- 原文格式：O
- 会议级别：全国
摘要
脱落酸(ABA)在调控植物生长发育和植物对非生物胁迫应答中的反应方面起着重要作用。9-顺式-环氧类胡萝卜素双氧合酶(NCED)作为ABA生物合成过程中的限速酶,其催化的氧化裂解反应是生成ABA的关键步骤。本文在姜花中克隆得到一个NCED基因的cDNA全长序列并进行了表达分析。序列分析表明:该基因属于RPE65家族,开放阅读框长1884bp,推测其编码一个含627个氨基酸的氨基酸序列,分子量为68.4kDa、等电点(pI)为6.01。其氨基酸序列与小麦的TaNCED有66%的一致性,与拟南芥AtNCED4和琴叶拟南芥AlNCED4有54%、47%的一致性。对该基因蛋白进行疏水性预测,结果表明其整个多肽链表现为亲水性。蛋白二级结构预测发现α-螺旋、延伸链和无规则卷曲是NCED的主要二级结构。亚细胞定位预测结果推测该基因可能定位于叶绿体。HcNCED基因的荧光定量表达(RT-QPCR)分析表明:HcNCED在花蕾期表达量最低,随后逐步上升至盛开期最高,衰老期表达量下调,表达规律与姜花花香释放规律一致。研究结果为进一步深入研究HcNCED在姜花花香形成和调控中的作用奠定了基础。
Abscisic acid(ABA) plays an important role in the regulation of plant growth and the response to abiotic stresses.Oxidative cleavage of cis-epoxycarotenoids catalyzed by 9-cis-epoxycarotenoid dioxygenase(NCED),the rate-limiting enzyme,is the critical step in the biosynthesis of ABA in higher plants.For further studying the relationship between fragrance and abscisic acid,a NCED gene was cloned and expressed in Hedychium coronarium.Sequence analysis showed that it belonged to RPE65 superfamily,contained an open reading frame of 1884 bp,and encoded a peptide of 627 amino acids,with molecular weight of 68.4 kDa,isoelectric point(pI) of 6.01.The amino acid sequence oi HcNCED shared the homology of 66%with TaNCED,AtNCEM and AlNCEDA are 54%、 47%,respectively.A hydropathy plot of the gene showed that the entire polypeptide was hydrophilic.Protein secondary structure prediction suggested that the main secondary structure of NCED,which concluded the alpha helix,the extended chain and the random coil.Subcellular localization prediction indicated that the gene may be located in the chloroplast.Fluorescence quantitative(RT-QPCR) analysis of HcNCED showed that HcNCED expressed low in bud stage,peaked at blooming stage,then decreased in senescence.The expression pattern was consistent with the release of the floral fragrance of Hedychium coronarium.The results laid a foundation for further study on the role of HcNCED in the formation and regulation of the floral fragrance of Hedychium coronarium.
Abscisic acid(ABA) plays an important role in the regulation of plant growth and the response to abiotic stresses.Oxidative cleavage of cis-epoxycarotenoids catalyzed by 9-cis-epoxycarotenoid dioxygenase(NCED),the rate-limiting enzyme,is the critical step in the biosynthesis of ABA in higher plants.For further studying the relationship between fragrance and abscisic acid,a NCED gene was cloned and expressed in Hedychium coronarium.Sequence analysis showed that it belonged to RPE65 superfamily,contained an open reading frame of 1884 bp,and encoded a peptide of 627 amino acids,with molecular weight of 68.4 kDa,isoelectric point(pI) of 6.01.The amino acid sequence oi HcNCED shared the homology of 66%with TaNCED,AtNCEM and AlNCEDA are 54%、 47%,respectively.A hydropathy plot of the gene showed that the entire polypeptide was hydrophilic.Protein secondary structure prediction suggested that the main secondary structure of NCED,which concluded the alpha helix,the extended chain and the random coil.Subcellular localization prediction indicated that the gene may be located in the chloroplast.Fluorescence quantitative(RT-QPCR) analysis of HcNCED showed that HcNCED expressed low in bud stage,peaked at blooming stage,then decreased in senescence.The expression pattern was consistent with the release of the floral fragrance of Hedychium coronarium.The results laid a foundation for further study on the role of HcNCED in the formation and regulation of the floral fragrance of Hedychium coronarium.
引文
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8.Schwartz S H,Tan B C,Gage D A,Zeevaar J A,Mccarty D R.1997.Ecific oxidative cleavage of carotenoids by vpl4of maize[J].Science,276:1872-1874.
9.Wang X Q,Zhang CH,Wang Y J.2012.Isolation and expression of 9-cisepoxycarotenoid dioxygenase gene in tree Peony[J].Acta Horticulture Sinica,39(10):2033-3044.
10.Xian,L H.,Sun,P.P.,Hu,S.S.,Wu,J.,Liu,J.H.2014.Molecular cloning and characterization of CrNCEDl,a gene encoding 9-cis-epoxycarotenoid dioxygenase in Citrus reshin,with functions in tolerance to multiple abiotic stresses[J].Planta 239,61-77.
11.Yu Y.T.,Wu Z.,Lu K.,Bi C.,Liang S.,Wang X.F.,and Zhang D.P..2015.Overexpression of the MYB37 transcription factor enhances abscisic acid sensitivity,and improves both drought tolerance and seed productivity in Arabidopsis thaliana[J].Plant Mol.Biol.,90(3):267-279.
12.Yue YC,Yu RC,Fan YP.2015.Transcriptorae profiling provides new insights into the formation of floral scent in Hedychium coronarium[J].BMC GENOMICS,DOI:10.1186/sl2864-015-1653-7.
13.Zheng C.,Halaly T.,Acheampong A.K.,Takebayashi Y.,Jikumaru Y.,Kamiya Y.,and Or E..2015.Abscisic acid(ABA)regulates grape bud dormancy,and dormancy release stimuli may act through modification of ABA metabolism[J].J.Exp.Bot,66(5):1527-1542.
2.Arc E,Sechet J,Corbineau F,Rajjou L,and Marion-Poll A.2013.ABA crosstalk with ethylene and nitric oxide in seed dormancy and germination[J].Front Plant Sci.,63(4):1-19.
3.Giraudat J,Parcy F,Bertauche N.1994.Current advances in abscisic acid action and signaling[J].Plant Mol Bio,26:1557-1577.
4.李瑞红,范燕萍.白姜花不同开花阶段的香味组分及其变化[J].植物生理学通讯,2007,43(1):175-180.
5.Melcher K.,NgLM.,Zhou X.E.,Soon F.F.,Xu Y.,Suino-PoellellMelcher K.,Ng L M.,Zhou X.E.,Soon F.F.,Xu Y.,Suino-Poellell K.M.,Park S.Y.,Werner J.J.,Fujii H.,Chinnusamy V.,Kovach A.,Li J.,Wang Y.H.,Li J.,Peterson F.C.,Jensen D.R.,Yong E.L,Volkman B.F.,Cutler S.R.,Zhu J.K.,and Xu H.E..2009.A gate-latch-lock mechanism for hormone signaling by abscisic acid receptors[J].Nature,462(7273):602-608.
6.Nishimura N.,Hitomi K.,Arvai A.S.,Rambo R.P.,Hitomi C.,Cutler S.R.,Schroeder J.I.,and Getzoff E.D..2009.Structural mechanism of abscisic acid binding and signaling by dimeric[J].PYRlScience,326(5958):1373-1379.
7.Qin,X.,Zeevaart,J.A.D..1999.The 9-cis-epoxycarotenoid cleavage reaction is the key regulatory step of abscisic acid biosynthesis in water-stressed bean Proc[J].Nat.Acad.Sci.U.S.A.96,15354-15361.
8.Schwartz S H,Tan B C,Gage D A,Zeevaar J A,Mccarty D R.1997.Ecific oxidative cleavage of carotenoids by vpl4of maize[J].Science,276:1872-1874.
9.Wang X Q,Zhang CH,Wang Y J.2012.Isolation and expression of 9-cisepoxycarotenoid dioxygenase gene in tree Peony[J].Acta Horticulture Sinica,39(10):2033-3044.
10.Xian,L H.,Sun,P.P.,Hu,S.S.,Wu,J.,Liu,J.H.2014.Molecular cloning and characterization of CrNCEDl,a gene encoding 9-cis-epoxycarotenoid dioxygenase in Citrus reshin,with functions in tolerance to multiple abiotic stresses[J].Planta 239,61-77.
11.Yu Y.T.,Wu Z.,Lu K.,Bi C.,Liang S.,Wang X.F.,and Zhang D.P..2015.Overexpression of the MYB37 transcription factor enhances abscisic acid sensitivity,and improves both drought tolerance and seed productivity in Arabidopsis thaliana[J].Plant Mol.Biol.,90(3):267-279.
12.Yue YC,Yu RC,Fan YP.2015.Transcriptorae profiling provides new insights into the formation of floral scent in Hedychium coronarium[J].BMC GENOMICS,DOI:10.1186/sl2864-015-1653-7.
13.Zheng C.,Halaly T.,Acheampong A.K.,Takebayashi Y.,Jikumaru Y.,Kamiya Y.,and Or E..2015.Abscisic acid(ABA)regulates grape bud dormancy,and dormancy release stimuli may act through modification of ABA metabolism[J].J.Exp.Bot,66(5):1527-1542.