|本期目录/Table of Contents|

[1]张骥诚,梁嘉庆,俞锦锋,等.白骨壤重金属耐受性相关基因AmNramp3的克隆与分析[J].厦门大学学报(自然科学版),2017,56(03):331-338.[doi:10.6043/j.issn.0438-0479.201611033]
 ZHANG Jicheng,LIANG Jiaqing,YU Jinfeng,et al.Cloning and Analysis of Heavy Metal Toleranceassociated Gene AmNramp3 in Avicennia marina (Forsk.) Vierh[J].Journal of Xiamen University(Natural Science),2017,56(03):331-338.[doi:10.6043/j.issn.0438-0479.201611033]
点击复制

白骨壤重金属耐受性相关基因AmNramp3的克隆与分析(PDF/HTML)
分享到:

《厦门大学学报(自然科学版)》[ISSN:0438-0479/CN:35-1070/N]

卷:
56卷
期数:
2017年03期
页码:
331-338
栏目:
研究论文
出版日期:
2017-05-24

文章信息/Info

Title:
Cloning and Analysis of Heavy Metal Toleranceassociated Gene AmNramp3 in Avicennia marina (Forsk.) Vierh
文章编号:
0438-0479(2017)03-0331-08
作者:
张骥诚1梁嘉庆2俞锦锋1刘景春1卢豪良1严重玲1*
1.厦门大学环境与生态学院,福建厦门361102;2.四川农业大学生命科学学院,四川雅安625014
Author(s):
ZHANG Jicheng1LIANG Jiaqing2YU Jinfeng1LIU Jingchun1LU Haoliang1YAN Chongling1*
1.College of the Environment & Ecology,Xiamen University,Xiamen 361102,China;2.College of Life Science,Sichuan Agriculture University,Ya′an 625014,China
关键词:
白骨壤 重金属耐受性 Nramp
Keywords:
Avicennia marina heavy metal tolerance Cd Nramp
分类号:
Q 94
DOI:
10.6043/j.issn.0438-0479.201611033
文献标志码:
A
摘要:
白骨壤(Avicennia marina(Forsk.)Vierh.)作为红树林的先锋植物,广泛分布于我国南海沿岸,是一种研究红树植物重金属污染物耐受性分子机制很好的材料.以抑制消减杂交获得的白骨壤重金属耐受性相关基因AmNramp3片段为基础,设计基因特异性引物,利用cDNA末端快速扩增(RACE)技术克隆了其全长,长度为1 958 bp; 通过实时荧光定量PCR(qRT-PCR)检测了AmNramp3在重金属镉胁迫时白骨壤叶、茎和根中的表达情况,其表达量在叶中高于根和茎中,但在茎和根中有快速的响应.利用生物信息学方法预测并分析了该基因的开放阅读框,位置位于1~1 527 bp,编码508个氨基酸; 进而预测并分析了AmNramp3蛋白的结构,发现其氨基酸序列中不含有信号肽,说明其不是分泌蛋白; 但其具有很复杂的跨膜结构,是一种跨膜蛋白.综上,AmNramp3基因编码一种跨膜蛋白,在重金属镉胁迫下表达量快速变化,可能与镉的跨膜运输密切相关,这为后续分析该基因发挥功能的分子机制奠定了基础.
Abstract:
The halophytic Avicennia marina(Forsk.)Vierh. is one of the pioneer mangroves along the southern coast of China,which is a good material for understanding molecular mechanisms behind heavy metal pollutant tolerance in mangrove plants.In this study,based on the result of suppressive subtractive hybridization using rapid amplification of cDNA ends(RACE)technology,AmNramp3 gene was cloned using gene specific primers,with the full-length cDNA of 1 958 bp.We analyzed the expression of the AmNramp3 gene in leaf,stem and root after Cd treatment based on the quantative real-time PCR(qRT-PCR),and found the expression level in leaf was higher than in stem and root,while exhibited rapid response in stem and root.Additionally,the open reading frame(ORF)of the AmNramp3 gene was predicted and analyzed using bioinformatics,and results show that it is located in 1 bp to 1 527 bp and encodes 508 amino acids.Then the structure of the AmNramp3 protein was predicted and analyzed,and results show that the protein has no signal peptide,indicating that it is not a secretory protein.However,the AmNramp3 protein has a complex transmembrane structure,indicating that it is a transmembrane protein.Therefore,our analysis predicts that AmNramp3 encodes a transmembrane protein and the expression is rapidly changed under Cd stress,which is possibly related to the transmembrane transport function of heavy metal cadmium.This is helpful for us to understand the molecular mechanism of the AmNramp3 gene function in terms of heavy metal tolerance.

参考文献/References:

[1] 林鹏.红树林[M].北京:海洋出版社,1984.
[2] DUKE N C,MEYNECKE J O,DITTMANN S,et al.A world without mangroves?[J].Science,2007,317(5834):41-42.
[3] NATH B,BIRCH G,CHAUDHURI P.Trace metal biogeochemistry in mangrove ecosystems:a comparative assessment of acidified(by acid sulfate soils)and non-acidified sites[J].Science of the Total Environment,2013,463/464(5):667-674.
[4] MACFARLANE G R,BURCHETT M D.Photosynthetic pigments and peroxidase activity as indicators of heavy metal stress in the grey mangrove,Avicennia marina(Forsk.)Vierh.[J].Marine Pollution Bulletin,2001,42(3):233-240.
[5] HUANG G Y,WANG Y S.Physiological and biochemical responses in the leaves of two mangrove plant seedlings(Kandelia candel and Bruguiera gymnorrhiza)exposed to multiple heavy metals[J].Journal of Hazardous Materials,2010,182(1/2/3):848-854.
[6] BOEDDI B,ORAVECZ A R,LEHOCZKI E.Effect of cadmium on organization and photoreduction of protochlorphyllide in dark-grown leaves and etioplast inner membrane preparations of wheat[J].Photosynthetica,1995,31(3):411-420.
[7] GONZALEZ-MENDOZA D,MORENO A Q,ZAPATA-PEREZ O.Coordinated responses of phytochelatin synthase and metallothionein genes in black mangrove,Avicennia germinans,exposed to cadmium and copper[J].Aquatic Toxicology,2007,83(4):306-314.
[8] HUANG G Y,WANG Y S.Expression and characterization analysis of type 2 metallothionein from grey mangrove species(Avicennia marina)in response to metal stress[J].Aquatic Toxicology,2010,99(1):86-92.
[9] DUKE N C.A systematic revision of the mangrove genus Avicennia(Avicenniaceae)in Australasia[J].Australian Systematic Botany,1991,4(2):299.
[10] MACFARLANE G R.Leaf biochemical parameters in Avicennia marina(Forsk.)Vierh. as potential biomarkers of heavy metal stress in estuarine ecosystems[J].Marine Pollution Bulletin,2002,44(3):244-256.
[11] DIATCHENKO L,LAU Y F,CAMPBELL A P,et al.Suppression subtractive hybridization:a method for ge-nerating differentially regulated or tissue-specific cDNA probes and libraries[J].Proceedings of the National Academy of Sciences of the United States of America,1996,93(12):6025-6030.
[12] ROSE P P,HANNA S L,SPIRIDIGLIOZZI A,et al.Natural resistance-associated macrophage protein is a cellular receptor for sindbis virus in both insect and mammalian hosts[J].Cell Host & Microbe,2011,10(2):97-104.
[13] SASAKI A,YAMAJI N,YOKOSHO K,et al.Nramp5 is a major transporter responsible for manganese and cadmium uptake in rice[J].Plant Cell,2012,24(5):2155-2167.
[14] CAILLIATTE R,SCHIKORA A,BRIAT J F,et al.High-affinity manganese uptake by the metal transporter NRAMP1 is essential for Arabidopsis growth in low manganese conditions[J].Plant Cell,2010,22(3):904-917.
[15] MARTINEZ-CUENCA M R,LEGAZ F,FORNER-GINER M A,et al.Bicarbonate blocks iron translocation from cotyledons inducing iron stress responses in Citrus roots[J].Journal of Plant Physiology,2013,170(10):899-905.
[16] OOMEN R J,WU J,LELIEVRE F,et al.Functional characterization of NRAMP3 and NRAMP4 from the metal hyperaccumulator Thlaspi caerulescens[J].New Phytologist,2009,181(3):637-650.
[17] HOAGLAND D R,ARNON D I.The water-culture method for growing plants without soil[M].Berkeley:Circular California Agricultural Experiment Station,1950:1-31.
[18] YANG G,ZHOU R,TANG T,et al.Simple and efficient isolation of high-quality total RNA from Hibiscus tiliaceus,a mangrove associate and its relatives[J].Preparative Biochemistry and Biotechnology,2008,38(3):257-264.
[19] ZHANG Q,YAN C,LIU J,et al.Silicon alleviation of cadmium toxicity in mangrove(Avicennia marina)in relation to cadmium compartmentation[J].Journal of Plant Growth Regulation,2014,33(2):233-242.
[20] ZHANG Q,YAN C,LIU J,et al.Silicon alleviates cadmium toxicity in Avicennia marina (Forsk.)Vierh.seedlings in relation to root anatomy and radial oxygen loss[J].Marine Pollution Bulletin,2013,76(1/2):187-193.
[21] VERBRUGGEN N,HERMANS C,SCHAT H.Molecular mechanisms of metal hyperaccumulation in plants[J].New Phytologist,2009,181(4):759-776.

备注/Memo

备注/Memo:
收稿日期:2016-11-15 录用日期:2017-02-13
基金项目:国家自然科学基金(31530008,31370516)
*通信作者:ycl@xmu.edu.cn
引文格式:张骥诚,梁嘉庆,俞锦锋,等.白骨壤重金属耐受性相关基因AmNramp3的克隆与分析[J].厦门大学学报(自然科学版),2017,56(3):331-338.

Citation:ZHANG J C,LIANG J Q,YU J F,et al.Cloning and analysis of heavy metal tolerance-associated gene AmNramp3 in Avicennia marina(Forsk.)Vierh.[J].J Xiamen Univ Nat Sci,2017,56(3):331-338.(in Chinese)
更新日期/Last Update: 1900-01-01