龙眼水通道蛋白基因(DLPIP1)的克隆与表达分析Cloning and expression of longan aquaporin(DLPIP1)gene
陈虎;何新华;罗聪;邓立宝;胡颖;李明娟;杨丽涛;
摘要(Abstract)：
应用蛋白质组学研究低温胁迫下龙眼叶片蛋白质组变化时,发现PIP1蛋白在龙眼低温胁迫中上调表达。应用RACE技术克隆龙眼水通道蛋白基因全长cDNA,命名为DLPIP1,基因登陆号为JN572691,长度为1 132 bp,包括1个900 bp的开放阅读框,编码299个氨基酸序列,同源性分析表明,DLPIP1在21个不同植物中的一致性为90%~93%。应用生物信息学软件对DLPIP1氨基酸序列分析表明,含有7个跨膜区,有2个NPA单元,其氨基酸残基与MIP家族蛋白保守区序列完全一致。氨基酸序列比对发现,该序列与其他物种PIP质膜水通道蛋白氨基酸序列有很高的同源性。利用实时荧光定量技术对DLPIP1在低温胁迫下不同组织表达谱分析表明,DLPIP1在龙眼根、茎、叶中都有表达,在根中的表达量最高,其次是茎和叶。DLPIP1在低温胁迫时,随着低温胁迫时间的延长而发生变化。这说明DLPIP1蛋白在龙眼低温逆境过程中起作用。
关键词(KeyWords)： 龙眼;水通道蛋白;低温胁迫;克隆;表达
基金项目(Foundation): 国家科技支撑计划项目(2008BADB8B01,2008BADB8B02);; 广西自然科学基金项目(2011GXNSFA018115);; 广西研究生创新计划项目(GXU11T31077);; 广西高等学校优秀人才资助计划项目(桂教人201065)
作者(Author): 陈虎;何新华;罗聪;邓立宝;胡颖;李明娟;杨丽涛;
Email:
DOI: 10.13925/j.cnki.gsxb.2012.02.015
参考文献(References)：
- [1]FORREST K L,BHAVE M.Major intrinsic proteins(MIPs)inplants:a complex gene family with major impacts on plant phenotype[J].Funct Integr Genomics,2007,7:263-289.
- [2]JOHANSON U,KARLSSON M,JOHANSSON I,GUSTAVSSON S,SJOVALL S,FRAYSSE L,WEIG A R,KJELLBOM P.The com-plete set of genes encoding major intrinsic proteins in Arabidopsisprovides a framework for a new nomenclature for major intrinsic pro-teins in plants[J].Plant Physiology,2001,126:1358-1369.
- [3]CHAUMONT F,BARRIEU F,WOJCIK E,CHRISPEELS M J,JUNG R.Aquaporins constitute a large and highly divergent proteinfamily in maize[J].Plant Physiology,2001,125:1206-1215.
- [4]SAKURAI J,ISHIKAWA F,YAMAGUCHI T,UEMURA M,MAESHIMA M.Identification of 33 rice aquaporin genes and analy-sis of their expression and function[J].Plant Cell Physiology,2005,46(9):1568-1577.
- [5]TYERMAN S D,BOHNERT H J,MAUREL C,STEUDLE E,SMITH J A C.Plant aquaporins:Their molecular biology,biophysicsand significance for plant water relations[J].Journal of ExperimentalBotany,1999,50:1055-1071.
- [6]YANG L T,HONG L,TAKAHASHI Y,CHEN F,WALKERD MA,CINEROLO L E.Proteomic analysis of grapevine stem in re-sponse to Xylella fastidiosa inoculation[J].Physiological and Molec-ular Plant Pathology,2011,75:90-99.
- [7]YOU Xiang-rong.Analysis of differential proteomice on longan(Di-mocarpus longan Lour.)floral reversion and normal flowering[D].Fuzhou:Fujian Agriculture and Forestry University,2009.游向荣.龙眼成花转变与成花逆转的差异蛋白质组学研究[D].福州:福建农林大学,2009.
- [8]LUO Cong,HE Xin-hua,CHEN Hu,WEI Yong-li,LI Ming-juan.A high-efficient method of RACE technique for obtaining the gene5′end[J].Plant Physiology Journal,2011,47(4):409-414.罗聪,何新华,陈虎,韦泳丽,李明娟.一种高效获取基因5′末端的RACE方法[J].植物生理学报,2011,47(4):409-414.
- [9]LIVAK K J,SCHMITTGEN T D.Analysis of relative gene expressiondata using real-time quantitative PCR and the2-ΔΔCT method[J].Methods,2001,25:402-408.
- [10]SCHAFFNER A R.Aquaporin function,structure and expression:are there more suiprises to surface in water relations[J].Planta,1998,204:131-139.
- [11]JAVOT H,MAUREL C.The role of aquaporins in root water uptake[J].Annals of Botany,2002,90:301-313.
- [12]HENZLER T,STEUDLE E.Reversible closing of water channels incharainternodes provides evidence for acomposite transport mode ofthe plasma membrance[J].Journal of Experimental Botany,1995,46:199-209.
- [13]GUENTHER J F,CHANMANIVONE N,GALTOVIC M P,WAL-LACE I S,COBB J A,ROBERTS D M.Phosphorylation of soybeannodulin 26 on serine 262 enhances water permeability and is regu-lated developmentally and by osmotic signals[J].Plant Cell,2003,15:981-991.
- [14]JOHANSSON I,KARLSSON M,SHUKLA V K,CHRISPEELS MJ,LARSSON C,KJELLBOM P.Water transport activity of the plas-ma membrane aquaporin PM28A is regulated by phosphorylation[J].Plant Cell,1998,10:451-459.
- [15]CHEN Ru-gang,ZHU Wen-chao,GONG Zhen-hui,LI Da-wei,YIN Yan-xu,LU Ming-hui.Cloning and sequence analysis of theaquaporins gene CaAQP in pepper[J].Scientia Agricultura Sinica,2010,43(20):4323-4329.陈儒钢,朱文超,巩振辉,李大伟,尹延旭,逯明辉.辣椒水通道蛋白基因CaAQP的克隆与序列分析[J].中国农业科学,2010,43(20):4323-4329.
- [16]AROCAA R,TOGNONIB F,IRIGOYENA J J,SANCHEZ-DIAZM,PARDOSSIC A.Different root low temperature response of twomaize genotypes differing in chilling sensitivity[J].Plant PhysiolBiochem,2001,39:1067-1073.