王超;李昕荷;张瑞芝;韩悦;齐波;王玉成;

• 1:东北林业大学林木遗传育种国家重点实验室

摘要(Abstract)：

[目的]研究刚毛柽柳水通道蛋白ThPIP基因启动子的克隆及其活性分析。[方法]采用染色体步移技术,克隆到刚毛柽柳ThPIP基因起始密码子上游1 241 bp启动子序列,并通过PLACE和PlantCARE预测启动子序列中所包含的主要顺式作用元件。将该启动子替换pCAMBIA1301上的35S启动子,构建融合表达基因proThPIP∷GUS,通过基因枪瞬时转化烟草,并进行GUS组织化学染色。[结果]该启动子具有活性,能够驱动GUS基因的表达。[结论]为进一步鉴定该启动子中的顺式作用元件及相互作用的调控因子奠定基础,从而为揭示刚毛柽柳ThPIP基因的分子调控机制提供依据。

关键词(KeyWords)： 刚毛柽柳;ThPIP基因;启动子克隆;瞬时表达

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金青年科学基金项目(31300571);; 东北林业大学大学生创新训练计划项目(201310225087)

作者(Author): 王超;李昕荷;张瑞芝;韩悦;齐波;王玉成;

Email:

DOI:

参考文献(References)：

• [1]MAUREL C,VERDOUCQ L,LUU D T,et al.Plant aquaporins:membrane channels with multiple integrated functions[J].Annu Rev Plant Biol,2008,59:595-624.
• [2]KALDENHOFF R,FISCHER M.Aquaporins in plants[J].Acta Physiol,2006,187(1/2):169-176.
• [3]DANIELSON J,JOHANSON U.Unexpected complexity of the aquaporin gene family in the moss Physcomitrella patens[J].BMC Plant Biol,2008,8(1):45.
• [4]JOHANSON U,KARLSSON M,JOHANSSON I,et al.The complete set of genes encoding major intrinsic proteins in Arabidopsis provides a framework for a new nomenclature for major intrinsic proteins in plants[J].Plant Physio1,2001,126(4):1358-1369.
• [5]SAKURAI J,ISHIKAWA F,YAMAGUCHI T,et al.Identification of33 rice aquaporin genes and analysis of their expression and function[J].Plant Cell Physio1,2005,46(9):1568-1577.
• [6]CHAUMONT F,BARRIEU F,WOJCIK E,et al.Aquaporins constitute a large and highly divergent protein family in maize[J].Plant Physio1,2001,25(3):1206-1215.
• [7]GUPTA A B,SANKARARAMAKRISHNAN R.Genome-wide analysis of major intrinsic proteins in the tree plant Populus trichocarpa:characterization of XIP subfamily of aquaporins from evolutionary perspective[J].BMC Plant Biol,2009,9(1):134.
• [8]LUDEWIG U,DYNOWSKI M.Plant aquaporin selectivity:where transport assays,computer simulations and physiology meet[J].Cell Mol Life Sci,2009,66(19):3161-3175.
• [9]侯丙凯,夏光敏,陈正华.植物基因工程表达载体的改进和优化策略[J].遗传,2001,23(5):492-497.
• [10]HUA Z M,YANG X,FROMM M E.Activation of the NaCl-and droughtinduced RD29A and RD29B promoters by constitutively active Arabidopsis MAPKK or MAPK proteins[J].Plant Cell Environ,2006,29(9):1761-1770.
• [11]张娟,张道远,尹林克.刚毛柽柳基因组DNA提取和RAPD反应条件探索[J].西北植物,2003,23(2):253-256.
• [12]WANG C,GAO C,WANG L,et al.Comprehensive transcriptional profiling of NaHCO3-stressed Tamarix hispida roots reveals networks of responsive genes[J].Plant Mol Biol,2014,84(1/2):145-157.
• [13]HIGO K,UGAWA Y,IWAMOTO M,et al.Plant cis-acting regulatory DNA elements(PLACE)database[J].Nucleic Acids Res,1999,27(1):297-300.
• [14]LESCOT M,DE HAIS P,THIJS G,et al.PlantCARE,a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences[J].Nucleic Acids Res,2002,30(1):325-327.
• [15]ABE H,URAO T,ITO T,et al.Arabidopsis AtMYC2(bHLH)and AtMYB2(MYB)function as transcriptional activators in abscisic acid signaling[J].Plant Cell,2003,15(1):63-78.
• [16]URAO T,YAMAGUCHI-SHINOZAKI K,URAO S,et al.An Arabidopsis MYB homolog is induced by dehydration stress and its gene product binds to the conserved MYB recognition sequence[J].Plant Cell,1993,5(11):1529-1539.
• [17]XIE Z,ZHANG Z L,ZOU X,et al.Annotations and functional analyses of the rice WRKY gene super family reveal positive and negative regulators of abscisic acid signaling in aleurone cells[J].Plant Physiol,2005,137(1):176-189.
• [18]YANAGISAWA S.Dof1 and Dof2 transcription factors are associated with expression of multiple genes involved in carbon metabolism in maize[J].Plant J,2000,21(3):281-288.
• [19]ROSS E J H,STONE J M,ELOWSKY C G,et al.Activation of the Oryza sativa non-symbiotic haemoglobin-2 promoter by the cytokinin-regulated transcription factor,ARR1[J].J Exp Bot,2004,55(403):1721-1731.
• [20]辛鲁生,阳江华,唐朝荣.一种启动子克隆的改良Adaptor-PCR方法及在橡胶树上的应用实例[J].植物研究,2012,32(3):296-303.
• [21]杨维康,张道远,张立运,等.新疆主要柽柳属植物(Tamarix L.)的生态类型划分与生境相似性研究[J].干旱区地理,2004,27(2):186-192.
• [22]DIDHIOU C J,POPOVA O V,GOLLDACK D.Transcript profiling of the salt-tolerant Festuca rubra ssp.litoralis reveals a regulatory network controlling salt acclimatization[J].J Plant Physiol,2009,166(7):697-711.
• [23]PRZEDPELSKA-WASOWICZ E M,WIERZBICKA M.Gating of aquaporins by heavy metals in Allium cepa L.epidermal cells[J].Protoplasma,2011,248(4):663-671.
• [24]ZHU C,SCHRAUT D,HARTUNG W,et al.Differential responses of maize MIP genes to salt stress and ABA[J].J Exp Bot,2005,56(421):2971-2981.
• [25]MAHDIEH M,MOSTAJERAN A.Abscisic acid regulates root hydraulic conductance via aquaporin expression modulation in Nicotiana tabacum[J].J Plant Physiol,2009,166(18):1993-2003.