论文

富集小麦磷胁迫响应基因的cDNA差减文库构建和部分ESTs的功能鉴定

  • 龙素霞 ,
  • 路文静 ,
  • 谷俊涛 ,
  • 郭程瑾 ,
  • 肖凯
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  • 1. 河北农业大学 农学院, 河北 保定 071001;
    2. 河北农业大学 生命科学学院, 河北 保定 071001
龙素霞( 1984- ), 女, 河北邯郸人, 在读硕士, 主要从事作物高产与分子生物学研究.

收稿日期: 2008-10-10

  网络出版日期: 2014-10-14

基金资助

国家"973"项目(2007CB116209);河北省自然科学基金项目(C2007000476)

Construction of a cDNA Subtractive Library Enriched the Response Genes of Deficient-Pi Stress and Functional Identification of Some ESTs in the Library

  • LONG Su-xia ,
  • LU Wen-jing ,
  • GU Jun-tao ,
  • GUO Cheng-jin ,
  • XIAO Kai
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  • 1. College of Agronomy, Agricultural University of Hebei, Baoding 071001, China;
    2. College of Life Science, Agricultural University of Hebei, Baoding 071001, China

Received date: 2008-10-10

  Online published: 2014-10-14

摘要

植物在形态学和生化代谢水平上对低磷胁迫逆境的响应,是特异表达基因在时空上精细协同作用的结果.以前期工作中鉴定的磷高效小麦品种石新828为材料.构建了富集不同低磷处理时间点特异表达基因的根系cDNA差减抑制杂交文库.获得的克隆总数为2 682个.对随机选取的文库克隆研究发现,克隆中插入的片段长度为20~70 bp.测序结果和功能比对发现,具有功能比对结果的克隆比例为70%,其中部分分别与小麦、大麦、水稻、玉米和拟南芥等植物种属高度同源,参与转录调控、蛋白质合成和代谢等多个生物学过程.该富集低磷胁迫响应基因文库为进一步鉴定小麦响应磷胁迫逆境的基因调控网络和克隆重要的磷高效相关基因奠定了坚实的基础.

本文引用格式

龙素霞 , 路文静 , 谷俊涛 , 郭程瑾 , 肖凯 . 富集小麦磷胁迫响应基因的cDNA差减文库构建和部分ESTs的功能鉴定[J]. 华北农学报, 2009 , 24(2) : 12 -16 . DOI: 10.7668/hbnxb.2009.02.003

Abstract

The responses of plant to deficient-Pistress on phenotype and biochemical levels are the distinctly cooperative results resulting from the specific expressed genes at the spatiotemporal range. In this study,using Shixin828,the wheat cultivar with high phosphorus utilization efficiency identified previously to be the experimental material,a root cDNA selected2subtractive suppression library enriched the response genes of deficient2Pi stress at various time-points was constructed.The clone number in total was 2682 in this library.It was found that the insertion lengths changed from 350 bp to 750 bp,by analyzing the randomly selected clones in the library.The clones with putative functions and the clones with unknown functions were comprised of the percentage of 70 % and 30 %,respectively.The former,being homogenous to wheat,barley,rice,maize,and Arabidopsis,owned the functions in the plant by involving in the transcriptional regulation,protein synthesis,and biochemical metabolism.Therefore,the root cDNA selected-subtractive suppression library will be useful for further identification of the gene2regulatory network to respond to the deficient-Pi and cloning of important genes related to forming the high-phosphorus utilization efficiency.

参考文献

[1] Raghothama K G,Karthikeyan A S.Phosphate acquisition[J].Plant Soil,2005,274:37-49.
[2] Vance C P,Uhde-Stone C,Allan D L.Phesphorus acquisition and use:critical adaptations by plants for securing a nonrenewable resource[J].New Phytol,2003,157:423-447.
[3] Misson J,Raghothama K G,Jain A,et al.A genome-wide transcriptional analysis using Arabidopsis thaliana Affymetrix gene chips determined plant responses to phosphate deprivation[J].Proc Natl Sci USA,2005,102:11934-11939.
[4] Uhde-Stone C,Zinn K E,Mario R,et al.Nylon falter arrays reveal differential gene expression in proteoid roots of white lupin in response to phosphorus deficiency[J].Plant Physiol,2003,131:1064-1079.
[5] Xiao K,Bai G H,Carver B F.Nylon filter arrays reveal differential expression of expressed sequence tags in wheat roots under aluminum stress[J].J Integ Plant Biol,2005,47:839-848.
[6] Ouyang B,Yang T,Li H,et al.Identification of early salt stress response genes in tomato root by suppression subtractive hybridization and microarray analysis[J].J Exp Bot,2007,58:507-520.
[7] Wang X L,He R F,He G C.Construction of suppression subtractive hybridization libraries and identification of brown planthopper-induced genes[J].J Plant Physiol,2005,162:1254-1262.
[8] Hammond J P,Broadly M R,White P J.Genetic responses to phosphorus deficiency[J].Ann Bot,2004,94:323-332.
[9] Lambers H Y,Shane M W,Cramer M D,et al.Root structure and functioning for efficient acquisition of phosphorus:matching morphological and physiological traits[J].Ann Bot,2006,98:693-713.
[10] Lynch J P.Root architecture and plant productivity[J].Plant Physiol,1995,109:7-13.
[11] Yan X,Liao H,Beebe S E,et al.QTL mapping of root hair and acid exudation traits and their relationship to phosphorus uptake in common bean[J].Plant Soil,2004,265:17-29.
[12] Hill J O,Simpson R J,Moore A D,et al.Morphology and response of roots of pasture species to phosphorus and nitrogen nutrition[J].Plant Soil,2006,286:7-19.
[13] Ochoa I E,Blair M W,Lynch J P.QTL analysis of adventitious root formation in common bean under contrasting phosphorus availability[J].Crop Sci,2006,46:1609-1621.
[14] Johnson J F,Vance C P,Allan D L.Phosphorus deficiency in Lupinus albus.Altered lateral root development and enhanced expression of phosphoenolpyruvate carboxylase[J].Plant Physiol,1996,112:31-41.
[15] Neumann G,Maosonneau A,Martinoia E,et al.Physiological adaptations to phosphorus deficiency during proteoid root development in white lupin[J].Planta,1999,208:373-382.
[16] Smith S E,Smith F A,Jakobsen I.Mycorrhizal fungi can dominate phosphate supply to plants irrespective of growth Responses[J].Plant Physiol,2003,133:16-20.
[17] West H M,Fitter A H,Watkinson A R.Respouse of Vulpia ciliata ssp ambigua to removal of mycorrhizal infection and to phosphate application under field conditions[J].J Ecol,1993,81:351-358.
[18] Jones D L.Organic acids in the rhizosphere:a critical review[J].Plant Soil,1998,205:25-44.
[19] Chen D L,Delatorre C A,Backer A,et al.Conditional identification of phosphate starvation response mutants in Arabidopsis thaliana[J].Planta,2000,211:13-22.
[20] Nǖnberger T,Abel S,Jost W,et al.Induction of an extracellular ribonuclease in cultured tomato cells upon phosphate starvation[J].Plant Physiol,1990,92:970-976.
[21] Duff S M,Sarath G,Plaxton W C.The role of acid phosphatases in plant phosphorous metabolism[J].Physiol Plant,1994,90:791-800.
[22] Duff S M,Moorhead G B,Lofebvre D D,et al.Phosphate starvation inducible bypasses of adenylate and phosphate dependent glycolytic enzymes in Brassica nigras Suspension cells[J].Plant Physiol,1989,90:1275-1278.
[23] Ticconi C A,Delatorre C A,Abel S.Attenuation of phosphate starvation responses by phosphite in Arabidopsis[J].Plant Physiol,2001,127:963-972.
[24] Jiang C,Gao X,Liao L,et al.Phosphate starvation root architecture and anthocyanin accumulation responses are modulated by the gibberellin-DELLA signaling pathway in Arabidopsis[J].Plant Physiol,2007,145:1460-1470.
[25] Essigmann B,Gvler S,Narang R A,et al.Phosphate availability affects the thylakoid lipid composition and the expression of SQD1,a gene required for sulfolipid biosynthesis in Arabidopsis thaliana[J].Proc Nail Acad Sci USA,1998,950:1950-1955.
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