采用PCR方法分别从玉米郑单958基因组DNA及RNA中克隆了菌根共生磷酸盐转运蛋白基因ZmPT6，序列分析表明，ZmPT6基因全长cDNA1665bp，包括一个长97bp的内含子；编码554个氨基酸，属于Pht1家族成员，由12个疏水的跨膜结构域组成。并构建了35S启动子驱动的植物正义、反义及RNAi表达载体，为下一步玉米的遗传转化及ZmPT6基因的功能研究奠定了基础。
The arbuscular mycorrhizal symbiotic phosphate transporter gene ZmPT6 was cloned from Zea mays Zhengdan 958 genomic DNA and RNA by PCR method.Sequences analysis showed that ZmPT6 fulllength cDNA 1665bp,and contain a intron of 97bp.ZmPT6 belong to member of Pht1 family,encoded 554 amino acid,and consist of 12 putative hydrophobic transmembrane domains.Three plant transformation vectors,sense vector p2300-35S-ZmPT6,antisense p2300-35S-ZmPT6 r,and RNAi vector p2300-35S-ZmPT6 RNAi were constructed and so as to provide basis of further genetic transformation and function of ZmPT6 gene.
[1] Heckman D S,Geiser D M,Eiddel B R,et al.Molecular evidence for the early colonization of land by fungi and plants[J].Science,2001,293:1129-1133.
[2] Martin P.Arbuscular mycorrhiza:the mother of plant root endosymbioses[J].Nat Rev Microbiol,2008,6:763-775.
[3] Smith S E,Read D J.Mycorrhizal symbiosis[M].Academic Press,San Diego,1997.
[4] Bago B,Pfeffer P E,Shachar-Hill Y.Carbon metabolism and transport in arbuscular mycorrhizas[J].Plant Physiol,2000,124:949-958.
[5] Schachtman D P,Reid R J,Ayling S M.Phosphorus uptake by plants:From soil to cell[J].Plant Physiology,1998,116:447-453.
[6] Harrison M J,Dewbre G R,Liu J.A phosphate transporter from Medicago truncatula involved in the acquisition of phosphate released by arbuscular mycorrhizal fungi[J].Plant Cell,2002,14:2413:2429.
[7] Rausch C,Daram P,Brunner S,et al.A phosphate transporter expressed in arbuscule-containing cells in potato[J].Nature,2001,414:462-470.
[8] Paszkowski U,Kroken S,Roux C,et al.Rice phosphate transporters include an envolutionarily divergent gene specifically activates in arbuscular mycorrhizal symbiosis[J].Proceedings of the National Academy of Science,USA,2002,99:13324-13329.
[9] Nagy R,Karandashov V,Chague V,et al.The characterization of novel mycorrhiza-specific phosphate transporters from Lycopersicon esculentum and Solanum tuberosum uncovers functional redundancy in symbiotic phosphate transport in solanaceous species[J].Plant Journal,2005,42:236-250.
[10] Glassop D,Smith S E,Smith F W.Cereal phosphate transporters associated with the mycorrhizal pathway of phosphate uptake into roots[J].Planta,2005,222:688-698.
[11] Maeda D,Ashida K,Iguchi K,et al.Knockdown of an arbuscular mycorrhiza-inducible phosphate transporter gene of Lotus japonicus suppresses mutualistic symbiosis[J].Plant Cell Physiol,2006,47:807-817.
[12] Chen A Q,Hu J,Sun S B,et al.Conservation and divergence of both phosphate-and mycorrhiza-regulated physiological responses and expression patterns of phosphate transporters in Solanaceous species[J].New Phytologist,2007,173:817-831.
[13] Javot H,Penmetsa R V,Terzaghi N,et al.A medicago truncatula phosphate transporter indispensable for the arbuscular mycorrhizal symbiosis[J].Proceedings of the National Academy of Science,USA,2007,104:1720-1725.
[14] Nagy R,Vasconcelos M J V,Zhao S,et al.Differential regulation of five Pht1 phosphate transporters from maize (Zea mays L.)[J].Plant Biol,2006,8:186-197.
[15] Pao S S,Paulsen I T,Saier M H Jr.Major facilititator superfamily[J].Microbiology and Molecular Biology,1998,62:1-34.
[16] Wassenegger M,Pelissier T.A model for RNA-mediated gene silencing in higher plants[J].Plant Mol Biol,1998,37(2):349-362.
[17] Cogoni C,Macino G.Homology-dependent gene silencing in plants and fungi:A number of variations on the same theme[J].Curr Opin Microbiol,1999,2(6):657-662.
[18] Elbashir S M,Lendeckel W,Tuschl T.RNA interference is mediated by 21-and 22-nucleotide RNAs[J].Genes Dev,2001,15(2):188-200.
[19] Smith N A,Singh S P,Wang M B,et al.Total silencing by intron spliced hairpin RNAs[J].Nature,2000,407:319-320.
[20] Fire A,Xu S,Montgomery M K,et al.Potent and specific genetic interference by double stranded RNA in Caenorhabditis elegans[J].Nature,1998,391:806-811.
[21] Fitter A H.Darkness visible:reflections on underground ecology[J].J Ecol,2005,93,231-243.
