磷转运蛋白AsPT6对紫云英结瘤及固氮酶活性的影响
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摘要
采用组织定位、超表达及RNAi等方法对紫云英中的Pht1家族磷转运蛋白AsPT6进行了研究。时空表达结果显示,AsPT6基因在根、茎、叶中均有表达,接种根瘤菌后在根和根瘤中高量表达;化学组织定位分析表明,AsPT6在根和根瘤的维管组织及根和根瘤连接处均有表达,起到向根瘤转运磷的作用;超表达及RNAi结果显示AsPT6能显著影响紫云英的结瘤及固氮酶活性。这些研究结果表明,AsPT6参与根瘤形成和发育过程中磷的转运。
Phosphorus is an important element required for plant growth.The absorption and transportation of Pi are carried out by phosphate transporter in legumes.In this paper,phosphate transporter AsPT6,a new member of pht1 family,was identified in Astragalus sinicus.AsPT6 was expressed in leaves,stems and roots.The expression of AsPT6 was increased significantly in the roots and nodules when inoculated with Mesorhizobium huakuii 7653 R.The results of tissue localization showed that AsPT6 was expressed in the vascular tissue of roots,nodules,and the junctions between root and nodule,indicating that AsPT6 may play a role in transporting phosphate from the root to nodule.Overexpression and RNA silence of AsPT6 significantly affected the nodulation and nitrogenase activity of nodules.It is indicated that AsPT6 is involved in the transportation of phosphorus during the formation and development of nodules.
Phosphorus is an important element required for plant growth.The absorption and transportation of Pi are carried out by phosphate transporter in legumes.In this paper,phosphate transporter AsPT6,a new member of pht1 family,was identified in Astragalus sinicus.AsPT6 was expressed in leaves,stems and roots.The expression of AsPT6 was increased significantly in the roots and nodules when inoculated with Mesorhizobium huakuii 7653 R.The results of tissue localization showed that AsPT6 was expressed in the vascular tissue of roots,nodules,and the junctions between root and nodule,indicating that AsPT6 may play a role in transporting phosphate from the root to nodule.Overexpression and RNA silence of AsPT6 significantly affected the nodulation and nitrogenase activity of nodules.It is indicated that AsPT6 is involved in the transportation of phosphorus during the formation and development of nodules.
引文
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[22]AI P,SUN S,ZHAO J,et al.Two rice phosphate transporters,OsPht1;2and OsPht1;6,have different functions and kinetic properties in uptake and translocation[J].Plant J,2009,57(5):798-809.
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[24]QIN L,ZHAO J,TIAN J,et al.The high-affinity phosphate transporter GmPT5regulates phosphate transport to nodules and nodulation in soybean[J].Plant physiology,2012,159(4):1634-1643.
[25]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 physiology,2006,47(7):807-817.
[26]JONES D H,WINISTORFER S C.A method for the amplification of unknown flanking DNA:targeted inverted repeat amplification[J].Biotechniques,1993,15(5):894-904.
[27]SCOTTO-LAVINO E,DU G,FROHMAN M A.3′end cDNAamplification using classic RACE[J].Nat Protoc,2006,1(6):2742-2745.
[28]TATJEWSKI P.Iterative optimizing set-point control-the basic principle redesigned[J].IFAC proceedings volumes,2002,35(1):49-54.
[29]KARANDASHOV V,NAGY R,WEGM LLER S,et al.Evolutionary conservation of a phosphate transporter in the arbuscular mycorrhizal symbiosis[J].PNAS,2004,101(16):6285-6290.
[30]SHEN J,YUAN L,ZHANG J,et al.Phosphorus dynamics:from soil to plant[J].Plant physiology,2011,156(3):997-1005.
[31]QIN L,GUO Y X,CHEN L Y,et al.Functional characterization of 14Pht1family genes in yeast and their expressions in response to nutrient starvation in soybean[J].PLoS One,2012,7(10):e47726.
[2]GRAHAM P H,VANCE C P.Legumes:importance and constraints to greater use[J].Plant physiology,2003,131(3):872-877.
[3]LINDSTROM K,MURWIRA M,WILLEMS A,et al.The biodiversity of beneficial microbe-host mutualism:the case of rhizobia[J].Res Microbiol,2010,161(6):453-463.
[4]梅江鹏,李豪,张忠明.拟南芥中根瘤共生基因线路的搭建及其表达分析[J].华中农业大学学报,2018,37(2):28-33.
[5]杨倩倩,刘元,陈大松,等.蒺藜苜蓿中1个聚半乳糖醛酸酶基因突变体的筛选及共生固氮表型的初步鉴定[J].华中农业大学学报,2017,36(4):62-70.
[6]汪涛,冯勇,张忠明.百脉根LjSERKs基因家族在根瘤共生中的功能鉴定[J].华中农业大学学报,2018,37(2):9-15.
[7]DANIEL W I.Symbiotic dinitrogen fixation and host-plant growth during development of and recovery from phosphorus[J].Physiologia plantarum,1993,88(2):294-300.
[8]CORDELL D,DRANGERT J O,WHITE S.The story of phosphorus:global food security and food for thought[J].Global environmental change,2009,19(2):292-305.
[9]CALDERN-VZQUEZ C,RUAIRIDH J H S,HERRERA-ES-TRELLA L.Phosphate deprivation in maize:genetics and genomics[J].Plant physiology,2011,156(3):1067-1077.
[10]JAKOBSEN I.The role of phosphorus in nitrogen fixation by young pea plants(Pisum sativum)[J].Physiologia plantarum,1985,64(2):190-196.
[11]ISRAEL D W.Investigation of the role of phosphorus in symbiotic dinitrogen fixation[J].Plant physiology,1987,84:835-840.
[12]TANG C,HINSINGER P,DREVONJ J,et al.Phosphorus deciency impairs early nodule functioning and enhances proton release in roots of Medicago truncatula[J].Ann Bot,2001,88:131-138.
[13]YANG Y.The effect of phosphorus on nodule formation and function in the casuarina-frankia symbiosis[J].Plant soil,1995,176(1):161-169.
[14]ALMEIDA J F,HARTWIG U A,FREHNER M,et al.Evidence that P deficiency induces N feedback regulation of symbiotic N fixation in white clover(Trifolium repens L.)[J].JExp Bot,2000,51(348):1289-1297.
[15]DREW E A,MURRAY R S,SMITH S E,et al.Beyond the rhizosphere:growth and function of arbuscular mycorrhizal external hyphae in sands of varying pore sizes[J].Plant soil,2003,251:105-114.
[16]SMITH S E,SMITH F A,JAKOBSEN I.Mycorrhizal fungi can dominate phosphate supply to plants irrespective of growth responses[J].Plant physiology,2003,133(1):16-20.
[17]RAGHOTHAMA K G.Phosphate acquisition[J].Annu Rev Plant Physiol Plant Mol Biol,1999,50:665-693.
[18]郑璐,包媛媛,张鑫臻,等.植物磷转运蛋白基因的研究进展[J].生态环境学报,2017,26(2):342-349.
[19]STEPHEN R M,ANNE L R,EUGENE D,et al.Expression analysis suggests novel roles for members of the Pht1family of phosphate transporters in Arabidopsis[J].Plant J,2002,31(3):341-353.
[20]TIAN J,VENKATACHALAM P,LIAO H,et al.Molecular cloning and characterization of phosphorus starvation responsive genes in common bean(Phaseolus vulgaris L.)[J].Planta,2007,227(1):151-165.
[21]MARYAM N E,MIYAKO K,KIEN H N,et al.Adaptation of the symbiotic Mesorhizobium-chickpea relationship to phosphate deficiency relies on reprogramming of whole-plant metabolism[J].PNAS,2016,113(32):E4610-E4619.
[22]AI P,SUN S,ZHAO J,et al.Two rice phosphate transporters,OsPht1;2and OsPht1;6,have different functions and kinetic properties in uptake and translocation[J].Plant J,2009,57(5):798-809.
[23]陈丽玉,秦璐,赵静,等.豆科植物Pht1磷转运蛋白家族基因的研究进展[J].大豆科学,2015,34(6):1057-1065.
[24]QIN L,ZHAO J,TIAN J,et al.The high-affinity phosphate transporter GmPT5regulates phosphate transport to nodules and nodulation in soybean[J].Plant physiology,2012,159(4):1634-1643.
[25]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 physiology,2006,47(7):807-817.
[26]JONES D H,WINISTORFER S C.A method for the amplification of unknown flanking DNA:targeted inverted repeat amplification[J].Biotechniques,1993,15(5):894-904.
[27]SCOTTO-LAVINO E,DU G,FROHMAN M A.3′end cDNAamplification using classic RACE[J].Nat Protoc,2006,1(6):2742-2745.
[28]TATJEWSKI P.Iterative optimizing set-point control-the basic principle redesigned[J].IFAC proceedings volumes,2002,35(1):49-54.
[29]KARANDASHOV V,NAGY R,WEGM LLER S,et al.Evolutionary conservation of a phosphate transporter in the arbuscular mycorrhizal symbiosis[J].PNAS,2004,101(16):6285-6290.
[30]SHEN J,YUAN L,ZHANG J,et al.Phosphorus dynamics:from soil to plant[J].Plant physiology,2011,156(3):997-1005.
[31]QIN L,GUO Y X,CHEN L Y,et al.Functional characterization of 14Pht1family genes in yeast and their expressions in response to nutrient starvation in soybean[J].PLoS One,2012,7(10):e47726.