水稻铁氧化酶基因OsLPR5对拟南芥根系发育和养分利用的影响
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摘要
[目的]本文旨在研究水稻铁氧化酶基因OsLPR5对拟南芥根系发育和养分利用的影响,从而解析其作用。[方法]通过蛋白表达、纯化及酶活性测定,确定OsLPR5蛋白具有铁氧化酶活性;利用农杆菌介导花序侵染法创制OsLPR5转化拟南芥atlpr1突变体的转基因材料,通过根系形态鉴定和多种养分含量测定,研究OsLPR5对转基因植株根系发育及养分利用的影响。[结果]p GS-21a-OsLPR5融合蛋白酶活性明显高于对照;低磷条件下,与突变体相比,OsLPR5转化株主根变短,根系总磷、氮、钾、钠、铁、铜和锌等多种养分含量降低,趋于野生型; OsLPR5的表达显著回补了At LPR1基因的突变对主根表型和养分含量的影响,表明OsLPR5在植株根系构型和多种养分利用方面起到重要作用。[结论]OsLPR5蛋白具有铁氧化酶活性,在拟南芥转基因植株中参与低磷条件下根系构型和养分利用等重要过程。
[Objectives]This work is aimed at confirming the effects of rice ferroxidase gene OsLPR5 on root growth and nutrient utilization in Arabidopsis plants,so as to investigated its function.[Methods]We analysed the ferroxidase activity of OsLPR5 protein expressed in bacteria by specific activity analysis.We obtained transgenic Arabidopsis plants overexpressing OsLPR5 in the atlpr1 background using the Agrobacterium-mediated floral dip method,and examined the impact of OsLPR5 overexpression on root morphology and nutrient utilization known to be related to phosphorous nutrition or its regulation.[Results]Ferroxidase activity of p GS-21 a-OsLPR5 protein was significantly higher than the control.Under the condition of low phosphorus,the primary roots of OsLPR5 transformed lines became shorter,and the contents of total phosphorus,nitrogen,potassium,sodium,iron,copper and zinc decreased,which tended to wild type.The expression of OsLPR5 significantly compensated for the effect of At LPR1 mutation on primary root phenotype and nutrient content,indicating that OsLPR5 plays an important role in root architecture and nutrient utilization.[Conclusions]The results confirm that OsLPR5 protein probably has ferroxidase activity,and indicate that OsLPR5 is probably involved in processes,including root morpholog and nutrient utilization to environmental phosphorus stress.
[Objectives]This work is aimed at confirming the effects of rice ferroxidase gene OsLPR5 on root growth and nutrient utilization in Arabidopsis plants,so as to investigated its function.[Methods]We analysed the ferroxidase activity of OsLPR5 protein expressed in bacteria by specific activity analysis.We obtained transgenic Arabidopsis plants overexpressing OsLPR5 in the atlpr1 background using the Agrobacterium-mediated floral dip method,and examined the impact of OsLPR5 overexpression on root morphology and nutrient utilization known to be related to phosphorous nutrition or its regulation.[Results]Ferroxidase activity of p GS-21 a-OsLPR5 protein was significantly higher than the control.Under the condition of low phosphorus,the primary roots of OsLPR5 transformed lines became shorter,and the contents of total phosphorus,nitrogen,potassium,sodium,iron,copper and zinc decreased,which tended to wild type.The expression of OsLPR5 significantly compensated for the effect of At LPR1 mutation on primary root phenotype and nutrient content,indicating that OsLPR5 plays an important role in root architecture and nutrient utilization.[Conclusions]The results confirm that OsLPR5 protein probably has ferroxidase activity,and indicate that OsLPR5 is probably involved in processes,including root morpholog and nutrient utilization to environmental phosphorus stress.
引文
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[9]Ai P H,Sun S B,Zhao J N,et al.Two rice phosphate transporters,OsP ht1;2 and OsP ht1;6,have different functions and kinetic properties in uptake and translocation[J].Plant Journal,2009,57(5):798-809.
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[13]Chevalier F,Pata M,Nacry P,et al.Effects of phosphate availability on the root system architecture:large-scale analysis of the natural variation between Arabidopsis accessions[J].Plant Cell&Environment,2003,26(11):1839-1850.
[14]Smith S E,Dickson S,Smith F A.Nutrient transfer in arbuscular mycorrhizas:how are fungal and plant processes integrated?[J].Functional Plant Biology,2001,28(7):685-696.
[15]Lynch J P,Brown K M.Topsoil foraging:an architectural adaptation of plants to low phosphorus availability[J].Plant and Soil,2001,237(2):225-237.
[16]Zhang C,Simpson R J,Kim C M,et al.Do longer root hairs improve phosphorus uptake?Testing the hypothesis with transgenic Brachypodium distachyon lines overexpressing endogenous RSL genes[J].New Phytologist,2018,217(4):1654-1666.
[17]Ticconi C A,Lucero R D,Sakhonwasee S,et al.ER-resident proteins PDR2 and LPR1 mediate the developmental response of root meristems to phosphate availability[J].Proc Natl Acad Sci USA,2009,106(33):14174-14179.
[18]Cao Y,Ai H,Jain A,et al.Identification and expression analysis of OsL PR family revealed the potential roles of OsL PR3 and 5 in maintaining phosphate homeostasis in rice[J].BMC Plant Biology,2016,16(1):210.
[19]Müller J,Toev T,Heisters M,et al.Iron-dependent callose deposition adjusts root meristem maintenance to phosphate availability[J].Developmental Cell,2015,33(2):216-230.
[20]Li H J.Comparative study on determination of phosphorus content in two kinds of plants[J].Modern Agricultural Science&Technology,2012,11:16-17.
[21]Svistoonoff S,Creff A,Reymond M,et al.Root tip contact with low-phosphate media reprograms plant root architecture[J].Nature Genetics,2007,39(6):792-796.
[22]Shin H,Shin H S,Dewbre G R,et al.Phosphate transport in Arabidopsis:pht1;1 and pht1;4 play a major role in phosphate acquisition from both low-and high-phosphate environments[J].Plant Journal,2004,39(4),629-642.
[23]Nagarajan V K,Jain A,Poling M D,et al.Arabidopsis Pht1;5 mobilizes phosphate between source and sink organs and influences the interaction between phosphate homeostasis and ethylene signaling[J].Plant Physiology,2011,156(3):1149-1163.
[24]Daram P,Brunner S,Rausch C,et al.Pht2;1 encodes a low-affinity phosphate transporter from Arabidopsis[J].Plant Cell,1999,11(11):2153-2166.
[25]Liu T Y,Huang T K,Yang S Y,et al.Identification of plant vacuolar transporters mediating phosphate storage[J].Nature Communications,2016,7:11095.
[26]Solomon E I,Augustine A J,Yoon J.O2reduction to H2O by the multicopper oxidases[J].Dalton Transactions,2008,252(30):3921-3932.
[2]Raghothama K G,Karthikeyan A S.Phosphate acquisition[J].Plant and Soil,2005,274:37-49.
[3]Rausch C,Bucher M.Molecular mechanisms of phosphate transport in plants[J].Planta,2002,216(1):23-37.
[4]Jones D L,Rousk J,Edwards-Jones G,et al.Biochar-mediated changes in soil quality and plant growth in a three year field trial[J].Soil Biology&Biochemistry,2012,45:113-124.
[5]Marschner H.Mineral nutrition of higher plants[J].The Journal of Ecology,1988,76(4):1250.
[6]Richardson A E,Barea J M,McN eill A M,et al.Acquisition of phosphorus and nitrogen in the rhizosphere and plant growth promotion by microorganisms[J].Plant and Soil,2009,321(1):305-339.
[7]梁永书,周军杰,南文斌,等.水稻根系研究进展[J].植物学报,2016,51(1):98-106.Liang Y S,Zhou J J,Nan W B,et al.Progress in rice root system research[J].Bulletin of Botany,2016,51(1):98-106(in Chinese with English abstract).
[8]吴伟明,程式华.水稻根系育种的意义与前景[J].中国水稻科学,2005,19(2):174-180.Wu W M,Cheng S H.Significance and prospects of breeding for root system in rice(Oryza sativa)[J].Chinese Journal of Rice Science,2005,19(2):174-180(in Chinese with English abstract).
[9]Ai P H,Sun S B,Zhao J N,et al.Two rice phosphate transporters,OsP ht1;2 and OsP ht1;6,have different functions and kinetic properties in uptake and translocation[J].Plant Journal,2009,57(5):798-809.
[10]Ticconi C A,Abel S.Short on phosphate:plant surveillance and countermeasures[J].Trends in Plant Science,2004,9(11):548-555.
[11]Williamson L C,Ribrioux S P,Fitter A H,et al.Phosphate availability regulates root system architecture in Arabidopsis[J].Plant Physiology,2001,126(2):875-882.
[12]Hammond J P,White P J.Sucrose transport in the phloem:integrating root responses to phosphorus starvation[J].Journal of Experimental Botany,2008,59(1):93-109.
[13]Chevalier F,Pata M,Nacry P,et al.Effects of phosphate availability on the root system architecture:large-scale analysis of the natural variation between Arabidopsis accessions[J].Plant Cell&Environment,2003,26(11):1839-1850.
[14]Smith S E,Dickson S,Smith F A.Nutrient transfer in arbuscular mycorrhizas:how are fungal and plant processes integrated?[J].Functional Plant Biology,2001,28(7):685-696.
[15]Lynch J P,Brown K M.Topsoil foraging:an architectural adaptation of plants to low phosphorus availability[J].Plant and Soil,2001,237(2):225-237.
[16]Zhang C,Simpson R J,Kim C M,et al.Do longer root hairs improve phosphorus uptake?Testing the hypothesis with transgenic Brachypodium distachyon lines overexpressing endogenous RSL genes[J].New Phytologist,2018,217(4):1654-1666.
[17]Ticconi C A,Lucero R D,Sakhonwasee S,et al.ER-resident proteins PDR2 and LPR1 mediate the developmental response of root meristems to phosphate availability[J].Proc Natl Acad Sci USA,2009,106(33):14174-14179.
[18]Cao Y,Ai H,Jain A,et al.Identification and expression analysis of OsL PR family revealed the potential roles of OsL PR3 and 5 in maintaining phosphate homeostasis in rice[J].BMC Plant Biology,2016,16(1):210.
[19]Müller J,Toev T,Heisters M,et al.Iron-dependent callose deposition adjusts root meristem maintenance to phosphate availability[J].Developmental Cell,2015,33(2):216-230.
[20]Li H J.Comparative study on determination of phosphorus content in two kinds of plants[J].Modern Agricultural Science&Technology,2012,11:16-17.
[21]Svistoonoff S,Creff A,Reymond M,et al.Root tip contact with low-phosphate media reprograms plant root architecture[J].Nature Genetics,2007,39(6):792-796.
[22]Shin H,Shin H S,Dewbre G R,et al.Phosphate transport in Arabidopsis:pht1;1 and pht1;4 play a major role in phosphate acquisition from both low-and high-phosphate environments[J].Plant Journal,2004,39(4),629-642.
[23]Nagarajan V K,Jain A,Poling M D,et al.Arabidopsis Pht1;5 mobilizes phosphate between source and sink organs and influences the interaction between phosphate homeostasis and ethylene signaling[J].Plant Physiology,2011,156(3):1149-1163.
[24]Daram P,Brunner S,Rausch C,et al.Pht2;1 encodes a low-affinity phosphate transporter from Arabidopsis[J].Plant Cell,1999,11(11):2153-2166.
[25]Liu T Y,Huang T K,Yang S Y,et al.Identification of plant vacuolar transporters mediating phosphate storage[J].Nature Communications,2016,7:11095.
[26]Solomon E I,Augustine A J,Yoon J.O2reduction to H2O by the multicopper oxidases[J].Dalton Transactions,2008,252(30):3921-3932.