海水胁迫对菠菜(Spinacia olerancea L.)叶绿体活性氧和叶绿素代谢的影响

29 8 2009 M83 ACTAECOLOGICASINICAVol.29,No.8Aug., 2009htp//[ “SEnp V S[ “20050307031; 81 – S[ “BK2006140l 2008-05-09;  2008-12-10＊YT€Correspondingauthor.E-Z } k’SpinaciaoleranceaL. s8Ÿ„ s }†•Y孙 锦1, 2,贾永霞1,郭世荣1,＊,李 娟11. 2 jv\, 2 210095;2. 8 j S v’ ,  730070K1[Z  k’„ 0Z € 3| k‹, ZE, Z } k’SpinaciaoleranceaL. s8ŸROS„ s Chl}†•Y。TV ,Z } /, 2k’  s8 | 0O-2  3 q、V _H2O2„d „MDAc A 6,i Ok’4–v€ 3|;k’ s8   SOD、 F V APX、‡t GRŸ„ F AsA、˜‡tGSHc  € 3|,7V PODŸ€ 3|;;4J,MVB„ P2k’  s8 O-2 3 qF y、H2O2c 4、 VF,Ÿ b“4AsA A† k’ s8 ROS , MV/ V ]。Z } /, 2k’   s bChlb、 s aChla † -8 s Pchl、 ⅨMg-protoIX、ⅨProtoⅨ„ UⅢUroⅢc  A†,7‰ PBG„δ-[ ALA,Chl† sžE,i Ok’ sEv€ 3|;MVB„F  ƒ sE,7AsA†s Z } „MV/ET。Z } A4 k’  s ChlaseŸ7€ 3| •Y,MV k’ChlaseŸ•Yv€ 3|,ŒAsA2  ChlaseŸ  A•Y。  T ,Z }/,k’ s8 ROSChl}† MM1,‚YV s8  ] PChl†,7 O PChl†PBG_UroⅢ„ sE。 0Z € 3| s8 b“ROS1YVSOD„AsA-GSH“d, b“  ,h ROS s8  „Chl† sE,i OZ }  ChlaseŸ•Y l;7Z  k’ s8 b“ROS1G SOD„POD,ROS b“ K,V7 ROSv  , s8  „Chl† sE,i OZ } A4 ChlaseŸ,F  Chl†。1oMZ ;k’SpinaciaoleranceaL.; s8;Ÿ; s cI|1000-0933200908-4361-11 ms |Q143, Q945, Q948 DS M ’AEffectsofseawaterstressonmetabolismofreactiveoxygenspeciesandchlorophylinchloroplastsofspinachSpinaciaoleranceaL.SUNJin1, 2, JIAYong-Xia1, GUOShi-Rong1,＊, LIJuan11ColegeofHorticulture, NanjingAgriculturalUniversity, Nanjing210095, China2VegetableInstitute, GansuAcademyofAgriculturalSciences, Lanzhou730070, ChinaActaEcologicaSinica, 2009, 2984361～ 4371.AbstractTheefectsofseawaterstresonmetabolismofreactiveoxygenspeciesROSandchlorophylChlinchloroplastoftwospinachcultivars, YuanyespinachseawatersensitivecultivarandHelanNo.3 seawatertolerantcultivarwereinvestigatedbyhydroponics.TheresultsshowedthatproductionrateofsuperoxideradicalO-2, contentofhydrogenperoxideH2O2andmalonaldehydeMDAinchloroplastoftwospinachcultivarswereremarkablyincreasedbyseawaterwithhigherrateincv.Yuanye;Underthestresofseawater, theactivityofanti-oxidationenzymesuchassuperoxidedismutaseSOD, ascorbicacidperoxidaseAPXandglutathionereductaseGR, andthecontentofhtp//antioxidantsuchasascorbicacidAsAandglutathionereducedGSHinchloroplastofcv.Yuanyewerelowerthanthoseofcv.HelanNo.3, whileperoxidasePODactivityinchloroplastofcv.Yuanyewashigherthanthatofcv.HelanNo.3;ThesupplementofmethylviologenMV, thephoto-oxidant, enhancedtheproductionrateofO-2, thecontentofH2O2 andMDAinchloroplastoftwospinachcultivars, whiletheadditionofAsA, theeliminatorforROS, decreasedROSlevelandaleviatedoxidizationofplasmolemma.ThecontentofchlorophylbChlb, chlorophylaChlaandprecursorofChlsuchasprotochlorophylPchl, Mg-protoporphyrinⅨ Mg-ProtoIX, protoporphyrinⅨ ProtoⅨ anduroorphyrinogenⅢ UroⅢwasremarkablydecreased, butthecontentofporphobilinogenPBGandδ-aminolevulinicacidALAwasincreasedbythestressofseawater, whichleadtoinhibitionforChlsynthesis, andtheinhibitionwasaggravatedbyMVandeliminatedbyAsA.ChlorophylaseChlaseactivityintheleavesofcv.YuanyewasimprovedwhileitwasnotinfluencedintheleavesofHelanNo.3underseawaterstress.TheChlaseactivityintheleavesofcv.YuanyewasmoregreatlyafectedbyMVtreatmentthanthatofHelanNo.3, however, ChlaseactivityoftwocultivarswasnotinfluencedbyAsA.TheseresultssuggestthatROSiscloselyrelatedtoChlmetabolism, whichnotonlyinjureplasmolemmabutalsoinhibittheprocesoftransationofPBGtoUroⅢ asaresultofChldecomposition.Inseawatertolerantcultivarcv.HelanNo.3, theROSeliminationismainlydependedonSODandAsA-GSHsystem, whichcouldaleviateoxidationinjuryofROStochloroplastmembraneandinhibitionofChlsynthesis, andChlaseactivitywaslessinfluencedbyseawaterstress;whileinseawatersensitivecultivarcv.Yuanye, itismainlydependedonSODandPOD, whichhaslimitedabilitytoeliminateROSandleadtoaccumulatemassROStosevereoxidationinjuryofchloroplastmembraneandinhibitionofChlsynthesis, andChlaseactivitywassignificantlyimprovedbyseawaterstress, whichenhancedChldecomposition.KeyWordsseawater;spinachSpinaciaoleranceaL.;chloroplast;reactiveoxygenspecies;chlorophyl s Chl  o K1;† ,‚ V[ lv ; ,7 O;“d;“dQ‹[ 1] 。8 Chlc  | Chl†† ™[ 2] 。Chl 3†[3]  B„„Cp,•YChl†,V7„ Chlc †;Chl† “ \†„;† Q‹ ˜,  s ChlaseChl†1“ M[ 4] ;ŸROS9 V„ Chl†[ 5],Yuzo[ 6] ROS V • Chl4- 6 ,V7F Chl†。[ 7] 、SO2[8] 、NO2[ 9]、 s} [5] 、 š [10]Hq/,Chl†ROS}†1;} /,ROS Ž•Chl†„E]Chl†[E]‚ 。 s8  3ROS1 [11],} / s8 ROSv  [ 12, 13] , s8  3VQ‹,;† †,•Y ;†T。Z } /,k’SpinaciaoleranceaL. Chlc †[ 14], ;† q†› † Chlc † [ 15] 。yN,Z } /k’ s8ROSChl}†1“, Z } T 3 3•Y 1• IN。 k ZE, Z } k’ s8ROSChl}†•Y,[   s8ROSChl}†1“,Z  T4 ‚G 。1 ‹ ZE1.1 k‹ kk’ Z  k’„ 0Z € 3|[ 14];Z |1S  Z,9c 26.64gL-1,pH7.8, 0FNa349.98 mmolL-1;Cl-410.14 mmolL-1;Mg240.20 mmolL-1;SO2-4 21.15mmolL-1;Ca27.61 mmolL-1;K6.83 mmolL-1。1.2  k2007 M811 2 jv1 e i ›。k’ ] -[14] , ks2†s›①Z } k’ s8ROS}†•Y2 sY 2 ,1/2 HoaglandAvu,EC0.14 dsm-1,pH7.3„c40Z 1/2 HoaglandA u,EC1.77dsm-1,pH7.6,sYV UYk’v;H€ 3|v;YSk’ ;HS€ 3| 。104362 3   29  htp//,|  c40Z 1/2HoaglandA, 3„6 |1 3/45 4 |›œ s8,M1S;②Z } /J,MV„ F AsAk’ Chl}†•Y。2 sY 8 1/2 HoaglandAv u,CK;1/2 HoaglandA1μmolL-1 MVCKMV;1/2 HoaglandA5mmolL-1 AsACKAsA;1/2 HoaglandA1μmolL-1MV5mmolL-1 AsACKMVAsA;c40Z 1/2 HoaglandAT;c40Z 1/2HoaglandA1μmolL-1 MVTMV;c40Z 1/2HoaglandA5mmolL-1 AsATAsA;c40Z 1/2HoaglandA1μmolL-1 MV5mmolL-1 AsATMVAsA。Z  2,sY1 μmolMV„5mmolL-1 AsA ,MV  H8s ”0.05Tween20 ,vF8s ”0.05Tween20 b  ;24hMV B5mmolL-1 AsA , 6B  b  ,72 h |1 3/45 ›Chl -8c „ s8 | 0O-2  3 q、V _H2O2„d „MDAc [ChlaseŸ,i9 O-2 3 q、H2O2„MDAc MM /v。  2[ k 3Q,› †  † 。1.3 SZE1.3.1 ›œ s84 |• ITakeda[ 16]ZE。  s8B,Percol k41500g/0 2～3min,|4 | s8Mžœ, P s8 irž1mg.ml-1, s8›œŸ,›œ qrž90。1.3.2 O-2 3 q、  SODŸ、V PODŸ0.05molL-1、pH7.8 APBS d s82～5,‘proteinc  I ” E;O-2 3 qS[ 17]ZE,†nmolmin-1mg-1Chl;SODŸS[ 18]ZE,[]‹ NBT;50 1 †U,[Umin-1g-1proteinV U;PODŸ7 pE[ 19],[1 min X‘b470 nm/ODMB  †U,†Umin-1g-1protein。 1.3.3 H2O2c 0.2molL-1HClO4 d s83,•vUchida[ 20]ZE,[μmolmg-1ChlV U。1.3.4 MDAc 5 oY TCA d s83,•vHeath„Packer[ 21]ZE,[μmolmg-1ChlV U。1.3.5 F V APX、‡t GRŸ0.05 molL-1K2HPO4-KH2PO4 ApH7.0 d s82 ～ 5, APXŸNakano„Asada[22]ZE, Ÿ[μmolAsAmin-1g-1proteinV U;GRŸ•vFoyer„Haliwel[ 23]ZE, Ÿ[μmolNADPHmin-1g-1proteinV U。1.3.6 F AsAc 8s ”1∶51 5TCA d s8,•vD[ 24] ZE,[mgg-1ChlV U。1.3.7 ˜‡tGSH„˜‡tGSSGc 5   d s83,•vElman[ 25]ZE,†[mmolg-1ChlV U。1.3.8 Chl† -8c  s aChla、 s bChlbc d[4 |E[26] ,†V Umgg-1.FW;ⅨProtoⅨ、 ⅨMg-ProtoIX、 s PchlHodgins„VanHuysteeZE[27], UⅢUroⅢ„‰ PBGBogoradZE[ 28],δ-[ ALA•v ZE[29] ,† V Uμgg-1FW。1.3.9 ChlaseŸ[k’ChlQ‹,•vMargaret[ 30]ZE,†μmolChlmin-1g-1FW。4363 8  Z } k’SpinaciaoleranceaL. s8Ÿ„ s }†•Y htp//1.4 ” sk ” MicrosoftExcel„SPSS10.0 qs,DuncansE›1 。2 Ts2.1 Z } k’ s8O-2 3 q、H2O2„MDAc •YZ } /, k’  s8O-2 3 qm1、H2O2c m1 3 6,6{/†Œ AM‹v,i Ok’ 6–v€ 3|。  6k’sY440.98„32.82,€ 3|sY415.35„20.93。Z } /,2k’  s8MDAc m1M pO-2 3 q„H2O2c M 。 Vn,Z } /,k’ s8O-2 3 qF y,H2O2v  , VMDAc 9F, s8  sž  ];Œ 0Z € 3| O-23 q„H2O2c  ,MDAc M‹ , s8  ] l。m1 Z } k’ s8O-2 3 q、H2O2„MDAc •YFig.1 EfectsofseawaterstressonproductionrateofO-2 , contentofH2O2 andMDAinchloroplastsofspinachYk’v,controlofYuanyespinach;YSk’40Z i , 40concentrationofseawatertreatmentofYuanyespinach;H€3|v,controlofHelanNo.3;HY€ 3|40Z i , 40concentrationofseawatertreatmentofHelanNo.3;mM]3 VUs‚APD0.05,SameletersmeantnosignificantdiferenceatP0.05levelwithinthecharts2.2 Z } k’ s8SOD、PODŸ•YZ } 3,k’„€ 3| s8 SODŸm2sY1M‹v421.35„14.11;} 6,k’1vA†49.78,7€ 3|1vA430.12。Z } /,k’ s8 PODŸm2 6,} 31v454.38,} 61v4118.23;7€ 3|} 3 vA489.49,} 6 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3| UroⅢc †–lk’,sY†12.50„25.34, Z } /2k’ Chl† sE,i O€ 3| sElk’。vHq/,MV Chl -8c •YZ  M ,ŒMVE]vZ  ;AsA Pk’Chl -8c  € 3|•Y A;MVAsA P2k’  Chlb、Chla、Pchl、Mg-protoIX、ProtoIX、UroⅢc  †BMV  4,7 PPBG„ALAc †。Z } /, Chl -8c •YvM 。 Vn,Z } Pk’ Chl† sžE,€ 34366 3   29  htp//| sElk’;MVB„F   sE,AsA†s ƒET。2.7 Z } /MV、AsAk’ ChlaseŸ•YZ } A4k’ ChlaseŸm6,7€ 3|  A•Y。v„Z }/,MV„MVAsA A4k’ ChlaseŸ,7AsA   A•Y。€ 3|vHq/,MV、AsA„MVAsA ChlaseŸ  A•Y;Z  /,MV„MVAsAA4ChlaseŸ,7AsA   A•Y。V1 Z } /MV、AsAk’  s † -8c •YTable1 EffectsofMVandAsAonChlprecursorscontentofspinachleavesunderseawaterstress TreatmentsChlbmgg-1 FWY HChlamgg-1 FWY HPchlμgg-1FWY HMg-ProtoIXμgg-1FWY HProtoⅨμgg-1FWY HUroⅢμgg-1FWY HPBGμgg-1 FWY HALAμgg-1FWY HCK 0.202b 0.350a 0.537b 0.815a 0.379b 0.475b 0.250b 0.265b 0.646b 0.639b 13.423b14.831a10.558d10.161f0.782d 0.835fCKMV 0.144d 0.250d 0.428e 0.695d 0.263d 0.402d 0.141d 0.197d 0.398d 0.520d 3.624e 6.607e12.806c12.145d3.454a 5.543aCKAsA 0.253a 0.358a 0.648a 0.804a 0.418a 0.480b 0.309a 0.267b 0.721a 0.753a 15.613a15.079a11.346d10.825ef2.483b 1.546eCKMVAsA 0.157c 0.301c 0.467cd0.772b 0.301c 0.470b 0.192c 0.245c 0.488c 0.553cd 6.593d 9.409d11.219d11.484de3.031ab 4.948bT 0.165c 0.296c 0.482c 0.727bc0.265d 0.436c 0.141d 0.241c 0.365e 0.578c 10.022c12.977b13.633bc13.600c1.523c 1.298dTMV 0.106e 0.256d 0.322f 0.679d 0.209e 0.303f 0.118f 0.211d 0.316f 0.467e 4.321e 7.220e15.