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:/www.ecologica.cn “:SEnp V S “(20050307031); 81 S “(BK2006140)l :2008-05-09; :2008-12-10YTCorrespondingauthor.E-mail:srguonjau.edu.cnZ k(SpinaciaoleranceaL.)= s8= s Y孙 锦1, 2,贾永霞1,郭世荣1,李 娟1(1. 2 jv, 2 210095;2. 8 j S v , 730070)K1:Z =k 0Z 3| k, ZE, Z k(SpinaciaoleranceaL.)= s8(ROS)= s (Chl)Y。TV ,Z /, 2k = s8 =| 0(O-2 ) 3 q、V _(H2O2)d= (MDA)c A 6,i O=k4v 3|;=k= s8 = (SOD)、 F V (APX)、!t (GR) F (AsA)、!t(GSH)c ( 3|,7V (POD) 3|;4J,(MV)B P2k = s8 =O-2 3 qF y、H2O2c 4、 VF, b“4AsA A k= s8 =ROS , MV/ V 。Z /, 2k = = s b(Chlb)、= s a(Chla)# -8= s (Pchl)、 (Mg-protoIX)、(Proto) U(Uro)c A,7 (PBG)- (ALA),Chl sE,i O=k sEv 3|;MVBF sE,7AsAs Z MV/ET。Z A4 =k= = s (Chlase)7 3| Y,MV) =kChlaseYv 3|,AsA2 = Chlase AY。 T ,Z /,k= s8 =ROSChl MM1,YV= s8 PChl,7 O PChlPBG_Uro sE。 0Z 3|= s8 b“ROS1YVSODAsA-GSH“d, b“ ? ,h ROS= s8 Chl sE,i OZ ChlaseY l;7Z =k= s8 b“ROS1G SODPOD,ROS b“ ? K,V7 ROSv ,= s8 Chl sE,i OZ A4 Chlase,F Chl。1oM:Z ;k(SpinaciaoleranceaL.);= s8;= s cI|:1000-0933(2009)08-4361-11 ms |:Q143, Q945, Q948 DS M :AEffectsofseawaterstressonmetabolismofreactiveoxygenspeciesandchlorophylinchloroplastsofspinach(SpinaciaoleranceaL.)SUNJin1, 2, JIAYong-Xia1, GUOShi-Rong1, LIJuan11ColegeofHorticulture, NanjingAgriculturalUniversity, Nanjing210095, China2VegetableInstitute, GansuAcademyofAgriculturalSciences, Lanzhou730070, ChinaActaEcologicaSinica, 2009, 29(8):4361 4371.Abstract:Theefectsofseawaterstresonmetabolismofreactiveoxygenspecies(ROS)andchlorophyl(Chl)inchloroplastoftwospinachcultivars, Yuanyespinach(seawatersensitivecultivar)andHelanNo.3 (seawatertolerantcultivar)wereinvestigatedbyhydroponics.Theresultsshowedthatproductionrateofsuperoxideradical(O-2), contentofhydrogenperoxide(H2O2)andmalonaldehyde(MDA)inchloroplastoftwospinachcultivarswereremarkablyincreasedbyseawaterwithhigherrateincv.Yuanye;Underthestresofseawater, theactivityofanti-oxidationenzymesuchassuperoxidedismutase(SOD), ascorbicacidperoxidase(APX)andglutathionereductase(GR), andthecontentofhtp:/www.ecologica.cnantioxidantsuchasascorbicacid(AsA)andglutathionereduced(GSH)inchloroplastofcv.Yuanyewerelowerthanthoseofcv.HelanNo.3, whileperoxidase(POD)activityinchloroplastofcv.Yuanyewashigherthanthatofcv.HelanNo.3;Thesupplementofmethylviologen(MV), thephoto-oxidant, enhancedtheproductionrateofO-2, thecontentofH2O2 andMDAinchloroplastoftwospinachcultivars, whiletheadditionofAsA, theeliminatorforROS, decreasedROSlevelandaleviatedoxidizationofplasmolemma.Thecontentofchlorophylb(Chlb), chlorophyla(Chla)andprecursorofChlsuchasprotochlorophyl(Pchl), Mg-protoporphyrin (Mg-ProtoIX), protoporphyrin (Proto )anduroorphyrinogen (Uro)wasremarkablydecreased, butthecontentofporphobilinogen(PBG)and-aminolevulinicacid(ALA)wasincreasedbythestressofseawater, whichleadtoinhibitionforChlsynthesis, andtheinhibitionwasaggravatedbyMVandeliminatedbyAsA.Chlorophylase(Chlase)activityintheleavesofcv.YuanyewasimprovedwhileitwasnotinfluencedintheleavesofHelanNo.3underseawaterstress.TheChlaseactivityintheleavesofcv.YuanyewasmoregreatlyafectedbyMVtreatmentthanthatofHelanNo.3, however, ChlaseactivityoftwocultivarswasnotinfluencedbyAsA.TheseresultssuggestthatROSiscloselyrelatedtoChlmetabolism, whichnotonlyinjureplasmolemmabutalsoinhibittheprocesoftransformationofPBGtoUro asaresultofChldecomposition.Inseawatertolerantcultivar(cv.HelanNo.3), theROSeliminationismainlydependedonSODandAsA-GSHsystem, whichcouldaleviateoxidationinjuryofROStochloroplastmembraneandinhibitionofChlsynthesis, andChlaseactivitywaslessinfluencedbyseawaterstress;whileinseawatersensitivecultivar(cv.Yuanye), itismainlydependedonSODandPOD, whichhaslimitedabilitytoeliminateROSandleadtoaccumulatemassROStosevereoxidationinjuryofchloroplastmembraneandinhibitionofChlsynthesis, andChlaseactivitywassignificantlyimprovedbyseawaterstress, whichenhancedChldecomposition.KeyWords:seawater;spinach(SpinaciaoleranceaL.);chloroplast;reactiveoxygenspecies;chlorophyl= s (Chl) o K1; , V lv ; ?,7 O;“d#;“dQ 1 。8 =Chlc | %Chl 2 。Chl 33 BCp,YChl,V7 Chlc ;Chl ; Q , = s (Chlase)Chl1“ M 4 ;(ROS)9 V Chl 5,Yuzo 6 ROS V ? Chl4- 6 ,V7F Chl。 7 、SO28 、NO2 9、 s 5 、 10Hq/,ChlROS1; /,ROS ChlEChl#E 。= s8 3ROS1% 11, /= s8 =ROSv 12, 13 ,= s8$ ? 3VQ,; ,Y ;T。Z /,k(SpinaciaoleranceaL.)= Chlc 14, ; q Chlc 15 。yN,Z /k= s8ROSChl1“, Z T 3 3Y 1 IN。 k ZE, Z k= s8ROSChlY, = s8ROSChl1“,Z T4 G 。1 ZE1.1 k kk Z =k 0Z 3| 14;Z |1S Z,9c 26.64gL-1,pH7.8, 0F:Na+:349.98 mmolL-1;Cl-:410.14 mmolL-1;Mg2+:40.20 mmolL-1;SO2-4 :21.15mmolL-1;Ca2+:7.61 mmolL-1;K+:6.83 mmolL-1。1.2 ) !k2007 M811 2 jv1 e i =。k -14 , ks2s:Z k= s8ROSY:2 sY !2) ,1/2 Hoagland!A(vu,EC0.14 dsm-1,pH7.3)c40%Z 1/2 Hoagland!A() u,EC1.77dsm-1,pH7.6),sYV U:Y:=kv;H: 3|v;Y+S:=k) ;H+S: 3|) 。104362 3 29 htp:/www.ecologica.cn?,|) c40%Z 1/2Hoagland!A,) 3?6? |1 3/45= 4 |= s8,M1S;Z /J,(MV) F (AsA)k= ChlY。2 sY !8) :1/2 Hoagland!A(v u,CK);1/2 Hoagland!A+1molL-1 MV(CK+MV);1/2 Hoagland!A+5mmolL-1 AsA(CK+AsA);1/2 Hoagland!A+1molL-1MV+5mmolL-1 AsA(CK+MV+AsA);c40%Z 1/2 Hoagland!A(T);c40%Z 1/2Hoagland!A+1molL-1 MV(T+MV);c40%Z 1/2Hoagland!A+5mmolL-1 AsA(T+AsA);c40%Z 1/2Hoagland!A+1molL-1 MV+5mmolL-1 AsA(T+MV+AsA)。Z ) 2?,sY1 molMV5mmolL-1 AsA= ,MV H8s 0.05%Tween20 ,vF8s 0.05%Tween20 b ) ;24hMV) B5mmolL-1 AsA= , 6B b = ,72 h |1 3/45= Chl -8c = s8 =| 0(O-2 ) 3 q、V _(H2O2)d= (MDA)c #Chlase,i9 O-2 3 q、H2O2MDAc M(M=) /v)。 2 k (3Q, 。1.3 S#ZE1.3.1 = s84 | ITakeda 16ZE。 = s8B,Percol k41500g/0 23min,|4 |= s8M, P= s8 ir1mg.ml-1,= s8, qr90%。1.3.2 O-2 3 q、 (SOD)、V (POD)0.05molL-1、pH7.8 A(PBS) d= s825,(protein)c I E;O-2 3 qS 17ZE,nmolmin-1mg-1Chl;SODS 18ZE, :(NBT);50% 1 (U),Umin-1g-1proteinV U;POD7 !pE 19,1 min Xb470 nm/ODMB (U),Umin-1g-1protein。 1.3.3 H2O2c 0.2molL-1HClO4 d= s83,vUchida 20ZE,molmg-1ChlV U。1.3.4 MDAc 5% oY (TCA) d= s83,vHeathPacker 21ZE,molmg-1ChlV U。1.3.5 F V (APX)、!t (GR)0.05 molL-1K2HPO4-KH2PO4 A(pH7.0) d= s82 5, APXNakanoAsada22ZE, molAsAmin-1g-1proteinV U;GRvFoyerHaliwel 23ZE, molNADPHmin-1g-1proteinV U。1.3.6 F (AsA)c 8s 151 5%TCA d= s8,vD 24 ZE,mgg-1ChlV U。1.3.7 !t(GSH)!t(GSSG)c 5% d= s83,vElman 25ZE,mmolg-1ChlV U。1.3.8 Chl -8c = s a(Chla)、= s b(Chlb)c d4 |E26 ,V Umgg-1.FW;(Proto)、 (Mg-ProtoIX)、= s (Pchl)HodginsVanHuysteeZE27, U(Uro) (PBG)BogoradZE 28,- (ALA)v ZE29 , (V Ugg-1FW。1.3.9 ChlasekChlQ,vMargaret 30ZE,molChlmin-1g-1FW。4363 8 :Z k(SpinaciaoleranceaL.)= s8= s Y htp:/www.ecologica.cn1.4 sk MicrosoftExcelSPSS10.0 qs,DuncansE1 。2 Ts2.1 Z k= s8O-2 3 q、H2O2MDAc YZ /, k = s8O-2 3 q(m1)、H2O2c (m1) 3? 6,6?/ AMv,i O=k 6v 3|。 ) 6?=ksY440.98%32.82%, 3|sY415.35%20.93%。Z /,2k = s8MDAc (m1)M? pO-2 3 qH2O2c M 。 Vn,Z /,k= s8O-2 3 qF y,H2O2v , VMDAc 9F,= s8 s ; 0Z 3| O-23 qH2O2c ,MDAc M ,= s8 l。m1 Z k= s8O-2 3 q、H2O2MDAc YFig.1 EfectsofseawaterstressonproductionrateofO-2 , contentofH2O2 andMDAinchloroplastsofspinachY:=kv,controlofYuanyespinach;Y+S:=k40%Z i) , 40%concentrationofseawatertreatmentofYuanyespinach;H:3|v,controlofHelanNo.3;H+Y: 3|40%Z i) , 40%concentrationofseawatertreatmentofHelanNo.3;mM3 VUsA(PD0.05),SameletersmeantnosignificantdiferenceatP0.05levelwithinthecharts2.2 Z k= s8SOD、PODYZ 3?,=k 3|= s8 =SOD(m2)sY1Mv421.35%14.11%; 6?,=k1vA49.78%,7 3|1vA430.12%。Z /,=k= s8 =POD(m2) 6, 3?1v454.38%, 6?1v4118.23%;7 3| 3? vA489.49%, 6? A,1v421.21%。 Vn,Z /, 0Z 3| S SOD,O-2 b“ ? v=k,7POD) 6?/,H2O2 b“ ? l=k。2.3 Z k= s8 =APX、GRYZ /,=k= s8APX(m3)A/,3?6?sY v66.90%75.76%,7 3|= s8APXA 6,3?6?sY v486.03%75.52%;GR(m3)APXM ,=k= s8GR v,7 3| v4。 Vn,Z /, 3|= s8 APXGR,H2O2 b“ ? ;7=k= s8APXGR/,H2O2 b“ ? ,V7H2O2= s8 =v 。2.4 Z k= s8AsA、GSHc GSH/GSSG1YZ /,=k= s8AsAc (m4) 3?6?sY vA36.14%48.19%,7 3| 3?vsA,6? vA25.52%;GSH(m4)AsA4364 3 29 htp:/www.ecologica.cnM ,=k= s8 =GSHc ) 3?6?sY v21.61%37.06%,7 3|3?vsA,6? v19.97%;=k= s8GSH/GSSG1(m4) 3?6?sY vA72.22%74.06%,7 3|) 6?vsA。Vn,Z /, 3|= s8AsAGSHc Al=k,i OGSH/GSSG1M A, GSHGSSGW 。2.5 Z /MVAsAk= s8O-2 3 q、H2O2MDAc MYm5TA U,Z /2k = s8O-2 3 q、H2O2MDAc M (v1;vHq/,MV) k= s8O-2 3 q、H2O2MDAc M9v1,AsA) Ml1,7MV+AsA) BMV) ;Z /,MV、AsAMV+ASA) f vM 。 Z Pk= s8 =ROS 4, VF;MV) B PROS9F, VF4365 8 :Z k(SpinaciaoleranceaL.)= s8= s Y htp:/www.ecologica.cn,7AsA V b“sROS, P V。2.6 Z /MV、AsAk= Chlb、Chla# -8Pchl、Mg-protoIX、Proto、Uro、PBGALAcYV1TV ,Z P2k = Chlb、Chla、Pchl、Mg-protoIX、ProtoIX、Uroc (1Mv,7PBGALAc 6, 3|= Uroc l=k,sY12.50%25.34%, Z /2k Chl sE,i O 3| sEl=k。vHq/,MV) Chl -8c YZ ) M ,MVEvZ ) ;AsA) P=kChl -8c 3|Y A;MV+AsA P2k = Chlb、Chla、Pchl、Mg-protoIX、ProtoIX、Uroc BMV) 4,7 PPBGALAc 。Z /,) Chl -8c YvM 。 Vn,Z Pk= Chl sE, 34366 3 29 htp:/www.ecologica.cn| sEl=k;MVBF sE,AsAs ET。2.7 Z /MV、AsAk= ChlaseYZ A4=k= Chlase(m6),7 3| AY。vZ /,MVMV+AsA) A4=k= Chlase,7AsA AY。 3|vHq/,MV、AsAMV+AsA) (Chlase AY;Z ) /,MVMV+AsAA4Chlase,7AsA AY。V1 Z /MV、AsAk= = s -8c YTable1 EffectsofMVandAsAonChlprecursorscontentofspinachleavesunderseawaterstress) TreatmentsChlb(mgg-1 FW)Y HChla(mgg-1 FW)Y HPchl(gg-1FW)Y HMg-ProtoIX(gg-1FW)Y HProto(gg-1FW)Y HUro(gg-1FW)Y HPBG(gg-1 FW)Y HALA(gg-1FW)Y 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 0.835fCK+MV 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 5.543aCK+AsA 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 1.546eCK+MV+AsA 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 1.298dT+MV 0.106e 0.256d 0.322f 0.679d 0.209e 0.303f 0.118f 0.211d 0.316f 0.467e 4.321e 7.220e15.