Through Thick and Thin Life is like a long trip, We should study hard and explore all the time. In my opinion, we must make a lot of decisionss in our life. some decisions are very easy to make, the others are difficult. however, no matter how difficult it is, we should learn to choose something what we want. Facing to difficult problem, we should learn how to deal with it all the time. Don ’t give up ! Show your power to your family or friends. If you insist to do it , you will be successful finally. On the other hand , facing to the easy problem, we should know everything is not easy what you want. Nobody can guess the next problem when to face. We should cherish the time and deal with the problem seriously an the time. Believe yourself! The wonderful life will come. In a word we should believe ourselves and consider seriously to make decision in our life!
()自然科学版广西师范大学学报 第 21 Juan 第 3 期 V o l. 21 N o. 3 2003 Nian 9 月 Sep tem ber 2003 JOU RNAL O F GUAN GX INORMAL UN IV ERS ITY DO ES ADA PTA T ION TO IN CR EA SED CON CEN TRA T ION S O F N aC l . M OD IF Y TH E R ESPON SE O F POR PH YR ID IUM SPTO O TH ER ?ENV IRONM EN TAL STR ESSES 1 2, -L IU X iaocan Av igad Von shak (1. , , 541004, ;Co llege of L ife SciencesGuangx i N o rm al U n iversityGu ilin Ch ina 2. , , M icroalgal B io thechno logy L abo rato rythe Jacob B lau stein In stitu te fo r D esert R esearch 2, 84990, )BenGu r io n U n iversity of the N egevSede Boker Israel :. AbstractW ith an a im to sub stan t ia te the hypo thesis that under one k ind of stress P orp hy rid ium spcells , m igh t deve lop a comm on sen so r and a respon sive m echan ism effective to o ther environm en ta l stresses ƒ. 1. 5 , P orp hy rid ium spcu ltu res w ere expo sed to m o lL N aC l t ill the cells reached to a new steady state , (4 ?). then fu rther sub jected to pho to inh ib it io no r ch illing tem peratu re C U nder bo th pho to inh ib it io n and , 2ch illing tem peratu rea decline in the pho to syn thetic activity of salt p restress cu ltu re w as found that p ro 2 .ceeded faster and reached to a low er level than in the con t ro l cu ltu re : . ; ; ; Key wordsP orp hy rid ium spsalin ity stresspho to inh ib it io nch illing tem peratu re : 945. 78 : : 1001- 6600 (2003) 03- 0072- 05CLC num berQ D ocum en t codeA Art icle ID 1 In t roduct io n . , P orp hy rid ium sphas attracted m uch atten t io n due to the econom ica l and pha rm aceu t ica l im po r2 [ 1, 3 ] tance of it s po lysaccharides and essen t ia l fatty acidsand the un icellu la r natu re and the sim p licity fo r [ 4 ]. . studying pho to syn thesis and ab io t ic stress adap ta t io n sP orp hy rid ium spis relatively stab le over a [ 5 ](20, 160 ?) , (2, 10) . w ide range of tem peratu res C pH values and salin it iesIn m any cases in teract io n be tw een these environm en ta l param eters p lays a m a jo r ro le in the ab ility of the cells to acclim ate o r [ 6 ]. adap t to the environm en ta l stressesIt has been suggested that algal and p lan t system s have deve lop ed . , a comm on sen so r and respon se m echan ism to environm en ta l stressT herefo readap t ing to one environ2 [ 7 ].m en ta l stress m ay resu lt in a better respon se and the ab ility to adap t to ano ther environm en ta l stress Ch lo rop lasts are p rim ary redox sen so rs of environm en ta l change that act together w ith o ther signal [ 8, 9 ]. t ran sduct io n pathw ays to elicit app rop ria te physio lo gical and m o lecu la r respon sesBohnert and [ 10 ] cow o rkerssuggested that adju stm en t of p lan t w ater statu s is a comm on featu re of acclim at io n to all of .these im po rtan t environm en ta l stresse s T h is study w as to sub stan t ia te the hypo thesis that a comm on sen so r and a respon se m echan ism have .deve lop ed in o rder to adju st to vario u s environm en ta l stresse s 2 M aterials and M ethods 2. 1 Growth cond it ion and sa l in ity trea tm en t [ 11 ] 2. 1. 5 ƒP orp hy rid ium spcu ltu res grow n in artificial seaw aterw ith o r w ithou t m o lL N aC l w ere in : 2003202224Rece ived da te : Founda tion itemSuppo rted by the Foundat io n of Bonna T erra : 2—) , , , , ,. .(1963B iographyL IU X iao can fem alebo rn in M ashanGuangx iL ectu rer of Guangx i N o rm al U n iversityM sC ?(25 ) , cubated in a gyrato ry shaker at a con stan t tem peratu re C and illum inated by f luo rescence lam p s - 2 - 1(2, 110 ??. 1% ) 100GRO L uxat pho ton f lux den sity of Λm o lm sA ir en riched w ith CO 2 w as sparged .in to the incubato rs 2. 2 Respon ses of sa lt adapted cells to the stress of photo inh ib it ion or ch ill ing tem pera ture 2. 2. 1 Pho to inh ib it io n (A lgal cells in the lo g phase of grow th w ere harvested and resu spended in fresh m edium A SW and + 1. 5 ƒ) 20 ƒA SW m o lL N aC l respectivelyw ith the addit io n of mm o lL N aHCO 3 to f inal ch lo rophyll a [ 6 ]- 2 - 1 25 ƒ. 150 ??concen t rat io n of m gL T heir pho to syn thetic activity w as m easu red under Λm o lm sas 0.t im e T he cu ltu res w ere p laced in a doub le jacket the rm o regu la ted glass vessel and then illum inated by a - 2 - 1(220, 230 , 1 000 ) 2 500 ??. h igh in ten sity ha lo gen lam p O SRAM V W at Λm o lm sA t t im e in ter2 ƒ, , (+ 1. 5 valssam p les w ere draw n ou tdilu ted w ith fresh m edium A SW and A SW m o lL N aC l ) 20 ƒ5 ƒ. respectively w ith the addit io n of mm o lL N aHCO 3 to m g LT heir pho to syn thetic activity w as - 2 - 1150 ??.m easu red under Λm o lm s 2. 1. 2 V ariab le ch lo rophyll f luo rescence " PS activity w as dete rm ined fo llow ing the variab le f luo rescence param eters of F o and Fm and the [ 12 ](- ) ƒ= ƒ.rat io of Fm F oFm F v Fm w as calcu la ted 2. 1. 3 Ch illing tem peratu re (+Exponen t ia lly grow ing cells w ere harvested and resu spended in fresh m edium A SW and A SW 1. 5 ) 20 ƒƒm o lL N aC l respectivelyw ith the addit io n of mm o lL N aHCO 3 to f inal ch lo rophyll a concen t ra2 - 2 - 1 ƒ??5 . 150 0. t io n of m gLT heir pho to syn thetic activity w as m easu red under Λm o lm sas t im e T he - 2 - 1 150, 170 ??sam p les w ere then incubated in ice w ater at ligh t in ten sity of Λm o lm sand taken ou t at . ƒ5 vario u s t im e in tervalsO xygen evo lu t io n rate w as m easu red and F v Fm w as reco rded after m in dark 25 ?.adap ta t io n at C 3 R esu lt s 3. 1 .In teract ion of sa l in ity and HPFD on Porp hyrid ium sp . In o rder to evaluate any po ssib le m odified respon ses of salt adap ted P orp hy rid ium spto pho to inh i2 , 1. 5 ƒ. , b it io n stresscells w ere f irst grow n in m o lL N aC lA f ter being acclim ated to salt stressas indicated , by estab lish ing a con stan t lo garithm ic grow ththe cells w ere sub sequen t ly expo sed to h igh irradiance (25 ?2). stress at op t im al tem peratu re C T he elicited respon se w as est im ated by m easu ring the ligh tdep en2 ƒ(. 1).den t O 2 evo lu t io n rates and F v Fm of the cu ltu res F ig 1 () , () 2 500 A s show n in F igu re A expo sing the cu ltu res to h igh pho to f lux den sity H PFD of Λm o l - 2 - 1 ??, . m sresu lted in a decline in their pho to syn thetic activityT h is reduct io n p roceeded faster and at a . , low er level of activity in the salt stress cu ltu re than in the con t ro l cu ltu reCon sisten t lythe m ax im al " (ƒ) (. 1) pho tochem ica l efficiency of PS F v Fm of salt adap ted cells F igB at differen t t im e in tervals w as ƒlow er than that of con t ro l although a sim ila r decline tendency of F v Fm w as found in bo th con t ro l and . . salt t reated cu ltu re sIt is thu s qu ite clear that P orp hy rid ium spcu ltu res grow n at h igh salin ity are m o re .sen sit ive to pho to inh ib it io n due to their reduced ab ility to u t ilize the ligh t energy ab so rbed 3. 2 (4 ?) .In teract ion of sa l in ity and ch ill ing tem pera ture Con Porp hyrid ium sp In o rder to dete rm ine w hether acclim at io n to h igh salin ity stress conferred p ro tect io n to the cells , . from ch illing tem peratu reP orp hy rid ium spcells grow n in con t ro l and salty m edium w ere expo sed to - 2 - 1 4 ?160 ??C at Λm o lm sand then their pho to syn thetic activity as w ell as pho tochem ica l efficiency of ("ƒ) .PS F v Fm w ere dete rm ined . 1 () " (ƒ) F igEffect of H PFD on oxygen evo lu t io n rate A and the m ax im al pho tochem ical efficiency of PS F v Fm () . 1. 5 ƒB of P orp hy rid ium spgrow n in con t ro l and mo lL N aC l respectively ?2 () , (4 ) , A s can be seen in F igu re A w hen cells are expo sed to ch illing tem peratu re C their pho to2 . syn thetic activity decline sT h is reduct io n p roceeded faster and reached a low er level of activity in the . , (" ƒ saline grow n than in the con t ro l cu ltu reM eanw h ilethe m ax im al pho tochem ica l efficiency of PS F v ) (. 2) .Fm of salt adap ted cells F igB at differen t t im e in tervals declined sim ila rly to that of the con t ro l . 2 (4 ?) () F igEffect of ch illing temperatu re C on the oxygen evo lu t io n rate A and m ax im al pho tochem ical efficien2 " (ƒ) () . 1. 5 ƒcy of PS F v Fm B of P orp hy rid ium spgrow n at con t ro l and mo lL N aC l respectively 4 D iscu ssio n 4. 1 . In teract ion of sa l in ity stress and photosyn thet ic photo inh ib it ion on P orp hyrid ium spcells . 2O u r experim en ts show ed that salt adap ted P orp hy rid ium spcells had a low er ligh tsatu rated rate of ; 2pho to syn thesisth is suggests that salin itystressed cells m igh t be con siderab ly m o re sen sit ive to pho to in2 [ 13 ] . 2h ib it io n than con t ro l cellsPatrick et alrepo rted that salin itystress of pho to syn thetic cells p redispo ses . the pho tochem ica l apparatu s to pho to inh ib it io nT he sam e phenom ena have been also found in Sp iru lina [ 14 ] [ 15 ]. p la tensisand green algae Ch lam y d om onas reinhard tii A lthough the underlying cau ses of th is salt 2, stressinduced su scep t ib ility to pho to inh ib it io n are unknow nit has been found that dam age by bo th ex2 [ 16 ]" . (680) cess ligh t and saline stress are associated w ith dam age to the react io n cen ter P of PS T h is . m ay indicate that stressed P orp hy rid ium spcells have a low er p ro tein syn thesis capacity and thu s a .slow er repair m echan ism 412 . In teract ion of sa l in ity and ch ill ing stresses on Porp hyrid ium spcells (25 ??) (4 ) A sh if t from op t im al grow th tem peratu re C to ch illing tem peratu res C w as found in th is study to cau se m o re p ronounced inh ib it io n of pho to syn thetic oxygen evo lu t io n activity in the salt t reated 2. . , P rop hy rid ium spcells than in the nonsalt t reated cu ltu re sSalin ity stressunder w h ich the cells su r2 , . vivedobv io u sly exacerbated the dep ressio n of pho to syn thesis cau sed by ch illing tem peratu reCh illing [ 17 ] tem peratu re has been p roved to cau se a drastic decrease in pho to syn thetic activity in p lan tsand in [ 18 ] [ 12 ] , , som e algal species such as Ch lorella ellip soid ea Ch lamy d om onas reinhard iiSp iru lina p la tensisand [ 19 ]. . U lva rotund ta T h is phenom enon can be also investigated in ou r resu lt sIt is w ell know n that ligh t , p roduces A T P and NAD PH by pho tochem ica l electron t ran spo rtw h ile the Calvin cycle p rovides their . 2m a jo r sinkA s tem peratu re decreases the activity of the Calvincycle enzym es as w ell as o ther so lub le en2 [ 17 ], .zym es decreaseresu lt ing in a decrease of the u t ilized pho ton s and an increase in pho to inh ib it io n , 4 ?" (ƒ) How everat C the m ax im al pho tochem ica l efficiency of PS F v Fm of bo th salt t reated and 2. nonsalt t reated cu ltu res decreased in a sim ila r w ay in ou r experim en tT h is suggests that the d im in ished . effect of salin ity on P orp hy rid ium sppho to syn thesis inh ib it io n induced by ch illing stress w as no t reflect2 2ƒ. " ed in the t im eco u rse changes of F v Fm valueSalt stress p er se has lit t le direct effect on PS pho to2 , "chem istrybu t ch illing tem peratu re p lays a crucial ro le in the effect of salt stress on PS pho tochem 2 , ƒistry in P orp hy rid ium cellsalthough the F v Fm rat io of the salt stressed cu ltu re m easu red befo re expo 2 (. 2).su re to ch illing tem peratu re w as sign if ican t ly low er than the con t ro l cu ltu re F igB [ 16 ][ 17 ] " . , PS has lo ng been con sidered as the p rim ary target of pho to inh ib it io nN everthelessSono ike 2I , , found that in ch illsen sit ive p lan tsthe t rea tm en t of ch illing tem peratu re affects PS rather than PS ". :References [ 1 ] () , . . . . : A rad M alisSPo lysaccharides of red m icroalgaeA Cohen ZChem icals from m icroalgaeC L ondonT aylo r and , 1999. 282—291.F rancis L td [ 2 ] , , , . L ap ido t M R aveh D Sivan A et a lStab le ch lo rop last t ran sfo rm at io n of the un icellu lar red alga P orp hy rid ium species . , 2002, 129: 7—12.J P lan t Physio l [ 3 ] , , , . . H uheihelM Ishanu V T al J et a lA ctivity of P orp hy rid ium sppo lysaccharide again st herpes sim p lex viru ses in vitro . , 2002, 50: 189—200.and in vivo J J B io chem B iop hys M ethods [ 4 ] . . . 2. : , V on shak APo rphyridium A M ichael AM icro a lgal b io techno logy C Cam b ridgeCam b ridge U n iversity P ress 1988. 123—134. [ 5 ] , , , . . , U cko M Cohen EGo rdin H et a lR elat io n sh ip betw een the un icellu lar red alga P orp hy rid ium spand its p redato r . . , 1989, 55 (11): 2 990—2 994.the dinoflagellate Gym nod in ium spJ A pp l Environ M icrob io l [ 6 ] , , , . V on shak A Kancharak sa N Bunnag B et a lRo le of ligh t and pho to syn thesis on the acclim at io n p rocess of the . , 1996, 8: 119—124.cyanobacterium Sp iru lina p la tensis to salin ity stressJ J A pp l Phyco l [ 7 ] , . : 2. , 2001, Kn igh t H Kn igh t M RA b io t ic stress signaling pathw aysspecificity and cro sstalk J T rends in P lan t Science () 6 6: 262—267. [ 8 ] , 2, . : A nderson J M Park YIChow W ST he grand design of pho to syn thesisacclim at io n of the pho to syn thetic apparatu s . , 1995, 46: 129—139.to environm en tal cuesJ Pho to syn th R es [ 9 ] ,, , . H uner N P A M axw ell D PGray G R et a lSen sing environm en tal temperatu re change th rough im balance betw een ; " . , 1996, 98: 358—364.energy supp ly and energy con sump t io nredox state of pho to system J Physio l P lan t [ 10 ] , , . . , 1995, 7: 1 099—1 111.Bohnert H J N elson D EJen sen R GA dap tat io n s to environm en tal stressesJ P lan t Cell [ 11 ] , , . . , 1963, 16:Jones R Speer H Ku ry WStudies on the grow th of red alga P orp hy rid ium cruentum J Physio l P lan t 636—643. [ 12 ] , . To rzillo GV on shak AEffect of ligh t and temperatu re on the pho to syn thetic activity of the cyanobacterium Sp iru li2 —. , 1994, 6: 399403.na p la tensis J B iom ass B io energy [ 13 ] , . 2Patrick J N A nastsio s MSalin itystress enhances pho to inh ib it io n of pho to syn thesis in Ch lamy d om onas reinhard tii . , 1989, 134: 619—622.J J P lan t Physio l [ 14 ] ,22. . () , V on shak A M ingT ao ZA dap tat io n of Sp iru lina p la tensis to salin itystressJ Comp B io chem Physio l Part A 1998, 120: 113—118. [ 15 ] , . Ro sa L F rancisco GIn teract io n betw een saline stress and pho to inh ib it io n of pho to syn thesis in the freshw ater green . , 1999, 37 (7ƒ algae Ch lamy d om onas reinhard tii im p licat io n s fo r glycero l pho top roduct io nJ J P lan t Physio l B io chem ) 8: 623—628. [ 16 ] . . , 1984, 35: 15—44. Pow els S BPho to inh ib it io n of pho to syn thesis induced by visib le ligh t J A nnu R ev P lan t Physio l[ 17 ] . I "Sono ike KT he differen t ro les of ch illing temperatu res in the pho to inh ib it io n of pho to system and pho to system . : , 1999, 48: 136—141.J J Pho tochem Pho tob io l BB io l [ 18 ] , , . . , , . O qu ist GGreer D O gren EL igh t stress at low temperatu re A Kyle D O sm ond CA rn tzen CPho to inh ib it io n . : , 1987. 67—87.C Am sterdamE lsevier Science Pub lishers B V [ 19 ] . 22. , F rank lin LT he effects of temperatu reacclim at io n on the pho to inh ib it io n respon ses of u lvaro tundata b lid J P lan2 ta 1994, 192: 324—331. Gao盐环境下紫球藻对其他逆境的适应性研究 1 2Liu晓灿, Av igad Von shak ()1. Guang西师范大学 生命科学学院, 广西 桂林 541004; 2. Ben2古里安大学 沙漠研究所, 斯德 泊可, 84990, Yi色列 摘 要: 紫球藻在盐逆境De胁迫下, 所产生的感应反映和适应机理有Ke能使其适应其他的逆境. 在此以生长在 1. 5 ƒHuan境下并达到对数期的紫球藻为试验材料, Guan察测定光抑制或低温对光合作用等主要生理 mo lL N aC l Sheng化参数的影响, 发现: 经过盐处理的紫Qiu藻在光抑制或低温逆境下, 光合效力下降Geng快更低. 关键词: 紫球藻; Yan胁迫; 光抑制; 低温 ()Ze任编辑 马殷华
() 植 物 学 报 2000 , 42 4:358 - 366 2 + Di温逆境中不同抗寒性植物细胞内 Ca细定Ping衡的 11121报令成报报报李报Hong王报德报Paul H L 报 2(1 . Zhong科院植物究国学研所 , 北京 100093 I; 2 . aboratory of Plant Hardiness , Department of Horticultural Science and Plant Biological Sciences L )Program University of Minnesota , St . Paul , MN 55108 , USA, ( ( ) Zhai要 : 报报报胞化报察揭示学 ,Bu抗寒的玉米 Zea mays L . cv. Black Mexican SweetHe抗寒的小偃麦 Triticum sect .2 + ) Trititrigia mackeyBao胞在 26 ?报浮培报报 ,报志 CaDing位的报酸报淀物主要分布在液泡沉内 ,报Bao报和报胞核中 很少报到 2 + 2 + CaChen 淀 ; 报 志 Ca- ATPase Huo 性 反 报 的 磷 酸 报 沉 淀 Wu 丰 富 地 分 布 在 报 膜 上 , Bao 示 报 两 报 植 物 报 Mo2 + Ca- ATPase在 26 ?Pei报中有着报高的活性 。报二者的报胞在当 4 ?Di培报温 1 h 和 3 h 后 ,二者Bao胞报和报胞核 中2 + 2 + 2 +De Ca报度都明报地增加 ,但报膜 Ca- ATPase Huo性有明报改没报 。在报报延报低培报温后 ,Er者之报的 Ca 2 + 2 + Ca- ATPase : ,Ca- ATPase ,Shui平和 活性报生明报的区报 在不抗寒的玉Mi报胞中 报膜 活性报著降低 甚至完全失2 + 2 + Huo ,报胞增加的内 Ca水平不降低 ,不Neng恢报 Ca的报定平衡 ,报胞的精报报受Po构坏 ; 抗寒的小偃报胞报麦相 反 ,2 + 2 + () Zai 3 h 以后的低培报报程中温 至 3 d,Bao膜 Ca- ATPase 仍然保持高活Xing ,报胞增加的内 Ca报度迅速降低 ,Hui 报 2 + 2 + 2 + Ca的报Ding平衡 ,并提高抗寒力 。报些报果表明 ,Zhi物在低件温条下 ,报膜 Ca- ATPase Huo性的报持亦报胞即内 Ca 报定平Heng的恢报其抗寒性有着密切的报系 与,报于Bao在逆境中生存起着重要作用它 。2 + 2 + 2 + () : Ca; Ca- ATPase ; Ca; ; ; Xi细细 报膜 的报定平衡 文章细号 植物Kang寒性 : 057727496 玉米 2000Xiao中细分细号 : Q945. 文献细细细 : 78A04203582092 + Ca - homeosta sis Differs Bet ween Plant Species with Different d- tolerance at 4 ? Col Chilling 1 1 2 1 1 J IAN Ling- Cheng, SUN Long- Hua, L I J i- Hong, WANG Hong, SUN De-Lan, 2Paul H L I (1 . Institute of Botany , The Chinese Academy of Sciences , Beijing 100093 , China ; 2 . L aboratory of Plant Hardiness , Department of Horticultural Science and Plant Biological Sciences )Program , University of Minnesota , St . Paul , MN 55108 , USA Abstract : A comparative study was carried out on the EM- cytochemical localization of calcium and 2 + ( Ca- ATPase activity in the suspension- cultured cells between the chilling- sensitive maize Zea mays L . cv. Black ) ( ) Triticum Trititrigia Mexican Sweetand chilling- insensitive Trititrigia sect . mackeyat 4 chilling. ? When maize and Tyititrigia cells were cultured at 26 ?, electron microscopic observations revealed that the electron- dense calcium antimonate deposits , an indication of the calcium localization , were localized mainly in the vacuoles , and few was found in the cytosol and nuclei . The electron- dense cerium phosphate deposits , 2 + ( ) an in2 dication of Ca- ATPase activity , were abundantly distributed on the plasma membrane PM. 2 + When the cells from both species were cultured at 4 ?for 1 and 3 h , an elevation of Calevel in the cytosol and nuclei was observed , whereas the cerium phosphate deposits on the PM showed no quantitative difference from those of the 26 ?- cultured cells , indicating that the enzymatic activities were not altered during these chilling 2 + 2 periods. However , there was a distinct difference in the dynamics of the Cadistribution and the PM Ca Received : 1999209217 Accepted : 2000201219Foundation item : The National Natural Science Foundation of China () 39870371. Calcium plays an important role as a active at all . The cellular structures showed damage . secondOn the contrary , the low temperature- induced increase in messenger in plant growth and development , and in and adaptation of plants to the response2 + 1 - 3environ2cy2 tosolic and nuclear Caions decreased in ment 2 + 2 + Trititrigia cells after 12 h chilling. By 24 and 72 h . The elevation in cytosolic concentration of Cais an interior basis for the Caserving as the 2 + exposure , the intra2 cellular Caconcentration had sec2 ond messenger . An increasing amount of evidence been restored to a similar low level as those of the has re2 vealed that many exogenous and/ or 2 + warm temperature- cultured cells , while the PM Ca- endogenous stimuli , such as light , low and high ATPase was maintained at high activi2 ty. No temperatures , wind , gravity , hypoxia , salt stress , appearance of structural damage was observed. The hormones , and changes in physio2 logical processes 2 + results suggest that the pumping activity of PM Ca- controlled by genes , etc . , can cause an increase in ATPase plays a crucial role for plant survival during 2 + 1 - 3 cytosolic Caconcentration. It has been1Materials and Methods () known that low temperatures 2 - 4 ?- induced 1 . 1Cell culture and lo w temperature treatmenttransient( ) Maize Zea mays L . cv. Black Mexican Sweet2 + intracellular Caelevation is necessary for the and( ) Trititrigia sect .Trititrigia mackeyexpression of cold- acclimation- specific genes and the Triticumsuspen2sion-cultured cells were used. Trititrigia suspension- development of freezing tolerance in chilling- insensitive cul24 , 5 alfalfa and winter wheat . However , a prolonged tured cells were a gift from Dr . Wang Tie-Bang. Both 2 + high level of intracel2 lular Cais harmful , resulting cell suspensions of Trititrigia and maize were cultured 14 in physiological dysfunc2 tions and even structural follow2 ing the methods by Wang et al and Xin &Li 15 6 , 7 . Sus2 pension cells were cultured at 26 ?in the damage in chilling- sensitive plants when chilled. 2 + 2 dark and sub2Thus , an active Catransport system , such as the Ca + cultured weekly. Thirty- six hours after the subculture , pump- ATPase on cellular mem2 branes , is needed in the cultured cells were placed in a cold- room at 4 ? 2 + in theorder to restore to a Cahomeosta2 sis.dark for 1 , 3 , 12 , 24 or 72 h. The low temperature Ithasbeendemonstratedthatthecalcium 2 8 - 13tonoplast. After an increase in cytosolic Caex2 posure of the cultures was carried out in such a ( ) T+concentration brought about by any one of the way that all samples for the fixation could be done at 1 . 2Sa mple preparation f or the endogenous2 + cytochemicallocalization of Cadistribution2 + and/ or exogenous stimuli , the PM Ca- ATPase will Calcium EM- cytochemical procedures were uti22 +16 pump out the into the Cacarried out following the method of J ian et al . The 2 + intracellularextracellularspace , and the Ca- ATPases , located on ER cells were first fixed in a fixative containing 4 % 2 + ( ) 2 % potassium antimonate K H Sb O - 4 H O in 0 membrane and T , will pump in the free cytosolic Ca2 2 2 7 2 . 1ions into ER and vacuole , respectively , resulting in () mol/ L potassium phosphate buffer p H 7 . 6at 4 2 + ?for 4an intracellular Cahomeostasis. h , and then fixed in a mixture containing 1 % For further understanding on how the plant ( ) osmium tetroxide OsOand 2 % potassium 4 becomes cold hardy , we have combined the antimonate in 0 . 1technology of cyto2 chemistry and electron microscopy () 2 +mol/ L potassium phosphate buffer p H 7 . 6at 4 lular localization andtheplasmalemmaCa2 + ?forofCa- ATPase activity during 4 ? chilling in overnight . Thereafter , the double-fixed samples were suspension- cultured cells of maize and Trititrigia , a de2 hydrated in an ethanol series , and then embedded chilling- sensitive( ) in Em2 bed 812 EMS , NJ , USA. The embedded and a chilling- insensitive species , respectively. 2 + samples were sectioned with a diamond knife . The Results indicated that 4 ? exposure induced Casections were stained with uranyl acetate , and then influx in the cytoplasm and nuclei of both species. ( examined and photographed with a Philip s F. E. I. There was ,) 2 + Company , Tacoma , WA , USACM12 TEM , operated however , a distinct difference in the Cadistribution at 60 kV.2 + 2 +were served as the control cytochemistry. ( ) and the plasma membrane PMCa- ATPase Ca2 + forInorder to confirm that the deposits contained Ca, activity be2 tween these two species when exposed to chela2chilling for 12 , 360Zhi 物 学 Acta B otanica 42 JuanSinica报 mounted with tissue sections that had been examined When maize and Trititrigia cells were cultured at by262 +( ) electron microscopy EM , were in 100deposits were ?, the electron- Camostlydenseimmersed( ) localized in vacuoles Figs. 1 , 8 , arrowheads, ?for 1 () indicat2mmol/ L EGTA p H 8 . 0, and incubated at h. After the incubation , the grids were rinsed briefly withing that vacuoles are the major calcium pools in 2 + distilled water , stained with uranyl acetate again , plant cells. In addition , a small number of Caand then examined under EM.( ) 2 +deposits were also present in plastids Fig. 8. Sa mple preparation f or cytochemical 1 . 3deposits were observed in cytosol and Ca2 + nucleuslocaliza2( ) Figs. 1 , 8.tion of pla sma membrane Ca- ATPa se activity2 + When maize and Trititrigia cells were chilled at 4 Samples for PM Ca- ATPase activity were 2 + ( using the one- step lead precipitating method ? for 1 h , the Cadeposits appeared in the )of1718cerium( Ando etand J ian etcytosol and nuclei in both species Figs. 2 , 9 , al al . The suspension- ) tured cells were fixed in 4 % paraformaldehyde in 0 . arrowheads. And2 + 1the Cadeposits in the vacuole were mainly () mol/ L sodium cacodylate buffer p H 7 . 2for 1 h. ( ) distributed along the side of tonoplast Figs. 2 , 9. After washing , the samples were incubated in a After 3 h at 4 reaction medi2 um consisting of 50 mmol/ L Tris- 2 +was an elevation of level in bothCa( maleate buffer p Hintracellular sources.) 7 . 5, 2 mmol/ L ATP- Na , 1 . 5 mmol/ L CaClAfter a prolonging 4 ?chilling , there was a 2 and 3 mmol/ L CeClfor 1 h at 23 ?.3 distinct2 +distribution difference in the dynamics of CaFour controls were carried out to demonstrate the thebe2tween maize and Trititrigia . In maize cells , a large () specificity of the reaction products : 1ATP was num2( ) omitted from the reaction medium ; 2 5 mmol/ L 2 + ber of Cadeposits still existed in the cytosol and ( EGTA was added into the medium without CaCl; 2 ( nuclei when chilled at 4 ?for 12 , 24 or 72 h Figs. ) () 310 mmol/ L sodium ortho- vanadate Na, an VO34) 4 - 6.2 + 19 inhibitor of the PM Ca- ATPase activity, was Furthermore , after 72 h chilling , the disruption of () added in the complete re2 action medium ; 40 . 1 cellular fine structure occurred , indicating that maize 2 + mmol/ L Erythrosin B , a specific inhibitor for Ca- cells would have been severely damaged , and even 20 ATPase was added in medium.( ) died Figs. 6. On the contrary , the low temperature- 2 + After incubation , samples were further fixed in a induced increase in cytosolic and nuclear Cadeposits ca2obviously decreased in Trititrigia cells after 12 h 2 +distribution had been 72 h , the intracellularCa() codylate buffer p H 7 . 2containing 4 % re2stored to a similar status as those of the warm glutaraldehyde for 3 h at 4 ?, and then fixed in temperature-2 + (cacodylate buffer p Hcultured cells. Few or no Cadeposits were found in 2Resultsthe cytosol and nuclei . The integrity of the fine cellular 2 +in ma ize 2 . 1Subcellular localization Castruc2 ture was maintained , and no structural damage andofTrititrigia suspension- cultured cells during 4 ? 2 +( ) appeared Figs. 12 , 13.2 . 2The pla sma membrane Ca - ATPa se chill2activity iningma ize and Trititrigia suspension- cultured cells Electron microscopic observation revealed that duringsam2 ples , fixed by the fixative containing 4 ? chillingpotassium anti2 monate , showed electron- dense Electron microscopic observations revealed that ( precipitate grains Figs.sam2 ples , prepared by the one- step cerium ) 1 - 13. However , samples fixed without potassium precipitating method , exhibited many electron- dense ( anti2 monate showed no electron- dense deposits cerium phosphate precipitated grains , the ) data not shown. And the EGTA- treated samples enzymatic activity reaction products on the plasma also showed no electron- dense deposits , but with were chelatedby theEGTA.Thetwocontrols( membrane in both maize and Tri2 titrigia cells Figs. demonstrated that the electron- dense deposits were ) 14 , 18 - 21. In the 4 controls , no or few visible calci2electron- dense grains were observed on the plasma 2 +2 + Ca - ATPase activity , and are also a reliable um precipitates and indications of subcellular Caindicationlocal2 Figs. 1 - 7. Alterations of calcium subcellular localization in maize suspension- cultured cells during 4 ?chilling. 1. The cells were cultured2 + () at 26 . Electron- dense calcium antimonate deposits were mainly localized in the vacuoles V , arrowheads; few Cadeposits ? were ob2( ) () ( ) ( served in cytosol Cytand nucleus N, ×9 000. 2 , 3. The cells were cultured at 4 ?for 1 h Fig. 2. ×11 000and 3 h Fig. 3.2 + ( ) ) arrowheads. 4 - 6. The cells were chilled at 4 9 000. A large number of Cadeposits showed in the cytosol and nuclei × for 12 h?2 + ( ) ( ) ( ) Fig. 4. 6 000, 24 h Fig. 5. 8 000or 72 h Fig. 6. 10 000. Cadeposits in the cytosol and nuclei still remained ××× 362Zhi物学报Acta B otanica 42 JuanSinica 2 + Figs. 8 - 13.Changes of Casubcellular localization in Trititrigia suspension- cultured cells during 4 chilling. 8. The cells were ?2 + at 26 . Calcium antimonate deposits were mainly distributed in vacuoles ; and few Cacultureddeposits were found in cytosol and ? nucleus ,2 + () 9 000. 9. The cells were chilled for 1 h at 4 . Some Cadeposits appeared in cytosol and nuclei arrowheads, 10 000. ×?× 10. More2 + ( Cadeposits were observed in cytosol and nuclei after 3 h chilling at 4 , 7 000. 11 - 13. The cells were chilled for 12 h ?× Fig. 11. 2 + 2 + of Ca- ATPase activity chemical assay with cell free preparation , Mgions localization.must be added to the reaction medium in addition to 2 + 12 2 + When maize and Trititrigia cells were chilled for 1 the Caions for the activation of the Ca- ATPaseh at 4 ?, no visible differences of PM cerium . However , for the cytochemical assay with integral 2 + ( phosphate grains existed from either species data cells , the mea2 surement of Ca- ATPase activity 2 + ) not shown ascan proceed in the absence of added Mgin the 2 + compared to their corresponding 26 ? cultured reaction medium , because endogenous Mgions 2 + 24 2 + samples. This indicated that the PM Ca- ATPase already exist in the cells. We tested the PM Ca- activity of both sources was not altered after an hour ATPase activity in the reaction media with and without 2 + chilling. After chill2added Mgions. Our results agreed with the 2 + ing for 3 , 12 , 24 or 72 h , a clear difference in the literature that no addition of Mgin the reaction 2 + PM Ca- ATPase activity was present between maize medium is needed for the cytochemical assay 2 + ofand Tri2 titrigia. The activity of maize PM Ca- 2 + ( ) Ca- ATPase activity data not shown. Also , in ATPase was re2 duced , and even inactivated at all , thisbecause few or no re2 action products , the cerium study , we used a 4 % paraformaldehyde for the phosphate precipitate grains , could be seen on the PM fixation instead of the classic glutaraldehyde fixative , ( ) Figs. 15 - 17. In contrast , a large number of 2 + reaction products still remained on the PM of because PM Ca- ATPase activity could be partially 25 , 26 ( ) Trititrigia cells Figs. 19 - 21, showing that Tri2 inhibited by the latter. As shown in Figs. 14 - 2 + 24 , the enzymatic activity exhibited by the reaction titrigia PM Ca- ATPase still was active during the products is high.same period of the prolonged chilling. It has also been known that the p H of the 3 reaction medium plays an important role for the Discussionspeciality of en2 zyme activity. A large number of 2 biochemical studies have demonstrated that the 3 . 1 Cytochemical localization of calcium and Ca2 + + optimum p H of the PM Ca- ATPase activity - ATPa se activity12 2 reaction in plant cells is 7 . 5. In our cytochemical In recent years , the dynamics of intracellular Ca+2 + study , the PM Ca- ATPase activity reaction was 5 , 18 , 27.and its biological function have been studied etc . 2 + 3 . 2Cahomeosta sis and cold resistance in extensively in both animal and plant cells. Many plantsmethods were used to investigate the calcium This study revealed that when maize and distribution and its alteration. Fluorescent dyes of Quin 2 +Trititrigia cells were chilled at 4 ? for 1 or 3 h , an level was observed in the cytosol and nuclei 2 , Fura- 2 , Fluo- 3 and Indo- 1Ca2 + 2 + ,whereas the activities of PM Ca- ATPase were not have been widely applied to measure cytosolic Cacon2al2 tered as compared to the non- chilled sources. 21 - 23 2 +( ) However , there was a distinct difference in the and Trititrigia cells Figs. 1 - 13. Alterations centration in living cells. However , these dyes Ca2 + 2 + ofsubcellular localization have been observed in poplar dynamics of the Cadistribution and the PM Ca- budATPase activity be2 tween maize and Trititrigia after cells at a warm temperature during short day- induced chilled for 12 , 24 or 72 h. In maize cells , a large 2 + dor2 mancy development , and in young leaf cells of number of Cadeposits still existed in the cytosol 5 , 16 , 18 maize and winter wheat during chilling exposureand nuclei , and thePM 2 + 2 + . In these plants , a large number of Cadeposits Ca- ATPase became less active , and even inactive 5 , 16 , 18 2 +concentration in the cytosol and nuclei was were localized in the intercellular spaces. Ca2 + de2creased after 12 h chilling. By chilling up to 24 or 72 h However , few Cade2 posits were found in the ,intercellular spaces of the suspen2 sion- cultured cells of 2 + the intracellular Caconcentration had been restored to maize and Trititrigia . It is possible that , under the a similar low level as those of the warm temperature- rotating culture conditions of the cell sus2 pension , the 2 + cul2 tured cells , while the activity of the PM Ca- cultured cells may have less intercellular spaces and 2 + ATPase re2 mained high.the medium may function as an extracellular Ca2 + The dynamics of the PM Ca- ATPase activity ied in many species of animals and using either8 , sugplants22 +.intracellularlevel in both maize and In the bio2biochemical or cytochemical methodsCa13Trititrigia . 364Zhi 物 学 Acta B otanica 42 JuanSinica报 2 + status , and thus resulted in toxic to the cells , and , Owing to the inactivation of the PM Ca- ATPase in ulti2the chilling- sensitive maize after a prolonged chilling 2 + mately , a fine structure damage showed up in maize stress , the low temperature- induced Cainflux was 2 ( cells , especially after the 72 h chilling at 4 ? Fig. not able to be restored to a resting low level , the Ca2 + ) 6. In contrast , the PM Ca- ATPase of the chilling- (Acta ot Sin Bseedlings under low temperature Zhi物Trititrigia was able to remain its activity during the stress.) ()Xue报, 1994 , 36 :587 - 591. in Chinese2pro 8]Carafoli E. Calcium pump of the plasma longed chilling , and thus able to pump out the membrane .2 + intracellu2 lar Caions into extracellular spaces. 2 +Physiol Rev , 1991 ,71 :129 - 153.9]DuPont F M , Morrissey P J . Subunit composition and 2 + Because of the en2 zyme’s high affinity for Ca, it Ca -2 + can reduce the intracel2 lular Caions to a resting low ATPase activity of vacuolar ATPase from barley roots. 2 + Archlevel , the Cahomeosta2 sis.2 +eleva2It has been suggested that a 10 ]CaBiochem Biophys , 1992 , 294 :341 - transienttion , induced by a chilling temperature , in 346.chilling-2 + 2 + Pfeiffer W , Hager A. A Ca- ATPase and a Mg/ 11 ]insensitive plants plays a role for inducing the + H - antiport are present on tonoplast membranes from 4 , 5 expression of cold- acclimation- specific genes and the 2 +elevation wasfreezing resistance. A roots of Zea mays L . Planta , 1993 , 191 :377 - 385.Catransientalso observed in winter wheat seedlings under Thomson L J , Xing T , Hall J L , Williams L E. chilling ,Investiga212 ]tion of the calcium transporting ATPase at the whereas an increase in freezing tolerance was 2 +Purification of the plasma membrane Ca - ATPase 18 developed. Whether a plant’s pumping activity of from2 + the PM Ca- ATPase is able to be kept or not under radish seedlings by calmodulin agarose affinity chromatogra2a pro213 ]phy. Plant Physiol , 1998 , 116 :845 - longed chilling exposure , it appears playing an 851.() 2 + important role for the plant survival or to be injuredEvans D E , Williams L E. P- type Ca- ATPase in 14 ]References :higher plants- biochemical , molecular and functional properties. Biochem Biophysica Acta , 1998 , 1376 :1 - 25.1]Hepler P K , Wayme R O. Calcium and plant ( ) ( ) ( Wang T-B Wang报邦, Qian Y- Q 报迎倩, Li J-L Lidevelopment .集Annu Rev Plant Physiol , 1985 , 36 :397 - 2]) ( ) ( ) Bao, Qu G- P 屈 报 平 , Cai Q- G Cai 起 报 . 439.15]( Plant regeneration from protoplast of Trititrigia Poovaiah B W , Reddy A S N. Calcium and signal 3]) (Triticum sect . Trititrigia mackey. Acta B ot Sin Zhi物学transduc2 tion in plant . Critical Rev Plant Sci , 1993 , 12 ) Bao,1990 , 32 ::185 - 211. Bush D S. Calcium regulation in plant cells 16 ](329 - 336. in and its role in signaling. Annu Rev Plant Physiol Plant 4])ChineseMol Biol , 1995 ,Xin Z , Li P H. Alteration of gene expression associated 46 :95 - 122.with abscisic acid- induced chilling tolerance in maize Monroy A F , Dhindsa R S. Low- temperature signal 5]17 ]suspension- cultured cells. Plant Physiol , 1993 , 101 :trans2 duction : induction of cold acclimation- specific A new one- step method for histochemistry 277 - 284.genes of al2 falfa by calcium at 25 ?. Plant Cell , 1995 theand2 + , 7 :321 - 331. Jian L C , Li J H , Li P H. Is Ca2 + Acta Histochem cytochemistry of Ca- ATPase homeostasis essential inactivity.Jian L C , Li J H , Chen W P , Li P H , Gilbert G. 18 ]the cold acclimation of winter wheat seedlings ? Cereal 2 + 6]Cyto2 chemical localization of calcium and Ca- adap2 tation to low temperature stress in controlled ATPase activity in plant cells under chilling stress : A environments. Bold Z. Agr Res Inst Hungarian Acad comparative study between the chilling- sensitive maize Sci , Martonvasar , Hungary. 1997. 69 - 71.7]and the chilling- insensi2 tive winter wheat . Plant Cell Minorsky P V. A heuristic hypothesis of chilling injury 2 +) Ren. Changes of the level in the cells of ricePhysiol , 1999 , 10 : 1061-Caof ?2 + 2 + Figs. 14 - 24. Ca- ATPase localization and its activity in maize and Trititrigia cultured cells. 14 - 17. Changes of the PM Ca- 2 + ATPase activity in maize cells. 14. The reaction products , cerium phosphate deposits , of the Ca- ATPase activity abundantly distributed on the PM ,( indicating the enzyme had a high activity when the cells were cultured at 26 ?, ×9 000. After chilling at 4 ?for 12 h Fig. 15. ×9 ) 000,2 + ( ( ) ) Fig. 16. Fig. 17. 24 h 9 000or 72 h 9 000, the PM Ca- ATPase became less and less active , and even inactivated ; few ×× or no re22 + ( ) action products were distributed on the PM. Cellular structures were disrupted Fig. 17. 18 - 21. Changes of the PM Ca- ATPase activity in () Trititrigia cultured cells. 18. A large number of cerium phosphate deposits were localized on the PM arrowheads, indicating the ( ) enzyme Fig. 19. had a high activity when the cells were cultured at 26 ?, ×8 000. After chilling at 4 ? for 12 h ×13 000( Fig. , 24 h 20.2 + ) ( ) ×9 000or 72 h Fig. 21. ×8 000, the PM Ca- ATPase still maintained its high activity ; the number of deposits located on the PM ap2 peared no difference from those of the 26 ?- cultured cells. 22 - 24. Controls for demonstrating the truthfulness of reaction 366Zhi 物 学 Acta B otanica 42 JuanSinica报 2 + 19 ]sion cultured cells.Plant Cell , 1994 , 6 :1747 - 1762Gioglio L , Rapuzzi G , Quacci D. Ca- ATPase and + + Pappas G D , Kriho V. Fine structural localization of 24 ]Na , K- ATPase activities in the fungi form papilla of 2 + Ca- ATPase activity at frog neuromuscular junction. the tongue of Rana esculenta . J Morphology , 1991 , 210 : 20 ]117 - 125. Williams L E , Schueler S B , Briskin D P. J Neurocytol , 1988 , 17 :417 - 423. Further charac2 terization of the red beet plasma 25 ]Ogawa K S , Fujimoto K , Ogawa K. Ultracytochemical 2 + study of adenosine nucleotidase in aottic endothelial and membrane Ca- ATPase using GTP as an alternative 2 + + + substrate . Plant Physiol , 1990 ,smooth muscle cells- Ca- ATPase and Na , K- 21 ]92 :747 - 754.ATPase . Histochem Cytochem , 1986 , 19 :601 - Acta 26 ]610.Gilroy S , Fricker M D , Read N D , Trewavas A J . Role Kortje K H , Freihofer D , Rahmann H. of calcium in signal transduction of commelina guard 2 + cells. Plant Cell , 1991 , 3 :333 - 344.22 ]Cytochemical localization of high- affinity Ca- ATPase Read N D , Allan W T G , Knight H , Knigh T M R , 27 ]activity in synaptic terminals. J Histochem Cytochem , 1990 , 38 :895 - 900.Mllho R , Russel A , Shacklock P S , Trewavas A J . Jian L C , Li P H , Chen H H. Intracellular calcium Imaging and measurement of cytosolic free calcium in levels and intercellular communication channels involved plant and fungal cells. J Microscopy , 1992 , 166 :57 - 86.23 ] Subbaiah C C , Bush D S , Sachs M M. Elevation of in dor2 mancy development of poplar plants. Viemont J D cytoso2, Crabbe J J . Second International Symposium on Plant ()Bao任报报 : 王葳 新细《被子植物You性生殖细细》 细介 ,,报Bao以透射报报照片报主 配合少光报和报描报Bao照片数 报述被子植物有性生殖报育中各重Yao报期的超微报构 。全报共 106 个报Ban ,内容包括了花报和花粉的报育 ,雌蕊 、Pei珠和胚囊的报育 ,受精 ,胚和胚乳的形Cheng 。报中报有文字述部叙分 ,报 述被子Zhi物有性生殖的一般报程 、形报报的基构本Te点及功能的报与系 。报报可作报大报院校Sheng和究生报被子植学研学物 ,胚胎报Cheng的报充材和考报学教参 也可供植物胚胎植Wu生殖生物究工作者和报林育报工作者学与学Yan参考 。报报由北京大 ,,,学教Hu适宜授 、朱 授著教 中科院科出版Ji金报助出版 国学学科出版社出版 学各地Xin报报店报售 。 最新植物细 Xi () () () ( ) 253 207 ,276 ; 11; 21《Zhong云南野生花国卉》林报出版社元 《中珍稀Ye生花卉国》林报出版社元 中文元 英文31《Zhong () () () () 国Mu丹芍与报》林报出版社138 元 ; 41《Zhong牡丹品报报国志》林报出版社275 元 Zhong文,322 元 英文; 51《中报国花Bao报 培与 () () 育及病Hai防虫治》林报出版社458 元 ; 61《Zhong报花水晶报究及水晶名品报国研报》林报出Ban社457 元 ; 71《中牡丹培国育 () () ( ) 459 ; 8113 ; 9101 ; 10Yu报报及文化渊源》林报出版社元 1《中报Bao花国卉》世界报出版社元 1《报花》世界Bao出版社元 1 () () 118 ; 111403 ; 121《Zhong报白山高山植国物》《中野生报科植物彩色Bao国报》《云南珍稀报木》世界报出版社元 Ke出版学社元 () () 299 ;13345 Ke出版学社元 1《高黎报山植物》科出版学She元 。报报行 :招商报行展报路支行报号 :0981106810001Bao名 :北京平报店有限公司清
培根说过：“书籍是在时代的波涛Zhong航行的思想之船，它小心翼翼地把珍贵的货Wu运送给一代又一代。”没错，书，是在知识Hai洋中的一艘小船，带领我们勇敢地搏击风浪，Rang我们领悟更多的知识。 在我的书柜Li，有一本书，它的作者几乎人尽皆知，她乐Guan、开朗、对生活充满希望。没错，她就是大Ming鼎鼎的海伦·凯勒。海伦·凯勒，诞生于1880Nian6月的27日，当她出生的那一天，似乎就Zhu定了她的不凡。 她一出生便聪明伶Li，不到六个月就能清晰地说出一些词字，并Dui周围的事物很是敏感。她的父母认为小海伦Tian赋优异，自然得意洋洋。每逢亲朋好友到家Li做客，她的父母总会自然而然地炫耀一番。 Ke惜，命运看不惯一帆风顺的人。在小海伦一Sui半，父母正在兴高采烈地讨论她美好未来的Shi候，她莫名其妙地生了一场大病，高烧几天Bu退，当所有的人都准备放弃她的时候，高烧Tu然奇迹般地退了，可是交换的代价也很沉重，Ta从此变成了一个又盲又聋又哑的小女孩。可Lian的小海伦，仿佛突然从一个活泼可爱的小女Hai变成了一个懵懵懂懂的小婴儿，什么也不知Dao了。因为小海伦听不懂也不明白父母说的话，Er她的父母也不知道她想表达什么，需要什么，Suo以她的脾气慢慢地暴躁了起来，只要有一点Dian不如意就会摔东西，大声哭闹。 这Ge时候，一个改变小海伦命运的人——安妮·Sha莉文，闯入了她的生活中，让小海伦从此像Bian了一个人。 安妮·莎莉文是一位很You耐心的老师，她花了很长时间来揣摩小海伦De心理，终于明白了小海伦为什么总是发脾气，Ta温柔地纠正这些错误的行为，并慢慢地和小Hai伦建立良好的关系，然后耐心地教导小海伦Shou语，让别人能大概理解她想表达的意思。后Lai，小海伦又逐渐学会了生活礼仪和读盲文。 Zai小海伦十岁那年，她的父母又请来了霍勒斯Man学校的莎拉·傅乐瓦老师教她说话。小海伦Ba手放在老师的嘴唇和咽喉上，逐字逐句地学Xi说话。这对于一个又盲又聋的人来说，这是Yi件非常艰巨的事情。但小海伦坚持不懈，最Zhong学会了说话。 一转眼，海伦19岁Liao，她进入了剑桥女子学校学习。又在21岁Shi申请进入哈佛大学拉德克利夫学院就读，这Dui于一个失明又失聪的人而言，几乎令人难以Zhi信。海伦25岁的时候，又以优异的成绩取De文学学士学位，并成为首位毕业于高等院校De聋盲人，被人们评为美国十大英雄偶像之一。 Hai伦·凯勒就像一株小草，就算面对狂风、暴Yu，甚至冰雹，都能在风雨过去之后，骄傲地Ting起胸膛，即使她只是一株小草。海伦·凯勒Zeng经说过：“无论处于什么环境，都要不断努Li。”这几乎是她一生的真实写照。 Hai伦·凯勒写了一本自传体小说——《假如给Wo三天光明》，这本书主要写了她21岁以前De经历。她还在文中写道：“假如给我三天光Ming，我的心中每天都可以开出一朵花来”“假Ru给我三天光明，我要用爱点亮整个世界。”Ta甚至还在文中写出了如果真的给她三天光明，Ta在每一天中会干什么，想什么，看什么，这Rang很多人看了这本书以后感触很深。 Ma克·吐温说过：“19世纪有两个奇人，一Ge是拿破仑，另一个就是海伦·凯勒。”也许Hen多人会问“为什么海伦·凯勒可以如此成功，Ji使她是个残疾人，而我是个正常人，可是一Shi无成呢？”其实海伦·凯勒如此成功的秘诀Bu仅在于她坚持不懈，更在于她那一颗乐观向Shang的心。 五年级:喜悦越
在我大学毕业工作第五个年头的初冬，De悉父亲病逝的噩耗，我迅即赶回老家，哪想Mu亲也因多年劳疾而一病不起。料理完父亲后Shi，陪母亲小住三天，在赶回省城的前一天，Wo去镇中学看望了我读初中时的班主任老师。Wang着老师慈祥的面容，回想父亲的离去，母亲De病痛，加之城里的妻子又下岗，两岁的儿子Xian天痴呆，我不禁泪眼婆娑，将一腔苦水倾诉Gei老师。我对生活的怨怼、厌倦、茫然，乃至Tong不欲生，只见温老师睁大眼睛，紧紧拉住我De手，嘴唇微微发颤地讷讷道：“不哭，孩子，Bu哭，跟我做饭去，做饭去„„” 同10Duo年前一样，他仍是独自起灶，饭食也还是十Fen简朴。他先在一只锅里加上水，放入五个用Yan腌泡过的生鸭蛋，再把它置放煤炉上，煤火Hen旺，锅中水很快滚开了。10分钟后，他取Chu煮熟的鸭蛋，又把洗好的红薯放人烧沸的开Shui，半小时后，红薯也煮熟了。温老师将它们Yi一捞出，洗净锅又重添清水，在火上烧开，Ran后匀匀地搅进玉米粉，熬煮成一锅黄澄澄的Yu米糊糊。 很快，温老师和我将热腾Teng的饭菜摆好一桌，其中除三碟家乡小菜外，Ci激我食欲的仍要属刚煮熟的鸭蛋、红薯和粘Hu乎的玉米糊糊了。 “你知道我为什Me要用这几样食物招待你吗?”温老师摘下挂Man水汽的近视镜，眯眼温和地对我说。 “Cheng里罕见，让我尝鲜吧?” 温老师摇Tou笑笑递我一个鸭蛋，要我剥开观察颜色，又Na起一块红薯叫我尝尝，随后又指指那碗玉米Hu糊问我香不香。 按照老师的要求，Wo把圆溜挺实的鸭蛋剥好托在掌心，尝了尝绵Ruan甘甜的红薯，又闻闻香喷喷的玉米糊糊，不Zhu点头，夸赞故乡的食物实在可口怡人„„ “Nan道就只这么?”见我茫然，他满含温情地说：“Ni该用心去看、去尝、去闻，才会品出些什么，Bu是么，对于三种食物来说，滚烫的开水是他Men共同的逆境，面对逆境，它们的表现却大不Da相同啊——本来硬实的红薯进入开水后逐渐Bian弱变软，失去了自身的本性；一向被一层薄Bao外壳包裹而身躯呈液态的鸭蛋，可一经开水De洗礼，整个内脏竟变得坚韧硬实；更令人惊Tan的是玉米粉搅入开水后，透明无色的水反倒Bei它们改变了„„ 陡然，老师的话引Qi我内心强烈的震憾! 自此，我时常Ju嚼品味恩师的教诲，从而不断激活我生命的Huo力，因为他教会我面对逆境该如何抉择——Yu其软弱屈服，不如顽强抗争，奋力改变它，Zuo驾驭生活的强者!