为了认识不同植物种类和采后果蔬过氧化氢酶(CAT)的生理特性,测定了40种植物叶片和30种采后果蔬的CAT活性和过氧化氢(H2O2)含量,分别从不同光合碳代谢类型和不同植物器官的角度进行了分析比较。结果表明,C3植物叶片的CAT活性大于C4和CAM植物,C4植物叶片的CAT活性大于CAM植物,叶片的CAT活性反映了不同光合碳代谢类型所具有的生理生态特性。叶菜类的CAT活性最大,其次是花菜类,结球叶菜类和果实类的CAT活性大于鳞茎菜类、地下茎菜类和地下根菜类,采后果蔬的CAT活性一般与植物器官的生理生态特性相关。
In order to understand the physiological properties of catalase(CAT) activity among different plant species,we measured CAT activity and hydrogen peroxide content in 40 plant leaves and 30 postharvest fruits and vegetables.The results show that CAT activity in leaves of C3 plants is higher than that of C4 and CAM plants,and CAT activity in C4 plants is higher than that in CAM plants.Leaf CAT activity reflects the physiological and ecological properties of plants in different photosynthetic metabolic types.CAT activity of leaf vegetables is the highest,followed by flower vegetables,and it is higher in corm leaf vegetables and fruit vegetables than in the bulbous vegetables,subterranean stem vegetables and subterranean root vegetables.CAT activity is closely related to physiological and ecological properties of plant organ.
全 文 :第29卷第6期
2010年12月
生态科学
EcologicalScience
29(6):573-578
Dec.2010
李哲,黄磊植物叶片和采后果蔬的过氧化氢酶活性阴.生态科学,2010,29旧:573—578.
LIZhe,HUANGLei.Catalasectivityinplantleavesandpostharvestfruitsandvegetables阴.EcologicalScience,2010,29(6):568—572.
植物叶片和采后果蔬的过氧化氢酶活性
李 哲,黄 磊
天津理工大学化学化工学院,天津300384
【摘要】为了认识不同植物种类和采后果蔬过氧化氢酶(CAT)的生理特性,测定了40种植物叶片和30种采后果蔬的CAT
活性和过氧化氢(H202)含量,分别从不同光合碳代谢类型和不同植物器官的角度进行了分析比较。结果表明,C3植物叶
片的CAT活性大于C4和CAM植物,C4植物叶片的CAT活性大于CAM植物,叶片的CAT活性反映了不同光合碳代谢类
型所具有的生理生态特性。叶菜类的CAT活性最大,其次是花菜类,结球叶菜类和果实类的CAT活性大于鳞茎菜类、地下
茎菜类和地下根菜类,采后果蔬的CAT活性一般与植物器官的生理生态特性相关。
关键词:植物叶片;采后果蔬;过氧化氢酶;过氧化氢
doi:10.3969/j.issn.1008.8873.2010.06.014中图分类号:Q945.79文献标识码:A文章编号:1008.8873(2010)06.573-06
Catalaseactivityinplantleavesandpostharvestfruitsandvegetables
LIZhe,HUANGLei
School/ChemistryandChemicalEngineering,TumjinUnivers砂ofTechnology,Ttanjin300384,China
Abstract:Inordertounderstandthephysiologicalpropertiesofcatalase(CAT)activityamongdifferentplantspecies,wemeasuredCAT
activityandhydrogenperoxidecontentiII40plantleavesand30postharvestfruitsandvegetables.TheresultsshowthatCATactivityin
leavesofC3plantsishigherthanthatofCAandCAMplants,andCATactivityinCAplantsishigherthanthatinCAMplants.LeafCAT
activityreflectsthephysiologicalande o ogicalpropertiesofplantsindifferentpho osyntheticmetabofictypes.CATactivityofleaf
vegetablesisthehighest,followedbyflowerv getables,anditishigherincormleafvegetablesandfruitvegetablesthani thebulbous
vegetables,subterraneans mvegetablesdsubterraneanrootvegetables.CATactivityscloselyrelatedophysiologicaland
ecologicalpropertiesofplantorgan.
Keywords:plantle ves;postharvestfruitsandvegetables;catalase;hydrogenperoxi e
收稿日期:2010-06-04收稿,2010-07.19接受
作者简介:李哲(1988—),女,学士,从事生物化学及生理方面的研究,E-mail:lizhelizhelizhe@126.com
万方数据
574 生态科学EcologicalScience 29卷
1引言(Introduction)
过氧化氢酶(catalase。CAT,EC.1.11.1.6)广泛存
在于植物细胞,其功能是催化活性氧过氧化氢
(H202)分解为H20和02,防止或减轻H202的直
接或间接损害。植物细胞内的H202一方面作为信号
分子,在气孔运动、生长发育调控、环境胁迫应
答、细胞程序性死亡等过程中起重要作用,另一方面
环境或细胞内信号分子的变化也会使H202在细胞内
发生累积,H202积累过多会造成各种氧化破坏,加
速细胞衰老或者造成细胞毒害【I巧】。
植物细胞内的CAT通过调控H202水平,参与细
胞建成【61、光合成【7-8】、抗逆【9m】、抗病【111、衰老
u2。”j等多种生理过程和生命现象。CAT活性与植物
本身具有的生理生态特性、受环境胁迫程度等有密切
联系,在不同植物种类、植物器官之间有较大差异
[14l。认识不同植物种类和采后果蔬CAT的生理特
性,可以为探讨植物生态适应、指导作物育种等提
供重要基础生理信息。本研究对40种植物叶片和30
种采后果蔬的CAT活性和H202含量进行测定,从不
同光合碳代谢类型和不同植物器官的角度进行了分
析比较。
2材料与方法(Materialsandmethods)
植物按不同光合碳代谢类型分为C3、C4和CAM
植物3大类(表1)。取样叶片为各种植物正常生长
的功能叶,取样重复3次。采后果蔬据食用植物器官
的不同分为叶菜类、花菜类、结球叶菜类、果实类、鳞
茎莱类、地下茎菜类和地下根菜类(表2),供试样品为
各种采后新鲜果蔬的食用部分,取样重复3次。
取供试植物样品2g,迅速保存于一80"C冰箱,用
于CAT活性和H202含量测定。CAT活性测定参照
“和Matsui(2001)[151的方法,在4"C低温条件下,
冷冻样品用50mmol·L1磷酸缓冲液(pH7.0)8mL
磨碎,12000xg离心20mim,取上清液1mL与pH
7.0的50mmol·L-1磷酸缓冲液(pH7.O)2mL混合
后,在24℃·l大气压条件下,加入0.88mol·L~H202
2mL反应1min,测量产生的02体积,02的1cm3
相当于0.041mmol,据2H202_2H20+02算出CAT
活性(H202mmol·g-lFW-minq)。
H202含量测定参照Okuda等(1991)【16J的方法,
冷冻样品用0.2N高氯酸10mL磨碎,15000xg离
心10min,用4N氢氧化钾将上清液的pH值调至7.5
表1C3、C4和CAM植物材料
Table1C3,C4andCAMplantsmaterials
万方数据
6期 李哲,等:植物叶片和采后果蔬的过氧化氢酶活 575
表2采后果蔬材料
Table2Postharvestfruitandvegetablemat rials
后再次离心,取上清液2mL过AGl-X8柱(1.0cm
i.d.x5cm,Bio-Rod)滤去植物色素,用蒸留水6mL
冲洗,合并收集液。收集液2mL中加入12.5mM3.
二甲氨基苯甲酸O.8mL、1.4mM3.甲基.2.苯并嚷唑
啉酮腙盐酸盐水合物160此和0.26units-mL-I过氧化
物酶40此,25℃水浴10min后,测定波畏590hill
的吸光度。
3结果与分析(ResultsandAnalysis)
3.1植物叶片
C3、C4和CAM植物叶片的CAT活性如图l,
C3植物最大,平均值4.7H202mmol·g~FW·min一,
为C4植物的3.7倍、CAM植物的9.1倍,其次是C4
植物,平均值1.3H202mmol·g-iFW·min~,为CAM
植物的2.5倍,CAM植物的平均值为O.5H202mmol
-g-1FW·minl。H202含量如图2,C3、C4植物平均
值1.8、1.9rtrnol-g—FW,低于CAM植物平均值3.1
0mol·g~FW。
3.2采后果蔬
各种采后果蔬的CAT活性如图3,叶菜类最大,
平均值5.3H202mmol·g-iFW·minl,其次是花菜类,
平均值3.1H202mmol·g-iFW·minl,结球叶菜类、果
实类的平均值分别为1.6、1.1H202mmol·g-lFW·
min~,鳞茎菜类、地下茎菜类、地下根菜类平均值分
别为0.8,0.8、0.7H202mmol’g-iFW‘min一。H202
含量如图4,以叶菜类最高,其次是花菜类,结球
叶菜类和果实类较低,鳞茎菜类、地下茎菜类和地
下根菜类最低。
4讨论(Discussion)
4.1植物叶片的CAT活性
植物细胞内CAT的基因表达及活性大小受底物
H202诱导,在H202较多时CAT的基因表达较多、
活性上升【17-1810叶细胞内的CAT多存在于进行光呼
吸代谢的过氧化酶体,用于分解光呼吸代谢中乙醛酸
氧化反应形成的H202【4】oC3植物叶片的光呼吸代谢
水平较高,而C4、CAM植物几乎不进行光呼吸,所
以C3植物叶片的CAT活性表现出大于C4、CAM植
物(图1)。
CAT还存在于叶细胞的线粒体和细胞质中,线
粒体内的呼吸作用是一系列电子(e-)传递和H+的氧
化还原反应,在e.传递过程中会发生e_遗漏【5】,呼吸代
谢旺盛的细胞遗漏的r也较多。遗漏的f会和02
万方数据
!Z! 生查型堂里塑!竺曼堕璺!塑堡璺堡 12鲞
植物种序号(见表1)Plantspeciesnumbcr(Table1)
图1(23、C4和CAM植物叶片的过氧化氢酶活性,图中竖线表示误差(胪3)
Fig.1CatalaseactivityofC3,C4andCAMplantleaves.VerticalbarsndicateSE(n=3)
植物种序号(见表1)Plantspeciesnumber(Table1)
图2(23、C4和CAM植物叶片的过氧化氢含量,图中竖线表示误差(垆3)
Fig.2HydrogenperoxidecontentofC3,C4andCAMplantleaves.VerticalbarsndicateSE(n-3)
采后果蔬序号(见表2)Postbarvestfruitsandvegetablesnumber(Table2)
图3采后果蔬的过氧化氢酶活性,图中竖线表示误差(以=3)
Fig.3Catala靶activityofpos harvestfruitsandvegetables.VerticalbarsindicateSE(,F3)
万方数据
6期 李哲,等:植物叶片和采后果蔬的过氧化氢酶活 577
采后果蔬序号(见表2)Postharvestfruitsandvegetablesnumber(Table2)
图4采后果蔬的过氧化氢含量,图中竖线表示误差(垆3)
Fig.4Hydrogenperoxidecontentofpostharvestfruitsandvegetables.VerticalbarsindicateSE(舻3)
反应形成活性氧超氧阴离子(02:.),02:-在超氧化物
岐化酶作用下转变为H202,02-和H202可通过
Haber-Weiss、Fenton反应形成氧化毒性更高的活性
氧羟自由基(·OH)[19】。植物细胞中的多种氧化还原
反应伴随有活性氧形成,呼吸等生理代谢旺盛的细胞
质中H202也较多。C4植物叶片的CAT活性大于
CAM植物(图1),可能与C4植物叶片细胞的多种
氧化还原反应、呼吸等生理代谢较CAM植物旺盛有
关。另外,含叶绿体多的叶片栅栏细胞比其他细胞更
容易形成活性氧,栅栏细胞在C4植物叶片所占比例
大于CAM植物;CAM植物叶片细胞昼夜循环利用
C02,叶片细胞内C02浓度高,能够抑制由于e-过剩
形成活性氧:CAM植物的叶表皮细胞一般具有发达
蜡质层,也可减轻由强光造成的e.过剩形成活性氧。
植物叶片细胞内活性氧形成的多少与其生理生
态特性密切相关,虽然C3、C4植物叶片细胞内活性
氧的形成较多,但较高的CAT活性(图1)使非逆
境条件、正常生长情况下的H202维持在一个较低水
平(图2),这可能与有利于其旺盛的光合成【7堪J、呼
吸等生理代谢进行有关。
可以看出,不同光合碳代谢类型C3、C4和CAM
植物间的生理生态特性规律与叶片细胞CAT活性有
密切联系,或者说叶片CAT活性反映了不同光合碳
代谢类型所具有的生理生态特性,一般植物叶片的
CAT活性具有C3植物大于C4和CAM植物、C4植
物大于CAM植物的规律。
4.2不同采后果蔬的CAT活性
采后果蔬器官的呼吸代谢在不断进行,呼吸代谢
水平一般以叶片、花器官较高,其次是叶球、果实,
鳞茎和地下根茎较低,如在20"(2条件下韭菜‘201、青
花菜贮11、桃[22】、番茄翰、洋葱【231、马铃薯【2卅的呼吸
速率分别为2.8、4.0~7.3、1.6、0.5、0.1~0.3、O.1C02
mmol-kg~FW·h~。呼吸代谢旺盛的植物器官活性氧
产生较多,其CAT活性受底物H202诱导表现较大。
本研究观测到采后果蔬的CAT活性以叶菜类和花菜
类最大,其次是结球叶菜类和果实类,鳞茎菜类、地
下茎菜类和地下根菜类较小(图3),与呼吸代谢水
平表现一致。CAT活性大的果蔬,H202含量也较高
(图4),说明其采后受活性氧胁迫程度高,与叶菜
类、花菜类、结球叶菜类和果实类、鳞茎菜类、地下
茎菜类和地下根菜类的耐贮藏性由小到大的表现一
致。
但也有一些组织构造和化学组分特殊的果蔬,如
叶球疏散、叶绿素含量较多的LactucasativaL.VaE
longifoliaLam.(结球生菜),果实种腔大、叶绿素含
量多的CapsicumannuumL.var.grossum(甜椒),果
实种腔大的CucumissativusL.(南瓜),以及韧皮部
发达的DaucuscarotaL.(胡萝卜)等,它们的CAT
活性大于同类果蔬(图3)。而有机酸含量高的Citrus
sinensisL.(柑橘)、酚类化合物含量多的Musa
acuminata(香蕉)和Nelumbonuclfera(莲藕)等CAT
活性低于同类果蔬,可能与细胞内的高酸性环境、多
量酚类化合物有助于抑制、消除活性氧有关[251。
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万方数据
植物叶片和采后果蔬的过氧化氢酶活性
作者: 李哲, 黄磊, LI Zhe, HUANG Lei
作者单位: 天津理工大学化学化工学院,天津,300384
刊名: 生态科学
英文刊名: ECOLOGICAL SCIENCE
年,卷(期): 2010,29(6)
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6.Gichner T DNA damage induced by indirect and direct acting mutagens in catalase-deficient
transgenic tobacco cellular and acellular comet assays 2003
7.Dat J F;Pellinen R;Beeckman T;Van De Cotte B Changes in hydrogen peroxide homeostasis trigger an
active cell death process in tobacco[外文期刊] 2003(4)
8.Neill S.Desikan R.Hancock J Hydrogen peroxide signaling 2002
9.Leon J;Lawton M A;Raskin I Hydrogen peroxide stimulates salicylic acid biosynthesis in tobacco[外
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10.Thannickal V J.Fanburg B L Reactive oxygen species in cell signaling 2000
11.Thannickal V J;Fanburg B L Reactive oxygen species in cell signaling 2000
12.de Marco A.Roubelakis-Angelakis K A The complexity of enzymic control of hydrogen peroxide
concentration may affect the regeneration potential of plant protoplasts 1996
13.Neill S;Desikan R;Hancock J Hydrogen peroxide signaling[外文期刊] 2002(5)
14.Shikanai T.Takeda T.Yamauchi H.Sano S Tomizawa K Yokota A Shigeoka S Inhibition of ascorbate
pemxidase under oxidative stress in tobacco having bacterial catalase in chloroplasts 1998
15.Gichner T DNA damage induced by indirect and direct acting mutagens in catalase-deficient
transgenic tobacco cellular and acellular comet assays[外文期刊] 2003
16.Vandenabeele S.Vanderauwera S.Vuylsteke M.Rombauts S Langebartels C Seidlitz H K Zabau M Van
Montagu M Inze D Van Breusegem F Catalase deficiency drastically affects gene expression induced by
high light in Arabidopsis thaliana 2004
17.Salah N;Miller N J;Paganga L;Tijburg L Bolwll G P Rice-Evabs C Polyphenolic flavanols as
scavengers of aqueous phase radical sandals,and as chain-breaking antioxidants[外文期刊] 1995
18.Engel N.Schmidt M.Lutz C.Feierabnd J Molecular identification,heterologous expression and
properties of light-insensitive plant catalases 2006
19.Nourian F;Ramaswamya H S;Kushalappa A C Kinetics of quality change associated with potatoes
stored at different temperatures[外文期刊] 2003(1)
20.Luna C M.Pastori G M.Driscoll S.Groten K Bemard S Drought controls on H2O2
accumulation,catalase(CAT)activity and CAT gene expression in wheat 2005
21.Downes K;Chope G A;Terry L A Postharvest application of ethylene and 1-methylcyclopropene either
before or after curing affects onion(Allium cepa L.) bulb quality during long term cold storage[外文
期刊] 2010(1)
22.Leon J.Lawton M A.Raskin I Hydrogen peroxide stimulates salicylic acid biosynthesis in tobacco
1995
23.Okuda T;Matusda Yamanaka A;Sagisaka S An abrupt increase in the level of hydrogen peroxide in
leaves of winter wheat is caused by cold treatment[外文期刊] 1991
24.Dat J F.Pellinen R.Beeckman T.Van De Cotte B Changes in hydrogen peroxide homeostasis trigger an
active cell death process in tobacco 2003
25.Li J;Matsui S Effects of chilling on antioxidative enzymes in leaves of Cattleya and Cymbidium
(in Japanese) 2001
26.Reilly K.Han Y.Tohme J.Beeching J R Isolation and characterisation of a cassava catalase
expressed during post-harvest physiological deterioration 2001
27.Engel N;Schmidt M;Lutz C;Feierabnd J Molecular identification,heterologous expression and
properties of light-insensitive plant catalases[外文期刊] 2006(4)
28.Song X.Zhao F Research progress on catalase in plants 2007
29.Vandenabeele S;Vanderauwera S;Vuylsteke M;Rombauts S Langebartels C Seidlitz H K Zabau M Van
Montagu M Inze D Van Breusegem F Catalase deficiency drastically affects gene expression induced by
high light in Arabidopsis thaliana[外文期刊] 2004(1)
30.Li J.Matsui S Effects of chilling on antioxidative enzymes in leaves of Cattleya and Cymbidium
(in Japanese) 2001
31.Shikanai T;Takeda T;Yamauchi H;Sano S Tomizawa K Yokota A Shigeoka S Inhibition of ascorbate
pemxidase under oxidative stress in tobacco having bacterial catalase in chloroplasts[外文期刊]
1998(1/2)
32.Okuda T.Matusda Yamanaka A.Sagisaka S An abrupt increase in the level of hydrogen peroxide in
leaves of winter wheat is caused by cold treatment 1991
33.Luna C M;Pastori G M;Driscoll S;Groten K Bemard S Drought controls on H2O2
accumulation,catalase(CAT)activity and CAT gene expression in wheat 2005
34.Guan L M.Scandalios J G Hydrogen-peroxide-mediated catalase gene expression in response to
wounding 2000
35.Brennan T;Frenkel C Involvement of hydrogen peroxide in the regulation of senescence in pear 1997
36.刘大永.王维香 过氧化氢对水稻幼苗中CAT和POD活性的影响团 1998(3)
37.Watada A E;Ko N P;Minott D A Factors affecting quality of fresh-cut horticultural products[外文期
刊] 1996(2)
38.Aasada K The water-water cycle in chloroplasts:Scavenging of active oxygens and dissipation of
excess photons 1999
39.Techavuthiporn C;Nakano K;Maezawa S Prediction of ascorbic acid content in broccoli using a model
equation of respiration[外文期刊] 2008
40.Wu C.Du X.Wang L.Wag W Zhou Q Tian X Effect of 1-methylcyclopropene on postharvest quality of
Chinese chive scapes 2009
41.Wu C;Du X;Wang L;Wag W Zhou Q Tian X Effect of 1-methylcyclopropene on postharvest quality of
Chinese chive scapes[外文期刊] 2009(3)
42.Techavuthiporn C.Nakano K.Maezawa S Prediction of ascorbic acid content in broccoli using a model
equation of respiration 2008
43.Cheng Y;Song C Hydrogen peroxide homeostasis and signaling in plant cells[期刊论文]-Science in
China(Life Sciences) 2006(1)
44.Watada A E.Ko N P.Minott D A Factors affecting quality of fresh-cut horticultural products 1996
45.Aasada K The water-water cycle in chloroplasts:Scavenging of active oxygens and dissipation of
excess photons[外文期刊] 1999(0)
46.Downes K.Chope G A.Terry L A Postharvest application of ethylene and 1-methylcyclopropene either
before or after curing affects onion(Allium cepa L.) bulb quality during long term cold storage 2010
47.刘大永;王维香 过氧化氢对水稻幼苗中CAT和POD活性的影响团[期刊论文]-作物学报 1998(03)
48.Nourian F.Ramaswamya H S.Kushalappa A C Kinetics of quality change associated with potatoes
stored at different temperatures 2003
49.Guan L M;Scandalios J G Hydrogen-peroxide-mediated catalase gene expression in response to
wounding[外文期刊] 2000
50.Salah N.Miller N J.Paganga L.Tijburg L Bolwll G P Rice-Evabs C Polyphenolic flavanols as
scavengers of aqueous phase radical sandals,and as chain-breaking antioxidants 1995
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