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,2016,36(7):1391-1398
犃犮狋犪犅狅狋.犅狅狉犲犪犾.犗犮犮犻犱犲狀狋.犛犻狀.
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ijklmno
(AsA)pq、rslmnotuv(DHAR)bwvxy、lmnoz{|$v(APX)bwvxy
Q}APX,~|。
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犛狋狌犱狔狅狀犃狊犮狅狉犫犻犮犃犮犻犱犪狀犱犚犲犾犪狋犻狏犲犈狀狕狔犿犲狊犃犵犪犻狀狊狋
犆犪犱犿犻狌犿犛狋狉犲狊狊犻狀狋犺犲犚狅狅狋狊狅犳犞犻犮犻犪狊犪狋犻狏犪犔.
RUIHaiyun1,2,ZHANGXingxing2,ZHUANGKai2,SHENZhenguo2,ZHANGFenqin3
(1TaizhouUniversity,Taizhou,Jiangsu225300,China;2ColegeofLifeSciences,NanjingAgriculturalUniversity,Nanjing
210095,China;3ColegeofHexi,Zhangye,Gansu734000,China)
犃犫狊狋狉犪犮狋:Twovarietiesof犞犻犮犻犪狊犪狋犻狏犪L.withdifferentCdtolerances(CdtolerantvarietyL3,Cdsensi
tivevarietyZM)werestudiedtocompareconcentrationsofascorbicacid (AsA),activitiesofde
hydroascorbatereductase(DHAR)isoenzymesandascorbateperoxidase(APX)isoenzymes,andAPX
geneexpressioninresponsetoCdtreatmentintheroots.Resultsshowedthat:(1)Cdtreatmentsignifi
cantlyincreasedtheconcentrationsofAsAanddehydroascorbate(DHA)intherootsofbothL3andZM.
CdtreatmentalsotendedtoincreaseratioofAsA/DHAinL3,buttodecreasetheratioinZM.Atthe
sameCdtreatment,L3rootshadhigherAsAconcentrationandAsA/DHAratiothanthatofZM.(2)
FourisoformsofDHARwereobtainedinL3andZMrootsbynativePAGEandtheiractivitiesgeneraly
increasedwithincreasingCdconcentration.DHAR1wasobtainedonlyinL3andDHAR4onlyinZM.At
equivalentCdconcentrations,thetotalactivityofDHARwashigherinrootsofL3thanthatinZM.(3)
ElevenAPXisoformswereobtainedinbothL3andZMrootsbynativePAGE.APX1,2,4wereinduced
byCdtreatmentonlyinZMandtheactivityofAPX8wasmoresignificantlyinducedinL3thanthatin
ZM.AnAPXgenewasclonedandsubjectedtoRTPCRanalysis.Thegeneexpressionlevelwasupregu
latedunderCdstressintherootsofbothvarieties.TheseresultssuggestedthatconcentrationofAsA,ac
tivitiesofDHARandAPX,andAPXgeneexpressionlevelincreasedinresponsetocadmiumstressinthe
rootsoftwo犞.狊犪狋犻狏犪varieties.ItispossiblethatthemoreeffectiveAsArecycleintherootsofL3con
tributedtoitshigherAsAconcentrationthanthatinZM,andthiscouldberelatedtoitsstrongerprotec
tionfromROSgenerationandfinalycontributedtoitshighertolerancetoCdstress.
犓犲狔狑狅狉犱狊:犞犻犮犻犪狊犪狋犻狏犪L.;cadmiumtolerance;ascorbicacid;ascorbateperoxidase;dehydroascorbatere
ductase
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[3]。
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(AsA)Æ
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Ò{|]rslmno
(DHA),DHA×zrsl
mnotuv
(DHAR)tu]AsA。9&ö,AsA
pq·:#$;
=>
、
?,*Z@
ef"l·A
[67]。BurkeyB[8]CD,E{ef
gXyVWFU(AsApq[\VW,X
yVW·¿À;¶AsAG´µ。SharmaZ
Dubey[9]&öHef£¤´IhiAPXxy,
bJbwvýþKC@L
。
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·:#$;Yef"l·A
,
=,MNOAsA
Pþ Q Ý LR S T o1,4÷ U,V + ù W
(犜狉犻狋犻犮狌犿犱狌狉狌犿cvCreso)AsA PþXO,Ye
fÃ@{|ef©¥ÁÂ
[10]。
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DHARbwvb|,¼ÌÖcd#$lmno
±²eçYefÉA
。
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1.1 UVWX>YZ
YZ^_RSTUhi1YXyVWL3Z
1Y[\VW ZM。Wûj3% H2O2 kÍ
15min,$lmû´nopqr,sW16h,®tu
vw20℃~25℃xC。3dr,yz{|}~
Ìêhi®tup2.5LHoagland
,
ti10。=g:
/
d25℃/20℃,;d60%~80% ,«
J12h,«Ïd400μmoL· m
-2·s-1。
3d1,pHÊ5.5。
YZÌ
:2RSTUVWti7dr,$
25μmol·L
-1
YA
(
YQCdCl2·2.5H2O
O
)。
A3È,A
3d1
。
úâA0、2、4、6、8、10、12、14d$
jk@{¬
。
YZ
:0(Control)、10、25、50μmol·L
-1
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A
(
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$YZ
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. @¡r
,-80℃¢%£@A
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ti3dr,®tuO25μmol·L
-1
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J]0、12、24、48、96h。§
V ¨¡
,-80℃¢%$jk
RNA:。
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1.2.1 89_`aR úâ25μmol·L
-1
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A0、2、4、6、8、10、12、14d,qhi:j{r,Ô
jÿ
,
Ól©´úª«È
。
1.2.2 bcdef ðî¬y¬®
SchützendübelB[11]¯°±²(Evansblue)ÓÔ
g
。
10¥³«#$1cm:jj´,0.025%
2931 ! " # $ % & 36µ
(W/V)¯ °±²¶·¸30min,lmû´pq
15minno¹º»¸r,1%SDS(W/V)50%(V/
V)¼½B¾þ¿À,50℃´Á15minr,14000×
gm!15min,¹600nmgÓ«。
1.2.3 cghijFkljmn[ îïz{|
ýÃ|%Ä® YamamotoB[12]fg。5
2cm:jj´sSchiffÅÆ (0.5gÇyVÈ,
0.5gK2S2O5,100mL1mol·L-1 HCl,100mL
H2O)·¸60min,*r$0.5% (W/V)
K2S2O5 ¶ (¶0.05mol·L-1 HCl)pq,
ÉʸËÌÉÝÍ
ÎÏgÐÑ
、
Ò
。
1.2.4 犃狊犃o 犇犎犃pq AsA pq¬®
KampfenkelB[13]fg。1g³«#$jk,O
Ó@5 mL6% TCA ÔÁB¾,4 ℃ m!
(12000×g,20min),¹]Õ。 AsAp
q¬
:
0.2mLÕ(;0.2mL6%
TCA),O0.2mL10mmol·L-1DTTZ0.4mL
0.2mol·L-1PBS(pH7.4),Öú¦¿,42℃æç
15minr×O0.2mL0.5% NØ,ÙÚÛ
ÜÝ
(NEM)、1 mL10%TCA、0.8 mL42%
H3PO4、0.8mL4%Þßà、0.4mL3% FeCl3,á
¨âãä妿
,42℃æç40min,¬525nm
Ó«
。
tuæ AsA ¬JQ0.2mLH2O Z
0.2mLPBSúâ±ç DTTZ NEM,èbÂ。
AsA pq7ltuæ AsA pq]{|æ
AsA,rslmno(DHA)pq。AsA ¶Â
6% TCA,bg#éê*。
1.2.5 犇犎犃犚o犃犘犡Frfst ëì:}a
íîxyïðñÛÝòó
(nativePAGE)®
Laemmli[14]fg4℃g¼ô。úmóZõó
(
apSDS)ö¼÷,÷<øOBqë
ìð
,
Áp]pH8.3TrisHCl,ù¬ú
30mA。¬û,¼ôvxy·¸。
DHARnativePAGEü$4%õóZ
10%úmó。DHARxy·¸® Aravind
B
[15]
fg
。
5ròósp4mmol·
L-1GSH,2mmol·L-1DHA0.1mol·LPBS
(pH6.2),ýþÂÿ20min。´qr,p
0.1%FeCl3 Z0.1% K3[Fe(CN)6]0.1mol·
L-1 HCl¶!ÿ,"²¸#$Â%D&²¸
。
(´)q
,
*+,-
。
APXnativePAGEü$4%õóZ8%
úmó
。APXxy·¸ü$ RaoB[16]f
g
。
òó.p2mmol·L-1 AsA50mmol·
L-1PBS!ÿ30min,/p4mmol·L-1AsA
Z2mmol·L-1 H2O2 50mmol·L-1PBS(pH
7.0)!ÿ20min。Áppq1minr,p
28mmol·L-1 TEMEDZ2.45mmol·L-1 NBT
PBS(pH7.8),88ý0,·¸10~15min。
´q
,
12æç
。
*+,-
。
1.2.6 犃犘犡,uvwFxy\q犘犆犚Oz
NCBI3456RSTU§U+#$ APX,
~78
。
9$DNAMAN6.0:¼ô;o78
;
,
j93WU+#$:TU(犘犻狊狌犿狊犪狋犻狏狌犿)、JU
(犌犾狔犮犻狀犲犿犪狓)、<=>?(犕犲犱犻犮犪犵狅狋狉狌狀犮犪狋狌犾犪)
d¢@78
,
ABä$¼ôPCRCX,©¥`R
STU犃犘犡D78。7°4TU&
ð犘狊犃犘犡01,~(78E]X62077.1);Fo
78Ìêy]94%。YJª«¬qPCRæçG
$ SYBR Premix Ex 犜犪狇 (TliRNaseH Plus,
Takara)ÅÆH,G$7500RealTimePCR kü
(AppliedBiosystems,USA)Ä,~q
|
。PCR§$犃犘犡 áIyä$],J×:5′GCT
GGTGTTGTTGCTGTTG3′;æ×:5′CCCTCT
GGTGGTGGATGG3′。Actin#]÷®,Actin(7
8E GU946218.1)áIyä$:J×:5′TCTG
GTGATGGTGTGAGTC3′;æ×:5′ACAGCAA
CATAGGCAAGC3′。PCR :95 ℃,30sÂ
Ù
;
r95℃,5s;60℃,34s,40½¾。<
A,~q4;
,
QActin]÷®,;
q]1,©¥Ar
G$SPSS13.0(SPSSInc.,Chicago,U.S.
A.)Ë9A:h¼ôfLúMZLSDõÈ
¶
,
abNOLI
(犘<0.05)。G$
QuantityOne(BioRad):¼ôòóÏd
¬qúM
。
2 4úM
2.1 CDEB2}4567~89_`FG
2RSTUVWL3ZZM j25μmol·
L-1YAg@{PQ=*1§。YZR¯
1~14d÷,ZM:jS{@{qZjk«ÈX
{qT1ùL3,VWLIA4dQr
¥
´µ
,
ÄWLID%UAJV
{WXXOY|
,
ÕQ:jS{D©¦Z%
,
=A14dJ,VWZM :jS{qZjk«È
X{qúâb[L3
663%Z41%。Å\,
39317[ 345,B:RSTUjklmno}³v;Yefeç
abNOAJË./VW0.05´µ%
yLI;Ë]µË±éL;gb
*1 25μmol·L
-1CdAg2RSTUVWL3ZZM:jS{Zj«ÈX{
Thediferentnormallettersindicatedsignificantdiferenceamongcultivarsortreatmenttimesat0.05level;Dataaremeans±SD(n=3);
Thesameasbelow
Fig.1 Effectsof25μmol·L
-1Cdtreatmentonrootelongationandincreaseofrootfreshweightintwo犞.狊犪狋犻狏犪
varieties,L3andZM
dYefg
,
Y[\VWZM jk@{
¶XYVWL3¢¥¦
]Ê。
2.2 CDE2}4567~8Fcghi
jobc
G$SchiffÅÆ;YA7d2RSTU
VWj´îïz{|cd¼ô`ýÃ|%Ä
,
SchiffÅÆ4îïz{|Ã@^_|P$æç@
þ`ȸ$ð
,
=*2§。,;
(0μmol·L
-1Cd)g,2RSTUVWj
´abÐÑa¥`ȸ
;
10~50μmol·L
-1
Y
Ag
,L3ZZM j´c%D`ȸ,Y
AdXOj´`ȸWX°
,
îïz{
|cdWXO&
;
bYAdg
,ZM ¶L3
j´·¸¦&
。
d°YefgZMj´îïz{|
cd¦eÈ
,
¢¥`L3¦eÈ{|ef。
Evansblue(¯ °±²)Æfghy·_,ij
´·ÒEvansblue·þ²¸,d°j´&;Ev
ansblue×h,ðî¬y¢¥jm,·¸k&
j´&ðîñòcdkJ
。
YAg2
RSTUVWj´EvansblueÓÔqúM°
(
*3),10~50μmol·L
-1
YA7dr,2RS
TUVWj´ð°
ñò
,
ñòcdY
AdXO
XO
;
bYAdg
,
ZMj´ðîñòcd
JL3。
2.3 CDEB2}4567~89犃狊犃、犇犎犃
pqo犃狊犃/犇犎犃
FG
1
,<dYA7dr2RST
UVWjkAsAZDHApqc¶;(0μmol·
*2 abdYefg2RSTUVWL3ZZM
j´îïz{|Schiff·¸
Fig.2 Histochemicaldetectionoflipidperoxidation
bySchiffsreagentintheroottipsoftwo犞.狊犪狋犻狏犪
varieties,L3andZM
*3 abdY;2RSTUVWL3ZZMj´¯
°±²ÓÔle
Fig.3 QuantitativeassayofEvansblueuptakeinthe
roottipsoftwo犞.狊犪狋犻狏犪varieties,L3andZM
4931 ! " # $ % & 36µ
v1 CDE2}4567~89犃犛犃、犇犎犃pqo犃狊犃/犇犎犃
Table1 TheconcentrationsofAsA,DHAandtheratioofAsA/DHAintherootsoftwo犞.狊犪狋犻狏犪varietiesL3
andZMunderCdstress
VW
Cultivar
Yd
Cdconcentration/
(μmol·L-1)
lmnopq
AsAcontent
(μmol·g-1)
rslmnopq
DHAcontent
(μmol·g-1)
lmno
/
rslmno
AsA/DHA
L3
0 0.30±0.03e 0.37±0.02c 0.83±0.12bcd
10 0.92±0.03cd 0.53±0.13bc 1.82±0.44a
25 1.18±0.01b 0.69±0.10b 1.72±0.24a
50 1.63±0.19a 1.41±0.19a 1.17±0.20b
ZM
0 0.42±0.07e 0.41±0.03c 1.01±0.19bc
10 0.83±0.02d 1.39±0.06a 0.60±0.04d
25 0.97±0.05c 1.51±0.10a 0.65±0.07cd
50 0.79±0.04d 1.48±0.42a 0.58±0.10d
ã
:
]3ÈYZµ±éL(n=3)
Note:Valuesaremeans±SD(n=3)ofthreedifferentexperiments
L-1Cd)
;L3jkAsA/DHA¶;
,
ZMjkAsA/DHA¶;
6
。
bYAdg
,L3jk AsA
pqZAsA/DHA
JZM。QÂ
d°
,
YefgXyVWL3Ǹ¶[\VWZM
¦ÏÃ@AsAZ¿À{|tumn·A。
2.4 CDEB2}4567~89犇犎犃犚
AFG
;YA7d2RSTUVWjkŶ
yëì:$¼ôxyZDHARxy·¸,Å
\4DHARbwv(*4)。Äobwv
xy U Y A d
,
DHAR1L3jk
,DHAR4ZM
jk
,DHAR2ZDHAR3L3jkx
yZM。¼ÌÖ;òóÏd¼ô¬qúM
,
10、25Z50μmol·L
-1
YAg
L3ZZMjkDHARbwv xyUY
Ad
。
25Z50μmol·L
-1
YAg
,L3jkDHAR xyúâÆZM
1.25Z1.16p,
ZM。
2.5 CDEB2}4567~89犃犘犡A
o,uvwFG
;YA7d2RSTUVWjkŶ
yëì:$¼ôxyZ APXxy·¸,Å
\11APXbwv(*5)。,APX5、6、
9、10、112VWjka¢Yef£
¤
;APX1、2、4ZMjk¢Yef
£¤
,
èL3jka¢Yef£¤;APX3
2VWjk¢Yef£¤,èVWq
*4 YefgrRSTUVWL3ZZMjk
DHARbwvb
Fig.4 ChangesofDHARisozymeprofileunder
increasingCdconcentrationintherootsofthetwo
犞.狊犪狋犻狏犪varieties,L3andZM
*5 abdYefg2RSTUVWL3ZZM
jkAPXbwvb|
Fig.5 ChangesofAPXisozymeprofileunder
increasingCdconcentrationintherootsofthetwo
犞.狊犪狋犻狏犪varieties,L3andZM
59317[ 345,B:RSTUjklmno}³v;Yefeç
*6 25μmol·L
-1
YAg2RSTUVWL3Z
ZMjkAPX,~ª«¬qPCRúM
Fig.6 RealtimeRTPCRanalysisofAPXgenein
responseto25μmol·L
-1Cdtreatmentintherootsof
two犞.狊犪狋犻狏犪varieties,L3andZM
LI
;APX7、8Æ2:;APXbwv,
2VWUYAd,
APX8L3jk¶ZM¦
。¼ÌÖ
;òóÏd¼ô¬qúMCD
,10、25Z
50μmol·L
-1
YAgL3ZZM jk APXb
wv xy¶;
,
L3jkúâ
Æ;1.06、1.09Z1.16p;ZMjkúâ
Æ;1.16、1.21Z1.26p;bYAd
g
,L34ZMjkAPX xys¸
LI。
bJ
,
¼ÌÖ×zª«¬q RTPCRtÑ`Y
efg2RSTUVWjk1&ð APX
,~PQ
,
CDYef£¤`ÄÌ APX,
~®¯
(
*6)。,YefA12hJ,
2RSTUVWjk APX,~uÂv
,
r APX,~´µYefAJ
V{¢À;q2.0p´µ,
VWs¸
LI
。
3 d w
Yefle#$Jé@{Cÿ
[1718],
ÌxY
;j]ʶºÂÿeÈ
,
jS{;YefÎ[
[19];
Yefté¤ê#${|ef
。
0YZ
,
Y;RSTUVWZM j@{]Êcd
JVWL3;ZM j´îïz{|Zðîñòcd
K
JL3。Äoy°VWZM ;Yef
L3¦[\。
bJ
,
0BCYefG2RSTUVWj
kAsApqc
,bYAdg
L3jk AsA pqZM,DHA pq6ZM,
AsA/DHAT1JZM,Äd°AsA®4
`YefgRSTUjkl{|eç
,L3jkÇ
¸¦Ïl{|
、
¢À{|tumn·A
,
ÄÅ·
ÆL3¶ZMǸ¦ÏYXyÈ;u~ËÌ。
#$Ý÷ AsA´µz AsAPþ、½
¾
、
ÂZ®{B¾|
,
AsA½¾#$ef
eçåU}È;#$
[20]。AsA{|þ
DHAÌf©ÅDHAR-|gtu]AsA,~
Ìf © Å Q ) C ´ Â þ 2,3 o[21]。
DHAR-|DHAtu,ÅQ7DHAúÂ,»
¼AsA½¾/@。0YZ2RSTUVW
jk¬¥4DHARbwv,Äobwv
xycYAd
,
d°RS
TUjkDHARxy¢Yef
£¤。b
YAdg
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[1] OLMOSE,MART?NEZSOLANOJR,PIQUERASA,犲狋
犪犾.Earlystepsintheoxidativeburstinducedbycadmiumin
culturedtobaccocels(BY2line)[J].犑.犈狓狆.犅狅狋.,2003,
54(381):291301.
[2] P?REZCHACAMV,RODR?GUEZSERRANOM,MOLI
NAAS,犲狋犪犾.Cadmiuminducestwowavesofreactiveoxy
genspeciesin犌犾狔犮犻狀犲犿犪狓 (L.)roots[J].犘犾犪狀狋犆犲犾犾犈狀狏犻
狉狅狀.,2014,37(7):16721687.
[3] GARNIERL,SIMONPLASF,THULEAUP,犲狋犪犾.Cad
miumaffectstobaccocelsbyaseriesofthreewavesofreac
tiveoxygenspeciesthatcontributetocytotoxicity[J].犘犾犪狀狋
犆犲犾犾犈狀狏犻狉狅狀.,2006,29(10):19561969.
[4] DENGX,XIAY,HU W,犲狋犪犾.Cadmiuminducedoxidative
damageandprotectiveeffectsofNacetylLcysteineagainst
cadmiumtoxicityin犛狅犾犪狀狌犿狀犻犵狉狌犿L.[J].犑.犎犪狕犪狉犱.犕犪
狋犲狉.,2010,180:722729.
[5] DOM?NGUEZDM,GARC?AFC,RAYAAC,犲狋犪犾.Cad
miuminducedoxidativestressandtheresponseoftheantioxi
dativedefensesystemin犛狆犪狉狋犻狀犪犱犲狀狊犻犳犾狅狉犪[J].犘犺狔狊犻狅犾.
犘犾犪狀狋犪狉狌犿,2010,139(3):289302.
[6] CHENZ,GALLIEDR.Increasingtolerancetoozonebyele
vatingfoliarascorbicacidconfersgreaterprotectionagainsto
zonethanincreasingavoidance[J].犘犾犪狀狋犘犺狔狊犻狅犾.,2005,
138(3):16731689.
[7] ELTAYEBAE,KAWANON,BADAWIGH,犲狋犪犾.Over
expressionofmonodehydroascorbatereductaseintransgenic
tobaccoconfersenhancedtolerancetoozone,saltandpolyeth
yleneglycolstresses[J].犘犾犪狀狋犪,2007,225(5):12551264.
[8] BURKEYKO,EASONG,FISCUSEL.Factorsthataffect
leafextracelularascorbicacidcontentandredoxstatus[J].
犘犺狔狊犻狅犾.犘犾犪狀狋犪狉狌犿,2003,117:5157.
[9] SHARMAP,DUBEYRS.Involvementofoxidativestress
androleofantioxidativedefensesysteminfrowingriceseed
lingsexposedtotoxicconcentrationsofaluminum[J].犘犾犪狀狋
犆犲犾犾犚犲狆.,2007,26:20272038.
[10] PARADISOA,BERARDINOR,DEPINTO MC,犲狋犪犾.
Increaseinascorbateglutathione metabolism aslocaland
precocioussystemicresponsesinducedbycadmiumindurum
wheatplants[J].犘犾犪狀狋犆犲犾犾犘犺狔狊犻狅犾.,2008,49(3):
362374.
[11] SCHTZENDBELA,POLLEA.Plantresponsestoabi
oticstresses:heavymetalinducedoxidativestressandpro
tectionbymycorrhization[J].犑.犈狓狆.犅狅狋.,2002,53:
13511365.
[12] YAMAMOTO Y,KOBAYASHIY,MATSUMOTO H.
Lipidperoxidationisanearlysymptomtriggeredbyalumi
num,butnottheprimarycauseofelongationinhibitionin
pearoots[J].犘犾犪狀狋犘犺狔狊犻狅犾.,2001,125(1):199208.
[13] KAMPFENKELK,VANMONTAGUM,INZED.Extrac
tionanddeterminationofascorbateanddehydroascorbate
fromplanttissue[J].犃狀犪犾.犅犻狅犮犺犲犿.,1995,225(1):
165167.
[14] LAEMMLIUK.Cleavageofstructuralproteinsduringthe
79317[ 345,B:RSTUjklmno}³v;Yefeç
assemblyoftheheadofbacteriophageT4[J].犖犪狋狌狉犲,1970,
227(5259):680685.
[15] ARAVINDP,NARASIMHA M,PRASADV.Modulation
ofcadmiuminducedoxidativestressin犆犲狉犪狋狅狆犺狔犾犾狌犿犱犲犿
犲狉狊狌犿byzincinvolvesascorbateglutathionecycleandgluta
thionemetabolism[J].犘犾犪狀狋犘犺狔狊犻狅犾.犅犻狅犮犺.,2005,43
(2):107116.
[16] RAOM V,PALIYATHC,ORMRODDP.UltravioletB
andozoneinducedbiochemicalchangesinantioxidanten
zymesof犃狉犪犫犻犱狅狆狊犻狊狋犺犪犾犻犪狀犪[J].犘犾犪狀狋犘犺狔狊犻狅犾.,1996,
110(1):125136.
[17] MOHAMED A A,CASTAGNA A,RANIERIA,犲狋犪犾.
Cadmiumtolerancein犅狉犪狊狊犻犮犪犼狌狀犮犲犪rootsandshootsisaf
fectedbyantioxidantstatusandphytochelatinbiosynthesis
[J].犘犾犪狀狋犘犺狔狊犻狅犾.犅犻狅犮犺.,2012,57:1522.
[18] PODAZZAG,ARIASM,PRADOFE.Cadmiumaccumula
tionandstrategiestoavoiditstoxicityinrootsofthecitrus
rootstockCitrumelo[J].犑.犎犪狕犪狉犱.犕犪狋犲狉.,2012,215:
8389.
[19] GUO Y T,MARSCHNER H.Uptake,distribution,and
bindingofcadmiumandnickelindifferentplantspecies[J].
犑.犘犾犪狀狋犖狌狋狉.,1995,18(12):26912706.
[20] STEVENSR,PAGED,GOUBLEB,犲狋犪犾.Tomatofruit
ascorbicacidcontentislinkedwithmonodehydroascorbate
reductaseactivityandtolerancetochilingstress[J].犘犾犪狀狋
犆犲犾犾犈狀狏犻狉狅狀.,2008,31(8):10861096.
[21] NOCTOR G,FOYER C H.Ascorbateandglutathione:
Keepingactiveoxygenundercontrol[J].犃狀狀狌.犚犲狏.犘犾犪狀狋
犘犺狔狊犻狅犾.犘犾犪狀狋犕狅犾.犅犻狅犾.,1998,49:249279.
[22] CHENZ,GALLIEDR.IncreasingvitaminCcontentof
plantsthroughenhancedascorbaterecycling[J].犘.犖犪狋犾.
犃犮犪犱.犛犮犻.犝犛犃,2003,100(6):35253530.
[23] YINL,WANGS,ELTAYEBAE,犲狋犪犾.Overexpressionof
dehydroascorbatereductase,butnotmonodehydroascorbate
reductase,conferstolerancetoaluminumstressintransgenic
tobacco[J].犘犾犪狀狋犪,2010,231(3):609621.
[24] QUEVALG,HAGERJ,GAKIREB,犲狋犪犾.Whyarelitera
turedataforH2O2contentssovariable?Adiscussionofpo
tentialdifficultiesinthequantitativeassayofleafextracts
[J].犑.犈狓狆.犅狅狋.,2008,59(2):135146.
[25] RUIH,CHENC,ZHANGX,犲狋犪犾.Cdinducedoxidative
stressandlignificationintherootsoftwo犞犻犮犻犪狊犪狋犻狏犪L.va
rietieswithdifferentCdtolerances.[J]J.犎犪狕犪狉犱.犕犪狋犲狉.,
2016,301:304313.
[26] HOREMANSN,FOYERCH,ASARDH.Transportand
actionofascorbateattheplantplasmamembrane[J].犜狉犲狀犱狊
犘犾犪狀狋犛犮犻.,2000,5(6):263267.
[27] SHIMAOKAT,YOKOTAA,MIYAKEC.Purificationand
characterizationofchloroplasthydroascorbatereductasefrom
Spinachleaves[J].犘犾犪狀狋犆犲犾犾犘犺狔狊犻狅犾.,2000,10:1110
1118.
[28] SHIGEOKAS,ISHIKAWAT,TAMOIM,犲狋犪犾.Regula
tionandfunctionofascorbateperoxidaseisoenzymes[J].犑.
犈狓狆.犅狅狋.,2002,53(372):13051319.
[29] FECHTCHRISTOFFERSM M,BRAUN HP,LEMAI
TREGUILLIERC,犲狋犪犾.Effectofmanganesetoxicityon
theproteomeoftheleafapoplastincowpea[J].犘犾犪狀狋犘犺狔狊犻
狅犾.,2003,133(4):19351946.
[30] PIGNOCCHIC,FOYERCH.Apoplasticascorbatemetabo
lismanditsroleintheregulationofcelsignaling[J].犆狌狉狉
犗狆犻狀.犘犾犪狀狋犅犻狅.,2003,6(4):379389.
[31] GREENMA,FRYSC.VitaminCdegradationinplantcels
viaenzymatichydrolysisof4OoxalylLthreonate[J].犖犪
狋狌狉犲,2005,433(7021):8387.
[32] HOUDE M,DIALLOA.Identificationofgenesandpath
waysassociatedwithaluminumstressandtoleranceusing
transcriptomeprofilingofwheatnearisogeniclines[J].犅犕犆
犌犲狀狅犿犻犮狊,2008,9:400412.
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