免费文献传递   相关文献

Study on Ascorbic Acid and Relative Enzymes Against Cadmium Stress in the Roots of Vicia sativa L.

箭舌豌豆根系抗坏血酸及相关酶对镉胁迫的响应



全 文 :书!"#$%&
,2016,36(7):1391-1398
犃犮狋犪犅狅狋.犅狅狉犲犪犾.犗犮犮犻犱犲狀狋.犛犻狀.
  !"#$:10004025(2016)07139108               犱狅犻:10.7606/j.issn.10004025.2016.07.1391
%&(
:20160303;)*&%+(:20160616
,-./

()*+%,-
(31160053,31560072);./%012,-(TZXY2015JBJJ005)
0123

345
(1969-),6,12,789,:;<=#$>?@ABC。Email:njrtz@sina.com
456789:;<=>?@ABCDEFGH
!"#
1,2,
$%%
2,
& 2,()*2,$+,3
(1./%0,DE./225300;2FGHIJ% @K+%%0,FG210095;3L!%0,MNOP734000)
I J:QRSTU(犞犻犮犻犪狊犪狋犻狏犪L.)VWL3(XY)ZZM(Y[\)]^_,BC`abcdYefgRSTUh
ijklmno
(AsA)pq、rslmnotuv(DHAR)bwvxy、lmnoz{|$v(APX)bwvxy
Q}APX,~€‚|。ƒ„…†:(1)2‡RSTUVWjk AsAZrslmno(DHA)pqˆYefg
…‰Š‹
;AsA/DHAŒˆYXyVWL3Ž…‰Š‹,ˆ[\VWZM Ž…‰g;bY‘A’dg,L3j
kAsApqZAsA/DHAŒ…‰J“ZM。(2)2‡VWjkDHARxy”•–…†4—bwv—˜,™š
xy›œY‘A’dŠ‹Š‹

žŽDHAR1ŸˆL3…†,DHAR4ŸˆZM …†;bY‘A’dg,V
WL3DHAR xyJ“VWZM。(3)2‡VWjk APXxy”•–…†11—bwv—˜,žŽ
APX1、2、4¡ˆ[\VWZMŽ¢Yef£¤,APX8ˆXyVWL3Ž¢¥Œ[\VWZM¦…‰£¤;§¨
©¥1‡RSTUAPX,~,ª«¬qRTPCRƒ„…†­,~®¯ˆL3ZZMjk›¢Y‘A£¤。BC


Yefg2‡RSTUVWjkAsApq,AsA±²³vDHARZ APXxyQ} APX®¯´µ
›…‰Š‹

YXyVWL3¶[\VWZM·¦¸¹º»¼AsA½¾,¿À¦‹AsA´µ,<¦¸¹ºÁÂ
Yef£¤Ã@{|ef

ÄÅ·ÆL3¶ZMǸ¦‹YXyÈ;ÉÊËÌ。
@KL

RSTU

YXy

lmno

lmnoz{|$v

rslmnotuv
MNOP$
:Q945.78 !QRST:A
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(CdtolerantvarietyL3,Cdsensi
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.AnAPXgenewasclonedandsubjectedtoRTPCRanalysis.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
  YÆÍyÎρÈ-ÐËÌ,~žÑÒ#$Ó
Ô

Õ¼ÌÖ×zØ$ÙÚÛÜÝÞß

~ࢥá
⁳ã

YaǸ{|tuxy

a·äåFen
tonZ HaberWeissæç,èYefé¤ê#$Ã
@ZëìJqxy{
(ROS)[12],ROSí£¤î
ïz{|

Ã@ðîñò

~à{|ñòÒó]Æ#
$YÍہ:;u~ËÌ
[3]。
ôõBC&ö`Y‘
Ag#${|efÃ@
[45]。
lmno
(AsA)Æ
#$Ý÷‹ødùúûl{|$ð

Æl{|kü
È;ýþÿú

Òó]Æ{|tuŽ!

ˆ#$
"l{|efŽÇ¸È;#$
。AsA%ˆ“#$


()Ý

*+ÝZð,ݎ

ÅQ×zlm
noz{|$vAPX-|#$./01234
H2O2 æç,5 H2O2 tu] H2O,6#$&
H2O2 ëì,78#${|efòÛ。AsA01
Ò{|]rslmno
(DHA),DHA×zrsl
mnotuv
(DHAR)tu]AsA。9&ö,AsA
pqŠ‹·:‹#$;
=>

?,*Z@
ef"l·A
[67]。BurkeyB[8]CD,E{ef
gXyVWFU(ŽAsApqŒ[\VW‹,X
yVW·¿À;¶‹AsAG´µ。SharmaZ
Dubey[9]&öHef£¤´IhiAPXxyŠ‹,
bJžbwvýþKC@L‚

K¸&ö AsA
·:‹#$;Yef"l·A

=,MNOAsA
Pþ  Q Ý LR S T o1,4÷ 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]。
0YZQŽ!"º[\]W#X^_

Ø
`a$#$RSTU
(犞犻犮犻犪狊犪狋犻狏犪L.)]YZ^
_

BC2‡abYXyRSTUVWˆYef
gjkAsApq、APXbwvbZ,~€Q}
DHARbwvb‚|,¼ÌÖcd#$lmno
±²eçYefÉA

1 ^_Zfg
1.1 UVWX>YZ
YZ^_RSTUhi1‡YXyVWL3Z
1‡Y[\VW ZM。Wûj3% H2O2 kÍ
15min,$lmû´nopqr,sW16h,®tu
vwŽ20℃~25℃xC。3dr,yz{|}~
Ìêhi®tup2.5LHoagland€‚ƒ
„Žƒ

…ƒ„ti10†。ƒ—‡=g:
ˆ

‰Šd25℃/20℃,;‹d60%~80% ,«
ŒJ12h,«ŒÏd400μmoL· m
-2·s-1。…
3dŽ1€‚,€‚pHÊˆ5.5‘’。
YZÌ
:2‡RSTUVWti7dr,$
25μmol·L
-1
Y‘A

YQCdCl2·2.5H2O“
”O•
)。
…‡‘A3È–,‘A‚…3dŽ1


ú∑A0、2、4、6、8、10、12、14d—˜$“
jk@{™š›¬

YZœ
:0(Control)、10、25、50μmol·L
-1

A

‘AfgbYZÌ
),
‘A7drԝjÿ,žŸ
$“YZ

.“‚ Ž@¡r
,-80℃¢%£@A
™š›¬

YZ¤

ti3dr,®tuO•25μmol·L
-1
CdCl2 €‚Ž‘A(‘AfgbYZÌ),…‡
‘AȖ3,‘A96h÷jk—˜,…‡˜V]4
¥i¦P˜

—˜J]0、12、24、48、96h。§
—˜VžŸ“‚ Ž¨¡
,-80℃¢%$“jk 
RNA:—。
1.2 [\./>]^
1.2.1 89_`aR úâˆ25μmol·L
-1

A0、2、4、6、8、10、12、14d,›qhi:j{r,ԝ
jÿ

Ól©´úª—«È

1.2.2 bcdef ðy›¬®Œ
SchützendübelB[11]¯°±²(Evansblue)ÓÔ
g

—10¥³«#$1cm:jj´,“0.025%
2931 ! " # $ % &                   36µ
(W/V)¯ °±²¶‚Ž·¸30min,lmû´pq
15minno¹º»¸r,“1%SDS(W/V)50%(V/
V)¼½ŽB¾þ¿À,50℃´Á15minr,14000×
gm!15min,—¹›600nmgÓ«。
1.2.3 cghijFkljmn[ îïz{|
ýÃ|%ě®Œ YamamotoB[12]fg。5
2cm:jj´s•SchiffÅÆ (0.5gÇyVÈ,
0.5gK2S2O5,100mL1mol·L-1 HCl,100mL
H2O)Ž·¸60min,*r$0.5% (W/V)
K2S2O5 ¶‚ (¶“0.05mol·L-1 HCl)pq,
ɓʸË̐ɁÝͅÎÏgÐÑ

Ҍ

1.2.4 犃狊犃o 犇犎犃pq AsA pq›¬®Œ
KampfenkelB[13]fg。—1g³«#$jk,O
•Ó@5 mL6% TCA ÔÁB¾,4 ℃ m!
(12000×g,20min),¹‚]՛‚。 AsAp
q›¬

—0.2mL՛‚(;Œ—0.2mL6%
TCA),O0.2mL10mmol·L-1DTTZ0.4mL
0.2mol·L-1PBS(pH7.4),Öú¦¿,42℃æç
15minrמŽO•0.2mL0.5% NØ,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,žèbÂ。
  AsA pq7ltuæ AsA pqŸ]{|æ
AsA,Ÿrslmno(DHA)pq。AsA ¶Â“
6% TCA,bg#šéê*。
1.2.5 犇犎犃犚o犃犘犡Frfst ëì:—}a
íîxyïðñÛÝòó”•
(nativePAGE)®Œ
Laemmli[14]fgˆ4℃g¼ô。úmóZ’õó

apSDS)ö¼”•÷,”•÷<øO•Bqë
ìð

”•Áp‚]pH8.3TrisHCl,ù¬”ú
30mA。”•¬û,¼ôvxy·¸。
DHARnativePAGEü$4%’õóZ
10%úmó。DHARxy·¸®Œ Aravind
B
[15]
fg

5”•ròós•p4mmol·
L-1GSH,2mmol·L-1DHA0.1mol·LPBS
(pH6.2)Ž,“ýþŠÿ20min。´qr,“p
0.1%FeCl3 Z0.1% K3[Fe(CN)6]0.1mol·
L-1 HCl¶‚Ž!ÿ,ˆ"²¸#$Â%D&²¸
—˜

(´)q

*+,-

APXnativePAGEü$4%’õóZ8%
úmó
。APXxy·¸ü$ RaoB[16]f
g

òó.ˆp2mmol·L-1 AsA50mmol·
L-1PBSŽ!ÿ30min,/ˆp4mmol·L-1AsA
Z2mmol·L-1 H2O2 50mmol·L-1PBS(pH
7.0)Ž!ÿ20min。Áp‚pq1minr,ˆp
28mmol·L-1 TEMEDZ2.45mmol·L-1 NBT
PBS(pH7.8)Ž,88ý0,·¸10~15min。
´q

12æç

*+,-

1.2.6 犃犘犡,uvwFxy\q犘犆犚Oz “
NCBI3456RSTU§ˆU+#$ APX,
~78

9$DNAMAN6.0:‡¼ô;o78Œ
;

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。›7ƒ„°™4TU&
ð犘狊犃犘犡01,~(78E]X62077.1);Fo
78Ìêy]94%。YJª«¬qPCRæç‚G
$ SYBR Premix Ex 犜犪狇 (TliRNaseH Plus,
Takara)ÅÆH,G$7500RealTimePCR kü
(AppliedBiosystems,USA)ě,~€q‚
|
。PCR§$犃犘犡 áIyä$],J×:5′GCT
GGTGTTGTTGCTGTTG3′;æ×:5′CCCTCT
GGTGGTGGATGG3′。Actin#]÷®,Actin(7
8E GU946218.1)áIyä$:J×:5′TCTG
GTGATGGTGTGAGTC3′;æ×:5′ACAGCAA
CATAGGCAAGC3′。PCR —‡:95 ℃,30sÂ
Ù

œr95℃,5s;60℃,34s,–40‡½¾。<‘
A,~€q4;ŒŒ

QActin]÷®,;Œ
€q]1,©¥‘Ar1.3 {|YZ
G$SPSS13.0(SPSSInc.,Chicago,U.S.
A.)Ë9‘A:‡h¼ôfLúMZLSDõȌ


abNO†LI…‰
(犘<0.05)。G$
QuantityOne(BioRad):‡¼ôòó—˜Ïd
¬qúM

2 ƒ„4úM
2.1 CDEB2}4567~89_`F€G
2‡RSTUVWL3ZZM jˆ25μmol·
L-1Y‘Ag@{PQ=*1§†。ˆYZR¯
1~14d÷,ZM:jS{@{qZjk«ÈX
{q›T1ù“L3,VWLIˆ‘A4dQrŸ
€¥…‰´µ

ÄWLID%œU‘AJV
{WXXOY|

ÕQ:jS{D©¦Z%

=ˆ‘A14dJ,VWZM :jS{qZjk«È
X{qúâŒb[L3…‰663%Z41%。Å\,
39317[             345,B:RSTUjklmno}³v;Yefeç
abNO†‘AJË./VWˆ0.05´µ%ˆ…‰yLI;ˍ]µ›Ë±šéL;gb
*1 25μmol·L
-1Cd‘Ag2‡RSTUVWL3ZZM:jS{Zj«ÈX{
Thediferentnormallettersindicatedsignificantdiferenceamongcultivarsortreatmenttimesat0.05level;Dataaremeans±SD(n=3);
Thesameasbelow
Fig.1 Effectsof25μmol·L
-1Cdtreatmentonrootelongationandincreaseofrootfreshweightintwo犞.狊犪狋犻狏犪
varieties,L3andZM
ˆŽdYef—‡g

Y[\VWZM jk@{
¶XYVWL3¢¥¦…‰]Ê。
2.2 CDE2}4567~8‚Fcghi
jobcƒ„
  G$SchiffÅÆ;Y‘A7d2‡RSTU
VWj´îïz{|cd¼ô`ýÃ|%ě

SchiffÅÆ4îïz{|Ã@^_|P$æç@
þ`ȸ$ð

ƒ„=*2§†。žŽ,ˆ;Œ
(0μmol·L
-1Cd)—‡g,2‡RSTUVWj
´abÐÑa¥`ȸ

ˆ10~50μmol·L
-1

A—‡g
,L3ZZM j´c%D`ȸ,œY‘
A’dXOj´`ȸWX°…

…†îïz{
|cdWXO&

ˆbY‘A’dg
,ZM ¶L3
j´·¸¦&

d°YefgZMj´îïz{|
cd¦eÈ

™¢¥`ŒL3¦eȁ{|ef。
Evansblue(¯ °±²)Æfghy·_,ij
´·ÒEvansblue·þ²¸,d°j´&;Ev
ansblue×h,žðy¢¥jm,·¸k&
†j´&ðîñòcdkJ

Y‘Ag2‡
RSTUVWj´EvansblueÓÔqúM°

*3),10~50μmol·L
-1
Y‘A7dr,2‡RS
TUVWj´ð¥°…ñò

ñòcdœY
‘A’dXO…‰XO

ˆbY‘A’dg

ZMj´ðîñòcd…‰J“L3。
2.3 CDEB2}4567~89犃狊犃、犇犎犃
pqo犃狊犃/犇犎犃…†F€G
  1ƒ„…†,<’dY‘A7dr2‡RST
UVWjkAsAZDHApqc¶;Œ(0μmol·
*2 ab’dYefg2‡RSTUVWL3ZZM
j´îïz{|Schiff·¸
Fig.2 Histochemicaldetectionoflipidperoxidation
bySchiffsreagentintheroottipsoftwo犞.狊犪狋犻狏犪
varieties,L3andZM
*3 ab’dY;2‡RSTUVWL3ZZMj´¯
°±²Óԁle
Fig.3 QuantitativeassayofEvansblueuptakeinthe
roottipsoftwo犞.狊犪狋犻狏犪varieties,L3andZM
4931 ! " # $ % &                   36µ
v1 ‡ˆ‰ŠCDE‹Œ2}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
Y’d
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

ƒ„]3È–YZµ›±šéL(n=3)
Note:Valuesaremeans±SD(n=3)ofthreedifferentexperiments
L-1Cd)…‰Š‹;L3jkAsA/DHAŒ¶;
Œ…‰Š‹

ZMjkAsA/DHAŒ¶;Œ
…‰6

ˆbY‘A’dg
,L3jk AsA
pqZAsA/DHAŒ›…‰J“ZM。Qƒ„


YefgXyVWL3Ǹ¶[\VWZM
¦ÏÃ@AsAZ¿À{|tumn·A。
2.4 CDEB2}4567~89犇犎犃犚ˆ
AŽF€G
  ;Y‘A7d2‡RSTUVWjkÅ¶
yëì:—$¼ôxy”•ZDHARxy·¸,Å
\4—DHARbwv—˜(*4)。Äobwv
xy › œ U Y ‘ A ’ d  Š ‹  Š ‹

ž Ž
DHAR1ŸˆL3jkŽ…†,DHAR4ŸˆZM
jkŽ…†
,DHAR2ZDHAR3ˆL3jkŽx
y‹“ZM。¼ÌÖ;òó—˜Ïd¼ô¬qúM
ƒ„…†

ˆ10、25Z50μmol·L
-1
Y‘Ag
L3ZZMjkŽDHARbwv xy›œUY‘
A’dŠ‹…‰Š‹

ˆ25Z50μmol·L
-1
Y‘Ag
,L3jkDHAR xyúâÆZM 
1.25Z1.16p,…‰‹“ZM。
2.5 CDEB2}4567~89犃犘犡ˆA
Žo,uvwF€G
  ;Y‘A7d2‡RSTUVWjkÅ¶
yëì:—$¼ôxy”•Z APXxy·¸,Å
\11—APXbwv—˜(*5)。žŽ,APX5、6、
9、10、11—˜ˆ2‡VWjkŽ›a¢Yef£
¤
;APX1、2、4—˜ˆZMjkŽ¢Yef…‰
£¤

èˆL3jkŽa¢Yef£¤;APX3—
˜ˆ2‡VWjk›¢Yef£¤,èVWq…
*4 Yefgr‡RSTUVWL3ZZMjk
DHARbwvb
Fig.4 ChangesofDHARisozymeprofileunder
increasingCdconcentrationintherootsofthetwo
犞.狊犪狋犻狏犪varieties,L3andZM
*5 ab’dYefg2‡RSTUVWL3ZZM
jkAPXbwvb‚|
Fig.5 ChangesofAPXisozymeprofileunder
increasingCdconcentrationintherootsofthetwo
犞.狊犪狋犻狏犪varieties,L3andZM
59317[             345,B:RSTUjklmno}³v;Yefeç
*6 25μmol·L
-1
Y‘Ag2‡RSTUVWL3Z
ZMjkAPX,~€ª«¬qPCRúM
Fig.6 RealtimeRTPCRanalysisofAPXgenein
responseto25μmol·L
-1Cdtreatmentintherootsof
two犞.狊犪狋犻狏犪varieties,L3andZM
‰LI
;APX7、8Æ2‡:;APXbwv—˜,
ˆ2‡VWŽ›œUY‘A’dŠ‹Š‹,ž
ŽAPX8ˆL3jkŽ¶ZMŠ‹¦…‰。¼ÌÖ
;òó—˜Ïd¼ô¬qúMCD
,10、25Z
50μmol·L
-1
Y‘AgL3ZZM jkŽ APXb
wv xy›¶;Œ…‰Š‹

ˆL3jkŽúâ
Æ;Œ1.06、1.09Z1.16p;ˆZMjkŽúâ
Æ;Œ1.16、1.21Z1.26p;bY‘A’d
g
,L34ZMjkŽAPX xys¸…‰LI。
bJ

¼ÌÖ×zª«¬q RTPCRtÑ`Y
efg2‡RSTUVWjkŽ1‡&ð APX
,~€PQ

CDYef£¤`ÄÌ APX,
~®¯€

*6)。žŽ,ˆYef‘A12hJ,
2‡RSTUVWjkŽ­ APX,~€uÂv

œr APX,~€´µœYef‘AJ
V{¢Àˆ;Œ€q2.0p‘’´µ,
VWs¸…‰LI

3 d w
Yefle#$Jé@{Cÿ
[1718],
ÌxY
;j]ʶºÂÿeÈ

jS{;YefÎ[
[19];
Yefté¤ê#${|ef

0YZ
Ž

Y;RSTUVWZM j@{]Êcd…‰
J“VWL3;ZM j´îïz{|Zðîñòcd
K…‰J“L3。Äoƒ„y°VWZM ;Yef
ŒL3¦[\。
bJ

0BCŽYefG2‡RSTUVWj
kAsApqc…‰Š‹,ˆbY‘A’dg
L3jk AsA pq‹“ZM,DHA pq6“ZM,
AsA/DHAŒT1J“ZM,Äd°AsA®4
`YefgRSTUjkl{|eç
,L3jkÇ
¸¦Ïl{|

¢À{|tumn·A

ÄÅ·
ÆL3¶ZMǸ¦ÏYXyÈ;u~ËÌ。
#$Ý÷ AsA´µ—z“ AsAPþ、½
¾

ÂZ®{B¾|

žŽAsA½¾ˆ#$ef
eçŽåU}žÈ;#$
[20]。AsA{|“þ
DHAÌf©ÅˆDHAR-|gtu]AsA,~
Ìf © Å Q ) C ´  þ 2,3œ  €  o[21]。
DHAR-|DHAtu,ÅQ7‚DHAúÂ,»
¼AsA½¾/@。0YZŽ2‡RSTUVW
jk–ƒ¬¥4—DHARbwv˜,Äobwv
xycœY‘A’dŠ‹…‰Š‹

d°RS
TUjkDHARxy¢Yef…‰£¤。b
Y‘A’dg

XyVWL3jkDHAR xy
‹“[\VWZM,ÄÅ·GL3jk¶ZM ¦¸¹
º»¼DHAtu]AsA,Ç¸¦‹AsA´µ
ZAsA/DHAŒ。ChenB[22]&ö,ˆ„…Z†
‡(ˆz€ùW DHAR,~Å:‹ DHARx
y

XO AsApqZ AsA/DHAŒ。YinB[23]
&öz€‰FŠDHAR,~„…#¥AsAp
qZAPXxyˆHefgXO,;HXy:‹。
Yefg

#$Ã@ROSéëì“ð,Ý[24],~
àð,ÝROS¹º;:‹#$YXy¸È;#
$

0YZQ[BCCD

Yefg2‡RST
UVWjk ROS:;ëì“&‹Zð,݌

[25]。AsAÆð,ݎÎÈ;l{|úû[23]。
èð,ݍŽAsA±²³v,ð,ݎ AsA{
|Ã@DHA;×zAsA/DHA®{Ý®{zð
î

*rˆ–ðݎDHARtu]AsA,AsA/
®{uð,Ý
[26]。


–ðݎDHARÅQ
bJtu–ðÝZð,ݎDHA]AsA。0Y


XYVWL3jk¶Y[\VWZM Ǹ¦
‹DHARxy,ÅQ¦¸¹º»¼–ðÝZð,
ݎDHAtu] AsA,¦¸¹º78Yef£
¤Ã@{|ef

<:‹L3;YefXy。
KQ

³“jk DHAR bwvBC&ö¶‚,
ShimaokaB[27]&öúm`‘F(ˆŽ’90%
DHAtuxyrW DHARbwv。0YZŽ
r‡RSTUVWjkDHARbwvLI…‰,Ä
oLI;™šDHAtu·Ale©“r&•
BC

AsAl{|:;”•Æ×zAPX-|AsA
4 H2O2 æç,5 H2O2 tu] H2O。0YZŽ2
‡abYXyRSTUVWjkAPXxycœ
6931 ! " # $ % &                   36µ
Y‘A’dŠ‹Š‹

d° APXxy¢Y‘
A£¤

‹B#$Ž
,APX%ˆõWbwv,h
ið APX、()Ý,ð APX Z_–ÝîƒP
APX、ÎÝ(hiØ^o½¾ÝZz{|$vÝ)î
ƒPAPXZ*+ÝîƒP APX[28]。0YZˆ2
‡RSTUVWjkŽ–ƒ¬¥11—APXbwv
˜

žŽAPX1、2、4ŸˆZMjk¢¥£¤,xy
΋bwvAPX8ˆL3jk¶ZM ¢¥¦…‰
£¤

—›2‡RSTUVWjk APX xy
˜*s¸…‰LI

èab APXbwv¢¥a
bcd£¤

Å·¤ê2‡RSTUVWjk-
|H2O2 ú·ALI。0YZŽ2‡RSTU
VWjkŽ1‡ðAPX,~ˆYefg…‰Â
v€

d° APXxyŠ‹4ž,~€XO
¸³

YefgRSTUjk APXxy%ˆ®¯
´µv

~,

º×z APX-| H2O2 úÂ、78#
$xy{òÛ
,AsAtÅ·×zž™zcle#
$YXy

™šQ[BCCDYefgZMj
´¶L3¦°…ºšð|[25],šð|zchpH2O2
®4›_$ð{|

›_$ð{|zcŽíÃ
@ŽÃ$›),

;ðîBƒœþñò
[29]。
AsAƸ¹›),¹ºÆ,L3YXyÅ·
4ž¿À¶‹AsA´µ,7‚`›),òÛ
¸³
。AsAtÅ#]vž~û;Ìk8È;
v»æçåv#$

le#$&úŸú|Z

,AsAtÆ#$&¡E®¤úû[3032]。~
à
,AsAtÅ·×zle&‹šð|Zš¢|z
c

le&úŸQ}j@{

<le#$Y
Xy

Äoc£;“r¼ÌÖ&•BC

!Q

[1] OLMOSE,MART?NEZSOLANOJR,PIQUERASA,犲狋
犪犾.Earlystepsintheoxidativeburstinducedbycadmiumin
culturedtobaccocels(BY2line)[J].犑.犈狓狆.犅狅狋.,2003,
54(381):291301.
[2] P?REZCHACAMV,RODR?GUEZSERRANOM,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):16721687.
[3] GARNIERL,SIMONPLASF,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):19561969.
[4] DENGX,XIAY,HU W,犲狋犪犾.Cadmiuminducedoxidative
damageandprotectiveeffectsofNacetylLcysteineagainst
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.,2010,180:722729.
[5] DOM?NGUEZDM,GARC?AFC,RAYAAC,犲狋犪犾.Cad
miuminducedoxidativestressandtheresponseoftheantioxi
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):289302.
[6] CHENZ,GALLIEDR.Increasingtolerancetoozonebyele
vatingfoliarascorbicacidconfersgreaterprotectionagainsto
zonethanincreasingavoidance[J].犘犾犪狀狋犘犺狔狊犻狅犾.,2005,
138(3):16731689.
[7] ELTAYEBAE,KAWANON,BADAWIGH,犲狋犪犾.Over
expressionofmonodehydroascorbatereductaseintransgenic
tobaccoconfersenhancedtolerancetoozone,saltandpolyeth
yleneglycolstresses[J].犘犾犪狀狋犪,2007,225(5):12551264.
[8] BURKEYKO,EASONG,FISCUSEL.Factorsthataffect
leafextracelularascorbicacidcontentandredoxstatus[J].
犘犺狔狊犻狅犾.犘犾犪狀狋犪狉狌犿,2003,117:5157.
[9] SHARMAP,DUBEYRS.Involvementofoxidativestress
androleofantioxidativedefensesysteminfrowingriceseed
lingsexposedtotoxicconcentrationsofaluminum[J].犘犾犪狀狋
犆犲犾犾犚犲狆.,2007,26:20272038.
[10] PARADISOA,BERARDINOR,DEPINTO MC,犲狋犪犾.
Increaseinascorbateglutathione metabolism aslocaland
precocioussystemicresponsesinducedbycadmiumindurum
wheatplants[J].犘犾犪狀狋犆犲犾犾犘犺狔狊犻狅犾.,2008,49(3):
362374.
[11] SCHTZENDBELA,POLLEA.Plantresponsestoabi
oticstresses:heavymetalinducedoxidativestressandpro
tectionbymycorrhization[J].犑.犈狓狆.犅狅狋.,2002,53:
13511365.
[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):199208.
[13] KAMPFENKELK,VANMONTAGUM,INZED.Extrac
tionanddeterminationofascorbateanddehydroascorbate
fromplanttissue[J].犃狀犪犾.犅犻狅犮犺犲犿.,1995,225(1):
165167.
[14] LAEMMLIUK.Cleavageofstructuralproteinsduringthe
79317[             345,B:RSTUjklmno}³v;Yefeç
assemblyoftheheadofbacteriophageT4[J].犖犪狋狌狉犲,1970,
227(5259):680685.
[15] ARAVINDP,NARASIMHA M,PRASADV.Modulation
ofcadmiuminducedoxidativestressin犆犲狉犪狋狅狆犺狔犾犾狌犿犱犲犿
犲狉狊狌犿byzincinvolvesascorbateglutathionecycleandgluta
thionemetabolism[J].犘犾犪狀狋犘犺狔狊犻狅犾.犅犻狅犮犺.,2005,43
(2):107116.
[16] RAOM V,PALIYATHC,ORMRODDP.UltravioletB
andozoneinducedbiochemicalchangesinantioxidanten
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):125136.
[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.,2012,57:1522.
[18] PODAZZAG,ARIASM,PRADOFE.Cadmiumaccumula
tionandstrategiestoavoiditstoxicityinrootsofthecitrus
rootstockCitrumelo[J].犑.犎犪狕犪狉犱.犕犪狋犲狉.,2012,215:
8389.
[19] GUO Y T,MARSCHNER H.Uptake,distribution,and
bindingofcadmiumandnickelindifferentplantspecies[J].
犑.犘犾犪狀狋犖狌狋狉.,1995,18(12):26912706.
[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):10861096.
[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.,1998,49:249279.
[22] CHENZ,GALLIEDR.IncreasingvitaminCcontentof
plantsthroughenhancedascorbaterecycling[J].犘.犖犪狋犾.
犃犮犪犱.犛犮犻.犝犛犃,2003,100(6):35253530.
[23] YINL,WANGS,ELTAYEBAE,犲狋犪犾.Overexpressionof
dehydroascorbatereductase,butnotmonodehydroascorbate
reductase,conferstolerancetoaluminumstressintransgenic
tobacco[J].犘犾犪狀狋犪,2010,231(3):609621.
[24] QUEVALG,HAGERJ,GAKIREB,犲狋犪犾.Whyarelitera
turedataforH2O2contentssovariable?Adiscussionofpo
tentialdifficultiesinthequantitativeassayofleafextracts
[J].犑.犈狓狆.犅狅狋.,2008,59(2):135146.
[25] RUIH,CHENC,ZHANGX,犲狋犪犾.Cdinducedoxidative
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.,
2016,301:304313.
[26] HOREMANSN,FOYERCH,ASARDH.Transportand
actionofascorbateattheplantplasmamembrane[J].犜狉犲狀犱狊
犘犾犪狀狋犛犮犻.,2000,5(6):263267.
[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):13051319.
[29] FECHTCHRISTOFFERSM M,BRAUN HP,LEMAI
TREGUILLIERC,犲狋犪犾.Effectofmanganesetoxicityon
theproteomeoftheleafapoplastincowpea[J].犘犾犪狀狋犘犺狔狊犻
狅犾.,2003,133(4):19351946.
[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):379389.
[31] GREENMA,FRYSC.VitaminCdegradationinplantcels
viaenzymatichydrolysisof4OoxalylLthreonate[J].犖犪
狋狌狉犲,2005,433(7021):8387.
[32] HOUDE M,DIALLOA.Identificationofgenesandpath
waysassociatedwithaluminumstressandtoleranceusing
transcriptomeprofilingofwheatnearisogeniclines[J].犅犕犆
犌犲狀狅犿犻犮狊,2008,9:400412.

!"

#$%
)  
8931 ! " # $ % &                   36µ