全 文 ：第 25卷　第 3期　　　　　　　　　　　　　植　　　物　　　研　　　究 2005年　7月
Vo .l 25　No. 3　　　　　　　　　　　　BULLETIN OF BOTAN ICAL RESEARCH July, 　2005
Foundat ion item:Supported by found ation for un iversity k ey science and technology by the m inistry of educat ion of Ch ina (02010)
Au thor in troduction:FAN Bao-Li(1976— ), Fem a le, Master, Ass is tan t, M ajor in cel l andm olecu lar b io logy.
*　Au thor for correspondence　E-m ai l:PYK cell@ eyou. com
Received date:2005 - 01 - 17
镉胁迫下三个萝卜栽培种蛋白质变化的双向电泳比较研究
范宝莉　王振英　彭永康*
(天津师范大学生物系 , 天津　300074)
摘　要　在 0. 5mmo l L -1 Cd2+处理时 ,小五缨(XWY)的发芽率下降至 92%,并且随着 Cd2 +浓
度的增加 ,发芽率逐渐降低 ,在 1和 5mmol L- 1 Cd2+浓度处理时 ,发芽率分别降至 83%和 67%。
象牙白 (XYB)则在 5mmol L - 1 Cd2+浓度时 ,发芽率下降。卫青 (WQ)在 0. 05mmol L -1 Cd2+
浓度时 ,发芽率已降至 83%, 1和 5 mmo l L - 1时则下降至 58%。幼苗生长也明显受 Cd2+处理影
响 ,在 0. 05mmol L - 1 Cd2+处理时 , 3种栽培萝卜的幼苗生长均受到明显影响 ,并且随 Cd2+浓度
的增加 ,生长受抑加重 。从发芽率和幼苗生长两种实验结果看 ,卫青对 Cd2 +最为敏感 。双向电泳
结果表明 , Cd2+处理后 3种栽培萝卜幼苗中蛋白质组分有明显变化 。小五缨中 , 0. 1 mmo l L -1
Cd
2 +处理后 ,有 5个蛋白质点消失 , 15个新的蛋白质点被诱导产生。象牙白中 , 2个蛋白质点消
失 , 1个蛋白质点含量减少 , 13个新的蛋白质点被诱导产生。卫青中 , 12个新的蛋白质点诱导产
生 ,但没有发现蛋白质点消失现象。 Cd2+处理后 , 3种栽培萝卜中 ,蛋白质合成的变化与幼苗生长
受抑存在明显相关性 ,这一实验结果对于探讨萝卜 Cd2 +害的生化机理是有重要意义的。
关键词　发芽率;双向电泳;蛋白质斑点;镉;萝卜
Protein changes induced by cadm ium stress in three rad ish cultivar seed lings:
a compar ison using two-dmi ensional gel electrophoresis
FAN Bao-Li　WANG Zhen-Ying　PENG Yong-K ang*
(Depa rtm en t of B io logy, T ianjin No rm al Unive rsity, T ianjin　300074)
Abstract　The germ ination rate was decreased to 92% when seeds w ere treated at 0. 5 mmo l L -1
Cd
2 +
and w as progressively decreased w ith h ighe rCd
2+
. 83% and 67% ge rm ination ra tesw ere observed
respective ly at 1mmo l L -1 and 5 mmo l L -1 Cd2+ treatment in XWY, bu t the low er germ ina tion ra te
w as observed w ith 5 mmo l L- 1 Cd2+ treatment in XYB. The ge rm ination ra te w as decreased to 83%
on ly at 0. 05 mmo l L- 1 Cd2+ and only 58% germ ination ra tew as observed when seedsw ere trea ted at
1 mmo l L -1 and 5 mmo l L- 1 Cd2+ inWQ. The effects o f Cd2+ on seed ling g row th we re obse rved in
th ree radish cu ltivars. Seed ling grow th dam age was much more serious w ith higher Cd
2+
concen tration.
WQ is the most sensitive to Cd
2+
treatm ent acco rding to the expe riment results w hich w ere obta ined by
testing germ ina tion ra te and seed ling grow th in three radish cultivars. 5 protein spots we re disappeared
and 15 new protein spo ts w ere induced in XWY seed ling w hich w ere treated by 0. 1 mmo l L- 1 Cd2+ ,
2 prote in spo tsw ere disappea red and 1 pro tein spotw as decreased in content, 13 new pro tein spots w ere
induced in XYB , 12 new pro tein spo tsw e re induced inWQ. A c lose positive re lationship w as observed
be tw een pro te in spo ts change and inhibited seedling grow th in the three radish cultiva rswh ich w ere trea-
ted by 0. 1 mmo l L - 1Cd2+. It may be importan t for understanding biochem ical mechanism of Cd2+
damage.
Key words　ge rm ination rate;tw o dimensional ge l electropho resis;p ro tein spo t;cadm ium;Raphanus
sa tivus L.
Cadm ium is the most serious pollution o f heavy
me tals in ag ricu ltura l soils, cadm ium-contam inated
crop so ils have been increased over 2 000 hm
2 ,
140 000 ton ag ricu ltura l products which w ere po lluted
w ith cadm ium were produced each year in C hina
[ 1, 2]
.
It makes agricu ltural y ield lowe r and affects human
health as they ente r the food chain. Fo r a long time,
particular atten tion w as paid to the effects o f so il cad-
m ium po llution on the crop y ie ld reduction and qua lity
decline
[ 3] , espec ially in the recen t years, many in-
vestigations have been m ade conce rning abou t the
physio log ica l, biochem ical and molecular biology
stud ies of the cadm ium toxicity effects in the c rop
grow th, for instance, H art et a l, K avita et al and R iv-
ea r et al have studied the regulation o f cadm ium ab-
sorbance and accumu lation in plan tsw hich w e re cu lti-
va ted a t differen t concentrations of cadm ium in durum
wheat cu ltivar and in th reeP isum sa tivum
[ 4 ～ 6]
. Zhou
et a l demonstrated tha t ultrastruc tura l damage w as
muchmore serious w ith increasing of Cd
2+
concentra-
tion in Alternantera philoxeroides
[ 7]
. M any expe ri-
men ts show ed that severalmetabolic pathw ay, such as
ch lo rophy ll pigment and protein synthesis
[ 8 ～ 16] , can
be seriously distu rbed by Cd
2+
exposure in plants,
despite many w orks general p lant response to Cd
2+,
the specific biochem ical bases o f plant adap tation and
tolerance to h igh Cd
2+
leve ls are still unclear, in
present investiga tion, we ana lyzed the re lationship be-
tw een the prote in pattern changes and plan t seed lings
tolerance to h igh Cd
2+
by 2D PAGE in three cu ltivat-
ed radishes. The a im w as to investigate the biochem i-
ca lmechan ism of Cd
2 +
damage in radish cu ltivars. It
may be useful to predict y ie ld reduction and qua lity
decline due to cadm ium po lluted in the so i.l
1　Materials and methods
1. 1　P lantmate ria ls and stress condition
Three rad ish (Raphanus sativus L. ) cultivars
(XWY , round rhizome, red. XYB , long rhizome,
white. andWQ , long rh izome, g reen)were used as
expe rim entma terials. 350 g ra ins of seed each radish
cu ltivars w ere su rface sterilized w ith a so lu tion of
0. 1% HgC l2 fo r 5 m in and we re then w ashed two
times w ith tap w a te r, and germ ina ted in petri dishes
on filter pape rmo istened w ith distilled w ate r fo r 24 h
a t room tempe rature. A fter that 24 h , the seedling s
we re treated w ith a so lu tion of CdC l2 a t 0. 05 ～ 5
mmol L- 1 for 72 h. A ll trea tmentswere performed at
23℃ in da rkness. Ge rm ina tion rate and bud height
were measured when seedling sw ere cultured fo r 72 h.
1. 2　2D po lyacry lam ide gel electropho resis
Tw o-dimensional po lyacry lam ide gel e lectropho-
resis w as pe rfo rmed according to the method of
O ’ Farrell et al[ 17] , w ith the use of amod ified proce-
dure:the one dimensional 3% po lyacry lam ide gel,
conta in ing 9 mol L - 1 urea 0. 98 g;2% nonion ic
de tergent NP40, 30% po lyacry lam ide 340 μL, am-
pho ly tes 30 μL (pH 3 ～ 10) and 60μL (pH 5 ～ 7)
;ddH2O 1. 09mL, TEMED 2 μL, 10%APS 10 μL,
we re each cast in g lass tubes(120mm ×3mm ) and
we re respec tively pre-electropho resed fo r 15 m in at
200V , 30 m in a t 300V and 60m in at 400 V. The
protein samples w ere disso lved in the samp le buffe r
which contained ddH2O 4. 0mL, 500 mmo l L -1
T ris-HC l(pH 6. 8) 1. 0 mL, g ly cerol 0. 8mL, 10%
SDS 1. 6 mL, β-mercaptothano l 0. 4mL, 10%(W /
V) b romophenol b lue 0. 2 mL. The first d imension
w as isoe lectrofocusing, 60 μL p ro tein samp les (4 ～
6mg mL - 1)were loca ted in each g lass tube;e lec-
trophoresis w as run fo r 14 ～ 16 h a t400V. A fter iso-
e lectrofocusing, the gelsw ere removed from the tubes
by shattering the g lass and p lacing in equilib ra tion
buffer wh ich con tained 6 mmo l L -1 T ris-HC l
(pH6. 8), β-mercapto thano l, 10% g lycero l, 2%
(W /V) SDS for 20m in. The second d imension w as
a 12. 5% SDS-PAGE tha twas pe rfo rmed acco rd ing to
the method of Laemm li
[ 18]
.
The tube gels w ere placed on top of second di-
2993期 范宝莉等:镉胁迫下三个萝卜栽培种蛋白质变化的双向电泳比较研究
mension gels, 1% agarose w as ove rlaye red and al-
lowed to po lyme rize. Cy lind rical gels we re run at a
constant voltage o f 80V for 5. 5 h in a B io-Rad uni.t
G els w ere stained w ith 0. 4% AgNO3 solution[ 19] .
Con tro ls and treated ge ls w ere analyzed by Ge lscan
GS800 densitometry and PDQuest softw are (B io-
Rad). To tal area and absorbance o f pro tein spots on
ge ls w ere determ ined, ad justed fo r backg round, and
assessed for any change in size.
Tab le 1　E ffec t o f different CdC l2 concen tra tions on
ge rm ina tion ra te in the three radish cu ltiva rs
C oncentration of
C dC l2(mm ol L - 1)
G erm ination rate (%)
XWY XYB WQ
C on trol 100 100 92
0. 05 100 100 83
0. 1 100 100 83
0. 5 92 100 67
1 83 100 58
5 67 67 58
　*　Date are based on a scoring of 350 grains of seed
2　Results
2. 1　E ffects o f C d2+ on germ ina tion rate and seed-
ling grow th in three radish cu ltivars.
G erm ination ra te:
Table 1 show s the effects o f various concentra-
tions of CdC l2 , the range from 0. 05mmol L - 1 to 5
mmo l L -1 , app lied a period o f 3d, on germ ination
ra te in three radish cultivars. Experiment data indica-
ted that the different e ffec ts o f Cd
2 +
on germ ination
ra te have been obse rved in the three rad ish cultivars,
ge rm ination rate o fXWY d id no t decrease at 0. 05 ～
0. 1mmo l L- 1 Cd2+ concentrations, but the germ i-
nation rate w as obviously decreased to 92% at 0. 5
mmo l L -1 Cd2+ concentration and ge rm ination rate
w as prog ressive ly decreased w ith increased Cd
2+
con-
centra tions, for instance, ge rm ination rate o f XWY
was 100% at 0. 05 ～ 0. 1mmol L - 1 Cd2+ concen-
trations, bu t it w as obv ious decreased to 83% and
67% a t 1 and 5mmo l L- 1 Cd2+ treatment, re spec-
tive ly. G erm ination rate decreased only when seeds
w ere trea ted by 5 mmo l L -1 fo r 3 d in XYB , this
fact show ed tha tXYB was appa rently mo re to lerant to
Cd
2+
than XWY. WQ is a radish cultivar w ith good
quality in China, but it w as the most sensitive to
Cd
2+
trea tmen t in three radish cultivars, germ ina tion
rate w as decreased to 83% a t on ly 0. 05 mmo l L -1
Cd
2+
trea ted and on ly 58% germ ination rate w as ob-
served at 1mmo l L -1 Cd2+ concentra tion.
Seed ling g row th:
Seed ling g row th w as inhibited in three radish
cu ltivars while treated by 0. 05mmo l L- 1 Cd2+ con-
centration, seed ling grow th w as progressively inh ibi-
ted w ith increased Cd
2+
concentration and seed ling
grow th w as complete ly inhibited in three radish culti-
vars a fte r 5mmo l L -1 of Cd2+ treatmen t(Fig. 1).
W e showed thatWQ was more sensitive to Cd
2+
treatment than XWY and XYB according to the expe r-
iment resu ltsw hich w ere ob tained by testing germ ina-
tion rate and seedling g row th in three radish cultivars.
F ig. 1　E ffect o f diffe rent CdC l2 concen tra tions on
seedling grow th in the three radish cu ltiva rs
2. 2　P ro tein changes
Pro te in compositions have been studied in the
three radish cultiva r seedlings wh ich we re treated at
various Cd
2 +
concentrations, ranged from 0. 05
mmol L- 1 to 5 mmo l L- 1 , by using two dimen-
sional ge l e lectrophoresis in o rde r to fu rthe r investiga-
tion the re lationsh ip between pro te in changes and
seedling g row th inh ib ition. Experimen t results indica-
ted that 2D pro tein patterns have been changed a l-
though the germ ina tion ra te has no t been affected in
the tw o radish cultiva rs which we re treated at 0. 05
mmol L- 1 Cd2+ concentration. But pro tein have
300 　　　　　　植　　物　　研　　究　　　　　　　　　　　　　　　　　　25卷
been changed in the three radish cultivar seed lings
w hich w ere treated at 0. 1 mmo l L -1 Cd2+ concen-
tra tion, a lso, inh ib ition o f seedling grow th w as ob-
served at this C d
2+
concentration.
F ig. 2　2D PAGE p ro te in patte rns ana ly sis of the seed lings treated w ith 0. 1 mm o l L - 1 CdC l2 in XWY
Fig. 3　2D PAGE pro te in patte rns analy sis of the seed lings trea ted w ith 0. 1 mm o l L - 1 CdC l
2
in XYB
Fig. 2a (contro l) and 2b ( trea tmen t) show ed 2
DE gels in XWY which w as trea ted by
0. 1mmo l L - 1 Cd2 + concentra tion, more than 300
pro tein spo ts we re reso lved on the ge ls, but five pro-
tein spo ts wh ich have mo lecu lar w eight and pI we re
22 KD /pI 6. 6, 18 KD /pI 7. 8, 16KD /pI 5. 2,
15 KD /pI 7. 6, 10 KD /pI 6. 4 respective ly w ere dis-
appeared (Fig. 2b) in comparison w ith con trol (Fig.
2a). 15 new prote in spo ts which have mo lecular
w e ight and pIw ere 128KD /pI 6. 9, 128KD /pI 7. 6,
98 KD /pI 6. 9 , 80 KD /pI 8. 6, 80 KD /pI 6. 6,
46KD /pI 8. 6, 43 KD / pI 8. 5, 38 KD /pI 8. 5, 33
KD /pI 8. 5, 32 KD /pI 8. 6, 13KD /pI 7. 2, 12 KD /
pI 6. 8, 10KD /pI 7. 8, 98KD /pI 6. 8, 6KD /pI 5. 7
we re induced respective ly in seedling s.
2 spots, 45 KD /pI 4. 8, 10 KD /pI 4. 6, were
disappeared. 1 protein spot decreased in con tent
(Fig. 3a) and 13 new protein spots, 110KD /pI 7. 2,
110 KD /pI 7. 4, 110 KD /pI 7. 6, 100 KD /pI 7. 8,
88KD /pI 6. 2, 26 KD /pI 7. 8, 23 KD /pI 6. 5, 22
KD /pI 7. 5, 18 KD /pI 7. 8, 15KD /pI 7. 8, 14 KD /
pI 7. 6, 10 KD /p I 7. 0, 8 KD /pI 5. 6, were in-
3013期 范宝莉等:镉胁迫下三个萝卜栽培种蛋白质变化的双向电泳比较研究
creased in XYB seed ling s which we re treated w ith
0. 1mmo l L - 1 Cd2+ fo r 3 d (Fig. 3).
F ig. 4 showed the change in pro te in spots in the
present o f 0. 1mmo l L -1 Cd2+ inWQ seedling s. 12
new pro te in spo ts wh ich havemo lecularw eigh t and pI
we re 24 KD /pI 8. 6, 19KD /pI 7. 8, 18KD /pI 5. 4,
17KD /pI 7. 8, 16 KD /pI 8. 1, 16 KD /pI 8. 0, 16
KD /pI 7. 7, 15 KD /pI 7. 0, 15KD /pI 7. 8, 15 KD /
pI 6. 7 , 10 KD /pI 6. 8, 8 KD /pI 6. 2 respec tive ly,
we re induced but w e have no t obse rved the p ro tein
F ig. 4　2D PAGE pro te in patte rns analy sis o f the seed lings trea ted w ith 0. 1 mm o l L - 1 CdC l2 inWQ
spot disappea red inWQ seedlings.
3　D iscussion
M any reports have been made in Cd
2 +
caused
pro tein changes in plan t seed lings, for instance, Hsu
et a l studied the changes in prote in and am ino acid
conten ts in tw o cultivars o f rice seed ling s w ith d iffe r-
ent apparent to lerance to cadm ium
[ 13] , expe riment re-
su lts indicated that 0. 5 mmol L -1 Cd2 + caused the
pro tein conten t decreased and protease activity, to tal
am ino acids increased in Cd
2+-sensitive rice seed-
ling s. Romer-puertas et al found cadm ium caused the
ox idative and mod ification o f pro te ins
[ 14] , by using
diffe rent antibodies, some of the ox id ized pro te ins
w ere identified as Rubisco, g lutathione, reducase,
manganese, supe roxide dismutase and catalase. Ueki
et a l identified a tobacco g ly cine-rich p ro tein, cdiG-
RP, spec ifically induced by low concen trations o f
cadm ium , cd iGRP may have a function fo r contro l
plant v iral sy stem ic movemen t
[ 15]
. F lem ing et al
found that 0. 5mmol L - 1 Cd2+ caused several stress
pro tein (SP) synthesis in Sago pondw eed[ 16] , the
changes in SP syn thesis may precede de tec table a lte r-
ations in grow th of aquatic p lants and, therefo re, may
be a potentially usefu l early b iomarker o f con tam inant
Cd
2+
stress. Kavita et al have successfu l iso lated a 18
KD cadm ium induc ib le pro tein comp lex from rice
seedlings expo sed for 20 days unde r 0. 5 mmo l L -1
Cd
2+[ 5]
. They have further characte rized th is p ro tein
comp lex and identified that this p ro tein complex helps
in sequestra tion of excess Cd
2 +
ions in rice plants.
In the p resent wo rk, we found tha t prote in spots
have obv ious been changed in the three radish cultiva r
seedlings afte r 0. 1mmol L - 1 Cd2+ trea tmen.t 12
new pro tein spo ts, which havemolecula rw eigh tbe low
24KD(8 ～ 24 KD) and most of them located in the
basic region (pI>7), were induced in WQ cultiva r
seedlings (F ig. 4). 15 new protein spo ts we re in-
duced in XWY cultivar seedlings, mo lecularw eigh t of
these increased pro tein spo ts located among 6 KD ～
128 KD, 5 out of 15 prote in spots located in be low
14KD, and 10 ou t o f 15 protein spo ts in wh ich have
mo lecular w eigh t from 31 KD to 97 KD , 5 p ro tein
spo ts we re disappeared in XWY cu ltivar seed ling s af-
ter C d
2+
trea tmen t(F ig. 2a a rrow ind ica ted). W hen
seedlings w ere treated w ith 0. 1 mmo l L - 1 CdC l2 ,
13 new p ro tein spotsw e re induced, tw o prote in spots
we re disappeared in comparison w ith contro l in XYB
(Fig. 3). Ou r expe riment resu lts show ed that a close
positive re lationship w as obse rved between changed
302 　　　　　　植　　物　　研　　究　　　　　　　　　　　　　　　　　　25卷
pro tein spo ts and inhibited seedling g row th in the
th ree radish cu ltivars we re treated w ith CdC l2.
In higher p lants, many Cd2 +-induced stress pro-
teins ranging from 10 ～ 70KDmay directly bind Cd i-
ons
[ 20]
and have been corre lated w ith plan t tolerance
to Cd
2+[ 21]
. F lem ing et al demonstrated tha t the
changes in Cd
2 +-stress prote in synthesis may precede
detectable alterations in g row th of plan ts
[ 16] , Radish
w as an important vege tables in Tian jin, WQ is a fa-
vo rable cu ltivar because of it’ s good quality. Recent
years, how eve r, y ield w as reducted and quality w as
declined due to Cd
2+
contam ina ted soils in T ianjin.
This study is one o f the first describing changes in
pro tein patterns caused by Cd
2 +
treated from radish
cu ltivars, it m ay be time-saving, econom ic analy sis
too l for early detecting Cd
2+
damage in yie ld and
qua lity o f rad ish vege tables.
Reference
1. Ye H aibo, Yang X iaoe, H e Bing, et a l. Response o f Sedum
A lfredii Hance towa rds Cd /Zn comp lex-po llution and accu-
mu la tion o f the heavy m eta ls. Journa l o f Ag ro-Env ironm ent
Science, 2003, 22(5):513 ～ 518
2. L i Kunguan, L iu Jianguo, Lu X iao long, et al. Up take and
distribu tion o f cadm ium in diffe rent rice cu ltiva rs. Journa l o f
Ag ro-Environment Science, 2003, 22(5):529 ～ 532
3. Das P, Sam an ta rays, Rou tG R. Studies on cadm ium toxic ity
in p lants:a review. Env iron Po llut, 1997, 98:29 ～ 36
4. H art J J, W elch RM , Norve llW A, et al. Characteriza tion o f
cadm ium bind ing, up take and translocation in in tac t seed-
ling s of bread and durum whea t cu ltivars. P lan t physio l,
1998, 116:1413 ～ 1420
5. Kav ita S, Dubey R S. A 18kDa cadm ium inducib le pro tein
comp lex:its iso lation and charac te riza tion from rice(Oryza
Sativa L. ) seed lings. Journa l o f P lan t Phy sio logy, 1998,
152:448 ～ 454
6. R ivera-Becerril F, C alantzis C , Turnau K, et al. Cadm ium
accumu la tion and buffe ring o f cadm ium-induced stress by
a rbuscular mycorrh iza in th reeP isum sativum L. Geno types.
Jou rna l of Experimenta l Bo tany, 2002, 53:1177 ～ 1185
7. Zhou H ongw ei, Shi Guox in, Xu Q insong. E ffect o f Cd2+ po l-
lu ted wa te r on the ac tivities of an tioxidant enzymes and u l-
trastructure in roo ts o f A lternan thera philoxe roides. P lant
Phy sio logy C omm un ica tions, 2003, 39(3):211 ～ 213
8. Ke ltjensW G, Beusichem M L. Phy to chela tins a s biom a rke rs
for heavy me ta l stress in m a ize(Z ea mays L. ) and w hea t
(Triticum aestivum L. ):Comb ined effe cts of copper and
cadm ium. P lant and So il, 1998, 203(1):119 ～ 126
9. Lag riffoul A, M ocquo t B, Mench M, et a l. Cadm ium toxic ity
effec ts on grow th, m ineral and chlorophy ll contents, and ac-
tiv itie s of stress re la ted enzym es in young m aize p lants(Zea
mays L. ). P lan t and Soil, 1998, 200(2):241 ～ 250
10. Khudsa r T, M ahm ooduzza far, Iqba M. Cadm ium-Induced
changes in leaf epiderm is, pho to syn thetic ra te and pigm ent
concen tra tions in Cajanus cajan. Bio log ia P lan ta rum , 2001,
44(1):59 ～ 64
11. M uhiling, K arl H , Lauch li, et a l. E ffec t o f bioaccum ula tion
o f cadm ium in biom ass produc tivity, e ssentia l trace e le-
m en ts, chlorophy ll biosynthe sis and m acrom o lecules o f
w hea t seedling s. B iologica l T race E lement Research, 2003,
92(3):257 ～ 274
12. Kevresom S, Pe trov ic N, Popovic M , et a l. N itrogen and
prote in m etabo lism in young pea plan ts a s a ffec ted by differ-
ent concentrations o f nicke l, cadm ium , lead and m o lybde-
num. Journa l o f P lant Nutrition, 2001, 24(10):1633 ～ 1645
13. H su Y T, Kao C H. Changes in prote in and am ino ac id
contents in two cultivars of rice seed lings w ith diffe rent ap-
paren t to lerance to cadm ium. P lant G row th Regu la tion,
2003, 40:147 ～ 155
14. Rom ero-pue rtasM C , Pa lm a J A, G om egM , et al. Sanda lio
L M. C adm ium causes the ox idative modification o f p ro te in
in pea plants. P lant, C ell＆Env ironm en t, 2002, 25(5):677
～ 687
15. Uek i S, Citov sky V. The system ic movement of a tobam o-
v irus is inhib ited by a cadm ium-induced g lycine-rich pro-
tein. Na ture cell b io logy, 2002, 4(7):78 ～ 87
16. F lem ingW J, G rossfe ld R M , S iesko M M. Stress p ro te in
synthesis and pe rox idase activ ity in a subm e rsed aqua tic
m acrophy te exposed to cadm ium. Env ironm ental Toxico logy
＆ Chem istry, 1997, 16(8):1755
17. O’ Farre l P H. H igh re so lutiono f two dim ensiona l e lectro-
pho resis o f pro te ins. J B io chem , 1975, 250:4007～ 4021
18. Laemm li U K. C leavage o f structure pro teins during the a s-
sem bly of the head of bac te riophase T4. Nature, 1972, 227:
680 ～ 685
19. W ary W , Boulikas T, W ary V P, et al. Silve r sta ining o f
prote ins in polyacry lam ide gels. Ana l B iochem , 1981, 118:
197 ～ 203
20. Sanita di, Toppi L, Gabbre lli R. Response to cadm ium in
h igher p lants. Environm en ta l and Experim enta l Bo tany,
1999, 41:105 ～ 130
21. H all J L. Ce llu larmechan ism fo r heavym eta l detoxifica tion
and to le rance. Journal of Experimenta l Botany, 2002, 53:1
～ 11
3033期 范宝莉等:镉胁迫下三个萝卜栽培种蛋白质变化的双向电泳比较研究