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Effects of elicitors on saffron pigment production in cell suspension cultures of Crocus sativus L.

诱导子对藏红花悬浮培养细胞生产藏红花色素的影响



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ChineseJournalofBioprocessEngineering
Vol.11No.3
May2013
doi:10.3969/j.issn.1672-3678.2013.03.004
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:2012-03-27
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îï ð,
(chitosan)、
 ñ,
(chitosanoligosaccharides,COS)、
òó6ôÉ
(methyljasmonate,MJ)、
õö6
(salicylicacid,SA)
b
Cu2+
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eOPîï
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 ñ,
(1~500mg/L)
b%&ˆd ð,
(≤10mg/L)、MJ(≤10μmol/L)、SA(≤10μmol/L)bCu2+
(≤1μmol/L)Cij1þÚ(EF;%%ˆd ð,(≥100mg/L)、MJ(≥100μmol/L)、SA(≥100μmol/L)b
Cu2+(≥10μmol/L)()Ñij1þ。5` øù„Cúû3ÿNÞªDøù*!:;,ì¥äøù„Ð+ˆd
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eij#$/

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ars
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 ð,b ñ,

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0+,1ˆd
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úû3ÿN
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:Q9431    
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HMNG
:1672-3678(2013)03-0018-06
Efectsofelicitorsonsafronpigmentproductionincelsuspension
culturesofCrocussativusL
SUNZhen,YUANLihong,WUPinmei
(ColegeofBiotechnologyandPharmaceuticalEngineering,NanjingUniversityofTechnology,Nanjing210009,China)
Abstract:Theefectsofchitosan,chitosanoligosaccharides(COS),methyljasmonate(MJ),salicylicacid
(SA),andcopperions(Cu2+)oncelgrowthandsafronpigmentproductionincelsuspensionculturesof
CsativuswereinvestigatedTheresultsshowedthatCOS(1-500mg/L),chitosan(≤10mg/L),MJ(≤10
μmol/L),SA(≤10μmol/L),andCu2+(≤1μmol/L)hadnosignificantefectsonsafroncelgrowth,
whereaschitosan(≥100mg/L),MJ(≥100μmol/L),SA(≥100μmol/L),andCu2+(≥10μmol/L)at
highconcentrationsobviouslyinhibitedthecelgrowthTheefectsofelicitorsweresignificantonsafron
pigmentbiosynthesis,andhadrelationswiththeconcentrationsofelicitorsandthetimingofelicitationMJ
wasthebestelicitor,thecontentofsafronpigmentreached2857mg/gDWbyadding100μmol/LMJon
day0,anditwashigherby1779% thanthecontrolCu2+wasthesecond,thecontentofsafronpigment
reached1982mg/gDW byadding500μmol/LCu2+onday4,anditwashigherby1082% thanthe
controlChitosanandCOSwerethethird,thecontentsofsafronpigmentreached1833mg/gDW and
1739mg/gDWbyadding100mg/Lchitosanandoligochitosanonday14,anditwashigherby691%and
690% thanthecontrol,respectivelyThelastoneisSA,thecontentofsafronpigmentreached1465
mg/gDWbyadding10μmol/LSAonday14anditwashigherby454% thanthecontrol
Keywords:celsuspensionculture;CrocussativusL;elicitor;safronpigment
  
™âT
(CrocussativusL)
ãä«âT

åâT

bcdäÏh‡æ

3L‡LŽ78I3

™âT
üçp¤‹dpתñ™âT!
(crocin)、
™â
Tè!
(picrocrocin)、
™âTW
(crocetin)
{’yé
ê
[13]
Fijbç|}

Æë65‚;A‚yéê‡

‘Y™âTQüçã‡

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Q™âTñòŦÐeçp”•
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。Chen
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[5]
Q™âTó

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Ö 
229
Ö¨©Q
¢“üæ&[‘
,^
¶mˆh¨©Q
Corm1,crocin1
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1677mg/g。
ö³÷{
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h™âTôõª«Ö-*Á–òó&•“™
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%‚çpX§Ð¨©òólvçp¨
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[7]
’ùúûÿüò
󨩢Ž(æ–‡\Å
10mg/L
àø÷

ýé
äþז=
3646mg/L,
g§3p3
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’fÿÿüò󨩢Ž(æ–‡\Å
100
mg/L
àø÷

1!Kˆ•×–=
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g§3p3
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à"÷5Åcd¦·#IX

á#¬Ð$%.

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çp×
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()WàáOt¹W5ÅÐe”±²
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ÖƧ]da&LMו“ijçp5‚
[10]。
Kang
{
[11]
’*+ò󨩢ñ­Ž(‡\Å
10μmol/L()WàáO 10mmol/Lt¹W,*
+”áO*+è”áh–J;。Cu2+
5
ÅÐ&[fghH–§!

_bcdA‚h‡
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¦·Ž(
Cu2+
ƒŽj}klШ
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­,-{
[12]
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.³ÿü¨©òó!
18
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400
μmol/LCuSO4,.³/ז= 719mg/L,g§3
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æ



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yz?²
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Cu2+
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ôL

1 
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11 
]^OP
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™âTÒ0üç

èY½12;

™âTñ
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(3000)
è’A6]78ïð

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è’Òø
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t¹W
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12 
ÌÍ7ÐÑYZ
121 
™âTôõª«6Lòó
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MS[14]
òóôÅôøòóô

c(
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:
÷
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9:ô;<
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=
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Ä`
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ñ>Y
100mL
12â

¥â
30mL,121℃
Å[
20min。
M
d¡ªY
(20±1)℃
?òó

¥
28d
’“6Lca

122 
™âT¨©ÿüòó
Q
1/2B5
[15]
òóô
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NaH2PO4‚–@µ)Åôøòóô,c(20g/L:÷,
05mg/L6 BA,20mg/L
AB

éW
(IAA),
pH58~60,
ñ>Y
250mL
12â

¥â
50mL,
121℃
Å[
20min。
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ôõª«

M
d–Å
12%),
Y
(20±1)℃,120r/min
l?¶·
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»¨©
(m2),Y60℃Ú°ÚÒ¯
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 ¤
3000g
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¤n!r¿

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ñ‚
60%
àärÏ

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 ¤™âTÒ0üç
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10mL
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(R2=09959),
’
0020~0104mg/mL
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500
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26d
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O6

(B)。
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1(A)
O6
1(B)
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àø
÷§™âT¨©&[œ
3文\àø
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1 
ÓÔ1ÅÓÕ1cÌÍ7ÎÏYZÐÑ5Ö
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(C,D)
;de
Fig.1 EfectsofchitosanandCOSongrowth(A,B)andpigmentbiosynthsis(C,D)ofCsativuscelsuspensioncultures
  
àø÷Oà"÷§™âTn!•“hSTK
d5‚‡\OŽ(Pwj°

6
1(C)
O6
1(D))。
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1(C)
O6
1(D)
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1~100mg/L
()”

™âTn!–Tàø÷‡\p39u(

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P

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’¨©òó!
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100mg/L
à
ø÷

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Å
1833mg(
Q
1g
Ò
¨©æ–;
),
g§3p3
691%。
à"÷æ–
‡\Å
1、10mg/L
P

Kàø÷hçp}@cq

P
à"÷æ–‡\Å
100mg/L
P

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100mg/L
à"÷

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1739mg,
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690%。
3文(500
mg/L)
àø÷Oà"÷§™âTn!•“j;Ë5‚

™âTn!–JsY§3

9Æàø÷
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àø÷Oà"÷5Åc
d¦™J#æ

ƒŽçIШ©h$%.

çpKÐa&LM÷°ôžh5=

kla&L
M×h•“
[8,18-21]。
àø÷áQp3™âT¨
©¢™âTn!h–

d‚ƒŽ(PwŨ©ò
ó!
14¨ ,
‚ƒ5‚æ–‡\Å
100mg/L。
à"
÷Kd‚ƒ5‚F%ôø÷2

ÁÁz§¨©&[
hSTj+-­

àø÷’kla&LMh2P

§¨©&[ijchcË5‚

ec3{’d:
Ш©òó¢_’
[22],
9à"÷§¨©&[d
œ;
MJ
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CK—
Ž(œ[×Øt
;0—
Ž(œ[œtéä
D
2 MJ
Å
SA
cÌÍ7ÎÏYZÐÑ5Ö
(A,B)
ÅÌÍ7Ò®¤m
(C,D)
;de
Fig.2 EfectsofMJandSAongrowth(A,B)andpigmentbiosynthsis(C,D)ofCsativuscelsuspensioncultures
22 
×Ø:ÙºÅÚÛ:cÌÍ7ÐÑ5֘Ì
Í7Ò®¤m;de
  
ñ­’¨©òó!
0、4、14、20
¨Ž(
MJ
O
SA,
>
MJ
‡\Å
0、01、1、10
O
100μmol/L,ñ
­QŽ(œ[×ØtOœtéäŧ3

>
SA

‡\Å
0、1、10、100
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500μmol/L,òó26dӔ
¨©

|}d§¨©&[O™âTn!•“ST

3@46
2。
‘6
2(A)
O6
2(B)
á7
:MJ
O
SA
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10μmol/L§™âT¨©&[œST
;MJ
O
SA
5‚‡\3Y
100μmol/L§™âT
¨©&[j
Æ
SA
cË5‚vY
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SA
5‚‡\Ü3

Ž(PwÜ,
cË5‚Üv

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¢á7

’
01~100μmol/L()”,™
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!•“K
MJ
Ž(Pwj°

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0¨
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(‡100μmol/LMJP,™âTn!–‚3,
=
2857mg(
Q
1g
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),
g§3p3í
1779%。
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MJ
r¿

|
Bžœté䧙âT¨©hST

u=í(
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3@I3œté䧙âT
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MJ
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1~10μmol/L()”,™âTn!–TU SA‡
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P
SA
‡\Å
100μmol/L
P

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2P_I3™âTn!
12 
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10μmol/LSAP,™âTn!–=ž
1465mg,
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454%。SA
‡\=
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mol/L
§™âTn!•“ij;
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Ž(PwÜ,
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MJ
O
SA
5ÅQSñX

‰K¨©wO¨©”
QS´p

ƒŽçI¨©h$%.

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 9klabLM×h•“
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!hçpX

K¢’c+\½c˙âT¨©&
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–

eK¹û{
[23-24]
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O
SA
‚ƒ5‚‡\OŽ(Pw^2
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‚ƒŽ(P
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¨

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100
μmol/L,9SA‚ƒŽ(PwŨ©òó!14¨,
‚ƒ5‚‡\Å
10μmol/L,Á MJçp}@è
Y
SA。
23 Cu2+
cÌÍ7ÐÑ5֘ÌÍ7Ò®¤m;
de
  
’¨©òó!
0、4、14、20
¨ñ­Ž(
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>
Cu2+
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O
1000μmol/L,
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Ӕ¨©

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!•“hST

3@46
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3(A)
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:1
μmol/LCu2+§™âT¨©&[œ;<5‚;Cu2+
‡\3Y
10μmol/L§™âT¨©&[ijcË5‚

Æ
Cu2+
‡\Ü3

Ž(PwÜ,

5‚Ü;<
。Cu2+
§™âTn!•“hST53
Å

’
1~500μmol/L()”,™âTn!–TU
Cu2+
‡\p39êëu(

P
Cu2+
‡\Å
500
μmol/LP,™âTn!–=ž‚3,ÖÆ Cu2+k
ln!•“}@KdŽ(Pwj°

’¨©òó
!
4¨
Ž(‡\Å
500μmol/LCu2+,™âTn!
–=ž
1982mg(
Q
1g
Ò¨©æ–;
),
g§3
p3
1082%。
P
Cu2+
‡\=
1000μmol/LP,
™âTn!–rs

‘Ÿá4

B3
Cu2+
‡\K
¢c˙âT¨©&[

Áƒ¢™âTn!h–
。Cu2+
5ÅÐ&[+fg
hYó§!

§Ð¨©&[Oa&LMij±²
h5‚
。Cu2+
klWeòóhШ©a&LM
ו“’LM
(Puerarialobata)
ÿüò󨩕
“»NN
[13]、
O\1‰
(Withaniasomnifera)
ÿüò
󨩕“PQ!
A[25]
O.³
(Campototheca
acuminata)
ÿüò󨩕“.³/
[12]
{78¢
_j)*

Rð’^2Ш©òóeQ¢
Cu2+
‚ƒ5‚‡\^2

ÁK
Cu2+
5ÅÐѨ©&[+
g‡\÷g

B3‡\h
Cu2+
ij%Ya&×h
•“

e›;3‡\h
Cu2+
?²5ÅçpX9Î

[26]。
D
3 Cu2+
cÌÍ7ÎÏYZÐÑ5Ö
(A)
ÅÌÍ7Ò®¤m
(B)
;de
Fig.3 EfectsofCu2+ongrowth(A)and
pigmentbiosynthsis(B)ofCsativus
celsuspensioncultures
3 
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b
àø÷

à"÷

()Wàá

t¹WO
Cu2+
’
†ºh5‚‡、
•†hŽ(PwáQj}p3™
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[1] EscribanoJ,AlonsoGL,PradosMC,etal.Crocin,safranaland
picrocrocinfromsafron(CrocussativusL.)inhibitthegrowthof
humancancercelsinvitro[J].CancerLet,1996,100:2330.
[2] AbdulaevFI,EspinosaAguireJJ.Biomedicalpropertiesof
safron and its potential use in cancer therapy and
chemopreventiontrials[J].CancerDetectPrev,2004,28:
426432.
[3] AnimeshD,SmitaM,GopalD,etal.Crocetininhibitspancreatic
cancercelproliferationandtumorprogressioninaxenograft
mousemodel[J].MolCancerTherap,2009,8(2):314323.
[4] 
àáâ

ShT

NU

™âTôõª«çpOmˆcË
[J].
A)&0ŽŽ)

’¢ÃŽ„
,2009,31(6):2126.
[5] ChenSA,WangXD,ZhaoB,etal.ScreeningofCrocussativus
L.caluslinesforcrocinproduction[J].ChinBulBotany,2004,
21(4):455460.
[6] 
ö³÷

©V

‚WX

¹¿Y–h™âT!&•“
[J].
«Z0ŽŽ)

’¢ÃŽ„
,2003,43(12):16091612.
[7] ChengX Y,Zhou H Y,CuiX,etal.Improvementof
phenylethanoidglycosidesbiosynthesisinCistanchedeserticola
celsuspensionculturesbychitosanelicitor[J].JBiotechnol,
2006,121:253260.
[8] FanGZ,LiXC,WangXD,etal.Chitosanactivatesdefense
responsesandtriterpenoidproductionincelsuspensioncultures
ofBetulaplatyphylaSuk[J].AfrJBiotechnol,2010,9(19):
28162820.
[9] KimSK,RajapakseN.Enzymaticproductionandbiological
activitiesofchitosanoligosaccharides(COS):areview[J].
CarbohydrPoly,2005,62:357368.
[10] ShimizuY,MaedaK,KatoM,etal.Methyljasmonateinduces
anthocyaninaccumulationinGynurabicolorculturedroots[J].In
VitroCelular& DevelopmentalBiology:Plant,2010,46:
460465.
[11] KangSM,MinJY,KimYD,etal.Efectsofmethyljasmonate
andsalicylicacidontheproductionofbilobalideandginkgolides
incelculturesofGinkgobiloba[J].InVitroCelular&
DevelopmentalBiology:Plant,2006,42:4449.
[12] 
­,-

[Ž.Cu2+
çpJ#§.³ÿüòó¨©.³/
&•“hST
[J].
ÈÉâ&Î
,2010,38(23):
1245712459.
[13] KorsangrusangS,SoonthornchareonnonN,ChintapakornY,etal.
Efectsofabioticandbioticelicitorsongrowthandisoflavonoid
accumulationinPuerariacandoleivar.candoleiandP.candolei
var.mirificacelsuspensioncultures[J].PlantCelTissueOrgan
Culture,2010,103(3):333342.
[14] MurashigeT,SkoogF.Arevisedmediumforrapidgrowthand
bioassayswithtobaccotissuecultures[J].PhysiolPlantarum,
1962,15:473497.
[15] GamborgOL,MilerRA,OjimaK.Nutrientrequirementsof
suspensionculturesofsoybeanrootcels[J].ExpCelRes,
1968,50:151158.
[16] AnaM S,ManuelC,AmayaZ,etal.Rapiddeterminationof
crocetinestersandpicrocrocinfromsafronspice(Crocussativus
L.)usingUVvisiblespectrophotometryforqualitycontrol[J].
AgricFoodChem,2008,56:31673175.
[17] 
­,-

[Ž,
]ß^

{

àø÷çpXkl.³òó¨
©.³/_`h78
[J].
ÈÉâ&Î
,2010,38(29):
1615116153.
[18] KomaraiahP,RamakishnaSV,ReddannaP,etal.Enhanced
productionofplumbagininimmobilizedcelsofPlumbagorosea
byelicitationandinsituadsorption[J].JBiotechnol,2003,101:
181187.
[19] PutalunW,LuealonW,DeEknamkulW,etal.Improvementof
artemisininproductionbychitosaninhairyrootculturesof
ArtemisiaannuaL[J].BiotechnolLet,2007,29:11431146.
[20] ZhangCH,FevereiroPS,HeGY,etal.Enhancedpaclitaxel
productivityand release capacity ofTaxuschinensiscel
suspensionculturesadaptedtochitosan[J].PlantSci,2007,172
(1):158163.
[21] ZhangHY,ZhaoXM,YangJL,etal.Nitricoxideproduction
anditsfunctionallinkwithOIPKintobaccodefenseresponse
elicitedbychitooligosaccharide[J].PlantCelReport,2011,30:
11531162.
[22] 
Ⱦ

n_;
,`
âa

àø÷§Qb¨©&[OÓKa&L
MhST
[J].
‹s0ŽŽ)

†Ž„
,2007,45(10):
10251029.
[23] 
¹û

°c

Ò4

{
.^
2‡\()Wàá§ÿüòóhd
@9e¨©•“9e>NNhST
[J].
fAÐ78

2008,30(5):586592.
[24] FürdenBV,HumburgA,FussE.Influenceofmethyljasmonate
onpodophylotoxinand6methoxypodophylotoxinaccumulationin
Linumalbum celsuspensioncultures[J].PhysiolBiochem,
2005,24:312317.
[25] BaldiA,SinghD,DixitV K.Dualelicitationforimproved
productionofwithaferinAbycelsuspensionculturesofWithania
somnifera[J].ApplBiochemBiotechnol,2008,151:556564.
[26] WeathersPJ,TowlerMJ,XuJF.Benchtobatch:advancesin
plantcelcultureforproducingusefulproducts[J].Appl
MicrobiolBiotechnol,2010,85:13391351.
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