免费文献传递   相关文献

Effects of culture conditions on biomass and active components ofadventitious roots culture in Panax ginseng

离体培养条件对人参不定根生长及其活性成分合成的影响



全 文 :·
WXYZN
·
”•–—˜™š›œ#ˆžŸ 
¡¢£¤¥£ƒ¦§
.7
1,2,
189
1,2,
*:
1,
;<

(1
]°¤k ¬$«k%¾¿k%


300072;
2
]°«¾¤k !¬’$#$­®¯


300457)

Žz
] 


™š›œ#ƒCD–—˜Cc†‡ƒðG

¡¢

­;<–()´¥œ+nԍ`‹Y€¤
llU‹

œËë}¬!

E–eUëŸXå5eU™š›œ#ƒˆž

š›Nï럚›î–¥£ƒ¦§

£¤

v
1L
–¨}¬ƒœ#F­>
20g
.š›œ#ƒ2­øyOBƒ„A6{

íîE–eUƒ_œ

š›œ#2­øyOB
GË_œßƒHI

š›î–x!øžHIœ²ˆ

œ-E–eU.mÕNïx!Jˆ§Rp

š›jNïx!íE
–eUƒ_œß

Ãĕu–¨$ƒî–YNï’!èŒ
40g·L-1
E–Ð!eU«ƒ„A6{

–¨$ƒXå5e
U™œ#ƒˆžëŸî–YN£Óuå‘Jƒ6¦§
,3/4MS
AJ­©œ#ƒˆžëŸN£

œ#$
ƒî–x!Zíî5eUƒ_œß

£V

}¬!

E–eU

Xå5eU‘+ˆ§¦§š›œ#ƒˆžëŸ ¡¢
£¤ƒ¥£Yuå


‘
] 
š›

œ#

}¬!

E–eU

Xå5eU

š›Nï

š›î–

’w)(
] 20090411

“”„Q
] 
%<

PJ

&(³KZLÛÜ
(2006BAI06A16
02)

•d}–
] 
œFM
,Tel/Fax:(022)87401895,Email:pharmgao
@tjueducn
  
š›
PanaxginsengCAMey
ÞJé&
Arali
accae
š›°înˆh×þŽ‰

N>,$Ò,

žwë…

{©âUL)

وš›¡COP

ÜF
©,ƒî>Qš›

š›ƒQˆ’Rwž

S
N¢u6

(+™ˆÃU’T£¶­UV

š›;
<–()ª œæˆžW®Kæ

œXòžÞ


–RwYäð±æ„ëZ…Zs[ƒ­î

%=
²©š›;<–ƒ`øPî
[13],
\%™š›
œ#|Jƒ>†‡ƒ/0
[46],
@ÞJ²© $š
›î–x!ƒ/0M]J`ø

ŸFŒ£Sµõš
›œ#n֕†ƒ¨,^

/0ë}¬!

E
–eUYXå5eU™š›œ#ˆžŸ NïY
£ƒ¦§

>­š›œ#”•–•†C
cš›¡¢£¤ƒˆ’^ëøYz›¨,

1 
0)
†=0)>Œéé
IBA5mg·L-1
Y
KT01
mg·L-1
ƒ
3/4MS
n•–¨$–7îƒš
›

‚ü6œFMÒÄ4

œ#

š›Nï
Re
™]â

*A
110754200421);
š›Nï
Rb1™]â

*A
110704200420);
š›Nï
Rg1™]â(*A
110703200425),
è_$%,∉æâr1

HPG280B
l]–[
(`
ga:\W×()
ÀbJKLM
);HNY82
ñCE

‚ü9bc9:
9%JKLM
);METTLERTOLEDODelta320pH
Z
(^
Pd½b

e­î9:LM
);SP2000UV
ñ
€
¯
J¤llUZ
(^
Pl`9:JKLM
);
Agilent1100
œ+nԍ`9

Û%GfgLM
)。
2 
Š‹
21 
}¬!™š›œ#ˆžƒ¦§
 
v
1cm
h
̓š›œ#ð}¬„
250mL
TÑD$7
–
30d,
TÑD=iJ
80mL
éé
50mg·L-1IBA
Y
01mg·L-1KT
ƒ
3/4MS
–¨

}¬!¤p>
10,15,20,25,30,35g·L-1。
–¨
pH
jkF>
60,
CEbn>
120r·min-1,
v;­Ù5ø

D

22 
E–Ð!eU™š›œ#ˆžŸ J+£
¤x!ƒ¦§
 
vl
16g(


š›œ#}¬
„E–Ð!eU¤p>
30,40,50g·L-1
ƒ
80mL
éé
50mg·L-1IBA
Y
01mg·L-1KT
ƒ
3/4MS
–¨$

–¨
pH
jkF>
60,
CEbn>
120
·31·
u
35
Cu

w
2010
n

o
                             
Vol.35,Issue 1
January,2010
r·min-1,
v;­Ù5ø

D

23 
5eU™š›œ#ˆžŸ J+£¤x!
ƒ¦§
 
vl
16g(


š›œ#}¬„Xå
5eU¤p>
1/2MS,3/4MS,MS,3/2MS
ƒ
80mL
éé
50mg·L-1IBA
Y
01mg·L-1KT
ƒ
MS

–¨$

–¨
pH
jkF>
60,
CEbn>
120
r·min-1,
v;­Ù5ø

D

24 
œ#ˆ‰!ƒ/
 
v–T„ƒš›œ
#á

mn»á%¢ƒ–¨

o©‚>Cc
È­È>F­

vÈâF­ƒœ#o©p[$

Œ
60℃
«p
24h,
. 2q¶r­

È 2­

½Z
q 2­øyOB

2­øyOB
=(
¼šœ#


}¬œ#2­
)/
}¬œ#2­

25 
š›î–x!Ÿ’!¤³
 
¤pÈá
02g
2qÅsƒš›œ#–‰Åt


100mL
Š
uTÑD$

é
20mL80%
ƒvf
,90℃
ûw¸Ï

h,
­Ù



x•ym

mz
80%
ĥvf{|

¾2ëÚ

mzÜ-mno©}D$

é
20mL
~

,100℃
ûw€à
1h,
x•âm

mz­Ùàá



¥½mn

”O¤”
,^
Gno©
100mL
!D
$

é~ûß¶±U



T†=âën

#
®Ô²Fú
[7],
Œ
480nm
ž—˜«/»lU

#®¸{Šã
A=00054C+00197,r=09997
Zqî–x!

š›î–’!

î–x!
×
¼šœ
#2­

–n•u

26 
š›Nïx!Ÿ’!/
 
¹·Èá
02g
2qÅsƒš›œ#–‰Åt


100mL
Š
uTÑD$

éÆ
20mL
Jf
,60℃
ûw
1h,
Y
ym

¼¯mn

mzÜ-mnxéÆä!ƒJf
­Ùûwàá

ym



¥½mn

mn~b2q
ë©
20mL
û‚Y$Ôf

¤nƒ„…á



¥½…áƒ$Ôfën

e¨¶2

†zë݌Jf
$

ß„
1mL,



022μmòõmÒâm,
T†=âën

`—˜

`ˆ>
kromasilC18
(46mm ×250mm,5μm)。ÏÂÔ Av‰,B
û

ŠU{‹ãÑ
0~9min,A26%,
Ïn
05mL
·min-1;9~10min,A26% ~18%,
Ïn
05mL·
min-1;10~15min,A18%,
Ïn
05mL·min-1;
15~16min,A18% ~35%,
Ïn
05~10mL·
min-1;16~30min,A35% ~40%,
Ïn
10mL·
min-1。
r/ž
203nm,
ˆ·
45℃,
CŽ•u
20
μL。¤pZqš›Nï Re,Rg1,Rb1ƒx!。;
š›jNï!


Re
!


Rg1!+NïRb1
!

š›Nï’!

jNï!
×
¼šœ#2­


–n•u

27 
‡Z¤³
 
Ÿ/01JB®è.
SPSS113
Œ˜CcŠj¤³

B®ë 珋x±s%&。
3 
´—
31 
}¬!™š›œ#ˆžƒ¦§
 
Ÿz{î
10,15,20,25,30,35g·L-1
}¬š›œ#

œË
 ™š›œ#ˆžƒ¦§
,´
—Jf
1。
f
1 
}¬!™š›œ#ˆžƒ¦§
}¬!
10g·L-1
.

œ­©œ#ƒˆž

}
¬!ý
10~20g·L-1
ƒâã$

íî}¬!ƒø
6

œ#ƒˆž|é

Å}¬!ƒ„
20g·L-1
.

–

€Rwœ#ƒ2­øyOBƒ„A
6{

Òíî}¬!øé

š›œ#ˆžæ„u
æ

–

€Rw 2­øyOBŽnß

32 
E–Ð!eU™š›œ#ˆžëŸNïYî
–x!ƒ¦§
 
Ÿz{œËë
30,40,50g·L-1
ô

€E–Ð!eUŠU

z{´—%²

–¨$ƒE
–eU™š›œ#ƒˆžŸ ¡¢£¤ƒ¥£è
JP6¦§

E–Ð!eU>
40g·L-1
.š›œ
#ƒˆžÉÃA

 2­øžOBƒ

Oë^

Å
E–Ð!eU‚ƒ.

š›œ#ƒˆžÉÜ
B

œƒE–Ð!eU™œ#ƒˆž|Jƒu
æù

E–Ð!eU™š›œ#$ƒî–x!
|J¦§

j•…©

œ#$ƒî–x!íî
–¨$E–Ð!eUƒø6ø6

@Þx!jp
½]ƒ„ˆ§¢®­
(P<005)。
ùÂ\ÃÄÐ!
œ#$ƒî–x!Ӝ#ƒˆž\¥]…œ

œËëÃĕu–¨$ƒî–’!ôQ(

´
—%²

E–Ð!eU>
40g·L-1
.Ãĕu–
¨$ƒî–’!Aœ

ƒ
23045mg·L-1,
J%
1。
·41·
u
35
Cu

w
2010
n

o
                             
Vol.35,Issue 1
January,2010
%1 
E–eU™š›œ#ˆžëŸî–¥£ƒ¦§
E–Ð!eU
/g·L-1
2­øžOB

O
î–Ð!¤B
/mg·g-1
!
/mg·L-1
30 179±0180a 1869±1920a 11938±14573a
40 306±0140b 2157±0320a 23045±0627b
50 207±0380ab 2772±1216a 13962±15968a
  
A
:5%
ˆ§û>

Ô-t¿%Xˆ§¢

%
2~4
-
)。
  
E–Ð!eU™š›œ#$Nïx!ƒ
¦§|èᲈ

J%
2。40g·L-1
E–Ð!e
UAJ­©œ#$Nï
Re
ƒ¥£

E–Ð!
eUßmÂ_œèœ­©‘ÕN£
Óuå


Rg1Œœ#$ƒx!ԙƒ,
eUƒE–’“ÀJ­©+ƒuå


Rb1
ŒeUE––«ƒœ#$x!ƒœ

Œ
40,50g·L-1
E–Ð!eU«”J²ˆjp

ù
Âvë^

¬Ã•Nïx!ÒY;>jNï
ƒx!CcœË
,´
—%²

íîE–Ð!eU
ƒ_œjNïx!G߃HI

-œ
ËŽ

ùÂvÃÄÐ!œ#$ƒNïx!Ó
œ#ƒˆž!\¥]…œ

œËëÃĕu
–¨$ƒNï’!ôQ(
,´
—ˆ&

œ

40g·L-1
E–Ð!eU«–.Nï’!


ƒ
9141mg·L-1。
%
2 
E–eU™š›N北ƒ¦§

/g·L-1

Re
/mg·g-1

Rg1
/mg·g-1

Rb1
/mg·g-1
jNï
/mg·g-1
jNï’!
/mg·L-1
30 2.65±0.250a 2.45±0.050a 4.25±0.550a 9.35±0.700a 59.70±5.950a
40 4.90±0.100b 1.45±0.050b 2.20±0.200b 8.55±0.100ab 91.41±3.205b
50 3.50±0.100c 0.85±0.200c 2.15±0.150b 6.50±0.200b 40.22±1.993a
33 
Xå5eU™š›œ#ˆžëŸNïYî
–x!ƒ¦§
 
Ÿz{œËë
1/2MS,3/4MS,MS,
3/2MS
ô

€Xå5eUŠU

z{´—%²


–¨$ƒXå5eU™š›œ#ƒˆžÉþëŸ
î–YNïƒx!JP6¦§

3/4MS
–¨AJ­©š›œ#ƒˆž

â

(1/2MS)
mÂâœ
(3/2MS)
ƒXå5eU虜
#ƒˆž]uæù

Xå5eU™š›œ#
$ƒî–¥£|J¦§

í$Xå5eU
ƒ_œ

œ#$ƒî–x!Gq•HI
,1/2MS
.
î–x!Aœ

ƒ
2259mg·g-1;
\ÃÄÐ!œ
#$ƒî–x!Ӝ#ƒˆž\¥]…œ¯ë
©


3/4MS
–¨–š›œ#.!


ƒ
15930mg·L-1,
J%
3。
%
3 
Xå5eU™š›œ#ˆžëŸî–¥£ƒ¦§
Xå5eU
/MS
–¨
2­øžOB

O
î–Ð!¤B
/mg·g-1
!
/mg·L-1
1/2 562±0595ab 2259±0448a 13561±5510a
3/4 906±1575a 1875±0064b 15930±27584a
1 735±1060ab 1754±0384bc 12573±11104ab
3/2 412±0115b 1734±0320c 6820±4160b
  
Xå5eU™œ-ÕNïx!ƒ¦§|œÔ
-

J%
4。

Re
Y
Rg1ƒx!“GHI¨Ÿ
‹

ƒƒXå5eUJ­©+,ƒ¥£


3/4MS
ƒ–¨.x!Aœ

 $
Re
x!œƒ
1175
mg·g-1;

Rb1ƒx!Œ3/4MSYMS–¨
–ƒœ#$x!ԙƒœ

ƒmƒœƒ
Xå5eU蜭©+ƒ¥£Yuå

Œœ-Xå
5eU—˜«–ƒœ#$jNïƒx!YNï
’!Jèᲈƒjp

ƒƒXå5eUJ­©
Nïƒuå
,3/4MS
–¨–.jNïx!YN
·51·
u
35
Cu

w
2010
n

o
                             
Vol.35,Issue 1
January,2010
ï’!胄A6{

¤p>
1590,13290 mg·L-1。
%
4 
Xå5eU™š›N北ƒ¦§
Xå5
/MS
–¨

Re
/mg·g-1

Rg1
/mg·g-1

Rb1
/mg·g-1
jNï
/mg·g-1
Nï’!
/mg·L-1
1/2 960±0500a 160±0100a 075±0050a 1195±0550a 7164±1805a
3/4 1175±0550ab 280±0900ab 135±0050b 1590±1500ab 13290±11100b
1 650±0100ac 090±0000ac 155±0050c 895±0150ac 6445±8350a
3/2 630±1500c 085±0150ac 080±0000a 795±1350ac 3088±2830c
4 
’
}¬!™;<–‰ˆžƒ¦§â®ÞEâ™
 ˆžwƒ¦§zsƒ

}¬!â?.

U–w
ԙƒž

}¬!â6.

–¨$º–‰Ð—˜
â

™Bˆžw¨Y

ý¦§ë–‰ƒˆžn
U

Ÿz{´—%²

Å}¬!â€.
(10g·
L-1),
œ#ˆžƒU–wƒž
,1
€–Rw 
2­øžOB|ԙƒ

@}¬!â6.
(25~
35g·L-1),
™Bˆžw¨Y

–¨$º–£¤—
˜â

]‹œ#ˆžnU“™
,1
€–Rw
ƒ2­øžOB|ԙƒ

Å}¬!àš.
(20
g·L-1),
š›œ#ƒˆžÉÃA

 2­øy
OBA6

–¨ƒç E–x!™Ž‰;žn›

ˆ‘’‰ƒ¥£ä‘J¦§

ŒŽ‰
;<–$

eUƒE–yyœëݎ‰;<
m$%ˆžƒ6®

œ–ò…œÓÔê|+™Ž
‰;
ªW–¨
ƒE–ç eUôêæŒ
30~50g·L-1。
Ÿz
{Œô€^ž=¹$NœËëç E–eU™š›
œ#ˆžëŸJ+£¤¥£ƒ¦§

>ðG–
—˜ëŸàœJ+‰Ðx!à†›œ

ñŸ
[8]
ä
²©2›œ#–ƒ`ø$Q

ƒE–eU
ÀJ­©ˆ‘’‰ƒ¥£

ôӟz{$ƒ
ƒE–eUÀJ­©N£ôƒ´Þ¨Ÿ
‹ƒ

Ÿz{M%²

œ#$ƒî–x!íî
–¨$E–eUƒø6ø6

@Þx!jp½
œ6
,¨
ŸŒ
20mg·g-1


ôÓ;<–š›
$%$î–x!¨Ÿ‹
[9]。
Ÿz{$!
ÓNï’!ô

€Q(Þ¢¥œëš›œ#ƒ
ˆžëŸî–YNïƒx!;ƒ

{´—¯×

E
–Ð!eU>
40g·L-1
.

!YNï’!è
ƒ„A6{

ªWš›œ#–ƒAàç E–
Ð!eU>
40g·L-1。
Xå5ƒeU™Ž‰”•–0)ƒˆžëŸ
J+‰Ðƒ¥£Yuå‘JP6ƒ¦§

ôÓ ¾
“–‰ƒ”×*UYÓÔêJ²

/0bs

Ó
Ô꾓]‹$%ßu¾“

ýF¡$%Ò^ƒ
”×Eø

C¦§$%øy
[10]。MS
–¨Xå
5x!ƒœ

ò!xά#l

eU|œ

  Þ
·CeUƒœ
[11],
ގ‰;<–$“A>s[
ƒ–¨Ò

ùÂë
MS
–¨>¨Ÿ–¨

E⾓ $Xå5ƒx!…œËXå5eU™š
›œ#ˆžŸ ¡¢£¤¥£ƒ¦§

±J`ø
È

eUƒXå5J­©ˆ‘’‰ƒu
å
[12]。GSivakumar[13]
ƒ/0%²
,3/4MS
–¨
AJ­©š›œ#ƒˆž

ƒƒXå5eU™
š›N£YuåJ­

ôӟz{ƒ´Þ
¨Ÿ‹ƒ

š›Nïޚ›ƒâ®J+£¤

|ޚ›Ð
!êæƒA­®Q(

Ÿz{$AœjNïx!œ
ƒ
1590mg·g-1,
œ©%=6îBFú$š›
;‰Nïx!ƒ`ø
,¨
Ÿ}ŸQš›$ƒN
ïx!

ô%²‘š›œ#–†Œˆ’š›N
JPƒbðF¡

W

Ÿz{M™š›
$ƒ¢â®J+£¤š›î–ƒ¥£3?Ccë
œË

´—%²

;<–ƒš›œ#$î–x

2%


Ÿ«nš›î–ƒ/0Z…Zæ„
­î

 ;½ƒSTYƒƒÞ¢. ŒDE“
$ˆ&s£ƒF¡

Ôkœ¤ƒv…

š›î–
ívŒ-,ûüÀ¥£–ƒ‚N

š›î
–ƒx!|ív£>?!š›Ð!ƒÛQ(

Ÿ
z{ƒ/0>Cè63ö–š›œ#

Àb
†Åg!âYG¦âà†ëøù®


—˜™š

[1] 
,õ/

š›§[;ê/0
[D]
ž
:¨
©Dª6
,2006
[2] LanghansováL,KonrádováH,VaněkTPolyethyleneglycol
·61·
u
35
Cu

w
2010
n

o
                             
Vol.35,Issue 1
January,2010
andabscisicacidimprovematurationandregenerationofPanax
ginsengsomaticembryos[J]PlantCelRep,2004,22:725
[3] MonteiroM,KeversC,DommesJ,etalAspecialroleforsper
midineintheinitiationphaseofsomaticembryogenesisinPanax
ginsengCAMeyer[J]PlantCelTissueOrganCult,2002,
68:225
[4] KimYS,YeungEC,HahnEJ,etalCombinedefectsofphy
tohormone,indole3butyricacid,andmethyljasmonateonroot
growthandginsenosideproductioninadventitiousrootculturesof
PanaxginsengCAMeyer[J]BiotechnolLet,2007,29:
1789
[5] KimJH,ChangEJ,OhHISaponinproductioninsubmerged
adventitiousrootcultureofPanaxginsengasafectedbyculture
conditionsandelicitors[J]AsiaPacJMolBiolBiotech,2005,
13(2):87
[6] BaeKH,ChoiYE,ShinCG,etalEnhancedginsenoside
productivitybycombinationofethephonandmethyljasmoantein
ginseng(PanaxginsengCAMeyer)adventitiousrootcultures
[J]BiotechnolLet,2006,28:1163
[7] 
ñ£«

‘²-

¬€>

ä

S6›î–x!/
[J]
ȉ
ӈ‰()`
,2005,24(5):7276
[8] 
ñŸ

­!(

œFM

ä

2›œ#”•–ƒ/0
[J]
$%$,23
,2006,31(17):1409
[9] 
Š®ª

;<–š›$%ӂ‘š›î–ƒ/0
[D]
ž

:¨
©6
,2005
[10] RobertW,WeiG,ScotHM,etalBlockingswelingactivated
chloridecurentinhibitsmouselivercelproliferation[J]J
Physio,2001,532:661
[11] 
­-

¯°

‘8±

Ž‰$%–()ӓ
[M]
7
8

Gqªyí
,2004:9596
[12] 
­!(

2›œ#;<–ƒ/0
[D]
‚ü

‚ü6

2006
[13] SivakumarG,YuKW,PaekKYProductionofbiomassand
ginsenosidesfromadventitiousrootsofPanaxginsenginbioreac
torcultures[J]EngLifeSci,2005,5(4):333
Efectsofcultureconditionsonbiomassandactivecomponentsof
adventitiousrootscultureinPanaxginseng
HUANGTao1,2,GAOWenyuan1,2,WANGJuan1,CAOYu2
(1ColegeofPharmaceuticalScienceandTechnology,TianjinUniversity,Tianjin300072,China;
2InstituteofBiologicalEngineeringofTraditionalChineseMedicine,TianjinUniversityofScience
andTechnology,Tianjin300457,China)
[Abstract] Objective:TooptimizethecultureconditionofadventitiousrootsofPanaxginseng.Method:Theadventitious
rootswereobtainedthroughtissueculturebymanipulationofinoculum,varioussucroseconcentrationsandsaltstrength.Thecontents
ofginsenosidesRe,Rb1andRg1weredeterminedbyHPLCwhilethecontentsofpolysaccharidesweredeterminedbyultravioletspec
trophotometry.Result:Themultiplicationofadventitiousrootsreachedthepeakwhentheinoculumwas20g·L-1.Theefectsofsu
croseconcentrationandsaltstrengthonadventitiousrootswereobserved.Thecontentsofpolysaccharideswerehigherwhenthemedium
containedmoresucrose.40g·L-1sucrosewasfavorableforrootsgrowthandbiosynthesisofRe,while30g·L-1wasfavorablefor
thebiosynthesisofRb1andRg1.3/4MSmediumwasbenefitforthegrowthofadventitiousrootsandthebiosynthesisofginsenosides.
Thecontentsofpolysaccharidesweredecreasedwiththeincreaseofsaltstrength.Conclusion:Theresultsshowedthatinoculum,vari
oussucroseconcentrationsandsaltstrengthhavesignificantinfluencesonadventitiousrootsgrowth,secondarymetaboliteandpolysac
charidesynthesisinP.ginseng.
[Keywords] Panaxginseng;adventitiousroots;inoculum;sucroseconcentration;saltstrength;ginsenosides;polysaccharides
doi:10.4268/cjcmm20100102

›œ-0
 
žŸ

·71·
u
35
Cu

w
2010
n

o
                             
Vol.35,Issue 1
January,2010