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Ergot Alkaloids Content of Symbiont of Epichlo gansuensis Achnatherum inebrians under Different Mn, Zn and Fe Conditions

Mn、Zn和Fe对醉马草内生真菌共生体麦角酰胺和麦角新碱含量的影响



全 文 :书!"#:10004025(2016)07142708               犱狅犻:10.7606/j.issn.10004025.2016.07.1427
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¡3、6{9žœ,Zn2+|] ?7.0mmol·L-1Ÿ¢£”`¤,¥|]3ž‡…†¦§¨©ª«|]œ;ª€
4„?3µ|]œŸ ©Zn2+yz¡21.0mmol·L-1œ¢£”`¤。(3)?wx|]œŸ,OEP€‚
ƒ‡…† ?Feyz¡6.0~12.0mmol·L-1Ÿ¢£”`¤,¥?3žœ‡…†¦§¨©ª«|]œ;€4
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yz›œ‡ Mn、Zn{Fe’“|]»¼©½¾~KLEMNOEP2€„
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犈狉犵狅狋犃犾犽犪犾狅犻犱狊犆狅狀狋犲狀狋狅犳犛狔犿犫犻狅狀狋狅犳犈狆犻犮犺犾狅犵犪狀狊狌犲狀狊犻狊
犃犮犺狀犪狋犺犲狉狌犿犻狀犲犫狉犻犪狀狊狌狀犱犲狉犇犻犳犳犲狉犲狀狋犕狀,犣狀犪狀犱犉犲犆狅狀犱犻狋犻狅狀狊
WANZhiwen,FENGJiangrong,WANGPing,LIChunjie
(KeyLaboratoryofGrasslandFarmingSystems,ColegeofPastoralAgricultureScienceandTechnology,LanzhouUniversity,
Lanzhou730020,China)
犃犫狊狋狉犪犮狋:AstudywasconductedtoinvestigatetheeffectsofdifferentconcentrationsofMn,ZnandFeon
variationsofergotalkaloidscontentinsymbiontof犈狆犻犮犺犾狅犵犪狀狊狌犲狀狊犻狊犃犮犺狀犪狋犺犲狉狌犿犻狀犲犫狉犻犪狀狊.There
sultsshowedthat:(1)ThecontentsofergineandergonovineunderMn2+5.0mmol·L-1hadapeakvalue
atthe3rdweek,whichhadsignificant(犘<0.05)differencewithotherMn2+treatments,butthecontents
ofergonovineunder0.01mmol·L-1hadapeakvalueatthe6thand9thweek.(2)Thecontentsofergine
hadmaximumunderZn2+7.0mmol·L-1atthe3rd,6thand9thweek,andthecontentsoferginewas
significantly(犘<0.05)higheratthe3rdweekthanthoseatthe6thand9thweek.(3)Thecontentsofer
gonovinehadmaximumunderFe2+21.0mmol·L-1atthe3rd,6thand9thweek.Thecontentsofergine
hadapeakvalueunder6.0~12.0mmol·L-1atthe3rd,6thand9thweek,andthecontentsofergine
wassignificantly(犘<0.05)higheratthe3rdweekthanthoseatthe6thand9thweek.Thecontentsof
ergonovinehadmaximumunderFe2+0.0mmol·L-1at3rdand6thweek,butunder0.03mmol·L-1had
apeakvalueat9thweek.So,highconcentrationsofMn,ZnandFeelementsishelpfultostimulatethe
drunkenhorsegrassinfectedbyendophyticfungitoaccumulatetowergotalkaloidforashorttimetreat
ment.
犓犲狔狑狅狉犱狊:犃犮犺狀犪狋犺犲狉狌犿犻狀犲犫狉犻犪狀狊;endophyte;ergine;ergonovine;Mn;Zn;Fe
  ~K(犃犮犺狀犪狋犺犲狉狌犿犻狀犲犫狉犻犪狀狊)ÓÔ"ÕÖ
,Ka‡×ØÙKÚÛ

gJÜ-ÝÝKÞ
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狀犪狋犺犲狉狌犿)ßàEKÜ#,?4á、âã、Läå、 !æ 、 çèéê ( ë ) ìí­î [1]。 JKLEMN (fungalendophyte)ÓÛ[Æï?JKÔðñò2 Î`2ìEóžô ,¯ wõ}öF#÷\‡Û[
MN
[2]。
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ZªùØúû‡犖犲狅狋狔狆犺狅犱犻狌犿 Þ,üý”4þ
MNóÿ!"

#hÛ¡LEMN+à?

@TAñB
C‡OEP

~KLEMNOEP¿E2i
G‡€E„ , D€‚ƒ{€4„ , E2E„S¡Û[ÀÁFGHI†‡ÌJKîLMN ©ÊË [1213]。 ¯ªOÓ~KPQ(R{SKT 8ÔهFGUV [1416]。 VW , €4„{€‚ ƒXY»ÛZ‡ÌN[¤ , O¿S¡\]^_‡E`abÌbÛcd7¼N

#E®71eG`†’“f 、 g 、 h 、 i 、 j {kéŠÑ , lG‘†’“m 、 n 、 o 、 p{qé 。 ‘†’“?#PLr,…†st
,¯
uÓ#vwE®w¿Îx‡?

}#PLE]Eyoz Q{|}iG‡SN 。 xt‘†’“Å#õ~#
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‚#$xBõ~#ƒk

EÓV¡?xB¸¹Ÿ,ƒ„‡…7{ƒ„†
‡E®;£v‡Á

ù!Añvw‡™k
。Mn、Zn
{Fe?#PL ˆ‰ßŠ‡‹ñ、ŒSN {ŽSNé , xE3‘†’“õ‘÷#$P L’“;” , •–—^ , †Ÿ˜é 。 ¯o†‡ Mn、Zn{ Feõ}#$Eٙ,ˆ‰#$vw E® [1721]。 ‘†’“evˆ‰š›œE]EyM , l ?#$PLE$‡’“ozÔY»iG‡S N 。 ‚m{o Æ‰¨é#$PL[žŸMEñ  $ , ¡[žŸ?#$PL‡¢?É£¤Z [22]。 E$„Ów¥‡E’“$ , ª…†‰E$V) ( ‚ .V¦ 、 \8™ 、 SK§$ 、 ‘E$ 、¨ ©ª«é ) { ¬E$V) ( ‚Œ² 、 pz 、 ­ ® 、¯ ìé ) » } [2325]。 ý&3 , JKLEMNԇE$„‰`† ’“»} , ‚ LEMN€[E$„‡ñ‰P’ “»} [26]; ­®Ô»°PÆ/N¨±²Ô€ E„‡…†
[27];N{CaO‰4!^K³±²
ÔE„…†»} [28];N、P、NP‹Z_´µ}~ K€‚ƒ{€4„…†O»ÛZ‡¶bS N [29]。 ¯}©‘†’“}€E„ñ’“ˆ
‰‡CD*·¸ù&3

¹ºé
[30]
} Mn、Zn{
Fe9:Ÿ~K‡E®{E]»+b¼v½¾,¯
¿}€4„{€‚ƒb¼bÛc‡ÀÁ

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ÜslÂ?wxyz‡ Mn、Zn{Fe|]Ÿtu
~KLEMNOEP‡„ÃÄ

Qo¨°ÅÆ
_ÇÀÁª€4„{€‚ƒ‡…†Èy

Š‹
Œ~KLEMNOEPE„³´Ž3‘† ’“‡”•yz–— , ¡ÃÈɨyz‡€‚ƒ {€4„½É”•¸¹ 。 1 ÊË{Õ! 1.1 WXYZ ~K)©2011à(+çèÖÌÍÎÏ (102°52′E,37°12′N)Ðѐ©^_`%ÒÔÓë, 2012à10ÔÕև)5℃]¢©bc2×K‰ KØK)?†ÙÚÀlÔÛ (^ _ ) )Üæm , 2013à3ÔÝÞ/N(E+){w/N(E)~Kß ë ,2014à7ÔàÕև~K)N©Üsl。3 ‘†/ޒ“ìአMnCl24·H2O、ZnSO4· 7H2O{Fe·citrate·3H2Oìâãs͇Aäå£。 1.2 YZ[\]^_ æçèéêë 、 šìíî‡/N~K)© 8241 ! " # % &                   36ï
2ð»300gñr>.?‡(òó∶ôõö=
3∶1)÷øùƒú(Gû15cm,üû10cm,ý
12cm)Ô,?^_`%ÒÔÓëþÆpm¸¹Ÿ

Œžô12hŒ²,pz(20±1)℃,Œ²ÿz
120μmol·m
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300mLHoagland=>Å。 KÁ~K%&?-20℃‡67685h,
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È50mg?@ð©1.5mL ‡eppendorfAۆÔ;£B1mLÃÈÅ(CHCl3∶ MeOH∶NH4OH=75∶25∶2),CDñE,mpF G¸¹ŸoH 。 ÅI15℃、10000r·min-1¸¹Ÿ AÛ5min,©QJKÔFG¸¹Ÿ+,L7ª:。 !µAۆ£B0.75mLMeOH∶CCl4(1∶2){ 0.25mL25mmol·L-1Móµ,CDñEÅ,©A Û3 (Beckman,Germany)Ô?21℃、12000r· min-1¸¹ŸAÛ5min,ÈšNÅ0.25mLI 0.22μmOû‡»3ÆoPQoP|1.5mLR_ _ÇS , N
[33]。
NAgilent1100series¨°ÅÆ
_Çgh
,ZORBAXXDBC18_ÇT,I§ÆI;
1mL·min-1b¼ÀÁ。I§Æ A ¡0.1mol·
L-1 NH4OAc,B¡øU/0.1mol·L-1 NH4OAc
(3/1)。V²95% AÅ3min、70% AÅ22min、
65%AÅ5min、95%AÅ10minI§ÆW‰œ
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ÀÁX®312nm,7YX®427nm[34]。
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2 ˜™‰ìâ
2.1 犕狀^_56789:;<=:>?@:b
DEFGHI
2.1.1 ?@ABEF 2{|]œ‡`®,~
KLEMNOEP€‚ƒ…†?wxyz‡
Mn2+|]¸¹Ÿ ab˜cÅ°£‡de,Р?
|]¶3žœ¢£”`¤,¥`ߦ§(犘<0.05)
¨©ª«|]œ

š2)。?Æx|]œL,O
EP€‚ƒ…†2{ Mn2+|]yz‡f¨b°
£Å˜c

¥œ|]œ ? Mn2+|]yz¡5.0
mmol·L-1œ¢£”`¤,ìá}²(CK)°£v
43.8%、26.5%{8.1%,<ªuœµ|] ‰}²
ù¦§Ø^_
(犘>0.05)。
2.1.2 ?@CDEF gš2¿h,2{|]œ
‡°£

~KLEMNOEP€4„…†?}
²{0.0mmol·L-1Mn2+|]Ÿaij°£‡d
e

?|]¶9žœ¢£”`¤,WœìáÓ|]¶
3žœ‡10.06k{2.76k(犘<0.05);?2.5
mmol·L-1Mn2+|]Ÿ,OEP€4„…†2{
œ‡`®almŸ˜de
,¯
n»¦§Èy
(犘>
0.05);?5.0{7.5mmol·L-1Mn2+|]Ÿ,OE
P€4„…†2{|]œ‡°£ab˜cŚ
f‡de
,¯
oŠ|]3žœ¦§­¨。?|]¶
3žœ,OEP€4„…†Š5.0mmol·L-1
Mn2+|]”¨,¥¦§¨©ª« Mn2+yz¬z(犘
<0.05),Ó}²‡4.53k;OEP€4„…† ?}²Ÿ¢£”`¤

¥¦
§¨©ª«|]
(犘<0.05)。
Šš˜™p‹

­¨yz‡ Mn2+}~KLE
k1 lmWX^_nopR4^_qr[28]
Table1 Traceelementsandtreatmentlevelsinthepot
‘†’“
Traceelement
|]¯)Treatmentlevel/(mmol·L-1)
Ⅰðò=>Å(}²)
Completesolution(control) Ⅱ Ⅲ Ⅳ Ⅴ
Mn 0.01  0 2.5 5.0 7.5
Zn 0.0008 0 7.0 14.0 21.0
Fe 0.03  0 6.0 12.0 18.0
92417ô       ;<=,é:Mn、Zn{Fe}~KLEMNOEP€‚ƒ{€4„…†‡ˆ‰
k2 st 犕狀^_u678`a?@AB4?@CDEFGvw
Table2 Theergineandergnovinecontentsin犃犮犺狀犪狋犺犲狉狌犿犻狀犲犫狉犻犪狀狊seedlingsunderdifferentMnconditions
€E$„
Ergotalkaloids
Mn2+yz
Mn2+concentration/(mmol·L-1)
|]œ Treatmenttime/week
3 6 9
€‚ƒ…†
Erginecontent
/(mg·kg-1)
0.01(CK) 100.54±7.79bA 58.41±5.07bcB 76.87±4.74abB
0 81.55±12.72bA 46.63±5.37cB 75.49±3.89abA
2.5 88.86±1.71bA 57.98±4.45bcB 65.36±5.15bB
5.0 144.60±3.50aA 73.88±2.05aC 83.08±1.84aB
7.5 101.32±2.77bA 62.42±4.01abC 82.97±7.63aB
€4„…†
Ergnovinecontent
/(mg·kg-1)
0.01(CK) 0.97±0.60bC 6.21±0.58aB 9.76±1.36aA
0 1.53±0.35bB 1.96±0.26bcB 4.22±0.29bA
2.5 2.89±0.11bA 2.76±0.10bA 2.67±0.03bA
5.0 4.39±1.10aA 1.56±0.02cB 3.55±0.27bAB
7.5 3.26±0.40bA 1.21±0.14cB 2.88±0.46bB
q

šÔZý¡) ¤±+rs(n=5);xԇwxßtuvšwxô|]^_¦§(犘<0.05),ڝ^_¦§
(犘<0.05)。Ÿx
Note:Dataarepresentasmean±SE(n=5);Differentnormalletterswithinthesamecolumnmeansignificantdifferenceamongtreat
mentsat0.05level,whiledifferentcapitalletterswithinthesamerowmeansignificantdiferenceamongstagesat0.05level.Thesameasbelow.
MNOEP€‚ƒ‡³´Y»ÛZ‡¶bSN

¥?›œL°™Éx

x?

›œ‡¨yz
Mn2+|]¿Š½¾OEPE€4„,<®œ
‡¨yz|]·w¼©€4„‡³´

2.2 犣狀2+ ^_5678`a?@:bDEFG
HI
2.2.1 ?@ABEF š3¦w,2{Zn2+|]
œ‡`®

~KLEMNOEP€‚ƒ…†
?0~14.0mmol·L-1Zn2+|]Ÿab˜cÅ°
£‡de

Ð?|]¶3žœ¢£”`¤,¥¦§
(犘<0.05)¨ ©ª«|]œ;L-1Zn2+yzœ,OEP€‚ƒ…†aijŸ˜
‡de

¥|]¶3žœ‡…†¦§(犘<0.05)¨
©ª«|]œ

?Æx|]œL

~KLE
MNOEP€‚ƒ…†2{Zn2+|]yz‡f
¨`ßab°£Å˜c‡de

?|]¶3、6{9
žœ

OEP€‚ƒ…† ?7.0mmol·L-1
Zn2+|]Ÿ¢£”`¤,ìá}²¦§°£v
84.0%、33.1% { 44.5%;? | ] ¶ 9 ž œ,
21.0mmol·L-1Zn2+yz|]‡€‚ƒ…†¦
§
(犘<0.05)c©ª«‡yz|],‰}²Æ¦
§ytv28.5%。
2.2.2 ?@CDEF Hš3l¿z~,2{
Zn2+|]œ‡`®,~KLEMNOEP€
4„…†?wxyz‡Zn2+|]Ÿ ij°£,Ð
?|]¶9 žœ¢£”`¤,¥`ߦ§(犘<
0.05)¨ ©ª«|]œ。~KLEMNOEP
€4„…†?|]¶3žœ2{Zn2+|]yz
‡f¨<°£

]yz°£ab˜cÅf¨‡de

?3µ|]œ
Ÿ

OEP€4„…† ?21.0mmol·L-1
Zn2+|]Ÿ¢£”`¤,ìá}²°£v3.66
k
、1.55k{1.29k。
Šš˜™p‹

›œL~KLEMNOE
PE€‚ƒ‡”{|Zn2+ yz¡7mmol·
L-1,o¨Îocyz õ}ªEw¼ˆ‰,<®
œLZn2+ |]}€‚ƒ‡³´°™w•;¯
Ó

®œ¨yz‡Zn2+|]»¼©OEP€4
„‡³´

2.3 犉犲^_5678`a?@:bDEFGHI
2.3.1 ?@ABEF 2{Fe|]œ‡`®,
~KLEMNOEP€‚ƒ…†?wxyz‡
Fe|]Ÿab˜cÅ°£‡de,Р?|]¶3
žœ¢£”`¤

Ð`ߦ§¨©ª«|]œ

š
4)。?Æx|]œL,OEP€‚ƒ…†2{
Fe|]yz‡f¨ab°£Å˜c‡de;?|]
¶3、6žœ,OEP€‚ƒ‡…† ?Feyz¡
6.0~12.0mmol·L-1Ÿ¢£”`¤,ìá}²
¦§°£v`}109.3%{204.2%;?|]¶9ž
œ

OEP€‚ƒ…†?Feyz¡6.0mmol·
L-1|]Ÿ¢£”`¤,‰xô}²Æ¦§°£
v113.9%。
0341 ! " #  % &                   36ï k3 st犣狀^_u678`a?@AB4?@CDEFGvw Table3 Theergineandergnovinecontentsin犃.犻狀犲犫狉犻犪狀狊seedlingsunderdifferentZnconditions €E„
Ergotalkaloids
Zn2+yz
Zn2+concentration/(mmol·L-1)
|]œ Treatmenttime/week
3 6 9
€‚ƒ…†
Erginecontent
/(mg·kg-1)
0.0008(CK) 100.54±7.79cA 58.41±5.07bB 76.87±4.74bB
0 144.51±14.90bA 67.08±7.07abB 69.03±3.67bB
7.0 185.03±14.79aA 77.77±10.49aC 111.10±5.75aB
14.0 172.89±10.49abA 59.09±5.88bB 79.88±0.89bB
21.0 168.77±10.33abA 73.64±1.84abB 54.98±3.62cB
€4„…†
Ergnovinecontent
/(mg·kg-1)
0.0008(CK) 0.97±0.60bC 6.21±0.58aB 9.76±1.36aA
0 2.07±0.14bB 3.86±0.29bA 4.02±0.29bA
7.0 2.44±0.07bC 4.18±0.12bB 4.99±0.14bA
14.0 2.67±0.21bB 5.05±1.21bAB 8.23±1.29abA
21.0 3.55±0.52aB 9.62±1.37aB 12.56±1.14aA
k4 st犉犲^_u678`a?@AB4?@CDEFGvw
Table4 Theergineandergnovinecontentsin犃.犻狀犲犫狉犻犪狀狊seedlingsunderdifferentFeconditions
€E$„
Ergotalkaloids
Feyz
Feconcentration/(mmol·L-1)
|]œ Treatmenttime/week
3 6 9
€‚ƒ…†
Erginecontent
/(mg·kg-1)
0.03(CK) 100.54±7.79bA 58.41±5.07cB 76.87±4.74dB
0 125.11±10.97bA 44.56±0.53cB 109.05±4.76cA
6.0 210.44±3.94aA 151.07±11.82aB 164.40±6.21aB
12.0 210.32±3.43aA 177.67±11.47aB 148.19±4.17abC
18.0 194.10±11.71aA 118.18±10.90bB 128.25±12.42bcB
€4„…†
Ergnovinecontent
/(mg·kg-1)
0.03(CK) 0.97±0.60bC 6.21±0.58cB 9.76±1.36aA
0 5.35±1.57aB 13.25±0.50aA 6.59±2.44abB
6.0 2.18±0.25bC 10.84±0.50abA 5.92±0.70abB
12.0 0.57±0.07bB 8.66±2.30bcA 6.11±0.96abA
18.0 0.80±0.02bC 8.92±0.79bcA 4.17±1.06bB
2.3.2 ?@CDEF š4¦w,2{|]œ‡


~KLEMNOEP€4„…†?}²
‹š#¡ij°£

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œyz|]¶3ž‡…
† ‹¦c©¶6、9ž。?|]¶3、6žœ,OE
P€4„…† ?Feyz¡0mmol·L-1|]
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