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Screening of Phosphate Solubilizing Endophytic Fungi from Ancient Sophora japonica Leaves and Their Potential for Plant Growth Promoting

古国槐叶片溶磷内生真菌的筛选及其促生潜力初探



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,2016,36(7):1456-1463
犃犮狋犪犅狅狋.犅狅狉犲犪犾.犗犮犮犻犱犲狀狋.犛犻狀.
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’“”$•”–
(SOD)y—“”$–
(POD)˜-r™š,›@gAstrnopqYœAž。Ÿ ¡¢:(1)DFdef£hi¤r55¥gA
st£
,28¥lmnopq,£nopq¦§r˜-t¥m12¥:ZG7¨©ª«¬­(犘犲狊狋犪犾狅狋犻狅狆狊犻狊sp.),
ZG9、ZG23、ZG32、ZG36yZG53¨®¯t(犉狌狊犪狉犻狌犿sp.),ZG15yZG34¨°±(犃狊狆犲狉犵犻犾犾狌狊sp.),ZG39
yZG51¨²³­(犃犾狋犲狉狀犪狉犻犪sp.),ZG42¨I±(犜狉犻犮犺狅犱犲狉犿犪sp.),ZG48¨´µt(犈狆犻犮狅犮犮狌犿sp.)。(2)°
±¶stZG15yZG34rnow·¸(犱)Y¹tº·¸(犇)r}»犱/犇¼½¾¿t¥,ŠCa3(PO4)2 rn
‚pqÀ§
(1238.28y941.22mg·L-1),ÁÂÃĽ¾¿t¥(犘<0.01)。(3)4¥gAstZG7、ZG9、ZG
15yZG48‡ˆÅdƒt‹10d‰,ŽD‹ÆÇÈrÉÊËÌ£ÍÎÏgAtÐ,£Ñ°±(犃狊狆犲狉犵犻犾犾狌狊狀犻
犵犲狉)ZG15ŽÃÄÒ½dƒt‹reŒÓÔ、Žn-‘ÓÔ、SOD、POD˜-(犘<0.05),DÕÖ×؋r
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犛犮狉犲犲狀犻狀犵狅犳犘犺狅狊狆犺犪狋犲犛狅犾狌犫犻犾犻狕犻狀犵犈狀犱狅狆犺狔狋犻犮犉狌狀犵犻犳狉狅犿犃狀犮犻犲狀狋
犛狅狆犺狅狉犪犼犪狆狅狀犻犮犪犔犲犪狏犲狊犪狀犱犜犺犲犻狉犘狅狋犲狀狋犻犪犾犳狅狉犘犾犪狀狋
犌狉狅狑狋犺犘狉狅犿狅狋犻狀犵
HOUJiaojiao,BUFangfang,YUZhongdong,KANGYongxiang
(ColegeofForestry,NorthwestA&FUniversity,Yangling,Shaanxi712100,China)
犃犫狊狋狉犪犮狋:Toexplorethephosphatesolubilizingabilityofendophyticfungiandtheirplantgrowthpromo
tingpotential,weevaluatedthephosphatesolubilizingcapabilityofendophyticfungifromancient犛狅狆犺狅狉犪
犼犪狆狅狀犻犮犪leavesinZhougongmiaobythemethodofdissolvedphosphoruscycleandmolybdenumbule,and
detectedtheeffectsofinoculatedsterilehostplantletonchlorophyl,solubleproteincontents,SODand
PODactivities.Theresultsshowedthat,therewere28outof55fungistrainsfromancient犛.犼犪狆狅狀犻犮犪
leaveswithphosphatesolubilizingcapability,andtwelvestrainsamongtwentyeightendophyticfungidis
playedstrongcapabilityofdissolvingphosphateofCa3 (PO4)2 byatestofphosphatesolubilizing
capability,theywere犘犲狊狋犪犾狅狋犻狅狆狊犻狊sp.(ZG7),犉狌狊犪狉犻狌犿sp.(ZG9,ZG23,ZG32,ZG36andZG53),
犃狊狆犲狉犵犻犾犾狌狊sp.(ZG15andZG34),犃犾狋犲狉狀犪狉犻犪sp.(ZG39andZG51),犜狉犻犮犺狅犱犲狉犿犪sp.(ZG42)and
犈狆犻犮狅犮犮狌犿sp.(ZG48).Twostrainswithstrongphosphatesolubilizingcapabilitywerethenpickedup,
ZG15andZG34,belongedto犃狊狆犲狉犵犻犾犾狌狊spp..Theirdiameterofphosphorussolubilizingcircle(犱)and
犱/犇 (犇,colonydiameter)werebothhigherthanthatofothers,andtheirsolubilizationofCa3(PO4)2
accountedfor1238.28mg·L-1and941.22mg·L-1,respectively,whichweresignificantlyhigherthan
thatofothersandcontrol(犘<0.01).Fourstrains(ZG7,ZG9,ZG15andZG48)fromdifferentgenera
wereinoculatedintoaseptic犛.犼犪狆狅狀犻犮犪seedlingsbottletoensuretheircontactuponwitheachother.
Mycelium wasobservedinfectingintothecortexcelsandaroundthesurfaceepidemicalcelsofroot
tissue;underthetreatmentby犃狊狆犲狉犵犻犾犾狌狊狀犻犵犲狉ZG15,犛.犼犪狆狅狀犻犮犪seedlingschorophylcontent,solu
bleproteincontent,SODandPODnetactivitieswerealobviouslyhigherthanthatofcontrol(犘<0.05),
so,ZG15couldmaintainhealthyseedlinggrowthandimproveseedlingresilience,whichshowedagreat
potentialforancient犛.犼犪狆狅狀犻犮犪protectionandapotentialapplicationasanorganismfertilizerforforesttree.
犓犲狔狑狅狉犱狊:ancient犛狅狆犺狅狉犪犼犪狆狅狀犻犮犪;endophyticfungi;phosphatesolubilizingfungi;asepticseedling;
growthpromoting.
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5g,MgSO4·7H2O0.25g,MgCl2·6H2O5g,
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ž16g,pH6.5~7.5,¢|•í–[£ž);MS
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ƒ„oj
|•í–£r(n-o°QRn‚

Õ½TrU¢w

now·¸
(犱)ytº·¸(犇)}»
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nopqW§
[10,27]。
†`aŸ 

¡1)ÃV,
Dã`Fdef£_hi¤55¥gAst,WX
¨ZG1~ZG55,<)o°Y¨ZðoUrj|·
¸¹

lmnowrnost¨28¥,£15¥
犱/犇≥0.5,3¥犱/犇 [¾0.4~0.5ï),10¥犱/
犇<0.4;t¥ZG34yZG15rnow·¸(犱)h
è66y60mm,now·¸ytº·¸r}»
(犱/犇)hè1.22y1.20,¼½¾¿t¥,Æâ
Púnopq§¾¿t¥

2.2 456789:;<=>p@B9:qF?
rs
  ŠŽÙÚÇ@r犱/犇≥0.4r12¥nost
ñ.NBRIP¢|•í–•í,€ò\¢£Žn
-oÓÔ

Ÿ ¡¢

]1),12¥nogAstŠ
Ca3(PO4)2 rn‚pq#§,NBRIP¢|•í–¹
×¢£moÓÔ½¾ŠÐ

moÓÔ<113.61
~1238.28mg·L-1ï),^ t¥noJ¨1.20%
~23.69%。£,t¥ZG15rò\¢£moÓ
t1 9:;<=>?G@uv(犱/犇≥0.4)
Table1 Preliminaryscreeningresultsofthephosphate
solubilizingendophyticfungi(犱/犇≥0.4)

Strain 犇
/mm 犱/mm d/D t¥Strain 犇
/mm 犱/mm 犱/犇
ZG7 58 23 0.40 ZG36 25 27 1.08
ZG9 32 33 1.03 ZG39 33 35 1.06
ZG13 33 33 1.00 ZG42 50 25 0.50
ZG15 50 60 1.20 ZG43 15 17 1.13
ZG18 36 37 1.03 ZG48 40 16 0.40
ZG23 23 23 1.00 ZG50 25 25 1.00
ZG26 17 15 0.88 ZG51 36 15 0.42
ZG32 20 22 1.10 ZG52 23 25 1.09
ZG34 54 66 1.22 ZG53 29 29 1.00
_
:犱.now·¸;犇.tº·¸
Note:犱.Diameterofphosphorussolubilizingcircle;犇.Colony
diameter.
8541 ! " # $ % &                   36`
Ô½a1238.28mg·L-1,noJ¨23.69%;ZG
34rnopq¯ï,ò\¢£moÓÔaÏ941.
22mg·L-1,noJ¨17.75%;t¥ZG15yZG
34rnopq¼ÂÃĽ¾¿t¥yŠÐ
(CK),ÁaÏÂÃğ·(犘<0.01)。bÙ,t¥
ZG53、ZG51、ZG48、ZG32、ZG42、ZG9y ZG23
rò\¢£moÓÔcÂÃľt¥ZG15y
ZG34,YÂÃĽ¾ŠÐ(犘<0.01)。
2.3 456789:;<=>?Gw_`
;nogAstr18SrDNAITS6îðhÃ
¹GenBank£rt¥îðñ.Ld-hô,ÁŸe
½¾%ŠfÝr12¥nogAstñ.Æâã€,
Ÿ gü

¡2):ZG7¨©ª«¬­(犘犲狊狋犪犾狅狋犻狅狆
狊犻狊sp.);ZG9、ZG23、ZG32、ZG36、ZG53¨®¯
t
(犉狌狊犪狉犻狌犿sp.);ZG15、ZG34¨°±(犃狊狆犲狉
犵犻犾犾狌狊sp.);ZG39、ZG51¨²³­(犃犾狋犲狉狀犪狉犻犪
sp.);ZG42¨I±(犜狉犻犮犺狅犱犲狉犿犪sp.);ZG48¨
´µt
(犈狆犻犮狅犮犮狌犿sp.)。
[…hi¡V¿ÀÂÃÄ
(犘<0.01)
]1 12¥nogAstrno 
Differentlettersmeansignificantdifferenceamongstrains
at0.01level
Fig.1 Thephosphatesolubilizingeffectoftwelve
endophyticfungistrains
2.4 456789:;<=>ga>bN?`x
jk[…no 

[…¶rgAstZG7
(犘犲狊狋犪犾狅狋犻狅狆狊犻狊sp.)、ZG9(犉狌狊犪狉犻狌犿sp.),ZG15
(犃狊狆犲狉犵犻犾犾狌狊sp.)yZG48(犈狆犻犮狅犮犮狌犿sp.),hÃ
ˆ¶dƒt‹
,10d‰tºÛ*•í–¡L,
d‹AÛÙÚ

ˆ?gAstrd‹Æþk3Ç
È£ŽÍÎÏgAsttÐ

]2),BC86þ
l¨ÉÊËÌ

<£mnËÌyopŽËÌ£Vò
qm[…çÜr86

¡¢ˆ?rgAstyd
ƒt‹ÆrstPrsr_AMr

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w¤#$rìíxy

™š#$rÛe”J
[28]。4
¥nogAstŠdƒt‹eŒÓÔr™š¿
À¢Ã

¡3):t¥ZG15Ýbrd؋Chla、
Chlb、Chl(a+b)ÓÔ¼½¾¿t¥Ýb,Rh
Ã}CKè£35.8%、23.8%y10.0%,¿ÀaÏ
Ãğ·
(P<0.05);t¥ZG7Ýb¯ï,zPChl
b¹CK¿À[ÃÄÙ,ChlayChl(a+b)¼Ã
Ľ¾CK(犘<0.05);t¥ZG9yZG48Ýbr
eŒÓÔ¹CKƒÃÄ¿À。bÙ,DChla/b
}»X{

\ˆ?t¥ZG9rd؋Chla/b}
»ÃĽ¾ CK(犘<0.05),ˆ?¿t¥Š
Chla/b}»rPv [¢Ã。†`a£nog
Astt¥ZG7yZG15ŽpN—Ò½dƒt
‹reŒÓÔX见ÛePv

2.5.2 ‚9ƒ„…†€ Žn-‘æ#$
|g«?|¹}~ö÷r–ÇTTh

·ˆ™š#
$|r$Pvh‚œ0ãp

9TËÌ

€”^?
”%vž7

lm7CrAbPv
[29]。4¥gAs
tŠdƒt‹Žn-‘Óԙš[…

¡
3)。£,ˆ?t¥ZG15yZG48rd؋Ž
n-‘ÓÔÃĽ¾CK(犘<0.05),RhÃ}
t2 12‡9:;<=>?Gw_`
Table2 Identificationofthe12phosphatesolubilizingendophyticfungistrains
t¥WXStrainNo. t?Strain t¥WXStrainNo. t?Strain
ZG7 ©ª«¬­犘犲狊狋犪犾狅狋犻狅狆狊犻狊sp. ZG36 ®¯t犉狌狊犪狉犻狌犿sp.
ZG9 ®¯t犉狌狊犪狉犻狌犿sp. ZG39 ²³­犃犾狋犲狉狀犪狉犻犪sp.
ZG15 °±犃狊狆犲狉犵犻犾犾狌狊sp. ZG42 I±犜狉犻犮犺狅犱犲狉犿犪sp.
ZG23 ®¯t犉狌狊犪狉犻狌犿sp. ZG48 ´µt犈狆犻犮狅犮犮狌犿sp.
ZG32 ®¯t犉狌狊犪狉犻狌犿sp. ZG51 ²³­犃犾狋犲狉狀犪狉犻犪sp.
ZG34 °±犃狊狆犲狉犵犻犾犾狌狊sp. ZG53 ®¯t犉狌狊犪狉犻狌犿sp.
95417^            9::,7:FdefnogAstrjkYœAÞqƛ
]2 nogAsttÐ|(‚èB)A.ZG7(犘犲狊狋犪犾狅狋犻狅狆狊犻狊sp.);B.ZG9(犉狌狊犪狉犻狌犿sp.);C.ZG15(犃狊狆犲狉犵犻犾犾狌狊sp.);D.ZG48(犈狆犻犮狅犮犮狌犿sp.)
Fig.2 Colonizationofmyceliumofendophyticfungiin犛.犼犪狆狅狀犻犮犪
t3 iˆ9:;<=>?56‰bTable3 Thephysiologicalindexesof犛.犼犪狆狅狀犻犮犪seedlingsinoculatedthephosphatesolubilizingendophyticfungi

StrainsNo.
eŒÓÔY}»
Chlorophylcontentandtheirratio/(mg·g-1)
Chla Chlb Chl(a+b) Chla/b
Žn-ÓÔ
Solubleprotein
content/(mg·g-1)
SOD˜-
SODactivity
/(U·g-1·min-1)
POD˜-
PODactivity
/(U·g-1·min-1)
CK 2.32±0.10c 1.22±0.07b 3.54±0.17c 1.90±0.03b 6.01±0.39c 275.87±27.69cd 1050.62±123.82c
ZG7 2.76±0.05b 1.33±0.07ab 4.09±0.12b 2.09±0.08ab 5.34±0.15c 303.95±40.77bc 1206.64±63.85c
ZG9 2.56±0.05bc 1.17±0.02b 3.74±0.07bc 2.19±0.02a 5.93±0.31c 231.13±34.77d 1487.92±143.10ab
ZG15 3.15±0.10a 1.51±0.05a 4.66±0.15a 2.09±0.05ab 6.84±0.59b 381.34±10.77a 1430.60±36.76b
ZG48 2.38±0.08c 1.15±0.09b 3.53±0.17c 2.08±0.11ab 7.71±0.25a 341.03±25.92ab 1693.39±160.41a
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