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Isolation and Screening of Phosphate Solubilizing Rhizobia of Sainfoin

溶磷红豆草根瘤菌的分离和筛选



全 文 :!24" !1#
 Vol.24  No.1
! " # $
ACTA AGRESTIA SINICA
   2016$  1%
  Jan.  2016
犱狅犻:10.11733/j.issn.10070435.2016.01.031
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犐狊狅犾犪狋犻狅狀犪狀犱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犚犺犻狕狅犫犻犪狅犳犛犪犻狀犳狅犻狀
LIJianfeng1,2,ZHANGShuqing1,2,SHIShangli2,
DUJianxiong3,LILixia1
(1.GuizhouNormalColege,InstituteofSoilandEnvironmentBioremediationinKarsthabitats,Guiyang,GuizhouProvince550018,China;
2.GansuAgricultureUniversity,KeyLaboratoryofGrasslandEcosystemofMinistryofEducation,Lanzhou,GansuProvince730070,China;
3.GuizhouUniversityofFinanceandEconomics,Guiyang,GuizhouProvince550025,China)
犃犫狊狋狉犪犮狋:Inordertoobtainphosphatesolubilizing犚犺犻狕狅犫犻狌犿strainsofsainfoin(犗狀狅犫狉狔犮犺犻狊狏犻犮犻犪犲犳狅犾犻犪).
WinogradskyN-freemediumandPKOmediumwereusedtoisolateandscreen犚犺犻狕狅犫犻狌犿strainsfrom
sainfoinrootnodules.Byanalyzingthecapabilitiesofsparinglyinorganicphosphorussolubilizing,IAAse
cretingandthenodulenitrogenaseactivityofscreenedstrains,threerhizobiastrainsRS1,RS8andRS14
wereobtained.AndthephosphatesolubilizingcapabilityofRS1strainreached6.98mg·L-1·d-1in14
days,andRS1withhigherIAAsecretingcapabilityandcouldsurviveatpHvaluesrangingfrom4to12,
andsalttolerancecapabilityat11%.Resultsshowedthatthescreeningmethodusedwassimpleandeffec
tive,andtheisolatedstrainscouldhavegoodprospectofapplication.
犓犲狔狑狅狉犱狊:犗狀狅犫狉狔犮犺犻狊狏犻犮犻犪犲犳狅犾犻犪;犚犺犻狕狅犫犻狌犿;Nitrogenfixation;Phosphorussolubilizing;IAA
HIJK
:20140826;LMJK:20150611
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Table1 TheabilitiesofinorganicphosphatesolubilizingandIAAsecretingofthestrains
Þr
Strains
Þrç”
Sourcesofstrains
D/d
ò©Ûj`
Psolubilization
capacity/mg·L-1·d-1
¤9Ÿu§¥
IAAsecreting
RS1
w<ºf-
犗狀狅犫狉狔犮犺犻狊狏犻犮犻犪犲犳狅犾犻犪 .
2.75±0.11a 6.98±0.27a +++
RS8

67

ºf-
犗狀狅犫狉狔犮犺犻狊狏犻犮犻犪犲犳狅犾犻犪犛犮狅狆.‘Gansu’
2.02±0.06c 5.49±0.20b ++
RS14

67

ºf-
犗狀狅犫狉狔犮犺犻狊狏犻犮犻犪犲犳狅犾犻犪犛犮狅狆.‘Gansu’
2.42±0.10b 3.89±0.13c ++
  ü:@j6@uU;Çg˜Øº…†(犘<0.05),œ@;“D/d”nÛj»ë,“D”Ûj¸"Â,“d”ÞP"Â
Note:Meanswithdifferentlowercasesineachcolumnmeansignificantdifferenceatthe0.05level,thesameasbelow;“D/d”means
Phosphate-dissolvingindex,“D”indicatePhosphatedissolvingringdiameter,“d”indicatecolonydiameter
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Table2 Nodulenumber,nodulefreshweightandnitrogenaseactivityofsainfoin
plantsinoculatedwithdifferentphosphatedissolvingrhizobia
#GÞr
Strains
÷r}•ë
Nodulenumber
}•¼•
Nodulefreshweight/mg
}•"Â
Nodulesdiameter/mm
}•êi½É
Nodulenitrogenaseactivity/μmol·g-1·h-1
RS1 21.60±3.21a 3.15±1.11ab 2.37±0.26a 4729±95a
RS8 7.60±3.51c 2.33±0.59b 2.20±0.22a 1040±73c
RS14 17.20±2.28b 3.48±1.75a 2.50±0.61a 2230±148b
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Table3 Physiologicalbiochemicalcharacteristicsofthestrains
é´µ Measuringitems
ÞrStrains
RS1 RS8 RS14
DæaR¿BetaD()Arabinose + + +
§e¿Rhamnose - + +
D¨¿Dribose + + +
^¿ Maltose + + +
ñ¿Synanthrin - - -
ÿ!¿Glucose + + +
f¿Fructose + + +
D¿Xylose + + +
„m¿Researcherssugar + - +
ÀåDextrin + + +
¾ÁStarch - - -
¾¿Sucrose + + +
G¿WCitrate - - -
=ÁðU½Catalasereaction + + +
¤32®„¿uŠProduction3ketolactose - - -
*kr].uŠ Methylenebluereductionreaction + + +
Ž¢r].uŠNilebluereductionreaction + + +
ÖW)].uŠNitratereductionreaction + - -
XðâŽcSalttolerance ≤10% ≤8% ≤11%
9ŸpH<=GrowthpHrange 4~12 5~12 4~11
BTBuŠBTBreaction
¤W
Producingacid
¤W
Producingacid
¤W
Producingacid
  ü:“+”g˜Þr§B9Ÿ$uŠd,“-”g˜Þr6§9Ÿ$uŠ€
Note:“+”meansstrainscouldgroworshowedpositivereaction;“-”meansstrainscouldnotgroworshowednegativereaction
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Table4 Antibioticsresistanceofthestrains
ø9uàF
Antibiotic
ÞrTø9uø
Antibioticresistanceofstrains/μg·mL-1
RS1 RS8 RS14
àKuStreptomycin ≤50 ≤70 ≤60
ÁÞuPolymyxin ≤15 ≤15 ≤10
ÂKu Neomycin ≤30 ≤30 ≤40
ó8Ku Kanamycin ≤30 ≤10 ≤40
ºKuErythrocin ≤200 ≤300 ≤250
XKuChloramphenicol ≤400 ≤350 ≤350
®Þ5Bacitracin ≤300 ≤300 ≤300
…3KuPenbritin ≤140 ≤110 ≤100
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1411

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