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Structure of Arbuscular mycorrhiza fungi community in soils with different salinity and influence factors

不同盐碱度土壤中AM菌根真菌群落结构组成及影响因子



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ChineseJournalofBioprocessEngineering
Vol.11No.3
May2013
doi:10.3969/j.issn.1672-3678.2013.03.011
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:2012-10-17
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(Arbuscularmycorhiza,AM)
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(Glomus)
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:1672-3678(2013)03-0059-06
StructureofArbuscularmycorrhizafungicommunityinsoilswithdifferent
salinityandinfluencefactors
LIDan1,2,GAOYongchao1,WANGJianing1,QIUWeizhong1,CHIJianguo1
(1.BiologicalResearchInstitute,ShandongAcademyofSciences,Jinan250014,China;
2.SchoolofPopulation,ResourcesandEnvironment,ShandongNormalUniversity,Jinan250014,China)
Abstract:InordertoexploretheArbuscularmycorhiza(AM)fungidiversityoftheplantrhizospherein
thesalinesoiloftheYelowRiverDelta,fourrhizospheresoilsamplesofSuaedaglaucaBgeandTamarix
chinensiswerecolectedfromGudongandGudaooilfieldsinDongyingCityforidentificationoftheAM
fungicommunities.TheresultsshowedthatGlomuswasthedominantspeciesinthesalinesoil
accompanyingwithmanyunidentifiedfungi.Theinvestigationofthemycorhizalfungicommunityin
diferentsoilsalinityshowedthattheAMfungaldiversityintherhizospheresoilofSuaedaglaucaBgewas
higherthanthatofTamarixchinensis,andGudonghigherthanGudao.Thecorelationanalysisshowedthat
theammoniumnitrogencontenthadasignificantnegativerelationshipwithAMfungaldiversity.
Keywords:Arbuscularmycorhizalfungi;biodiversity;DGGE;nestPCR
  
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NCBI
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÷£å
/%
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1 Unculturedeukaryote(AB534506.1) 99 +
2 UnculturedGlomus(AM946840.1) 100 + +
3 Scutelosporacalospora(AJ306445.1) 100 + + + +
4 Unculturedeukaryote(GU072180.1) 99 +
5 Fusariumoxysporum(HE586716.1) 99 + + + +
6 Telonemasubtilis(AJ564772.1) 100 + + +
7 Glomussp(FR715050.1) 99 +
8 unculturedeukaryote(AY789780.1) 100 +
9 1 unculturedeukaryote(AB534506.1) 99 + + +
9 2 Aspergilusawamori(GQ472777.1) 91 + + +
10 Circinelaumbelate(AF157128.1) 100 + +
11 unculturedcryptophyte(FJ410743.1) 100 +
12 Pinguiochrysispyriformis(AB058926.1) 96 + +
13 unculturedmarine(FR874662.1) 100 +
14 Cadophorasp(HM116764.1) 99 +
15 Alternariaalternata(JN088533.1) 99 + + + +
16 Chrysosporiumkeratinophilum(AY234824.1) 99 +
        
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2 
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Table2 Basicphysicalchemicalpropertiesofsampledsoils
RS ä 
pH Ât*/%
ÂF–


«Øc–

(μg·g-1)
ÂGØc–

(μg·g-1)
Âj}¬–

(μg·g-1)
ÂÍ}Ž–

(μg·g-1)

±s/­&[û
7.7 18.7 0.11 3.2 0.24 1950.0 9.2

±/­&[û
7.9 20.4 0.07 1.0 0.06 550.0 5.2

±®¯&[û
8.3 22.5 0.07 2.2 0.08 650.0 1.2

±s®¯&[û
8.1 21.1 0.07 4.4 0.45 1400.0 4.0
2.4.2 AM
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AM
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4。
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7

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AM
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Fig.4 Diversityindicesandspeciesrichnessofsoilsamples
36 
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Table3 PearsoncorrelationshipofdiversityindicesandspeciesrichnessofAM fungalwithsoilproperties

pH
ÂF– Ât* «Øc GØc j}¬ Í}Ž
H ns ns ns -0980 ns ns ns
4ns ns ns -0959 ns ns ns
    
T

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 P<005, P<001
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pH、
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AM
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[1] 
mIÃ

‚Ÿl

N12©F/û¢
AM
…[h„–¨„
[J].
&]óå
,2001,9(4):389392.
[2] 
µ¡

Ý;

y44

{

û{žX§«öF/û¢
VA
[a…
[hST
[J].
û{Ž)
,2008,45(4):758763.
[3] AliasgharzadehN,SalehRastinN,TowfighiH,etal.Occurence
ofArbuscularmycorhizalfungiinsalinesoilsoftheTabrizPlain
ofIraninrelationtosomephysicalandchemicalpropertiesofsoil
[J].Mycorhiza,2001,11:119122.
[4] 
½¤



ÄÅ

{

½¾¿¶5íò§û{H&ªŠh
ST
[J].
tû„âŽ)
,2010(8):229233.
[5] 
]ôc

ÇÖÆ

²q

ûodzH&§û{¢ú³KÝÞ
hrÏå78
[J].
Û܄ÈÎ
,2005,31(5):3839.
[6] 
Ò§

ÉÊ

¾1

ú³ÝÞû{[aFjœ78
[J].
ú³
K¨¢öˆ)
,2004,33(3):217218.
[7] 
·b¸

ËlÌ

Íh

{
.AM
…[
DNA
hp¤K
PCR
DGGE
ñí
[J].
[Ž)
,2005,24(4):564569.
[8] 
m`


PCRDGGE
œñí^2&ÜNÎ
AM
…[]ó
å
[D].
ϓÐ

l·,0Ž
,2009.
[9] AgwaH E,AbdelFatahGM.Arbuscularmycorhizalfungi
(Glomales)inEgypt:I.anecologicalviewofsomesaline
afectedplantsinthedeltaicMediteraneancoastalland[J].Acta
BiologicaHungarica,2002,44:117.
[10] AzizT,HabteM.InfluenceofinorganicNonmycorhizalactivity
nodulationgrowthofLeucaenaleucocephalainanoxisolsubjected
tostimulatederosion[J].CommunSoilSciPlantAnal,1989,71
(3/4):239251.
[11] ZandavaliRB,DilenburgLR,PauloVD.Growthresponsesof
Araucariaangustifolia(Araucariaceae)toinoculationwiththe
mycorihizafungusGlomusclarum[J].ApplSoilEcol,2004,25
(3):245255.
[12] 
yV%

‚+»

Ñ5«

{

¢–F/û{¢
AM
[h„–
¨„
[J].
Ò¸âŽÄŽ)
,1999,16(1):1620.
[13] 
A³u

Ò(Ó

ú³ÝÞ§û{c!ԈOGˆ5‚hS
T
[J].
âÕ&ØÛÜ
,1998,14(1):3942.
[14] 
+‘Ö

m×a

3{ÐhÔæYó
[M].2
„

ö

¢–
â&0Žæ„˜
,2001:160168.
[15] MarschnerH,DelB.Nutrientuptakeinmycorhizalsymbiosis
[J].PlantSoil,1994,59:89102.
[16] TanakaY,YanoK.Nitrogendeliverytomaizeviamycorhizal
hyphaedependsontheform ofN supplied[J].PlantCel
Environ,2005,28(10):12471254.
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