全 文 :!24" !1#
Vol.24 No.1
! " # $
ACTA AGRESTIA SINICA
2016$ 1%
Jan. 2016
犱狅犻:10.11733/j.issn.10070435.2016.01.008
ST$UV/J,-.2|u
Ä-¶
1,2,
ÅÆÇ
3,
ÈÉÊ
1,2,
ËÌÍ
1,2,
ÎÏÐ
1,2,
ÈÑÒ
2,4,
jÄÓ
1,2,
ÔÕÖ
1,2
(1.®()*+,()9\/01,® ®Y350003;2.®²_GH()Ãľ¨/0&!,® ®Y350003;
3.®()*+,9K¾¨/01,® ®Y350003;4.®()*+,ê?\]/01,® ®Y350003)
HIJK
:20150207;LMJK:20150707
NOPQ
:
¬*¾¿À´µ
(2012BAD14B15,2011BAD17B02);®ºà*/,1®»*/©´(2014R10177);®QÙУ
Y©´
(2012NZ0002)Å
5RST
:
?
(1975),È,@,ÊË,®/0¯,Ì«ÍÎ()9\6´µ9\/0,Email:mume19@126.com,<Ôc Author
forcorrespondence,Email:wengboqi@163.com
78
:
nWhAáBïDÙC¥|?9KTUV
,
AásÙC¥TÁ
,
»ÔdXåæ°â
、
Z©
`PCR(fluorescencequantitativePCR)6PCR-DGGEâ/06@@AAáBï`(CÚN`200,150,100,50kg
N·hm-2)DDE-(犘.犪犿犲狉犻犮犪狀狌犿×犘.狆狌狉狆狌狉犲狌犿)_ÙC?9KTUV。efgh,DÒ¸,AáBï§
sÙC?9K9K`N6C,òAáBï`?9K`N6CTí`
。AáBï]§
íîHÞ
16SrDNA®56ÞSSUrDNA®5ëA,&200kgN·hm-2Óù»16SrDNA®56SSUrDNA®5T?
9Kë`Ý
,
AáBï`ÙC&»16SrDNA®56SSUrDNA®5T?9Kë`
,I
êiÞ狀犻犳犎 T®5ëA。AáBïÉÃ-_D³´HÞ、Þ6êiÞL2TÙC?9KOP:ï
UV
,
L[\»16SrDNA®56SSUrDNA®5T?9K
I,ILß©ÄASÿ)»狀犻犳犎 ®5T?9KT
I
,
;<ÙC»16SrDNA®5、SSUrDNA®56狀犻犳犎 ®5T?9Kë`。
9:;
:
Aá
;
HÞ
;
Þ
;
êiÞ
;
®5ëA
;
?9K:
<=>?@
:S182 ABCDE:A AFG@:10070435(2016)01005407
犈犳犳犲犮狋狊狅犳犅犻狅犵犪狊犛犾狌狉狉狔犃狆狆犾犻犮犪狋犻狅狀狅狀狋犺犲犛狅犻犾犕犻犮狉狅狅狉犵犪狀犻狊犿狊狅犳
犘.犪犿犲狉犻犮犪狀狌犿×犘.狆狌狉狆狌狉犲狌犿犌狉犪狊狊犾犪狀犱
ZHONGZhenmei1,2,SONGYana3,HUANGXiusheng1,2,YOUXiaofeng1,2,WENGBoqi1,2,
HUANGQinlou2,4,CHENZhongdian1,2,FENGDeqing1,2
(1.AgriculturalEcologyInstitute,FujianAcademyofAgriculturalSciences(FAAS),Fuzhou,FujianProvince350003,China;
2.FujianEngineeringandTechnologyResearchCenterforHilyPrataculture,Fuzhou,FujianProvince350003,China;
3.Instituteofbiotechnology,FAAS,Fuzhou,FujianProvince350003,China;
4.InstituteofAnimalHusbandryandVeterinaryMedicine,FAAS,Fuzhou,FujianProvince350003,China)
犃犫狊狋狉犪犮狋:Inordertorevealtheeffectsofbiogasslurryapplicationonimprovingsoilfertility,thechangeof
soilmicroorganismsunderdifferentamountsofbiogasslurry(equalto200,150,100and50kgN·ha-2
pureN)applicationon犘犲狀狀犻狊犲狋狌犿.犪犿犲狉犻犮犪狀狌犿×犘犲狀狀犻狊犲狋狌犿.狆狌狉狆狌狉犲狌犿grasslandwasstudiedbychlor
oformfumigation,fluorescencequantitativePCRandPCRDGGEmethodsbasedonfieldexperiment.The
resultsshowedthatsoilmicrobialbiomassCandNof4treatmentssignificantlyincreasedbybiogasslurry
applicationcomparedtocontrol,andtherewasasignificantcorrelationbetweentheslurryapplicationa
mountandtheincrementofsoilmicrobialbiomassCandN.Biogasslurrysignificantlyincreasedtheabun
danceof16SrDNAandSSUrDNAgenewhichwassignificantlyrelatedwiththeapplicationamountof
biogasslurry,andthequantitiesof16SrDNAcontainingbacterialandSSUrDNAcontainingfungiwere
themaximuminthetreatmentofcontaining200kgN·ha-2biogasslurry,whiletheabundanceof狀犻犳犎
containinggenesignificantlydecreased.Asaresult,itcouldconcludethattheapplicationofbiogasslurry
hasnosignificanteffectonthecommunitydiversityofsoilbacteria,fungiandnitrogenfixingbacteria,but
promotesthegrowthofmicroorganismscontaining16SrDNAgeneandSSUrDNAgenes,inhibitsbacteri
!1# ??:AáDDE-_ÙC?9KOPTUV
algrowthcontaining狀犻犳犎gene,andchangesthenumberofmicroorganisms.
犓犲狔狑狅狉犱狊:Biogasslurry;Bacterial;Fungi;Azotobacter;Geneabundance;Microbialcommunitydiversity
ÀìêFDI)T×Ø°±,êFDI´µþ
ÀnÓ´µTÌ«ç
[1]。
ÏdGÁ°H
(
A
ÃÄ
)
ÓùDI¨IKÚß´d,-òtn
kT¾¨
,
L ùDI)´µ=Ä&°ì«
Tcd
。
AáÚGÁ°H_¤9TáNI/K
,
ì
»GED=
、
?`üu6JK9u
(
~JK
u
、
LMìW?
)
?9ýÉKF
,
ÚßGHFJÁ
ý
,
DsÙC¥
、
;ÅÙCepÞJÒ`Tc
d
[23]。
IÚ
,
èéAá&¸¬»`QF
、
EDü
u»`62j|GÁ°H=Ä&|dTë`
Y
、
Gà*CT?9K
,
JL§DÙCHl+6
ÒUV
。
ÙC?9KÚÙC&KFÚð6§`oØT!
¥
,
67ÙCJÁFT=
、
NIFTñ?=
Ä
,
äë`]Ú,`ÙCF`
、
¢£ÙC¥
6cK9¤¥TßM«»F
[34],
òÙC´µK
ØÝ
、
9KO*=Ä
[56]。
þJ/0g
h
,
A§íîÙC?9KTë`
,
;ÅÙCD=
ö
,
sÙC¥
[712],
Ic/0Òâ
(
RPDâ
)
TA
,
LÍMNQRADÙC?9K
]TUVS+LbA
;
AJ/0dPCRDGGE
ÒâÍ®5QRAáDÛËæ¸ÙC?9KO
PepTUV
[13],
IÚAÚ~;9KOP
ä
、
[\?9K
I\nsÙC¥T=Ä¢
6zQ
。
ðé.
,
»/0jABïTDE-
(犘.
犪犿犲狉犻犮犪狀狌犿×犘.狆狌狉狆狌狉犲狌犿)ÉÃ-_n/0D
R
,
/0ABï8½#Ø_ºCÉÃ-_ÙC
¥|?9K:T<ð
,
(LAáäd_
êÙC¥TÁù
,
nAáÏd*+ÿÏ
。
1 UVWXY
1.1 Z[\^_
/0éd¦§¦yR49\()*¾J
Aºp
(
.ì)¯S¬T!åæT?¨
),E
118°09′738″,N27°22′435″。YBC®_é2012$
Ƽ½d
,
ÓÖ300m2,¬*pÜ,
Û6
,
$R©Û17.5℃,s`OU,$s`
1800mm,òÒ1850~1980h,ï#W300dj
S
,
ÙCnØ_ºC
,
ÙC®»ùðF
:pH4.72,
ci0.79g·kg-1,cj0.38g·kg-1,ck56.38
g·kg-1,JÁF15.28g·kg-1,J:i 73.91
mg·kg-1,J:j41.51mg·kg-1。
1.2 jkÛµ
/0ÔdÀÁij
,
²15MÓÖn6m2(2m
×3m)Tu(g1),¿Mu20.5m×1.0mT
rùGJ12r‘V?6{’DE-(犘犲狀狀犻狊犲狋狌犿.
犪犿犲狉犻犮犪狀狌犿×犘犲狀狀犻狊犲狋狌犿.狆狌狉狆狌狉犲狌犿犆犞.‘Min
muVI’),uã%jrPr,%ù1m。AáB
ï-_ãvéàN»`(AáR©»i`n0.47
g·L-1,»j`n0.66mg·L-1,cgn2.10
mg·L-1),ÆÐCÚi`_j.nÿϲÂA
áBï@A
。
»/0}Ï9¤^wÐW6Bï²Â
nÚN`200kg·hm-2,t²5MÓù,T1:DÒ,
7Bï¶çQ
;T2:AáBï`nW6BïT25%;
T3:AáBï`nW6BïT50%;T4:AáBï
`nW6BïT75%;T5:W6ïX,¿Óù3
*
,
¿10dBïß(s6B,Rs_ÏÙCYx
34UB
)。
ôö?-h°¼½Bï
,11%?-Z
[92Z[Bï
,
%fÙ:
。
1.3 d2®¯
LGJuañÍD6f
,
jñÍD6fTæ
n&!
,
¿ÍD6f®ñº2Mf:,t4
Mf:
。
dÙCf:f0~5,5~10,10~15,15
~20,20~25cmÙTÙC,?Q,d>=â=f×
100g¼Ù:ÜÊ^CËÄd;Uf0~20,20~40
cmñÙTÙC:1d>=â=fK`déÙCù
ðF
、
cJÁi
(Totalorganicnitrogen,TON),c
JÁg
(Totalorganiccarbon,TOC)6?9K`N6
CTé。
1.4 ²jXY
1.4.1 @A½¾a×、ÀlmMCpNEbc
ÙC®»ùð^Þ~
[14]
TÒâ
,
ÙCpH
ÔdWA²â
(
Q
/
Ù=2.5/1)é;JÁFÔd
äWkù©âé
;
cN(TN)Ôd\â©iâÃ
©
;
cj
(TP)6ck(TK)ÔdlWsXWKðâ
é
;
ÙC?9K`C6 NÒ¢jy[15]TXå
æ°âé
。
55
! " # $ !24"
`1 WùXYjkÛµ`
Table1 Thedesigntableofrandompermutationtest
BCÓùij Arrangementoftheexperimenttreatment
!ß Thefirstline T5 T4 T3 T2 T1
!å Thesecondline T2 T5 T4 T1 T3
!ñ Thethirdline T1 T3 T5 T2 T4
1.4.2 @AÀlm^_`]a ÙCDNA
f
:
ÔdFastDNASPINKitForSoil(Q(BIO
gene)TBÜßÆ}ÏBÜßTÒâfÙC
:1?9KcDNA。
Z©`PCR:=¾ÏdHÞ16SrDNA®5、
ÞSSUrDNA®56êiÞ狀犻犳犎 ®5T8ºÊ
K
(
g2),jÜM®5TäF]DNAnFÏ1[16],
ÔdSYBR(PremixExTaqTM(9KÃĺp,
TakaRa,Yå)BÜßé ABIPRISM7500RealTime
PCRSystemiíTS\^DZ©`PCR=。
=¾j10±@A^ÙHÞ16SrDNA®5、
ÞSSUrDNA®56êiÞ狀犻犳犎 ®5äF]F
Ïéf
。
1J:1\3*,uÍÖ~g21
,
uN]n25L,&2L^Ù5±TDNAÂP
îuá23L,uá³´12.5LSYBR(犘狉犲犿犻狓犈狓
犜犪狇犜犕(2x),0.5LROXReferenceDyeII(50x),v、
_ÊKÜ1L5pmol·L-1,°_9KÃĺp,S
ª
)
68LQÚQ。
PCRDGGE:=¾ÏdHÞ16SrDNA®5、
ÞSSUrDNA ®56êiÞ狀犻犳犎 ®58ºÊ
K
[1719]
\PCRií(g2),uN]50μL,&2
μL^Ù5±TDNAÂPîuá48μL,uá³
´1μLTaqDNA`½(2.5U·L
-1,
á}9Kº
p
,
:;
),5μLdNTPs(2mmol·L
-1
Ü*®
,
Sª9
KÃĺp
,
Sª
),5μL10×PCRbufer(10±T
PCR á)(á}9Kºp,:;),v、_ÊKÜ4
μL(5pmol·μL
-1,
Sª°_9KÃĺp
,
Sª
)
6
29μLQÚQ。PCR¤KÀaô¿`bEB(cðì
d
)
µp׿
。
·_d40LTPCR¤K,=¾Ôd
@An30%~55%(HÞ)、40%~50%(Þ)6
40%~60%(êiÞ)T8%T`efghij(ð+
<Ün100%"u7mol·L-1640%(NÖ¸)T
A5mkgh
)
L1± TAE á&150V60℃
`b5h。`b_d10mLSyBRgreenI(Sigma)(1
±TAE^Ù10000±)¨ Wµýµp45min,dBio
Radl]^mÒ。
`2 犘犆犚Z[\,f]vNO
Table2 PrimersandPCRconditionsusedforthestudy
µF9K
Target
group
ÊK
Primer
lj
Sequence(5′-3′)
PCRuÍÖ
ThermalprofileforPCRDGGE
Z©`PCRuÍÖ
ThermalprofileforrealtimePCR
^}
Reference
HÞ
Bacteria
F341
F341GC
R534
CCTACGGGAGGCAGCAG
ATTACCGCGGGTGCTGG
94℃S<5min,94℃<1
min、55℃ïn2min672℃Á
o2min,35MGH,72℃10min
95℃S<30s,94℃<5s,
60℃ïnÁo34s,40MGH
Muyzer
犲狋犪犾.,
1993
Þ
Fungi
FF390
FR1FR1GC
CGATAACGAACGAGACCT
AICCATTCAATCGGTAIT
95℃S<8min,95℃<30
s、50℃ïn45s672℃Áo2
min,35MGH,72℃10min
95℃S<30s,94℃<30
s,50℃ïn45s,72℃Áo2
min,40MGH
Vainioand
Hantul,
2000
êiÞ
N2fixer
PolF
PolFGC
AQER
TGCGAYCCSAARGCBGACTC
GACGATGTAGATYTCCTG
94℃S < 5 min,94℃ < 1
min、55℃ïn1min672℃Á
o2min,35MGH,72℃10min
95℃S<30s,94℃<5s,
60℃ïnÁo34s,40MGH
Poly
犲狋犪犾.,
2001
ü:g=¾LF341,FR16PolFÊKT5′?îßMGCp
Note:indicateGCclamp(5′CCGCCGCGCGGCGGGCGGGGCGGGGGCACGGGG3′)wasattachedtothe5′endofprimersF341,
FR1andPolF
1.5 lm>r
dQuantityOne(BioRad)ÕÖDDGGEë[
&DNAÍÜTÂ6~A\ë;ð,²ß?9
KOP:»ëShannonWeaverindex(H′)=
((n/N)×ln(n/N)(n=¿MÜT~A,N=@ß
:11JÜ~AT6
)[20]。
ÏdSPSS18.0^²Õ
Ö\ÒØ
(ANOVA)=6=,^ ²Ò
ânLSD¸t,غ
QR犘<0.05。
2 pq>r
2.1 oQÏÓ、J,1CwN
ädAá_
,
DÒ¸
,0~20cm620~40
cmÙÙCài、àj、6àk©íîóI,IØ
65
!1# ??:AáDDE-_ÙC?9KOPTUV
º©6
,
nJÁF»`(
s
,
& T3, T46T5
séDÒ(g3)。
`3 N^oQÏÓ
Table3 Thebasicphysicalandchemicalpropertiesofsoil
Óù
Treatment
0~20cm 20~40cm
pH
pHvalue
ài TN
/g·kg-1
JÁF
Organic
matter
/g·kg-1
àj TP
/g·kg-1
àk TK
/g·kg-1
pH
pHvalue
ài TN
/g·kg-1
JÁF
Organic
matter
/g·kg-1
àj TP
/g·kg-1
àk TK
/g·kg-1
T1 4.45±0.24a 0.67±0.08a12.82±1.96c 0.34±0.04a 55.5±0.502b 4.36±0.04a 0.81±0.25a 11.95±2.46c 0.34±0.12a 55.5±0.502b
T2 5.12±0.31a 0.73±0.08a14.15±1.26bc0.43±0.07a 55.8±0.212b 4.58±0.11a 0.62±0.03a 13.90±1.66bc0.44±0.03a 55.8±0.212b
T3 4.74±0.23a 0.89±0.15a 17.20±4.17a 0.34±0.08a 56.7±0.472a 4.43±0.08a 0.72±0.10a 14.96±1.45ab0.42±0.05a 56.7±0.472a
T4 4.55±0.30a 0.82±0.042a15.11±0.95ab0.37±0.05a 57.6±0.556a 4.48±0.08a 0.90±0.11a 18.64±3.05a 0.49±0.02a 57.6±0.556a
T5 4.75±0.31a0.84±0.13a 17.10±1.67a 0.40±0.02a 56.3±0.272a 4.52±0.05a 0.73±0.20a 15.17±2.83ab0.37±0.06a 56.3±0.272a
ü:@j6@uU;Çg6@Óù%غ
(犘<0.05),@
Note:Differentlowercasesinthesamecolumnindicatesignificantdifferenceatthe0.05level.Thesameasbelow
AáïX-__ÙC?9K`C(SMBC)6N
(SMBN)~g41。0~20620~40cmÙ
ÙC?9K`N6C©ÀìAáBï`Tíîn
íî
,
& T5W6Bï(iCÚ`n:200kg·
hm-2)ÉÃ-_ÙC?9K9K` N6CÝs。
0~20cmÙ,T2~T5Óù?9K` N©
séDÒT1Óù,T46 T5ÓùA
sé T2
Óù
,
XÓùã%(ï
غ
;T46T5Óù
?9K`C
séDÒ6XÓù,n T46
T5ã%غAWd
QR。ghAáBï§
íî0~20cmÙÙC?9K`。20~40cmÙ
,T5Óù?9K` N
séDÒ6XÓ
ù
,
nT2~T5ÓùT?9K`C
séDÒ。
=gh
,
AáBïQR?9K`C6
NT]ë=¾n0.83360.824,©Wdú
QR
(犘<0.01)(g5)。
`4 c~_犛犕犅犖w犛犕犅犆
Table4 SMBNandSMBCofdifferentsoillayers
Óù Treatment
0~20cm 20~40cm
SMBN/mg·kg-1 SMBC/mg·kg-1 SMBN/mg·kg-1 SMBC/mg·kg-1
T1(CK) 9.02±1.81c 63.74±9.91d 7.25±0.90b 58.32±1.94b
T2 22.78±2.72b 136.40±43.81cd 8.58±1.55b 263.20±14.53a
T3 34.41±3.98ab 172.67±34.96c 15.98±7.64b 145.87±2.84a
T4 43.81±0.16a 377.59±88.44b 14.76±3.14b 117.12±1.394a
T5 38.92±12.71a 524.62±8.68a 35.28±9.86a 195.98±3.09a
2.2 J,Nø`
2.2.1 @AØÙ16SrDNAÚÛI ÏdZ
©`PCR¾¨DHÞ16SrDNA®5ëA\=
,
ef~ë11。0~5,5~10610~15cmÜ
Ùã%16SrDNA®5ëAغ6Y。ÀìAá
Bï`Tíî
,
ÜÙÙCHÞ16SrDNA®5ë
AAíî
。0~5,5~10610~15cmÙ,T5Ó
ùHÞ16SrDNA®5ëA
séT1(DÒ)|
XÓù
,T36T4ÓùA
séDÒ6T2Ó
ù
,0~5610~15cmÙT26T1ã%غA
Wd
QR
。
n15~20620~25cmÙ T4
6T5Óù
séT1,T26T3Óù,T26T3
T1ã%AWd
QR。ghAáBï_§í
î0~25cmÙÙCHÞ®5qëë,[\»16S
rDNA®5T?9KT
I,"#»16SrDNA®
5THÞë`íî
。
=gh
,
AáBïQ
RÙCHÞ16SrDNA®5ëAT]ën
0.968,Wdú
QR(犘<0.01)(g5)。
=1 ab16犛狉犇犖犃Nø`
Fig.1 Geneabundanceof16SrDNA
75
! " # $ !24"
=2 cb犛犛犝狉犇犖犃Nø`
Fig.2 GeneabundanceofSSUrDNA
=3 dKb狀犻犳犎 Nø`
Fig.3 Geneabundanceof狀犻犳犎
2.2.2 @AÜÙSSUrDNAÚÛI èë2L
ã
,
ÀìAáBï`Tíî
,
ÙCÞSSUrDNA
®5ëAAíîóI
。T46 T5ÓùÜÙÙ
CÞSSUrDNA ®5ëA
sé T1,T26
T3Óù,0~565~10cmÙT26T3ÓùA
séDÒT1Óù,n10~15,15~20620~25
cmÙT26T3ÓùDÒT1ã%غ6
。
ghAáBï_§íî0~25cmÙÙCÞ®
5qëë
,
Þc`íî
,
ò.Gíî:LG
gPónh
,
ÀìÙAíî.Gcd
óI
。
=gh
,
AáBïQRÙCÞ
SSUrDNA®5ëAT]ën0.972,Wdú
QR
(犘<0.01)(g5)。
2.2.3 @AÝÁÙ狀犻犳犎ÚÛI ÀìAáBï`
Tíî
,
ÙCêiÞ狀犻犳犎®5ëAóI(ë3)。
0~5cmÙ,ÜÓùÙCêiÞ狀犻犳犎®5ëA©
éDÒT1Óù,T5T4
éT36T2,T5
T4ã%غAW
QR。5~10610~15cmÙ
T5Óù
éDÒ,XÓùDҸغ6
。15~20620~25cmÙT46T5ÓùêiÞ
狀犻犳犎®5ëA
éDÒT1,&15~20cmÙ
T26T3DҸغ6
,20~25cmÙT3
Óù
séDÒ
,T2DÒT1ã%غ6
。
ghAáBï_§0~25cmÙÙCêiÞ
狀犻犳犎®5qëë,êiÞc`,ò.G:
L0~5cmÙgPÝnh
。=gh,A
áBïQRÙCêiÞ狀犻犳犎 ®5ëAT]ë
n-0.779,IöWd
QR(g5)。
2.3 J,-.adÏ
AáBïDÙC?9KOP:TUV~ë
4~61。èë4Lã,6@@ATAáBïDÜ
ÙHÞ:ï
UV
,T5Óù20~25cm
ÙHÞOP:
íî
。
n6@`TAáB
ïD0~25cmÜÙÞ、êiÞ:©ï
UV
(
ë56ë6),ghAáBï$_DÙC?
9KOPäïh
UV
。
=4 ab-.adϹl
Fig.4 Diversityindexesofbacterialcommunities
=5 cb-.adϹl
Fig.5 Diversityindexesoffungicommunities
=6 dKb-.adϹl
Fig.6 Diversityindexesofazotobactercommunities
85
!1# ??:AáDDE-_ÙC?9KOPTUV
`5 STefóôWJ,1fJ,Nø`29Ï
Table5 Therelationshipsbetweenbiogasslurryvolumeandmicrobialbiomassormicrobialgeneabundance
?9K`C
SMBC
?9K`N
SMBN
16SrDNA®5ëA
16SrDNAgeneabundance
SSUrDNA®5ëA
SSUrDNAgeneabundance
狀犻犳犎 ®5ëA
狀犻犳犎geneabundance
AáBïQR
Concentrationofbiogasslurry
0.833 0.824 0.968 0.972 -0.779
ü:g
(犘<0.05),gú
(犘<0.01)
Note:indicatesignificantcorrelationatthe0.05level,indicatesignificantcorrelationatthe0.01level
3
ÙC?9K³´HÞ
、
rHÞ
、
>fÞ
、
Q
?
ep?9K?.¨?9K|Þ
、
sà
(
rsõe
)、
_t?¨9K
[21],Alison6 Martiny[22]ceX
ÉT/0_°P
,
ÙC?9KOPäD N,P6
KÈP/0,n.G/0,5()øù¡äT
6@n+Lغ
。
.e
,
ÙC?9K:cJK
àF
、
ÙCF_ÙCøù¡ä?5uTUV
[2324],
ò}wÙC?9KcJKTUVÄAQ=}Ù
C
[13]。
.©?
[14]
/0i½
,
AáÓùLsuv
(犆犪狆狊犻犮狌犿犪狀狀狌狌犿)}wÙCHÞ、ÞT:
ȑ
,
n»/0Tefgh
,
AáBïÉÃ-_$
ã_
,
}ÙCHÞ
、
Þ|êiÞ:©ï
<ð
,
.L§Ú5n}ÙC?9KOPäT/
0ué}wÙC+T
。
ÙC?9Kä6ë`Ú,`ÙCF`
、
¢£
ÙC¥6cK9¤¥TßM«»F
[45],
nÙC
?9K67ÙCEDüuTÚð
,
J
Kã%]+LDEDüuThi^_
[25]。
Aá
&ì»N,P6 K?ED=、G?`üu(~
Fe,Zn,Cu,B,Mo?)6JK9u(~JKu、L
MìW?
)、
¢9u
、
½?9ýÉKF
[26],
Yn
LÛ[^_T=
。
èé/0TÙCtnw
x
,
-AáBïØ_ºC2
,
ÙC?9Khi^_
Aá&TED=
,
ÙC?9K9K`C6N
s
,
nÙC&TN,TP6TK»`(ï
<
ð
。
Z©`PCRTefAgh,AáBï[\Ù
C&»16SrDNA®5THÞ6»SSUrDNA®
5TÞY`
I
,
íî16SrDNA ®56Þ
SSUrDNA®5ëA,I)¡»狀犻犳犎 ®5Tê
iÞT
I
,
狀犻犳犎 ®5ëA
。Coelho
?
[27]
Lsy
(犆犺犻狀犲狊犲狊狅狉犵犺狌犿)ÙC&°P,狀犻犳犎
®5ëAÀiäd`Tíîn
,
.»/0
efß#
,
n Ogilvie?[28]6Juraeva?[29]T/0
z°P
,
iTäLíî狀犻犳犎 ®5TëA,.L
§iä`J
。
èéÙCêi?9KTÉ
cÙC C/N TUV[2526],ÙC C/N
[3031],
pH W[32]。»/01BïTAá
C/Nt,îSPÙCnØ_ºC,pH
(4.36~5.12),.L§Ú"#»/0êiÞ/)
TÌ«.5
。
4 p
0~20620~40cmÙÙCTN,TP6TK
»`ï
<ð
,
nÙC?9K`N6C©ÀìA
áBï`Tíîníî
,
ò?9K`C6NAá
BïQRú
(
g5),]ë=¾n
0.83360.824,&iCÚ`n200kg·hm-2T
W6Bï
(T5)ÙC?9K9K`N6CÝs。
T1DÒ¸,0~25cmÜÙT2~T5Ó
ùÙCHÞ16SrDNA®5ëA6ÞSSUrDNA
®5ëAYú=
íî
,
òÙCHÞ16SrDNA
®5ëA6ÞSSUrDNA®5ëAAáBï
QRTú
,
]ë=¾n0.9686
0.972。@2,0~5cmÙÙC狀犻犳犎 ®5ëAÀ
AáBï`Tíîn
,
XÙ7Ls×
AAáBï
(T5$T4)2ÙC狀犻犳犎 ®5ëA
,
@2AáBïQRÙCêiÞ狀犻犳犎 ®
5ëAgPnW
,
IöWd
QR
。
6@A
áBï`D0~25cmÜÙÞ、êiÞ:©
ï
UV
。
AB
[1] ª{z,ª|½,xµ,?.£ÂðDIêFÌ«J±=
é=/0
[J].JKEDý+i,2005,11(6):822
829
[2] eeV,{},~¾,?.AáäkDf¸ÙCùð8
6?9K|f^1FUV
[J].&()*+,2010,43
(22):46714677
[3] ,,,e,?.AáT¸¢_KÓùÏd|D
ÙC?9Kg
、
i½ÉTUV
[J].()Hl*++i,
2012,31(10):19651971
95
! " # $ !24"
[4] ¡,
N¶.ã6@ËÄÒâDÙC?9K6½ÉT
UV
[J].ÙC+i,2005,42(2):348351
[5]
Ø,eª,{õv,?.I4ËÄncÒâDÙC
|ù»zQ=Ïd:ÝTUV
[J].()ÃÄ+i,2005,21
(1):5256
[6] HenriksenT M,BrelandT A.Carbonmineralization,fungal
andbacterialgrowthandenzymeactivitiesasaffectedbycon
tactbetweencropresiduesandsoil[J].BiologyandFertilityof
Soils,2002,35(1):4148
[7] øÉ,Å,{ð,?.äAáDu^}wÙC9KÉ
TUV
[J].^àcK+i,2010,30(4):721726
[8] ª»R,x,&,?.6@JÁðäD§ãÙC
?9KÉTUV
[J].ÙC[9] ¼õ,/R,¹Å.AáD6råcÙC9K+FT
UV
[J].dHl9K+i,2011,17(3):384387
[10]Å,øÉ,{¾à,?.AáðäDu^}wÙC?
9Kë`6½ÉTUV
[J].d9\+i,2011,22(4):
10071012
[11]x*,,{kz.AáLåæSTd|ÙCF`
:
[J].&A,2009,27(1):4347
[12]AbubakerJ,CederlundH,ArthursonV,犲狋犪犾.Bacterialcom
munitystructureandmicrobialactivityindifferentsoilsamen
dedwithbiogasresiduesandcattleslurry[J].AppliedSoilE
cology,2013,72(10):171180
[13].©,P¹Y,{,?.AáädDuvTÜ :f|D
ÙC9K+8TUV
[J].&()*+,2013,46(3):507
516
[14]Þ~.ÙC()ð+=Òâ[M].:;:&()*¾O
Ó
,2000
[15]¢jy.ÙC?9K/0.ùÒâ[M].:;:s?öJO
Ó
,2010:73
[16]HeJZ,ShenJP,ZhangLM,犲狋犪犾.Quantitativeanalysesof
theabundanceandcompositionofammoniaoxidizingbacteria
andammoniaoxidizingarchaeaofaChineseuplandredsoilun
derlongtermfertilizationpractices[J].EnvironmentalMicro
biology,2007,9(9):23642374
[17]MuyzerG,DeWaalEC,UitterlindenAG.Profilingofcom
plexmicrobialpopulationbydenaturinggradientgelelectro
phoresisanalysisofpolymerasechainreactionamplifiedgenes
codingfor16SrRNA[J].AppliedandEnvironmentalMicrobi
ology,1993,59(3):695700
[18]VainioEJ,HantulaJ.Directanalysisofwoodinhabitingfun
giusingdenaturinggradientgelelectrophoresisofamplifiedri
bosomalDNA[J].MycologicalResearch,2000,104(8):927
936
[19]PolyF,MonrozierLJ,BalyR.ImprovementintheRFLP
procedureforstudyingthediversityofnifHgenesincommuni
tiesofnitrogenfixersinsoil[J].ResearchinMicrobiology,
2001,152(1):95103
[20]ZakJC,WiligMR,MoorheadDL,犲狋犪犾.Functionaldiver
sityofmicrobialcommunities:aquantitativeapproach[J].Soil
Biology&Biochemistry,1994,26(9):11011108
[21]JenkinsonDS,LaddJN.Microbialbiomassinsoil:Measure
mentandturnover[M].NewYork,1981:415471
[22]AlisonSD,MartinyJBH.Resistance,resilience,andre
dundancyinmicrobialcommunities[J].ProceedingsoftheNa
tionalAcademyofSciencesoftheUSA,2008,105(S1):11512
11519
[23]¹,.ÙC?9K:UV5u|/0ÒâTP
±
[J].9K:,2007,15(3):306311
[24]eª,ªÏ/,ª1L.ÙC?9KOPepUV5uTWX
[J].¡¢£Y
ersonsoilmicroorganismsAReview[J].SoilBiology&Bio
chemistry,2014,75(8):5463
[26]{44,x2.AáÓÏd/0\±[J].()*+,
2007,35(4):10891091
[27]CoelhoMRR,MarrielIE,JenkinsSN,犲狋犪犾.Molecularde
tectionandquantificationof狀犻犳犎genesequencesintherhizo
sphereofsorghum(犛狅狉犵犺狌犿犫犻犮狅犾狅狉)sownwithtwolevelsof
nitrogenfertilizer[J].AppliedSoilEcology,2009,42(1):4853
[28]OgilvieLA,HirschPR,JohnstonAW.Bacterialdiversityof
theBroadbalk‘classical’winterwheatexperimentinrelation
tolongtermfertilizerinputs[J].MicrobialEcology,2008,56
(3):525537
[29]JuraevaD,GeorgeE,DavranovK,犲狋犪犾.Detectionandquan
tificationofthe狀犻犳犎 geneinshootandrootofcucumber
plants[J].CanadianJournalof Microbiology,2006,52(8):
731739
[30]ªé,{/,e=,?.ñs¥-Ùêi®5
(狀犻犳犎)T:6]^°J/0[J].?9K+i,2005,45
(2):166171
[31]ë¹Â,ÃS,Ë+,?.ÍÖ6cKDÙ6Ùêi
?9KOP:TUV
[J].ÙC+i,2012,49(1):130
138
[32]ShuW,PabloGP,JunY,犲狋犪犾.Abundanceanddiversityof
nitrogenfixingbacteriainrhizosphereandbulkpaddysoilun
derdifferentdurationoforganicmanagement[J].WorldJour
nalofMicrobiologyandBiotechnology,2012,28(2):493503
(
%&( ,-.)
06