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Molecular Identification and Analysis of Phylogenetic Position for Two Truffles

2种块菌的物种分子鉴定与系统地位分析



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,2016,36(8):1560-1565
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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犕狅犾犲犮狌犾犪狉犐犱犲狀狋犻犳犻犮犪狋犻狅狀犪狀犱犃狀犪犾狔狊犻狊狅犳犘犺狔犾狅犵犲狀犲狋犻犮
犘狅狊犻狋犻狅狀犳狅狉犜狑狅犜狉狌犳犳犾犲狊
FANYongjun1,ZHAOYanling1,CHENXiuli1,YANWei2
(1DepartmentofBiologyScienceandTechnological,BaotouTeachersColege,Baotou,InnerMongolia014030,China;2For
estColege,InnerMongoliaAgriculturalUniversity,Hohhot010018,China)
犃犫狊狋狉犪犮狋:TwotrufflesweretakenasexperimentalmaterialswhichwerefromHelanmountainareainInner
Mongolia,andtheywereidentifiedbasingonthemorphologicalandmolecularmethods.Intheprocessof
experiment,theDNAsequenceswereblastedusingtheGenBank,andphylogenetictreeswereconstructed
byusingMega5.1software.Theresultsshowthattwostrainsofascomaarehypogeous.Thereinto,or
deraissalow,subsphaeroidal,toirregularshape,andwithmicrovilionsurface.Andtuberbisyelow
ishbrown,uniformtexture,withawartonsurface,andthereisaspecialaromaofdriedfruit.Through
theanalysisofmolecularbiologicalmethods,orderawasidentifiedasaspeciesof犜狌犫犲狉genus,tuberbis
犜狌犫犲狉犫狉狌犿犪犾犲,andthetuberbisidentifiedasanewrecordspeciesinInnerMongolia.
犓犲狔狑狅狉犱狊:犜狌犫犲狉;ITS;speciesidentification;phylogeneticposition
  c<(truffles)»Ãn‘<ÄÅ(Ascamycoti
na)c<+(Tuberaceae)c<»(犜狌犫犲狉),ÆQ…Ç
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50~100mg,[?LƒCTABpqr|DNAéå。
gef”PCRê>Öëçìd¼íîïð2
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Table1 Informationofsample
ÆÇ
Sample
4

¹’
Latitudeandlongitude
ùú
Altitude/m
ordera 38°51′40″/105°57′05″ 2548
tuberb 38°51′55″/105°54′46″ 2353
165185              123,è:2bcDNAmnwxy”6728mnuqr{ž,Û$
,kÜ}? Mega5.1~,[? NeighborJoining
p9{|p|ordera、tuberba28әp:Ó
qri†u€‚ƒq„…

‰t‡i7_Š

3 ‹ŒQi†
3.1 STU^_`Uab
<ordera@!1,A、BÖ;,n‘Œ_“J;
•–—

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–—

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n

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:æç<”©éåš,N?DNA,¹øùW
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3.3 >?efg7hi=9B
3.3.1 犖犲犻犵犺犫狅狉犑狅犻狀犻狀犵狋狉犲犲7hi k? Gen
BankamnqÎp:Órs‚ƒ(BLAST),Y?
›2%GtZ,Ny”au»Ãc<»
(犜狌犫犲狉)a
ITS`bmn,v½c<(犜狌犫犲狉犫狉狌犿犪犾犲)、wxc
<
(犜.犫狅狉犮犺犻犻)、yc<(犜.犪犲狊狋犻狏狌犿)、zêc<
(犜.犮犪犾犻犳狅狉狀犻犮狌犿),{¼bcA.orderaÉÎlm‹€(×45);B.ordera×n€£‹€!(×400)
C.tuberbÉÎlm‹€(×45);D.tuberb×n€£‹€!(×400)
!1 2bcA.Morphologicalfeatureofordera(×45);B.Microstructurefeatureoforderaspores(×400)
C.Morphologicalfeatureoftuberb(×45);D.Microstructurefeatureoftuberbspores(×400)
Fig.1 Morphologicalfeatureandmicrostructurefeatureoftwotruffles
2651 ! " # $ % &                   36|
A.ÆÇPCRä$þÿ‹Œ;B.Æǹ„+,#þÿ‹Œ;
a.ÆÇorderaaDNA}÷‹Œ;b.ÆÇtuberbaDNA}÷‹Œ
!2 ÆÇPCRä$™¹„+,#aøùþÿ‹Œ
A.Amplificationproductsofthesamples;B.Purificationof
amplificationproductsofthesamples;aisresultsofgel
electrophoresisofordera;bisresultsofGelelectrophoresis
oftuberb
Fig.2 ThecheckresultsofPCRproductsand
purificationproductsofthesamples
W2 jklmno
Table2 Thestateofreferencesequence
~
Number

Name
GenBank~
GenBanknumber
1 ½c<犜狌犫犲狉犫狉狌犿犪犾犲 FM205639.1
2 FM205699.1
3 FM205552.1
4 wxc<犜狌犫犲狉犫狅狉犮犺犻犻 JN392229.1
5 JN392228.1
6 AF003920.1
7 yc<犜狌犫犲狉犪犲狊狋犻狏狌犿 AY226036.1
8 HQ706000.1
9 HQ705997.1
10 zêc<犜狌犫犲狉犮犪犾犻犳狅狉狀犻犮狌犿 DQ478657.1
11 DQ478659.1
12 DQ478658.1
13 xc<犜狌犫犲狉犾犻狌犻 DQ898182.1
14 ”Åc<犜狌犫犲狉狕犺狅狀犵犱犻犪狀犲狀狊犲 DQ898186.1
15 DQ898187.1
16 É7‚b犆犺狅犻狉狅犿狔犮犲狊狏犲狀狅狊狌狊 JF300146.1
™Y?”Åc<
(犜.狕犺狅狀犵犱犻犪狀犲狀狊犲)mn2ã、
xc<
(犜.犾犻狌犻)mn1ã(2)[19]。Dv|Öh
3mnqrœOӃ„™É7‚a‰t

*#k?
Mega5.1~[?NeighborJoining(p9{|p)
|ordera、tuberba28әp:Óqri†u
€q„…

u…\u»Ãc<+
(Tuberaceae)a
犆犺狅犻狉狅犿狔犮犲狊狏犲狀狅狊狌狊dÉ7‚。
3.3.2 6p狅狉犱犲狉犪>?ef9B @!3Ö;,
w†;‡ˆ‰

!42
!3 ordera‚ƒq„…
Thevaluerepresentingthesupportrate,thesameas
Fig.3Thephylogenetictreeofordera
365185              123,è:2bc!4 tuberb‚ƒq„…
Fig.4 Thephylogenetictreeoftuberb
:q„…ÜJZŠ‹

½c<

wxc<

yc<

z
êc
‡ˆ‰’‘Ã96%。‡”,
Ӂc<

xc<ŽŒ¼

‡ˆ‰d86%。J
ZŠ‹”Åc<

xc
V
orderaQ½c<、wxc<Œd¼T,‡ˆ‰d
95%;p{½c‚‘˜

‡ˆ‰d61%;’É7‚犆犺狅犻狉狅犿狔犮犲狊狏犲
狀狅狊狌狊ž“。:Lj‰ž¶,¾¿‰tdc<»
a¼ãb

”ëq¼¿[Æst

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3.3.3 6p狋狌犫犲狉犫>?ef9B @!4,$q
„…Jid4T。‡”,gBCÆÇd¼;yc
<
(犜.犪犲狊狋犻狏狌犿)、zêc<(犜.犮犪犾犻犳狅狉狀犻犮狌犿)•d
¼

Ӂc<
(犜.狕犺狅狀犵犱犻犪狀犲狀狊犲)、xc<(犜.
犾犻狌犻)ŒdŽ¼,‡‡ˆ‰d92%,–Ž‡Mˆ
‚ž˜

Ž¼dÉ7‚
(outgroup)。gefÆ
ÇQÖk3½c<(犜.犫狉狌犿犪犾犲)Œd¼,‡ˆ
‰ž‘

K87%,–Ž‡Mˆ‚ž˜。N$‹—
‡lm%YS¾¿‰t$bd½c<

!4)。
4 ˜ ™
gBC”¹|ÆÇorderaqr€‚ƒq„
6[…i†

štorderadc<»a¼b,Q‡›
c,orderaQwxc<M
ˆ‚‡ˆ‰d61%;œÛlmij‹€o ,G
«ž9p:8‰

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?
[J].RäÇz],2011,(1):6367.
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[2] ¥¦,· ö,·w½,è.”c<9¾™‡tu¿µ
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,2003,16:7385.
XUAS,ZHENGWL.Researchcommentonfungioftuber
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