4
,t CO ,`zñ0N7ÆÙ3 DNAS¢=¹j \Ù¦¬`a båc !",->Eé<r^JEF!"$% 316022 /0 78K«FdÇéS{>S[\。[ [\_`ûl"lGVCO VkÈyAqr8 DNAeZ&C fgééééóÝ8vZé>%º>S。[ st20 UBÝy 18 UüD˯öKëß]^ NCBI xØC Blast (9C %8Z³10 UfgééBÝ8>SstCõÎ>Sst´x8¨>S 10 UééóÝBÝyhiy) äC jÝÓk;cUBÝr÷。[ sDNAe¨Çlm8Zï>S£¤)Ç$ùÊéégÝV%vZCL´ óÝ8S{>Sn²NÝ8©ðF:%8£¤Ñk123 DNAeS{>&ééCO Vk 45678 S917.4  9:;<= A  9>?8 0517-6611 2018 28-0092-04 IdentificationofMainEconomicCephalopodsandProductsbyDNABarcodingBasedonCO Gene ZHOUGuangdong DENG Shanggui HUOJiancong  SchoolofFoodandMedicine ZhejiangOceanUniversity Zhoushan Zhejiang 316022 Abstract Objective Toestablisharapidandefficientmethodfortheidentificationofcephalopodspecies. Method Inthispaper the COgenewasusedasthegeneralDNAbarcodingofthistest.Thecompositionofthefrozencephalopodsandproductswereidentifiedrespectively. Re sult SepecificstrapeswereamplifiedbyPCRin18outof20samples.Theidentificationresultsofthetenfrozencephalopodswereconsistentwith themorphologicalidentificationresults whilethetencephalopodproductsweremainlysquit andtwowereinconsistentwiththecommoditylabels twosamplesfailedtoverify. Conclusion DNAbarcodingisanefficientandconvenientmolecularidentificationtechnology whichcanbeeffec tivelyappliedtospeciesidentificationofcephalopodfrozenproductsandtheirrelativesingleproducts itprovidesbettertechnicalsupportforthe supervisionofrelatedproductsonthemarket. Keywords DNAbarcoding Speciesidentification Cephalopods COIgene CDEF \Ù¦1994 —), ¸Þ(½ÂÃ¥¦b¥¦[¨mNÝECop]ÇÈÉKê«ÃbHÁ«ÃÄÅmNÝECop¥¦GHIJ 2018-05-10 ¤,Z'(<1785Mygh6é<ÒºèÊ 768D¤,Z8ZÜ8B4(Hz{J@B¼ºJ @6Þ#×u6¤,Z<8|´ÞWn@ô-Ùx4 uG'(<¤,ZwX|´yiÙYÞÄtp v3¬í§7lèÊ7ò(GH²s(<60 ð&ZCh(s4¤,Z(<WÁÑbJJ6=¦ ²É3|´.~bPäé<ð÷8y6Z 3U¤,Z(<6EÖF4ÃJ)ODNA}bfÝÞ1îïðâ-x#h6@ÅjøÝ ÞÐ7DNA}bfÑÒB¬>@ZEU PaulHebert ¤ 1ÑVF²³7}ÔMÿF86w6X, Ô'#6Z]µ&axå6 DNAª5!ÝwæÒhj øF86¾ 1-2 DNA}bfOPÍÃ78w¢ È\ÝÞö÷lWß[÷F86w DNA¤ ðr DNA}bf-?MÖtsØÑOX-ø[ ÷À<6F8 DNA}bf3617&xJ6F 8jøÝÞ|3 2011 ï6é<rJ<¦ÍR~¼G w)ÿÝÞ3é<rJ<ÏT6ÐÑÝ6³ÙñÝÞ6 )*xQ)*¾_ôõ54àUJKtP_X ñ'(<6)*ÁÌ4Ý945 ZHt CO m ¦@s 32 8s^6ó¢nJ8ä8³ÝÕËÖ t)*âçÑ~1u7ʳb»EpqÙY×ZS@6 s^ÜÔGH DNA}bf¹ÞX-6jm4 CO m ¦6 DNA}bfÝÞ3s^`¹)x«6ÐÑÿ)* À CO ¤,Z6ßöGÑF,ÉtF8jPÓ50y-6¤,Zjø¾ é<I&ré<=¦¤<ÝÞw=m<1 KLMNO 1.1 KLMUe ô|¤,ZÀ<B$%Öönö R¤<¤,ZÉ<JZ+ùdZ@!$%6/'( >û³`Àô|¤,ZÀ<4-45℃XY5 W2Ñ+ù,ZÉ<4}~UW2Ñ('ê|#K|¢¦½`,6Jõì¢Ev êX c » ddH 2× TaqDNA ILDNAMarker DL2000 )、 GelRed ó`,VíVFíãXc»ghL V6 Merck »1.2 ÑÒMû¯ ô|Ë])V6 Sigma »NanoPho tometerTMl,óngh÷ø©V6 Implen »DYY-8C ÒA©b17©«2720 Ò PCR©6 Applied Biosystems » @£Jû6 Bio-Rad »1.3 NO 1.3.1 À<(ÌÍÍsGõ6Àt+,L1~L10 òb»}à6ô|¤,ZZ1~Z10 ¤,ZÉ<5 Z1~Z3 ô|L]Z4~Z6 L]iZ7 x]RZ8~ Z10 x]JÀ¤,ZÉ<5×Mxy6À<L]itÖtÍM ÌÍÑsÚ/Þe 5g À<±e°4 50mL Ë]¦ 5¬. 5mL MiNÌ 30s §4 45℃¬'«-5' «¬É 3h ÀË]¦ 10000r/min Ë] 5min §ÿ`°× M¬. 15mL70%ê|¨eNÌ 2min 10000r/min Ë] 5min ȽΠ2~3 Ŭ.('ê| 15mL NÌ 2min 10000r/min Ë] 10min ÿ`?e±4 65℃¬|Y WXYZ[\J.AnhuiAgric.Sci.2018 46 28 ): 92-95

t Ⅰ,‘zæ0ŁŒŽN7˘Ù3– DNA · ,t COⅠ,‘zæ0ŁŒŽN7˘Ù3– DNAS¢=„j \Ùƒ,‹‘a,båc (!",->EØ

  • Upload
    others

  • View
    0

  • Download
    0

Embed Size (px)

Citation preview

Page 1: t Ⅰ,‘zæ0ŁŒŽN7˘Ù3– DNA · ,t COⅠ,‘zæ0ŁŒŽN7˘Ù3– DNAS¢=„j \Ùƒ,‹‘a,båc (!",->EØ

,t

COⅠ,`zñ0���N7ÆÙ3�

DNAS¢=¹j

\Ù¦

,¬`a

,båc

 (!",->Eé<r^JEF

,!"$%

316022)

/0

 [78

]K«Fd�Ç8éé�S{>S[\

。[[\

]�_`ûl"�lGVⅠ(COⅠ)VkÈyAqr8

DNA��e

,Z&C

fgéé��éé�óÝ8vZé>%º>S

。[st

]20UBÝy

18U�üD˯öKë�ß

。]^

NCBIxØC

Blast�(9C

�%�8Z³��

,10Ufgéé�BÝ8>SstC�õÎ>Sst´x

,8¨>S

10Uéé�óÝBÝy�hiy�

,)

2äC

jÝÓk;c

,2UBÝ��r÷

。[s�

]DNA��e5´¨�Çlm8Zï>S£¤

,�)Ç$ùÊéé�gÝV%vZ�CL´

óÝ8S{>S

,y«n�²NÝ8©ð�F:%8£¤Ñk

123

 DNA��e

;S{>&

;éé�

;COⅠVk

45678

 S917.4  9:;<=

 A  9>?8

 0517-6611(2018)28-0092-04

IdentificationofMainEconomicCephalopodsandProductsbyDNABarcodingBasedonCOⅠGeneZHOUGuangdong,DENGShanggui,HUOJiancong (SchoolofFoodandMedicine,ZhejiangOceanUniversity,Zhoushan,Zhejiang316022)Abstract [Objective]Toestablisharapidandefficientmethodfortheidentificationofcephalopodspecies.[Method]Inthispaper,theCOIgenewasusedasthegeneralDNAbarcodingofthistest.Thecompositionofthefrozencephalopodsandproductswereidentifiedrespectively.[Result]SepecificstrapeswereamplifiedbyPCRin18outof20samples.Theidentificationresultsofthetenfrozencephalopodswereconsistentwiththemorphologicalidentificationresults,whilethetencephalopodproductsweremainlysquit,andtwowereinconsistentwiththecommoditylabels,twosamplesfailedtoverify.[Conclusion]DNAbarcodingisanefficientandconvenientmolecularidentificationtechnology,whichcanbeeffectivelyappliedtospeciesidentificationofcephalopodfrozenproductsandtheirrelativesingleproducts,itprovidesbettertechnicalsupportforthesupervisionofrelatedproductsonthemarket.Keywords DNAbarcoding;Speciesidentification;Cephalopods;COIgene

CDEF

 \Ù¦

(1994—),̧ ,��Þ(½

,ÂÃ¥¦b

,¥¦[¨

mNÝE�Cop

。]ÇÈÉ

,Kê

,«ÃbHÁ

,«Ã

ÄÅmNÝE�Cop¥¦

GHIJ

 2018-05-10

  ¤,Z'(<1785Mygh6��é<

,ÒºèÊ

768D

。¤,Z8ZÜ8

,B4(H

、z{J@B¼ºJ

@6Þ#

,×u6¤,Z<8|´Þ�Wn@ô­

。Ùx4

uG'(<

,¤,ZwX|´y

、�iÙY

、��Þ�Ätp

v

,�3¬í§

,7lèÊ7ò(�GH�²s(<60�

ð&�ZC�

,h(s4¤,Z(<WÁÑbJJ�6=¦

²É

。3|´.�

、1Ö~bPäé<ð&ö÷8�y6Z

3U

,¤,Z(<61Ö

、EÖ

、�F4ÃJ�)O�

DNA}bfÝÞ1îïðâ-x�#h6@ÅjøÝ

ÞÐ7

。DNA}bfÑÒB¬�>@ZEU

PaulHebert¤

,1��ÑVF²³7}�ÔM�ÿF86

、�w6

、X,

Ô'#6

、Z]µ�&axå6

DNAª5

,!ÝwæÒhj

øF86¾�

[1-2]。DNA}bfOPÍÃ�78�w¢�

È\�ÝÞ

,ö÷lWßkÀ[÷F86�w

DNA¤��

ð

,r

DNA}bf-?MÖtsØ

,ÑOwæX­H½ø[

÷À<6F8

[3]。DNA}bf36�17&x�J�6F

8jøÝÞ

,|3

2011ï�6é<rJ<¦ÍR~�¼G

w)ÿÝÞ3é<rJ<ÏT6ÐÑ

。Ý6³ÙñÝÞ6

)*xQ

,�)*¾_�ôõ54àUJKtPF¾_

,X

ñ'(<6)*ÁÌ4Ý945

[2,4]。ZHt

[5]bÑ

COⅠm¦@�s

328s^6ó¢n�J

、8�ä8³ÝÕ�ËÖ

t)*âç

,Ñ~1u7ʳb»EpqÙY�×ZS@6

s^Ü�ÔGH

DNA}bf¹ÞX­6j�

,m4

COⅠm¦6

DNA}bfÝÞ3s^`�¹)x«6ÐÑ

。ÿ)*

À

COⅠm¦��¤,Z6�ß

,�öGÑ�F

,s¤,Zä

�É<ÖtF8j�

,PÓ50wæy­6¤,Zjø¾

,�é<I&ré<��=¦¤<ÝÞw�BÍ=m<

1 KLMNO

1.1 KLMUe

 ô|¤,ZÀ<B$%Ö��önö�

R¤<

,¤,ZÉ<

(JZ

、+ùdZ

)@�!$%6/'(

>

、û&ä³`��

,Àô|¤,ZÀ<�4

-45℃XY5

W2�Ñ

,+ùd¤,ZÉ<�4��}~UW2�Ñ

('ê|

、#K|

、�¢¦

、�½

,̀,6Jõì¢Ev

êXc»�

;ddH2O、2×TaqDNA� I L

、DNAMarker(DL2000)、GelRed

ónÏ�

,̀,VíVFíãXc»�

ghL

K,V6

Merck»�

1.2 ÑÒMû¯

 ô|Ë])

,V6

Sigma»�

;NanoPhotometerTM

l,óngh÷ø©

,V6

Implen»�

;DYY-8CÒ�A©

,b1�7©�«

;2720Ò

PCR©

,�6

AppliedBiosystems

»�

;@£Jû=£

,�6

Bio-Rad»�

1.3 NO

1.3.1 À<(ÌÍ

。Í�

,sGõ6À<Öt+,

,L1~L10ò�b»}à6ô|¤,Z

;Z1~Z10�¤,ZÉ<

,�5

Z1~Z3�ô|L]�

,Z4~Z6�L]i

,Z7�x]R

,Z8~Z10

�x]J

À¤,ZÉ<5×Mxy6À<

(L]it

)ÖtÍM

ÌÍ

。ÑsÚ/Þe�

5gÀ<±e°�4

50mLË]¦

5

,¬.

5mL�Mi

,NÌ

30s§4

45℃¬�'«­5

,'

«¬É

3h;ÀË]¦

10000r/minË]

5min§ÿ`°×

M

,¬.

15mL70%ê|¨eNÌ

2min,10000r/minË]

5min,ȽÎ�

2~3Å

;¬.('ê|

15mLNÌ

2min,10000r/min

Ë]

10min,ÿ`?e

,±�4

65℃¬�|Y

   WXYZ[\

,J.AnhuiAgric.Sci.2018,46(28):92-95

Page 2: t Ⅰ,‘zæ0ŁŒŽN7˘Ù3– DNA · ,t COⅠ,‘zæ0ŁŒŽN7˘Ù3– DNAS¢=„j \Ùƒ,‹‘a,båc (!",->EØ

5|¾

,)O§W2�Ñ

[6]。

1.3.2 ¤,Zä�É<

DNA¤�

1.3.2.1 ¤,Z

。¤�¾�GÑy*�

[7]。ÑsÚ/Þe

�¤,ZÀ<

100mg4

1.5mLË]¦5

,¬.

400μL8Ã

e

(10mmol/LTris-HClpH 8.0,20mmol/LEDTA,400mmol/LNaCl,2%SDS)

§e°

;¬.

20mg/mLghL

K(20mg/mL)1μL,¬.Ë]¦5

,NÌ

30s,Ù@�ªªI

ÀË]¦6.ã�æ�`

,�4

60℃¬�'«­5

,'«¬

É

8~10h;ôÛ£à�§

,¬.

300μL6mol/LÔB

NaCl̈e§

,NÌ

30s,p§À��4

4℃ô|Ë])5

10000r/minË]

30min;�`?e£B6Ë]¦5

,¬.t

²E

(>N

620μL)6#¶|

,ªI�ª

,-20℃XY5¬

2h§

4℃ô|Ë])5P

10000r/minË]

5min;ÿ`

?e

,¬.

70%ê|¨e

1000μL,Ù@ªI§¬.

4℃ô

|Ë])P

10000r/minË]

5min;ÿ`?e§

,ÀË]¦

�4

65℃¬�|Y5|¾

;¬.

60μLTE¨e

,�4

-20℃XY5

1.3.2.2 ¤,ZÉ<

。�

50mgì¬×)O6À<4

1.5mLË]¦5

,�Ì9Tuô|¤,Z

DNA¤�¾�

1.3.3 DNAü�äã�÷ø

。�

DNAX�

2μL,P¬ËÅ

'���hsk

,�Ñl,óngh÷ø©÷øÀ<

DNAX�6i,ã�ä

A260/A280。�?

DNAX�m/ã� ¡

£

50~100ng/μL,W2�Ñ

1.3.4 PCR]µr÷&

。@k&N56&a

,��s(�

��F6Ë�²

COⅠGÑ�F

(�

1),sÀ<6

COⅠm¦&

aÖt

PCR]µ

[8],�sÿ�F(�]µ�p#v}D6

À<

,bÑ]ZË�²

COⅠGÑ�FÖtÖ79jø

。GÑ

�FB`,VíVFíãXc»�IJ

]

1 PCRÓ¼Ég

Table1 TheprimersforPCR

&,

No.�FЯ

Primername�F&a

Primersequence

1 LCO1490 5′-GGTCAACAAATCATAAAGATATTGG-3′

2 HCO2198 5′-TAAACTTCAGGGTGACCAAAAAATCA-3′

3 Fish-F 5′-TCAACCAACCACAAAGACATTGGCAC-3′

4 Fish-R 5′-TAGACTTCTGGGTGGCCAAAAGAATCA-3′

  PCR¶Ð²=

:25μL,2×TaqMasterMix(Dye)12.5μL,X�

DNA 1μL(<10ng),�、¶ _ � F K

1μL(0.4μmol/L),sÚ6HÛ'

9.5μL。PCR¶Ð}~

:94℃+'v

4min;94℃'v

30s,48℃$Ç

30s,72℃ò�

1min,35�>S

;72℃fò�

7min。PCR

]µ�¤§

,GHªXónÏ�6

1%LM�@£

�As

PCR(Fi,Ötö÷

,±À}D?'ô��97

6(Fa£`,VíVFíãÖt�¶H_÷&

1.3.5 -?ÌÍ

。ÀVF»�¶I60Ã-?Ñ

SeqMeaã

&¬@FR¸!B�¶_�F&a§ÖtlíPZ

,¹ÞÛ

�N�

680bp6

COⅠm¦&a

,±Ñ

GenBank-?M56

BlastíwÖtØs

,�?&aÙY�sÀ<6F8Ötj

,±rY<�FØs

,æ!À<F8r�F1¹7e

。[

÷À<68&aØsGH

ClustalXÖt

,±bÑ

MEGA5.0ÿ~�½8�r8³ÝÕ�Ë

,bÑ

NeighborJoining��5

Ö¢\

2 pqM6r

2.1 ÔÕ�N7¹j6r

2.1.1 ô|¤,Z

COⅠm¦&a]µ

。GHÿ)*+Ñ�

F]µ6h�ª5Û�>N�

680bp,[÷À<6

PCR(F

3

1%LM�@£`6}Dx�ô��?'97

(Í

1),þ

ôÿGÑ�Fs¤,Z6GÑvxy

,OX­Ñ4¤,Z

COⅠm¦6]µ

5

1 y�ÔÕ�N7

PCR+g«¾pq

Fig.1 ElectrophoreticdiagramofPCRproductsofdifferentfro

zencephalopods

2.1.2 ô|¤,Z

COⅠm¦÷&��@!

。Gõ6

10�
<�&2`x�:6ô|¤,Z

,B!",->EýÄ�Ü

ss�Ötb»Ejø

,p§bÑ

DNA}bfÝÞsjø

��Ötn%

。���ô

,DNA}bfjø��rb»Ej

ø��7e

,�

COⅠ&ar

GenBank-?M56F8ÙY�

�>4

91%(�

2)。  

GH

NeighborJoining��5֢\

,OsF8&au&

vxy6

COⅠ&aÖt�Z@!

,Xå4Ö79æ!ÿ&a

+sÐ6F8

,±½¾×u8À<Ð�1¹�Ô�X­S

@

。À

10.À<6&ar!

GenBank-?MU§6

30}

sÐF8&a�5Ö¢\

,P

1000ÅO-@wön

(Boot-strapstest)。

2O$

,×uÊ6¤,Z�ÔP�+Ê6

pvÙa�Z

,±�@�F>@w

,�5Q]ë6#æ-R

Q]

、S�T

、RQ]

、Q]Ùa�J7�>w

,xßw�¯

76%;̄Ì

68¤,ZÙa��G7�>w

。P`

108

¤,Z�ObJ/06@w

,þô

DNA}bfÝÞ�ÔÀ

P`F8wæj�

2.2 �N73�¹j6r

2.2.1 ¤,ZÉ<

COⅠm¦&a]µ

。¬ÉB¡¢1de

DNAÂÃ6�ô¦r

[9-10]。L]iÙñÉ<6U{ÌÍ}

3946Ó

28Ô

              \Ù¦Õ

 VÊ

COⅠVk8�p�­éé�V%óÝ

DNA��e>S

Page 3: t Ⅰ,‘zæ0ŁŒŽN7˘Ù3– DNA · ,t COⅠ,‘zæ0ŁŒŽN7˘Ù3– DNAS¢=„j \Ùƒ,‹‘a,båc (!",->EØ

~7l�

75~85℃,3~5min;JZtÙñÉ<6ÚÉÌÍ}

~7l�

80~90℃,15~20min[11-12]。¤,ZÉ<3¬íH

ã5O�9�

DNA�ÈÂÃ

。GHvnâç

,åç�6¬

É×,P'ô¤,Z6+XË�²

DNA,ÝÛç�6¬ÉÌ

Ís¤,Z6

DNA÷øx>

]

2 y�ÔÕ�N7´

BOLD�Õçz¹jpq

Table2 IdentificationofdifferentfrozencephalopodsinBOLDdatabase

À<

Sampleß*&EÐ

Latinname

5&EÐ

Chinesename

F8ÙY�

Speciessimilarity∥%

b»EjøÐ

Morphologicalidentificationname

jø1¹ÙÓ

Identifywhetheritmatches

L1 Todarodespacificus#æ-RQ]

97#æ-RQ] 1

L2 Illexargentinus�VRQ]

99Þ�VRQ] 1

L3 DosidicusenvironmentalRQ]

99RQ] 1

L4 OmmastrephesbartramiiQ]

99Q] 1

L5 Uroteuthischinensis56Ni�T

10056i�T 1

L6 Loligojaponica½6i�T

91½6i�T 1

L7 SymlectoteuthisoualaniensisS�T

99S�T 1

L8 SepiellamaindroniWy(��T

99Wy(��T 1

L9 OctopusocellatusåX

99åX 1

L10 Octopusvulgaris�X

99�X 1

5

2 ÔÕ�N7<�

COⅠðÖM

GenBank�ÕçG×z

COⅠðÖzß.Â

Fig.2 EvolutionarytreebasedonCOIsequencesoffrozencephalopodssampleandsequencesfromGenBank

2.2.2 ¤,ZÉ<

COⅠm¦÷&��@!

。3

GenBenk-

?M5s

10�À<6

COⅠm¦&aÖtØs

,���ôó

>8-¤,ZÉ<ð&rY<�FÓI

(�

3)。�5x]

ZÉ<ð&r�F×ÓI6ÚÛx�Q�

,23ÀQ]»£

1]Z��0�¬í6ÚÛ

;X

2.À<Ñ�J¨]µp#

v}D

,��JZÉ<

,O�1

DNAÂÃ�È��×e¤,

ZJ@

,kôi�B6å&a�FÖtjø

,X[Ö79v

nn%

3 p�M��

ÿ)*ö�)ô|¤,ZB¤,ZÉ<FZÀ<Öt

F8jø

,m4

COⅠm¦6

DNA}bf3ô|¤,ZF8

jø¾¿¹Þ)n«6­�

,>?@À<�ÔÔÞjøÞ8

6h�

,ÝX?@¤,ZÉ<×�ÔÔÞjø6h�

,w²

0¦@!ÌU

:①JZÉ<3¬íHã5kôÀ¤,ZÉJ

�Y

,Ó�eë¥B¡¢�ÑO�9s

DNA1Jx>6'

ô

,�Ûç�6y�ÛÚW9s

DNA6}àv1Jx>6÷

49           Ìͧ0¬Î

                         2018�

Page 4: t Ⅰ,‘zæ0ŁŒŽN7˘Ù3– DNA · ,t COⅠ,‘zæ0ŁŒŽN7˘Ù3– DNAS¢=„j \Ùƒ,‹‘a,båc (!",->EØ

ø

,AÍi�å&a�F

,bÑl}bfsÑ�J¨jø6

À<ÖtÖ79)*

[13];②�ÔbÑ

DNA}bfjø6[

÷À<m6Ü1Ñì¬í6ô|¤,ZrL]itÉ<

,�

J@x�97

,Ýx]J

、LTqt¤,ZÉ<5eX]�

Ò�t88J@

,AͧÓi�8È�Fs�Ötjø

;③

¤,ZÉ<¬íHã5«¬)PF×Päé<«¬êtF

i

,À<+ÌÍçÑ�À�}�@¬

,O�9Â5

DNA6¤

�¯�úÉ

PCR¶Ð

。¦~

,DNA}bfÝÞ3¤,Zé

<56ÐÑ237ø6Rcv

,+Ñ4¤,Z-¬í<�¬

í}~Ùsx��B

、J@976¤,ZÉ<

]

3 y��N73�´

BOLD�Õçz¹jpq

Table3 IdentificationofdifferentcephalopodproductsinBOLDdatabase

À<

Sample�FJ@

Labelcompositionß*EÐ

Latinname5&EÐ

ChinesenameF8ÙY�

Speciessimilarity∥%

jø1¹ÙÓ

Identifywhetheritmatches

Z1L]

OmmastrephesbartramiiQ]

991

Z2L]

Uroteuthischinensis56Ni�T

991

Z3L]

Todarodespacificus#æ-RQ]

971

Z4L]

DosidicusenvironmentalRQ]

991

Z5L]

Illexargentinus�VRQ]

991

Z6L]

— — — —

Z7x]

SepiellamaindroniWy(��T

991

Z8x]

HypophthalmichthysmolitrixZ]

100¹

Z9x]

OmmastrephesbartramiiQ]

99¹

Z10x]

— — — —

 -

:“—”�ÎÑ��¹p#v}D

 Note:“-”indicatesthatnospecificbandshavebeenobtained

  DNA}bfÝÞ3]

、̂ 、[tìZ1&56Ðѽ�

J�

。Eugenehkt

[14]3bÑ}bfs

918,(<6ö÷

5âç

25%6,(<23Y<�FMN6ç�

,±!�

DNA178OÀ,(<jøÞ86x�ìþy­6jøÝÞ

Hayet

[15]m4

DNA}bfÝÞs�b&!\[É<jø

5âç�578(<rY<�F×Ó

,�ô

DNA}bf1

78�ÔÑ4tÚ\[É<¬íBv!6ÝÞ

。ìZ(<

DNA}bfÝÞ6â-¹Ù4

DNA}bf-?M6×¾]

>

,~�1]Z

DNA}bf-?

,|3

2014ï

FISH-BOL-

?M~ëÇÇ)8Ô

102678]Z6

75249}

。DNA}b

fÝÞ!@Å'æs¤,ZÖtö÷

,Ú¸è�ÒÓ

,jø

��è¬�æ

,À�ÐÑ4&2`Y<�FrmF×Ó6¤

,Z(<1s'(<&2tÚ67&=B

��9:

[1]ZHANGJB,HANNERR.DNAbarcodingisausefultoolfortheidentificationofmarinefishesfromJapan[J].Biochemicalsystematics&ecology,2011,39(1):31-42.

[2]փ1

,Jç

,¥è

,>

.5¬�T

DNAX`éDERS`"

[J].45

FÖaUâd

,2013(6):99-106.[3]HEHERTPDN,CYWINSKAA,BALLSL,etal.BiologicalidentificationsthroughDNAbarcodes[J].ProceedingsoftherovalsocietyseriesB,2003,270(1512):313-321.

[4]XPÔ

,erê

,4[

.�T

DNAX`éDERSB23�Ä

[J].Çr

��Otë

,2011(1):28-30.[5]

úë

,M��

,Mg

,>

.A»ì^«

COⅠ1DNAX`é+�ía�Â

^_41DE

[J].,-bd

,2018,42(1):1-9.[6]

6ØO

,Mî

,!�

,>

.DNAX`éUV+�ïBC¡G^·412

3

[J].qGab

,2013,34(18):337-342.[7]ALJANABISM,MARTINEZI.UniversalandrapidsaltextractionofhighqualitygenomicDNAforPCRbasedtechniques[J].Nucleicacidsresearch,1997,25(22):4692-4693.

[8]FOLMERO,BLACKM,HOEHW,etal.DNAprimersforamplificationofmitochondrialcytochromecoxidasesubunitIfromdiversemetazoaninvertebrates[J].MolMarBiolBiotechnol,1994,3(5):294-299.

[9]ASLAN?,HAMILLRM,SWEENEYT,etal.Integrityofnucleargenomicdeoxyribonucleicacidincookedmeat:Implicationsforfoodtraceability[J].Journalofanimalscience,2009,87(1):57-61.

[10]ZHANGLW,WUQY.SinglegeneretrievalfromthermallydegradedDNA[J].JBiosci,2005,30(5):599-604.

[11]ùè.

.ð�ñ:-I$

[J].F¼ØUV

,2010(6):33-34.[12]

�ý

.�ïò:-I$

[J].F¼ØUV

,2010(20):35.[13]MEHRDADH,MALEXS,DANIELHJ,etal.Aminimalistbarcodecani

dentifyaspecimenwhoseDNAisdegraded[J].Molecularecologyresources,2006,6(4):959-964.

[14]EUGENEHKW,ROBERTHH.DNAbarcodingdetectsmarketsubstitutioninNorthAmericanseafood[J].Foodresearchinternational,2008,41(8):828-837.

[15]HAYEPA,SEGOVIANI,VERAR,etal.AuthenticationofcommercializedcrabmeatinChileusingDNABarcoding[J].Foodcontrol,2012,25(1):239-244.

檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿

檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿檿

殨殨殨

%3&'

  ]-

H�-

:�ÿÓa3£�âï��6=&5

,£gX

he=&6��ðÅ×54

hÅ

ð&&N,

:�ð&Óa3£�âïâ�6�?=&-

,×ö1£�Óa�Ñ-?6ð&

&Nö�¯

:/£�&6ö�0Áö�6&N-rÓa6â�&N-ÐØ

@A&N,

:�ð&Óa=&+�Ñ6�?@A&N-

,1ø,ÿÓaëE�/ã�BÆÇ�?ìÖ�¥67���

�&-

:�ð&Óa�7e=&æ��Ñ6@A&N-

�7-

:�ð&Óa�7e=&æ�öX6�7-

,1ø,ÿÓaëEV(�¥67���

5946Ó

28Ô

              \Ù¦Õ

 VÊ

COⅠVk8�p�­éé�V%óÝ

DNA��e>S