CN104630244A - COI gene sequence of species of coral reef benthos and species identification method utilizing COI gene sequence - Google Patents
COI gene sequence of species of coral reef benthos and species identification method utilizing COI gene sequence Download PDFInfo
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- CN104630244A CN104630244A CN201510037246.0A CN201510037246A CN104630244A CN 104630244 A CN104630244 A CN 104630244A CN 201510037246 A CN201510037246 A CN 201510037246A CN 104630244 A CN104630244 A CN 104630244A
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Abstract
The invention belongs to the field of biosystematics and molecular biologics, and in particular discloses a cytochrome C oxidase O gene I (COI) gene sequence of species of coral reef benthos and a species identification method utilizing the COI gene sequence. The standard sequences of the COI gene of different dominant species and common species of Hainan coral reef benthos are as shown in the nucleotide sequences of SEQ ID NO.1-14. On the basis of massive study and investigation, the standard COI gene sequences of 14 dominant species and common species of Hainan coral reef benthos are provided, and meanwhile by adopting the method, the species can be rapidly and effectively identified.
Description
Technical field
The invention belongs to taxology and biology field, specifically each species of a kind of karang bottom-dwelling cytochrome C oxidase I gene (COI) sequence and utilize the species identification method of COI gene order.
Background technology
Coral reef ecologic system is one of ecosystem that global species diversity is the highest, and the complicated and diversified coenosis perched in it also becomes the focus of marine biology and ecological study.Simultaneously, coral reef ecologic system is also most fragile in ocean, is vulnerable to the ecosystem of interference most, it is just constantly subject to the interference from each side pressure at present, the wherein Ocean acidification phenomenon that causes of global warming and mankind's activity, as discharged land-sourced pollutant, oceanographic engineering construction and fishing for collection activity etc., have impact on the growth and reproduction of reef-building coral greatly, directly cause the degeneration of coral reef ecologic system.The bottom-dwelling perched in karang is the important component part of coral reef ecologic system, in the health of maintenance, the adjustment ecosystem, play key player.The benthic diversity height of karang is proportionate with the health degree of coral reef ecologic system, and many scholars are using the Main Means of Investigation of biodiversity in reef as the change of monitoring coral reef ecologic system.Along with carrying out in a deep going way of China's karang child care work, in reef, coenosis investigation and ecological parameter more and more come into one's own.
Karang prerequisite that is ecological and biodiversity research is the precise Identification of marine organisms species.In karang, bottom-dwelling diversity is high, comprehensively China's South Sea karang Ecological Investigation data of carrying out in recent years, and in reef, benthic species number can reach kind more than 400, even and if dominant species in Schima superba also nearly more than ten is planted.So abundant species diversity brings huge sample survey work to ecological study, but the scholar being engaged in means of taxonomic research is less, the access approaches of the taxonomy bibliography of specialty is narrow, makes the sample survey operational difficulties based on traditional form heavy; Traditional biological authentication method too relies on subjective judgement, is especially embodied in the qualification of doubtful kind, causes the appearance of fuzzy qualification and mistake qualification, has a direct impact the exactness of the every data in marine ecology research.Karang bottom-dwelling DNA bar code rapid identification method can walk around this step of empirical morphologic observation, direct amplification and measure the COI gene order of unknown species, by the existing sequence information of comparison, simply and accurately identify the kind name of sample.But it is at present both at home and abroad very deficient about COI sequence information benthic in China Seas karang inside species gene pool or the gene sequence data delivered, even if dominant species in Schima superba and Common Species, sequence information that can be for reference is also considerably less, even if therefore investigators determine the COI sequence of unknown species, also have no way of contrasting and obtaining qualification result.As can be seen here, bottom-dwelling in mensuration China karang of system, especially the COI gene order of dominant species in Schima superba and common species is extremely necessary.
Summary of the invention
The object of the present invention is to provide cytochrome C oxidase I gene (COI) sequence of each species of a kind of karang bottom-dwelling and utilize the species identification method of COI gene order.
The technical solution used in the present invention is for achieving the above object:
A COI gene order for each species of karang bottom-dwelling,
The COI gene standard sequence of each sociales and Common Species in bottom-dwelling in the karang of Hainan:
1) peaceful drum shrimp (crustacean door, Alphaediae) Alpheus pacificus Dana, the COI gene standard sequence of 1852 is as shown in SEQ ID NO.1:
GCTTGGGCTGGTATAGTAGGAACCGCACTCAGTTTACTAATTCGAGCTGAACTTGGACAACCCGGTAGATTAATTGGAAATGACCAAATTTACAATGTAATCGTAACAGCCCATGCATTTGTAATAATTTTCTTTATGGTCATACCAATCATAATTGGAGGTTTTGGGAACTGACTGATTCCCCTTATACTAGGAGCGCCAGATATAGCCTTCCCACGTATAAATAATATAAGATTCTGACTTCTACCCCCCTCCCTCACCCTACTTCTATCTAGTGGCCTAGTAGAAAGGGGGGTCGGCACAGGATGAACTGTTTATCCACCACTATCAGCAGGTATTGCCCACGCGGGAGCATCAGTAGATCTGGGGATTTTCTCTCTCCATTTAGCCGGTGTCTCCTCAATCCTAGGAGCAGTCAACTTTATAACAACAGTTATTAATATACGAGCTACAGGCATAACTATAGACCGTATGCCACTGTTTGTATGAGCTGTATTCCTGACAGCCATTCTACTCCTCCTAAGGCTCCCAGTCCTAGCAGGAGCAATTACTATACTTCTAACAGACCGAAACCTAAATACAGCTTTCTTCGACCCCGCTGGAGGTGGAGATCCAATTTTATACCAACACCTATTTTGATTTTTTGGTCACC
GTCTCCACCTCCAGCGGGGTCGAAGAAAGCTGTATTTAGGTTTCGGTCTGTTAGAAGTATAGTAATTGCTCCTGCTAGGACTGGGAGCCTTAGGAGGAGTAGAATGGCTGTCAGGAATACAGCTCATACAAACAGTGGCATACGGTCTATAGTTATGCCTGTAGCTCGTATATTAATAACTGTTGTTATAAAGTTGACTGCTCCTAGGATTGAGGAGACACCGGCTAAATGGAGAGAGAAAATCCCCAGATCTACTGATGCTCCCGCGTGGGCAATACCTGCTGATAGTGGTGGATAAACAGTTCATCCTGTGCCGACCCCCCTTTCTACTAGGCCACTAGATAGAAGTAGGGTGAGGGAGGGGGGTAGAAGTCAGAATCTTATATTATTTATACGTGGGAAGGCTATATCTGGCGCTCCTAGTATAAGGGGAATCAGTCAGTTCCCAAAACCTCCAATTATGATTGGTATGACCATAAAGAAAATTATTACAAATGCATGGGCTGTTACGATTACATTGTAAATTTGGTCATTTCCAATTAATCTACCGGGTTGTCCAAGTTCAGCTCGAATTAGTAAACTGAGTGCGGTTCCTACTATACCAGCCCAAGCCCCGAAAATAAAGTATAAGGTGCCAATATCTTTATG
2) the COI gene standard sequence of smooth green crab (crustacean door, Xanthidae) Chlorodiella laevissima (Dana, 1852) is as shown in SEQ ID NO.2:
TAGCTGGAATAGTTGGAACATCACTTAGTTTAATTATTCGTGCCGAATTAGGTCAACCTGGAACTCTTATTGGAAATGATCAAATTTATAATGTTGTAGTTACAGCTCATGCTTTTGTTATAATTTTCTTTATAGTTATACCCATTATAATTGGAGGATTTGGTAATTGATTAGTACCCCTTATACTTGGAGCCCCTGATATAGCTTTCCCTCGTATAAACAACATAAGATTTTGACTTCTTCCTCCTTCCCTTACTCTTCTTTTAATAAGAGGAATAGTGGAAAGTGGGGTTGGTACCGGGTGAACAGTTTATCCTCCTCTTGCTGCCGCTATTGCTCATGCTGGAGCTTCAGTTGACATGGGTATTTTCTCTCTTCATTTAGCGGGTGTCTCTTCCATTCTAGGAGCGGTAAACTTCATAACTACCGTAATTAATATACGTTCATTTGGAATGACTATAGATCAAATGCCTCTTTTTGTTTGAGCAGTATTTATTACTGCTATTTTACTTCTCTTATCTCTTCCTGTTCTAGCAGGTGCAATCACTATACTTCTTACAGATCGAAATTTGAATACATCCTTTTTTGACCCAGCTGGGGGAGGTGATCCTGTTCTTTATCAACATTTATTTTGATTTTTTGGTCACC
ATCCCCCAGCTGGGTCAAAAAAGGATGTATTCAAATTTCGATCTGTAAGAAGTATAGTGATTGCACCTGCTAGAACAGGAAGAGATAAGAGAAGTAAAATAGCAGTAATAAATACTGCTCAAACAAAAAGAGGCATTTGATCTATAGTCATTCCAAATGAACGTATATTAATTACGGTAGTTATGAAGTTTACCGCTCCTAGAATGGAAGAGACACCCGCTAAATGAAGAGAGAAAATACCCATGTCAACTGAAGCTCCAGCATGAGCAATAGCGGCAGCAAGAGGAGGATAAACTGTTCACCCGGTACCAACCCCACTTTCCACTATTCCTCTTATTAAAAGAAGAGTAAGGGAAGGAGGAAGAAGTCAAAATCTTATGTTGTTTATACGAGGGAAAGCTATATCAGGGGCTCCAAGTATAAGGGGTACTAATCAATTACCAAATCCTCCAATTATAATGGGTATAACTATAAAGAAAATTATAACAAAAGCATGAGCTGTAACTACAACATTATAAATTTGATCATTTCCAATAAGAGTTCCAGGTTGACCTAATTCGGCACGAATAATTAAACTAAGTGATGTTCCAACTATTCCAGCTCAAGCACCAAAGATAAAATATAATGTTCCAATATCTTTATGT
3) the green crab of black finger (crustacean door, Xanthidae) Chlorodiella nigra (
1775) COI gene standard sequence is as shown in SEQ ID NO.3
TAGCTGGATAGTAGGGACTTCCCTCAGTTTAATCATCCGTGCTGAACTTGGTCAACCCGGAACCCTTATTGGAAATGATCAAATTTATAATGTTGTAGTTACAGCCCACGCTTTTGTTATAATTTTCTTTATAGTTATACCTATTATAATTGGAGGATTTGGTAATTGATTAGTTCCATTAATGCTTGGAGCCCCTGATATAGCCTTTCCTCGTATAAACAATATAAGATTTTGACTTTTACCTCCATCCCTAACTCTTCTTTTAATAAGTGGAATAGTTGAAAGAGGGGTTGGTACGGGATGAACTGTATATCCTCCTCTTGCCGCTGCTATCGCCCATGCAGGTGCTTCAGTTGATATAGGAATTTTCTCCCTTCACTTGGCAGGTGTATCATCAATTCTAGGGGCAGTAAATTTTATAACTACCGTTATTAATATACGTTCATTTGGTATAACTATAGACCAAATACCTCTTTTTGTTTGAGCAGTATTTATTACTGCTATTTTACTTCTACTTTCTCTTCCTGTGCTAGCAGGAGCTATTACTATGCTTCTTACAGATCGAAATTTAAATACATCTTTCTTTGATCCTGCTGGAGGAGGAGACCCTGTTCTCTACCAACACTTATTCTGATTTTTTGGTCACC
TCTCCTCCTCCAGCAGGATCAAAGAAAGATGTATTTAAATTTCGATCTGTAAGAAGCATAGTAATAGCTCCTGCTAGCACAGGAAGAGAAAGTAGAAGTAAAATAGCAGTAATAAATACTGCTCAAACAAAAAGAGGTATTTGGTCTATAGTTATACCAAATGAACGTATATTAATAACGGTAGTTATAAAATTTACTGCCCCTAGAATTGATGATACACCTGCCAAGTGAAGGGAGAAAATTCCTATATCAACTGAAGCACCTGCATGGGCGATAGCAGCGGCAAGAGGAGGATATACAGTTCATCCCGTACCAACCCCTCTTTCAACTATTCCACTTATTAAAAGAAGAGTTAGGGATGGAGGTAAAAGTCAAAATCTTATATTGTTTATACGAGGAAAGGCTATATCAGGGGCTCCAAGCATTAATGGAACTAATCAATTACCAAATCCTCCAATTATAATAGGTATAACTATAAAGAAAATTATAACAAAAGCGTGGGCTGTAACTACAACATTATAAATTTGATCATTTCCAATAAGGGTTCCGGGTTGACCAAGTTCAGCACGGATGATTAAACTGAGGGAAGTCCCTACTATTCCAGCTCAAGCTCCAAAAATAAAATATAGTGTTCCAATATCTTTATG
4) coral armor shrimp (crustacean door, armor shrimp section) Galathea coralliophilus Baba & Oh, the COI gene standard sequence of 1990 is as shown in SEQ ID NO.4
GAGCAGGATAGTAGGAACTTCTCTGAGACTTATTATTCGAGCTGAATTAGGACAACCAGGTAGATTAATTGGAGATGATCAAATTTATAATGTAGTTGTAACAGCTCATGCTTTCGTTATAATTTTCTTTATAGTTATACCAATTATAATTGGAGGATTTGGAAATTGACTTGTACCTTTAATATTAGGAGCGCCTGATATAGCTTTCCCTCGAATAAATAATATAAGATTTTGGCTTCTTCCGCCCTCTCTTACTTTACTACTCATAAGAGGAATAGTAGAAAGAGGTGTTGGTACAGGTTGAACTGTATACCCTCCTCTTGCAGCAGGAATTGCTCATGCAGGTGCATCTGTTGATATAGGAATTTTTTCTCTTCATCTTGCTGGGGTGTCATCAATTTTAGGGGCAATTAATTTTATAACAACAGTTATTAATATACGACCTAAAGGTATGACAATAGACCGTATACCTCTTTTTGTTTGATCTGTGTTTATTACCGCTATTTTATTACTTTTATCCTTACCTGTATTAGCAGGTGCTATTACAATGTTATTAACAGATCGAAACCTAAATACATCATTCTTTGATCCTGCTGGTGGTGGTGATCCTATTCTATATCAGCATTTATTCTGATTTTTTGGTCACC
CCCACCACCAGCAGGATCAAAGAATGATGTATTTAGGTTTCGATCTGTTAATAACATTGTAATAGCACCTGCTAATACAGGTAAGGATAAAAGTAATAAAATAGCGGTAATAAACACAGATCAAACAAAAAGAGGTATACGGTCTATTGTCATACCTTTAGGTCGTATATTAATAACTGTTGTTATAAAATTAATTGCCCCTAAAATTGATGACACCCCAGCAAGATGAAGAGAAAAAATTCCTATATCAACAGATGCACCTGCATGAGCAATTCCTGCTGCAAGAGGAGGGTATACAGTTCAACCTGTACCAACACCTCTTTCTACTATTCCTCTTATGAGTAGTAAAGTAAGAGAGGGCGGAAGAAGCCAAAATCTTATATTATTTATTCGAGGGAAAGCTATATCAGGCGCTCCTAATATTAAAGGTACAAGTCAATTTCCAAATCCTCCAATTATAATTGGTATAACTATAAAGAAAATTATAACGAAAGCATGAGCTGTTACAACTACATTATAAATTTGATCATCTCCAATTAATCTACCTGGTTGTCCTAATTCAGCTCGAATAATAAGTCTCAGAGAAGTTCCTACTATTCCTGCTCAGGCACCAAAAATAAAATAAAGTGTTCCAATATCTTTATG
5) the COI gene standard sequence of thick chela porcelain crab (crustacean door, Ci Xie section) Pachycheles sculptus (H.Milne Edwards, 1837) is carved as shown in SEQ ID NO.5
ACCTAAGAaGTGTTGAGGGAAaATGTTAGATTTACTCCTAAGAATATAACTGAGAAGTGAATTTTTAGCCATTTAGGGTTGACTGATATTCCGGAAAATAATGAGAACCAGTGAGCAATACCAGCAAAAATACCAAAGACAGCTCCTATTGATAAAACATAATGGAAATGGGCAACAACATAGTATGTGTCATGGAGAATTGTATCAATAGAAGAATTTGCAAGAACAACTCCCGTGAGTCCTCCTACAGTAAACAGAAAAATAAATCCTAGGGCCCAGATTATAGAGGGGGAGTAAGTTATTTGATTCCCCTGGAGAGTTCCTAGTCAACTAAAAATCTTAATTCCAGTTGGAATGGCAATAATCATGGTTGCAGATGTAAAGTAGGCTCGTGTATCAACATCTATACCAACAGTGAACATATGGTGAGCTCACACAATAAACCCTAAAATTCCAATTGCTAACATGGCGTAAATTATTCCAAGGGTTCCAAAAGATTCTTTTTTTCCTGATTCCTGGGTTACAATATGAGAGATTATTCCAAAAGCTGGTAAAATTAAAATATATACTTCAGGATGGCCGAAGAACCAGAACAAATGTTGGTAAAGGACGGGGTCTCCTCCCTCCTGCA
TTTGTTCTGGTTCTTCGGCCATCCTGAAGTATATATTTTAATTTTACCAGCTTTTGGAATAATCTCTCATATTGTAACCCAGGAATCAGGAAAAAAAGAATCTTTTGGAACCCTTGGAATAATTTACGCCATGTTAGCAATTGGAATTTTAGGGTTTATTGTGTGAGCTCACCATATGTTCACTGTTGGTATAGATGTTGATACACGAGCCTACTTTACATCTGCAACCATGATTATTGCCATTCCAACTGGAATTAAGATTTTTAGTTGACTAGGAACTCTCCAGGGGAATCAAATAACTTACTCCCCCTCTATAATCTGGGCCCTAGGATTTATTTTTCTGTTTACTGTAGGAGGACTCACGGGAGTTGTTCTTGCAAATTCTTCTATTGATACAATTCTCCATGACACATACTATGTTGTTGCCCATTTCCATTATGTTTTATCAATAGGAGCTGTCTTTGGTATTTTTGCTGGTATTGCTCACTGGTTCTCATTATTTTCCGGAATATCAGTCAACCCTAAATGGCTAAAAATTCACTTCTCAGTTATATTCTTAGGAGTAAATCTAACATTTTTCCCTCAACACTTCTTAGGTCTAAACGGAATACCACGTCGATACTCTGACTACCCA
6) coarse hair shell crab (crustacean door, Xanthidae) Pilodius scabriculus Dana, the COI gene standard sequence of 1852 is as shown in SEQ ID NO.6
GAGCTGGTATGGTCGGTACATCTCTAAGTCTTATTATCCGTGCTGAACTAGGACAACCCGGAACACTTATTGGAAATGATCAAATCTACAATGTTATTGTAACCGCTCATGCCTTTGTAATAATCTTTTTCATAGTTATACCTATTATAATTGGAGGATTTGGTAATTGACTTGTCCCATTAATATTAGGAGCTCCTGATATAGCCTTTCCTCGTATAAACAATATAAGATTTTGACTTCTTCCCCCATCCCTTACTCTTCTTCTTATAAGAGGTATAGTCGAAAGAGGAGTTGGTACAGGTTGGACCGTTTATCCACCTCTTGCTGCCGCTATCGCTCACGCTGGGGCCTCAGTAGATATAGGTATTTTCTCTCTCCATTTGGCTGGTGTATCCTCTATTTTAGGAGCAGTTAATTTTATAACTACCGTTATTAACATACGATCCTTTGGTATAACCATAGACCAAATACCTCTTTTTGTTTGAGCTGTATTTATTACCGCAATCCTGCTTTTACTTTCCTTACCTGTATTAGCAGGAGCTATTACTATGCTCCTTACTGACCGTAATTTAAACACATCCTTCTTTGATCCAGCCGGAGGTGGTGATCCTGTTCTTTATCAACATCTATTCTGATTTTTTGGTCACC
GTGAAGACAGGATCACCACCTCCGGCTGGATCAAAGAAGGATGTGTTTAAATTACGGTCAGTAAGGAGCATAGTAATAGCTCCTGCTAATACAGGTAAGGAAAGTAAAAGCAGGATTGCGGTAATAAATACAGCTCAAACAAAAAGAGGTATTTGGTCTATGGTTATACCAAAGGATCGTATGTTAATAACGGTAGTTATAAAATTAACTGCTCCTAAAATAGAGGATACACCAGCCAAATGGAGAGAGAAAATACCTATATCTACTGAGGCCCCAGCGTGAGCGATAGCGGCAGCAAGAGGTGGATAAACGGTCCAACCTGTACCAACTCCTCTTTCGACTATACCTCTTATAAGAAGAAGAGTAAGGGATGGGGGAAGAAGTCAAAATCTTATATTGTTTATACGAGGAAAGGCTATATCAGGAGCTCCTAATATTAATGGGACAAGTCAATTACCAAATCCTCCAATTATAATAGGTATAACTATGAAAAAGATTATTACAAAGGCATGAGCGGTTACAATAACATTGTAGATTTGATCATTTCCAATAAGTGTTCCGGGTTGTCCTAGTTCAGCACGGATAATAAGACTTAGAGATGTACCGACCATACCAGCTCAAGCTCCAAAGATAAAATATAGTGTACCAATATCTTTATG
7) the COI gene standard sequence of three seta thorn crab (crustacean door, burr crab section) Pilumnus trispinosus (Sakai, 1965) is as shown in SEQ ID NO.7
TTTACCTAGGAATGTTGTGGAAAAGGTTGTGTTAACTCCTACAAATATAGTGAAGAAATGAATTTTTATTCATCGTGGGTTTAATGATAGGCCTGTAAACAGGGGAAATCAGTGAGCGATTCCTCCGAAAATACCAAATACGGCACCTATTGATAAAACGTAGTGGAAATGGGCTACAACATAATATGTATCATGTAAAATAATATCAATTGAGGAGTTGGCTAATACTACTCCTGTTAATCCACCTATAGTAAATAAAAAAATAAACCCTAAGGCTCATATTAAGGAAGGTCTATAATTAATTTGAGTTCCATGAAGGGTTCTTAACCAACTGAAAATTTTAATTCCCGTGGGGACAGCAATGATTATTGTAGCAGAGGTGAAATAAGCTCGGGTATCTACATCTATTCCAACTGTGAATATATGGTGAGCTCAAACTACAAATCCTAAGATACCAATAGCTAATATAGCATAAATTATCCCTAAAGTGCCGAATGATTCTTTTTTTCCTGATTCTTGTCTTACAATATGGGAGATTATACCGAAGGCTGGAAGAATAAGAATATAAACTTCAGGGTGACCGAAGAATCAAAATAAATGTTGATAGAGGACAGGGTCTCC
TTGATTCTTCGGTCACCCTGAAGTTTATATTCTTATTCTTCCAGCCTTCGGTATAATCTCCCATATTGTAAGACAAGAATCAGGAAAAAAAGAATCATTCGGCACTTTAGGGATAATTTATGCTATATTAGCTATTGGTATCTTAGGATTTGTAGTTTGAGCTCACCATATATTCACAGTTGGAATAGATGTAGATACCCGAGCTTATTTCACCTCTGCTACAATAATCATTGCTGTCCCCACGGGAATTAAAATTTTCAGTTGGTTAAGAACCCTTCATGGAACTCAAATTAATTATAGACCTTCCTTAATATGAGCCTTAGGGTTTATTTTTTTATTTACTATAGGTGGATTAACAGGAGTAGTATTAGCCAACTCCTCAATTGATATTATTTTACATGATACATATTATGTTGTAGCCCATTTCCACTACGTTTTATCAATAGGTGCCGTATTTGGTATTTTCGGAGGAATCGCTCACTGATTTCCCCTGTTTACAGGCCTATCATTAAACCCACGATGAATAAAAATTCATTTCTTCACTATATTTGTAGGAGTTAACACAACCTTTTTTCCACAACATTTCCTAGGTTTAAATGGTATGCCACGACGATATTCGGACTACCCA
8) the COI gene standard sequence of special-shaped beans porcelain crab (crustacean door, Ci Xie section) Pisidia dispar (Stimpson, 1858) is as shown in SEQ ID NO.8
GGAATAGTTGGTACATCTCTTAGTTTAATTATTCGAGCTGAACTAGGTCAATTAGGAAGATTAATTGGTGACGATCAAATTTACAATGTTGTAGTCACTGCTCACGCTTTTGTTATAATTTTTTTTATAGTTATACCAATTTTAATTGGAGGATTCGGGAACTGACTAGTTCCTTTAATACTCGGAGCCCCTGATATAGCTTTTCCTCGAATAAATAATATAAGATTTTGACTTTTACCCCCAGCATTGTCACTTCTTTTAATAAGAGGATTAGTTGAAAGAGGAGTGGGAACGGGATGAACCGTTTACCCACCCTTAGCCTCAAATATCGCCCACGCTGGCGCTTCTGTAGACATAGGAATTTTTTCTCTCCACCTTGCAGGAGTTTCCTCAATTTTAGGAGCTGTAAATTTTATTTCTACTGTTATCAACATACGATCAGTTGGAATAACCTTTGACCGTATACCACTTTTTGTTTGATCGGTTTTTATTACAGCAATTTTACTTCTTTTATCTTTACCCGTTTTAGCTGGGGCAATTACCATACTTCTAACAGATCGAAACCTTAATACTTCATTTTTCGACCCAGCAGGGGGAGGAGATCCTATTCTTTATCAACACTTATTTTGATTTTTTGGTCACC
TCCTCCCCCTGCTGGGTCGAAAAATGAAGTATTAAGGTTTCGATCTGTTAGAAGTATGGTAATTGCCCCAGCTAAAACGGGTAAAGATAAAAGAAGTAAAATTGCTGTAATAAAAACCGATCAAACAAAAAGTGGTATACGGTCAAAGGTTATTCCAACTGATCGTATGTTGATAACAGTAGAAATAAAATTTACAGCTCCTAAAATTGAGGAAACTCCTGCAAGGTGGAGAGAAAAAATTCCTATGTCTACAGAAGCGCCAGCGTGGGCGATATTTGAGGCTAAGGGTGGGTAAACGGTTCATCCCGTTCCCACTCCTCTTTCAACTAATCCTCTTATTAAAAGAAGTGACAATGCTGGGGGTAAAAGTCAAAATCTTATATTATTTATTCGAGGAAAAGCTATATCAGGGGCTCCGAGTATTAAAGGAACTAGTCAGTTCCCGAATCCTCCAATTAAAATTGGTATAACTATAAAAAAAATTATAACAAAAGCGTGAGCAGTGACTACAACATTGTAAATTTGATCGTCACCAATTAATCTTCCTAATTGACCTAGTTCAGCTCGAATAATTAAACTAAGAGATGTACCAACTATTCCTGCTCATGCGCCAAAAATAAAATATAATGTTCCAATATCTTTATGT
9) the knurl palm closes the COI gene standard sequence of drum shrimp (crustacean door, Alphaediae) Synalpheus tumiolomanus (Paul'son, 1875) as shown in SEQ ID NO.9
ATTCGAGCCGACTTGGTCAACCTGGTAGCCTAATTGGGAACGACCAAATTTATAACGTAATTGTCACTGCACACGCATTTGTCATAATTTTCTTTATGGTTATACCTATTATAATCGGGGGATTCGGAAACTGACTAGTTCCCCTTATATTAGGAGCTCCAGATATAGCATTTCCCCGAATAAATAACATAAGATTTTGATTACTTCCTCCCTCTCTTACCCTCCTTTTATCTAGAGGAATAGTGGAAAGAGGAGTGGGAACTGGTTGAACAGTCTATCCACCTCTTGCAAGAGGAATTGCCCATGCAGGAGCTTCAGTTGACATAGGAATTTTTTCATTACATCTAGCAGGAGTCTCCTCAATTTTAGGAGCTGTTAATTTTATAACCACTGTAATCAACATACGAGCAGCAGGAATAACAATAGACCAAATACCACTTTTTGTATGAGCAGTTTTTTTAACTGCAATCTTATTATTATTAAGACTTCCAGTTTTAGCTGGAGCTATTACTATACTATTAACAGACCGAAATCTAAAACACATAC
CTAACTGGAGTCTTAATAATAATAAGATTGCAGTTAAAAAAACTGCTCATACAAAAAGTGGTATTTGGTCTATTGTTATTCCTGCTGCTCGTATGTTGATTACAGTGGTTATAAAATTAACAGCTCCTAAAATTGAGGAGACTCCTGCTAGATGTAATGAAAAAATTCCTATGTCAACTGAAGCTCCTGCATGGGCAATTCCTCTTGCAAGAGGTGGATAGACTGTTCAACCAGTTCCCACTCCTCTTTCCACTATTCCTCTAGATAAAAGGAGGGTAAGAGAGGGAGGAAGTAATCAAAATCTTATGTTATTTATTCGGGGAAATGCTATATCTGGAGCTCCTAATATAAGGGGAACTAGTCAGTTTCCGAATCCCCCGATTATAATAGGTATAACCATAAAGAAAATTATGACAAATGCGTGTGCAGTGACAATTACGTTATAAATTTGGTCGTTCCCAATTAGGCTACCAGGTTGACCAAGTTCGGCTCGAATTAAAAGTCTGAGAGAAGTACCAACTATTCCTGCTCACGCTC
10) carefully sting greatly ophiruid (Echinodermata, thorn ophiruid section) Macrophiothrix lorioli A.M.Clark, the COI gene standard sequence of 1968 is as shown in SEQ ID NO.10
TCTCAGCTAGAGTGTTGGGGAAATGTAAGGTTGACTCCAATAAACATTAAAAAGAAGTGGGTAAGTGAGCTGGCCCGATCAATTGTGGCACCGGTAAATAGAGAGAATCAGTGATTAAATCCGGCAAAAATTGCAAATACTGCTCCCATTGACAGTACATAGTGGAAGTGAGCCGTGACGTAATATGTATCGTGGAGGGCTACGTTTAAAGATGAGTTTGATAAAATTATTCCAGTTAAACCCCCTATGGTGAATAAAAATATGAATCCTAGGGCTCAATACAGAGAGGGGTGTGCCTTGTTTATGTGGAAGAATGCTCCCTGGAGTGTTGCAAGTCATCTAAAAACCTTGACCCCTGTCGGAATAGCTATTATCATTGTGGCTGCTGTAAAGTAGGCGCGTGTGTCCACGTCCATGCCTACGGTGAACATGTGGTGGGCTCAGACGATGAAGCCTAGTATTCCAATGGAGATCATTGCGTACATCATACCTAAATACCCGAAGGGGGTAGCCTTCCCTGTTCGGTTAGCCACTACGTGTGAAATAATCCCAAATCCTGGTAAAATTAAGATATAAACTTCGGGGTGCCCAAAAAATCAAAATAGGTGTTGAAATAGGATTGGGTCTCCTCT
GGGCACCCCGAGTTTATATCTTAATTTTACCAGGATTTGGGATTATTTCACACGTAGTGGCTAACCGAACAGGGAAGGCTACCCCCTTCGGGTATTTAGGTATGATGTACGCAATGATCTCCATTGGAATACTAGGCTTCATCGTCTGAGCCCACCACATGTTCACCGTAGGCATGGACGTGGACACACGCGCCTACTTTACAGCAGCCACAATGATAATAGCTATTCCGACAGGGGTCAAGGTTTTTAGATGACTTGCAACACTCCAGGGAGCATTCTTCCACATAAACAAGGCACACCCCTCTCTGTATTGAGCCCTAGGATTCATATTTTTATTCACCATAGGGGGTTTAACTGGAATAATTTTATCAAACTCATCTTTAAACGTAGCCCTCCACGATACATATTACGTCACGGCTCACTTCCACTATGTACTGTCAATGGGAGCAGTATTTGCAATTTTTGCCGGATTTAATCACTGATTCTCTCTATTTACCGGTGCCACAATTGATCGGGCCAGCTCACTTACCCACTTCTTTTTAATGTTTATTGGAGTCAACCTTACATTTTTTCCCCAACACTTCTTAGGCCTTGCAGGAATGCCACGGCGATACTCAGACTACCCAGAG
11) the COI gene standard sequence of spoke ophiruid (Echinodermata, spoke ophiruid section) Ophiactis savignyi (M ü ller & Troschel, 1842) is as shown in SEQ ID NO.11
AGGGTCTCCTCCCCCTGTTGGATCGAAGAAGGTAGTGTTAATTTTTCGGTCTGTGAGGAGCATGGTGATGGCTCCGGCTAGAACAGGTAAGGAGAGCAGGAGGAGGATTGTGGTTAACAAAATGGATCAAACGAAAAGAGGCATACGGTCCATGGTCATTCCAGGGGCCCGCATGTTAAGAATAGTGGTTATAAAGTTTATAGAGGCCATAATAGATGAGGCTCCGGCCAGGTGTAGAGAAAAGATCGCGAGGTCTACACACCCTCCTGCGTGGGCGGTTGGGCCAGATAAGGGCGGGTAAACTGTTCACCCGGTTCCTACTCCCCCTTCTTTGGCAGCCGAAGCTAAGAGTAAAATGAAGGCGGGAGGTACAAGTCAGAATCTCATTTTTTTCATACGTGGGAATGCCATATCTGGAGCTCCTAACATTAAAGGGACCAGTCAGTTTCCAAATCCT
GATATGGCATTCCCACGTATGAAAAAAATGAGATTCTGACTTGTACCTCCCGCCTTCATTTTACTCTTAGCTTCGGCTGCCAAAGAAGGGGGAGTAGGAACCGGGTGAACAGTTTACCCGCCCTTATCTGGCCCAACCGCCCACGCAGGAGGGTGTGTAGACCTCGCGATCTTTTCTCTACACCTGGCCGGAGCCTCATCTATTATGGCCTCTATAAACTTTATAACCACTATTCTTAACATGCGGGCCCCTGGAATGACCATGGACCGTATGCCTCTTTTCGTTTGATCCATTTTGTTAACCACAATCCTCCTCCTGCTCTCCTTACCTGTTCTAGCCGGAGCCATCACCATGCTCCTCACAGACCGAAAAATTAACACTACCTTCTTCGATCCAACAGGGGGAGGAGACCCTATTTTATTTCAACACTTGTTCTGATTTTTTGGTC
12) the COI gene standard sequence of coarse two finger squama worm (Annelita, Shuan Zhilin worm section) Iphione muricata (Lamarck, 1818) is as shown in SEQ ID NO.12
TGATCTGGATTATTGGGCACATCAATAAGTCTTTTAATTCGGGCCGAGCTCGGCCAACCAGGGTCCCTTCTAGGCAGAGACCAACTTTATAATACGATTGTTACGGCTCATGCTTTTCTTATAATCTTCTTTCTGGTAATACCAGTATTAGTAGGAGGATTTGGTAACTGACTTATCCCGTTAATACTAGGGGCTCCGGACATGGCTTTCCCACGTCTGAATAATATAAGATTCTGACTCTTACCCCCTTCACTTATCCTTCTTTTATCATCTAGTGCTGTTGAAAAAGGAGTTGGGACTGGCTGAACTGTCTACCCCCCTCTTGCTAGAAACATTGCTCATGCTGGACCATCTGTTGACCTAGCAATTTTTTCTTTACATATTGCAGGAGTCTCTTCAATTCTAGGTGCTCTAAATTTTATTACTACTGTAATTAATATACGATATAAAGGCCTCCGTCTAGAACGTGTCCCATTGTTTGTATGAGCCGCCAAGATTACTGCCATTCTTCTTTTACTTTCTCTCCCCGTCTTAGCCGGCGCTATTACTATACTTCTCACCGACCGTAATCTAAACACTGCTTTCTTTGACCCTGCTGGTGGTGGTGACCCCGTCCTTTATCAGCATTTATTTTGATTTTTTGGTCAC
CCACCACCAGCAGGGTCAAAGAAAGCAGTGTTTAGATTACGGTCGGTGAGAAGTATAGTAATAGCGCCGGCTAAGACGGGGAGAGAAAGTAAAAGAAGAATGGCAGTAATCTTGGCGGCTCATACAAACAATGGGACACGTTCTAGACGGAGGCCTTTATATCGTATATTAATTACAGTAGTAATAAAATTTAGAGCACCTAGAATTGAAGAGACTCCTGCAATATGTAAAGAAAAAATTGCTAGGTCAACAGATGGTCCAGCATGAGCAATGTTTCTAGCAAGAGGGGGGTAGACAGTTCAGCCAGTCCCAACTCCTTTTTCAACAGCACTAGATGATAAAAGAAGGATAAGTGAAGGGGGTAAGAGTCAGAATCTTATATTATTCAGACGTGGGAAAGCCATGTCCGGAGCCCCTAGTATTAACGGGATAAGTCAGTTACCAAATCCTCCTACTAATACTGGTATTACCAGAAAGAAGATTATAAGAAAAGCATGAGCCGTAACAATCGTATTATAAAGTTGGTCTCTGCCTAGAAGGGACCCTGGTTGGCCGAGCTCGGCCCGAATTAAAAGACTTATTGATGTGCCCAATAATCCAGATCATGTACCAAAAATGAAGTAAAGAGTACCAATATCTTTAT
13) pearl palpus rock clam worm (Annelita, Eunicidae) Leodice antennata Savigny in Lamarck, the COI gene standard sequence of 1818 is as shown in SEQ ID NO.13
GACAGCAGGGGGATAGCAGGGACAGCCATTAGAATGCAAATTCGTTTGGCCTTAGCCCAACCCGGCTCTCTCTTAGGAAATGACCAACTTTATAATACACTAGTAACCGCCCACGCTTTTTTAATAATTTTCTTTTTAGTAATGCCCGTAATAATTGGTGGATTTGGTAATTGACTTCTCCCCCTTATACTAGGGGCTCCAGATATAGCCTTCCCTCGAATAAATAATATAAGCTTCTGACTCTTGCCCCCCTCTCTAATTCTTTTATTAGCCTCTGCAGCAGCAGAAAAAGGAGCCGGGACAGGATGAACGGTTTACCCCCCCTTATCAAGAAATATTGCCCATGCCGGACCCTCTGTAGACCTAGCTATTTTTTCTCTGCACTTAGCAGGAGTATCTTCTATTCTAGGATCAATTAACTTTATTACAACCGCCTTCAACATACGATGAGAAGGAATGCGAGTAGAACGTATACCCTTATTTGCATGGGGGGTAGTTATTACTACTGTCCTTTTACTTTTATCTCTCCCGGTTTTAGCAGGAGCAATCACTATACTACTAACAGACCGAAACCTAAATACCTCATTCTTTGACCCAGCCGGAGGGGGGGACCCTATCTTATTCCAACACTTGTTCTGATTTTTTGGTCAC
CCCCTCCGGCTGGGTCAAAGAATGAGGTATTTAGGTTTCGGTCTGTTAGTAGTATAGTGATTGCTCCTGCTAAAACCGGGAGAGATAAAAGTAAAAGGACAGTAGTAATAACTACCCCCCATGCAAATAAGGGTATACGTTCTACTCGCATTCCTTCTCATCGTATGTTGAAGGCGGTTGTAATAAAGTTAATTGATCCTAGAATAGAAGATACTCCTGCTAAGTGCAGAGAAAAAATAGCTAGGTCTACAGAGGGTCCGGCATGGGCAATATTTCTTGATAAGGGGGGGTAAACCGTTCATCCTGTCCCGGCTCCTTTTTCTGCTGCTGCAGAGGCTAATAAAAGAATTAGAGAGGGGGGCAAGAGTCAGAAGCTTATATTATTTATTCGAGGGAAGGCTATATCTGGAGCCCCTAGTATAAGGGGGAGAAGTCAATTACCAAATCCACCAATTATTACGGGCATTACTAAAAAGAAAATTATTAAAAAAGCGTGGGCGGTTACTAGTGTATTATAAAGTTGGTCATTTCCTAAGAGAGAGCCGGGTTGGGCTAAGGCCAAACGAATTTGCATTCTAATGGCTGTCCCTGCTATTCCCCCTGCTGTTCCGAAGATAAAGTAAAGGGTTCCAATATCTTTAT
14) flap pine worm (Annelita, Eunicidae) Lysidice ninetta Audouin & Milne-Edwards, the COI gene standard sequence of 1833 is as shown in SEQ ID NO.14
AATTCTCATCCGCCTTGAATTAAGGCAACCTGGATCATTCCTCGGAAGGGACCACTTATACAACACAATTGTAACCGCACATGCTTTCCTAATAATTTTCTTCATAGTAATACCTATTATAATTGGTGGGTTTGGAAATTGACTCCTCCCCCTAATACTTGGCGCCCCAGACATAACATTCCCCCGAATAAATAATCTCAGATTCTGACTTCTACCACCAGCCTTATTACTCCTAATCTCTTCTGCCACAGTAGAAGGAGGTGCTGGCACAGGATGAACAGTTTACCCCCCCCTATCTAGAAACATTGCCCACAGAGGCCCCTCTGTAGACTTAGCTATCTTTTCCCTTCACTTGGCTGGTGCCTCATCCATTTTAGGATCAATTAATTTTATTGCCACCATAATTAATATACGCAACAAAGAAATACGCTTCGAACAAGTTACCTTATTCGTATGAGCAGCCATCATCACCACCATCCTCCTCCTTCTATCTTTACCCGTCCTAGCCGGCGCAATTACCATGCTCCTAACAGACCGAAATTTCAATACCGCTTTCTTTGATCCCTCTGGGGGCGGGGACCCAATTCTATTCCAACATCTATTTTGATTTTTTGGTCACC
TTGAAATTTCGGTCTGTTAGGAGCATGGTAAATGCGCCGGCTAGGACGGGTAAAGATAGAAGGAGGAGGATGGTGGTGATGATGGCTGCTCATACGAATAAGGTAACTTGTTCGAAGCGTATTTCTTTGTTGCGTATATTAATTATGGTGGCAATAAAATTAATTGATCCTAAAATGGATGAGGCACCAGCCAAGTGAAGGGAAAAGATAGCTAAGTCTACAGAGGGGCCTCTGTGGGCAATGTTTCTAGATAGGGGGGGGTAAACTGTTCATCCTGTGCCAGCACCTCCTTCTACTGTGGCAGAAGAGATTAGGAGTAATAAGGCTGGTGGTAAAAGTCAGAATCTGAGATTATTTATTCGGGGGAATGTTATGTCTGGGGCGCCAAGTATTAGGGGGAGGAGTCAATTTCCAAACCCACCAATTATAATAGGTATTACTATGAAGAAAATTATTAGGAAAGCATGTGCGGTTACAATTGTGTTGTATAAGTGGTCCCTTCCGAGGAATGATCCAGGTTGCCCTAATTCAAGGCGGATGAGAATTCTTATGGCTGTTCCTAAAAGGCCACCCCAAGTTCCAAGGATGAAGTAGAGTGTGCCAATATCTTTATG
A method for Rapid identification karang bottom-dwelling species, extracts the genomic dna of each species, with the mitochondrial COI gene sequence of the specific amplification of Auele Specific Primer about 450-650bp;
Primer sequence is:
Forward COI-1490:5 '-GGTCAACAAATCATAAAGATATTGG-3 '
Reverse COI-2198:5 '-TAAACTTCAGGGTGACCAAAAAATCA-3 '
Or,
Forward COI-f:5 '-CCTGCAGGAGGAGGAGAYCC-3 '
Reverse COI-a:5 '-AGTATAAGCGTCTGGGTAGTC-3 ';
Wherein, the Y in COI-f is C or T.
The nucleotide sequence of described specific amplification products and the standard C OI sequence alignment of above-mentioned species, difference be less than 3% can be considered same species.
The advantage that the present invention has is:
The present invention, on the basis of a large amount of research, provides the standard C OI gene order of bottom-dwelling 14 sociales and Common Species in the karang of Hainan.Meanwhile, can identify species fast and effectively according to method of the present invention.
Embodiment
Embodiment 1
The extraction of 1.DNA masterplate.
Get the muscle tissue (in group sociales or the more species sample of individual amount) of about the 100mg of detected sample, shred or grind evenly, put into 1.5mL centrifuge tube.Use marine animal tissue gene group DNA extraction kit (or effective ways of other any extraction DNA), step to specifications, extract the genomic dna of sample.The DNA of acquisition is dissolved in TE damping fluid or ultrapure water and (in lysate, adjusts about pH to 8.0 with NaOH), be placed in-20 DEG C of refrigerators and preserve.By carrying out 1.5% agarose gel electrophoresis to the DNA extracted, ethidium bromide (EB) dyeing and viewed under ultraviolet radiation, the extraction quality of inspection DNA.
2. the pcr amplification of testing sample COI gene order.The PCR reaction system of COI gene order is: cumulative volume 50 μ L.Wherein 10 × PCR damping fluid (containing Mg2+) 5 μ L, dNTP (each 10mM) 4 μ L, each 1 μ L of forward and reverse primer (10 μMs), Taq archaeal dna polymerase (5 units/μ L) 0.5 μ L, DNA profiling 1-5 μ L, all the other volume ultrapure waters are supplied.The PCR response procedures of COI fragment is: 94 DEG C of denaturation 10min, enters PCR circulation afterwards, 95 DEG C of sex change 30s, 48 DEG C of annealing 40s, and 72 DEG C extend 1min, altogether circulation 30-40 time, and last 72 DEG C extend 10min.After pcr amplification terminates, getting 3-5 μ L pcr amplification product, use 1.5% agarose gel electrophoresis, observe under ultraviolet light after EB dyeing, there is object band in inspection 450-650bp.
Primer sequence is:
Forward COI-1490:5 '-GGTCAACAAATCATAAAGATATTGG-3 '
Reverse COI-2198:5 '-TAAACTTCAGGGTGACCAAAAAATCA-3 '
As invalid in above-mentioned primer, then select primer:
Forward COI-f:5 '-CCTGCAGGAGGAGGAGAYCC-3 '
Reverse COI-a:5 '-AGTATAAGCGTCTGGGTAGTC-3 '
Wherein, the Y in COI-f is C or T.
3. sequencing.PCR primer is delivered to order-checking company, use ABI 3730xl automatic sequencer (Applied Biosystems) to measure sequence.In order to ensure the accuracy of sequence, every bar sequence all have employed positive and negative two-way primer and measures.The PCR purified product primer used that checks order is consistent with primer used during pcr amplification.Order-checking obtains this species specificity COI gene order.
4. the comparison of sequence.The peak figure obtained that checks order uses Chromas 2.23 software to carry out the inspection of naked eyes, gets rid of and has bimodal sequence.Use Cont igExpress (a component of Vector NTI Suite 6.0) to align reverse sequence afterwards to splice, indivedual uncertain site is checked and proofreaded simultaneously.Afterwards, the sequence of software Mega 6.0 pairs of amplification genes is used to compare to the sequence of similar species in table 1 sequence table SEQ ID NO.1-14.
After comparison terminates, more with the naked eye check with or without manifest error, whether room aligns.Check errorless after, clip head and the tail two ends primer sequence, obtain the sequence that length is equal.Calculate base difference between the two sequences ratio shared by number of different base (between the sequence) afterwards, two individualities of usual COI sequence difference within 3% can think same species.
Table 1
Claims (3)
1. a COI gene order for each species of karang bottom-dwelling, is characterized in that:
The COI gene standard sequence of each sociales and Common Species in bottom-dwelling in the karang of Hainan:
1) peaceful drum shrimp (crustacean door, Alphaediae) Alpheus pacificus Dana, the COI gene standard sequence of 1852 is as shown in SEQ ID NO.1:
GCTTGGGCTGGTATAGTAGGAACCGCACTCAGTTTACTAATTCGAGCTGAACTTGGACAACCCGGTAGATTAATTGGAAATGACCAAATTTACAATGTAATCGTAACAGCCCATGCATTTGTAATAATTTTCTTTATGGTCATACCAATCATAATTGGAGGTTTTGGGAACTGACTGATTCCCCTTATACTAGGAGCGCCAGATATAGCCTTCCCACGTATAAATAATATAAGATTCTGACTTCTACCCCCCTCCCTCACCCTACTTCTATCTAGTGGCCTAGTAGAAAGGGGGGTCGGCACAGGATGAACTGTTTATCCACCACTATCAGCAGGTATTGCCCACGCGGGAGCATCAGTAGATCTGGGGATTTTCTCTCTCCATTTAGCCGGTGTCTCCTCAATCCTAGGAGCAGTCAACTTTATAACAACAGTTATTAATATACGAGCTACAGGCATAACTATAGACCGTATGCCACTGTTTGTATGAGCTGTATTCCTGACAGCCATTCTACTCCTCCTAAGGCTCCCAGTCCTAGCAGGAGCAATTACTATACTTCTAACAGACCGAAACCTAAATACAGCTTTCTTCGACCCCGCTGGAGGTGGAGATCCAATTTTATACCAACACCTATTTTGATTTTTTGGTCACC
GTCTCCACCTCCAGCGGGGTCGAAGAAAGCTGTATTTAGGTTTCGGTCTGTTAGAAGTATAGTAATTGCTCCTGCTAGGACTGGGAGCCTTAGGAGGAGTAGAATGGCTGTCAGGAATACAGCTCATACAAACAGTGGCATACGGTCTATAGTTATGCCTGTAGCTCGTATATTAATAACTGTTGTTATAAAGTTGACTGCTCCTAGGATTGAGGAGACACCGGCTAAATGGAGAGAGAAAATCCCCAGATCTACTGATGCTCCCGCGTGGGCAATACCTGCTGATAGTGGTGGATAAACAGTTCATCCTGTGCCGACCCCCCTTTCTACTAGGCCACTAGATAGAAGTAGGGTGAGGGAGGGGGGTAGAAGTCAGAATCTTATATTATTTATACGTGGGAAGGCTATATCTGGCGCTCCTAGTATAAGGGGAATCAGTCAGTTCCCAAAACCTCCAATTATGATTGGTATGACCATAAAGAAAATTATTACAAATGCATGGGCTGTTACGATTACATTGTAAATTTGGTCATTTCCAATTAATCTACCGGGTTGTCCAAGTTCAGCTCGAATTAGTAAACTGAGTGCGGTTCCTACTATACCAGCCCAAGCCCCGAAAATAAAGTATAAGGTGCCAATATCTTTATG
2) the COI gene standard sequence of smooth green crab (crustacean door, Xanthidae) Chlorodiella laevissima (Dana, 1852) is as shown in SEQ ID NO.2:
TAGCTGGAATAGTTGGAACATCACTTAGTTTAATTATTCGTGCCGAATTAGGTCAACCTGGAACTCTTATTGGAAATGATCAAATTTATAATGTTGTAGTTACAGCTCATGCTTTTGTTATAATTTTCTTTATAGTTATACCCATTATAATTGGAGGATTTGGTAATTGATTAGTACCCCTTATACTTGGAGCCCCTGATATAGCTTTCCCTCGTATAAACAACATAAGATTTTGACTTCTTCCTCCTTCCCTTACTCTTCTTTTAATAAGAGGAATAGTGGAAAGTGGGGTTGGTACCGGGTGAACAGTTTATCCTCCTCTTGCTGCCGCTATTGCTCATGCTGGAGCTTCAGTTGACATGGGTATTTTCTCTCTTCATTTAGCGGGTGTCTCTTCCATTCTAGGAGCGGTAAACTTCATAACTACCGTAATTAATATACGTTCATTTGGAATGACTATAGATCAAATGCCTCTTTTTGTTTGAGCAGTATTTATTACTGCTATTTTACTTCTCTTATCTCTTCCTGTTCTAGCAGGTGCAATCACTATACTTCTTACAGATCGAAATTTGAATACATCCTTTTTTGACCCAGCTGGGGGAGGTGATCCTGTTCTTTATCAACATTTATTTTGATTTTTTGGTCACC
ATCCCCCAGCTGGGTCAAAAAAGGATGTATTCAAATTTCGATCTGTAAGAAGTATAGTGATTGCACCTGCTAGAACAGGAAGAGATAAGAGAAGTAAAATAGCAGTAATAAATACTGCTCAAACAAAAAGAGGCATTTGATCTATAGTCATTCCAAATGAACGTATATTAATTACGGTAGTTATGAAGTTTACCGCTCCTAGAATGGAAGAGACACCCGCTAAATGAAGAGAGAAAATACCCATGTCAACTGAAGCTCCAGCATGAGCAATAGCGGCAGCAAGAGGAGGATAAACTGTTCACCCGGTACCAACCCCACTTTCCACTATTCCTCTTATTAAAAGAAGAGTAAGGGAAGGAGGAAGAAGTCAAAATCTTATGTTGTTTATACGAGGGAAAGCTATATCAGGGGCTCCAAGTATAAGGGGTACTAATCAATTACCAAATCCTCCAATTATAATGGGTATAACTATAAAGAAAATTATAACAAAAGCATGAGCTGTAACTACAACATTATAAATTTGATCATTTCCAATAAGAGTTCCAGGTTGACCTAATTCGGCACGAATAATTAAACTAAGTGATGTTCCAACTATTCCAGCTCAAGCACCAAAGATAAAATATAATGTTCCAATATCTTTATGT
3) the green crab of black finger (crustacean door, Xanthidae) Chlorodiella nigra (
, 1775) COI gene standard sequence as shown in SEQ ID NO.3
TAGCTGGATAGTAGGGACTTCCCTCAGTTTAATCATCCGTGCTGAACTTGGTCAACCCGGAACCCTTATTGGAAATGATCAAATTTATAATGTTGTAGTTACAGCCCACGCTTTTGTTATAATTTTCTTTATAGTTATACCTATTATAATTGGAGGATTTGGTAATTGATTAGTTCCATTAATGCTTGGAGCCCCTGATATAGCCTTTCCTCGTATAAACAATATAAGATTTTGACTTTTACCTCCATCCCTAACTCTTCTTTTAATAAGTGGAATAGTTGAAAGAGGGGTTGGTACGGGATGAACTGTATATCCTCCTCTTGCCGCTGCTATCGCCCATGCAGGTGCTTCAGTTGATATAGGAATTTTCTCCCTTCACTTGGCAGGTGTATCATCAATTCTAGGGGCAGTAAATTTTATAACTACCGTTATTAATATACGTTCATTTGGTATAACTATAGACCAAATACCTCTTTTTGTTTGAGCAGTATTTATTACTGCTATTTTACTTCTACTTTCTCTTCCTGTGCTAGCAGGAGCTATTACTATGCTTCTTACAGATCGAAATTTAAATACATCTTTCTTTGATCCTGCTGGAGGAGGAGACCCTGTTCTCTACCAACACTTATTCTGATTTTTTGGTCACC
TCTCCTCCTCCAGCAGGATCAAAGAAAGATGTATTTAAATTTCGATCTGTAAGAAGCATAGTAATAGCTCCTGCTAGCACAGGAAGAGAAAGTAGAAGTAAAATAGCAGTAATAAATACTGCTCAAACAAAAAGAGGTATTTGGTCTATAGTTATACCAAATGAACGTATATTAATAACGGTAGTTATAAAATTTACTGCCCCTAGAATTGATGATACACCTGCCAAGTGAAGGGAGAAAATTCCTATATCAACTGAAGCACCTGCATGGGCGATAGCAGCGGCAAGAGGAGGATATACAGTTCATCCCGTACCAACCCCTCTTTCAACTATTCCACTTATTAAAAGAAGAGTTAGGGATGGAGGTAAAAGTCAAAATCTTATATTGTTTATACGAGGAAAGGCTATATCAGGGGCTCCAAGCATTAATGGAACTAATCAATTACCAAATCCTCCAATTATAATAGGTATAACTATAAAGAAAATTATAACAAAAGCGTGGGCTGTAACTACAACATTATAAATTTGATCATTTCCAATAAGGGTTCCGGGTTGACCAAGTTCAGCACGGATGATTAAACTGAGGGAAGTCCCTACTATTCCAGCTCAAGCTCCAAAAATAAAATATAGTGTTCCAATATCTTTATG
4) coral armor shrimp (crustacean door, armor shrimp section) Galathea coralliophilus Baba & Oh, the COI gene standard sequence of 1990 is as shown in SEQ ID NO.4
GAGCAGGATAGTAGGAACTTCTCTGAGACTTATTATTCGAGCTGAATTAGGACAACCAGGTAGATTAATTGGAGATGATCAAATTTATAATGTAGTTGTAACAGCTCATGCTTTCGTTATAATTTTCTTTATAGTTATACCAATTATAATTGGAGGATTTGGAAATTGACTTGTACCTTTAATATTAGGAGCGCCTGATATAGCTTTCCCTCGAATAAATAATATAAGATTTTGGCTTCTTCCGCCCTCTCTTACTTTACTACTCATAAGAGGAATAGTAGAAAGAGGTGTTGGTACAGGTTGAACTGTATACCCTCCTCTTGCAGCAGGAATTGCTCATGCAGGTGCATCTGTTGATATAGGAATTTTTTCTCTTCATCTTGCTGGGGTGTCATCAATTTTAGGGGCAATTAATTTTATAACAACAGTTATTAATATACGACCTAAAGGTATGACAATAGACCGTATACCTCTTTTTGTTTGATCTGTGTTTATTACCGCTATTTTATTACTTTTATCCTTACCTGTATTAGCAGGTGCTATTACAATGTTATTAACAGATCGAAACCTAAATACATCATTCTTTGATCCTGCTGGTGGTGGTGATCCTATTCTATATCAGCATTTATTCTGATTTTTTGGTCACC
CCCACCACCAGCAGGATCAAAGAATGATGTATTTAGGTTTCGATCTGTTAATAACATTGTAATAGCACCTGCTAATACAGGTAAGGATAAAAGTAATAAAATAGCGGTAATAAACACAGATCAAACAAAAAGAGGTATACGGTCTATTGTCATACCTTTAGGTCGTATATTAATAACTGTTGTTATAAAATTAATTGCCCCTAAAATTGATGACACCCCAGCAAGATGAAGAGAAAAAATTCCTATATCAACAGATGCACCTGCATGAGCAATTCCTGCTGCAAGAGGAGGGTATACAGTTCAACCTGTACCAACACCTCTTTCTACTATTCCTCTTATGAGTAGTAAAGTAAGAGAGGGCGGAAGAAGCCAAAATCTTATATTATTTATTCGAGGGAAAGCTATATCAGGCGCTCCTAATATTAAAGGTACAAGTCAATTTCCAAATCCTCCAATTATAATTGGTATAACTATAAAGAAAATTATAACGAAAGCATGAGCTGTTACAACTACATTATAAATTTGATCATCTCCAATTAATCTACCTGGTTGTCCTAATTCAGCTCGAATAATAAGTCTCAGAGAAGTTCCTACTATTCCTGCTCAGGCACCAAAAATAAAATAAAGTGTTCCAATATCTTTATG
5) the COI gene standard sequence of thick chela porcelain crab (crustacean door, Ci Xie section) Pachycheles sculptus (H.Mi lne Edwards, 1837) is carved as shown in SEQ ID NO.5
ACCTAAGAaGTGTTGAGGGAAaATGTTAGATTTACTCCTAAGAATATAACTGAGAAGTGAATTTTTAGCCATTTAGGGTTGACTGATATTCCGGAAAATAATGAGAACCAGTGAGCAATACCAGCAAAAATACCAAAGACAGCTCCTATTGATAAAACATAATGGAAATGGGCAACAACATAGTATGTGTCATGGAGAATTGTATCAATAGAAGAATTTGCAAGAACAACTCCCGTGAGTCCTCCTACAGTAAACAGAAAAATAAATCCTAGGGCCCAGATTATAGAGGGGGAGTAAGTTATTTGATTCCCCTGGAGAGTTCCTAGTCAACTAAAAATCTTAATTCCAGTTGGAATGGCAATAATCATGGTTGCAGATGTAAAGTAGGCTCGTGTATCAACATCTATACCAACAGTGAACATATGGTGAGCTCACACAATAAACCCTAAAATTCCAATTGCTAACATGGCGTAAATTATTCCAAGGGTTCCAAAAGATTCTTTTTTTCCTGATTCCTGGGTTACAATATGAGAGATTATTCCAAAAGCTGGTAAAATTAAAATATATACTTCAGGATGGCCGAAGAACCAGAACAAATGTTGGTAAAGGACGGGGTCTCCTCCCTCCTGCA
TTTGTTCTGGTTCTTCGGCCATCCTGAAGTATATATTTTAATTTTACCAGCTTTTGGAATAATCTCTCATATTGTAACCCAGGAATCAGGAAAAAAAGAATCTTTTGGAACCCTTGGAATAATTTACGCCATGTTAGCAATTGGAATTTTAGGGTTTATTGTGTGAGCTCACCATATGTTCACTGTTGGTATAGATGTTGATACACGAGCCTACTTTACATCTGCAACCATGATTATTGCCATTCCAACTGGAATTAAGATTTTTAGTTGACTAGGAACTCTCCAGGGGAATCAAATAACTTACTCCCCCTCTATAATCTGGGCCCTAGGATTTATTTTTCTGTTTACTGTAGGAGGACTCACGGGAGTTGTTCTTGCAAATTCTTCTATTGATACAATTCTCCATGACACATACTATGTTGTTGCCCATTTCCATTATGTTTTATCAATAGGAGCTGTCTTTGGTATTTTTGCTGGTATTGCTCACTGGTTCTCATTATTTTCCGGAATATCAGTCAACCCTAAATGGCTAAAAATTCACTTCTCAGTTATATTCTTAGGAGTAAATCTAACATTTTTCCCTCAACACTTCTTAGGTCTAAACGGAATACCACGTCGATACTCTGACTACCCA
6) coarse hair shell crab (crustacean door, Xanthidae) Pilodius scabriculus Dana, the COI gene standard sequence of 1852 is as shown in SEQ ID NO.6
GAGCTGGTATGGTCGGTACATCTCTAAGTCTTATTATCCGTGCTGAACTAGGACAACCCGGAACACTTATTGGAAATGATCAAATCTACAATGTTATTGTAACCGCTCATGCCTTTGTAATAATCTTTTTCATAGTTATACCTATTATAATTGGAGGATTTGGTAATTGACTTGTCCCATTAATATTAGGAGCTCCTGATATAGCCTTTCCTCGTATAAACAATATAAGATTTTGACTTCTTCCCCCATCCCTTACTCTTCTTCTTATAAGAGGTATAGTCGAAAGAGGAGTTGGTACAGGTTGGACCGTTTATCCACCTCTTGCTGCCGCTATCGCTCACGCTGGGGCCTCAGTAGATATAGGTATTTTCTCTCTCCATTTGGCTGGTGTATCCTCTATTTTAGGAGCAGTTAATTTTATAACTACCGTTATTAACATACGATCCTTTGGTATAACCATAGACCAAATACCTCTTTTTGTTTGAGCTGTATTTATTACCGCAATCCTGCTTTTACTTTCCTTACCTGTATTAGCAGGAGCTATTACTATGCTCCTTACTGACCGTAATTTAAACACATCCTTCTTTGATCCAGCCGGAGGTGGTGATCCTGTTCTTTATCAACATCTATTCTGATTTTTTGGTCACC
GTGAAGACAGGATCACCACCTCCGGCTGGATCAAAGAAGGATGTGTTTAAATTACGGTCAGTAAGGAGCATAGTAATAGCTCCTGCTAATACAGGTAAGGAAAGTAAAAGCAGGATTGCGGTAATAAATACAGCTCAAACAAAAAGAGGTATTTGGTCTATGGTTATACCAAAGGATCGTATGTTAATAACGGTAGTTATAAAATTAACTGCTCCTAAAATAGAGGATACACCAGCCAAATGGAGAGAGAAAATACCTATATCTACTGAGGCCCCAGCGTGAGCGATAGCGGCAGCAAGAGGTGGATAAACGGTCCAACCTGTACCAACTCCTCTTTCGACTATACCTCTTATAAGAAGAAGAGTAAGGGATGGGGGAAGAAGTCAAAATCTTATATTGTTTATACGAGGAAAGGCTATATCAGGAGCTCCTAATATTAATGGGACAAGTCAATTACCAAATCCTCCAATTATAATAGGTATAACTATGAAAAAGATTATTACAAAGGCATGAGCGGTTACAATAACATTGTAGATTTGATCATTTCCAATAAGTGTTCCGGGTTGTCCTAGTTCAGCACGGATAATAAGACTTAGAGATGTACCGACCATACCAGCTCAAGCTCCAAAGATAAAATATAGTGTACCAATATCTTTATG
7) the COI gene standard sequence of three seta thorn crab (crustacean door, burr crab section) Pilumnus trispinosus (Sakai, 1965) is as shown in SEQ ID NO.7
TTTACCTAGGAATGTTGTGGAAAAGGTTGTGTTAACTCCTACAAATATAGTGAAGAAATGAATTTTTATTCATCGTGGGTTTAATGATAGGCCTGTAAACAGGGGAAATCAGTGAGCGATTCCTCCGAAAATACCAAATACGGCACCTATTGATAAAACGTAGTGGAAATGGGCTACAACATAATATGTATCATGTAAAATAATATCAATTGAGGAGTTGGCTAATACTACTCCTGTTAATCCACCTATAGTAAATAAAAAAATAAACCCTAAGGCTCATATTAAGGAAGGTCTATAATTAATTTGAGTTCCATGAAGGGTTCTTAACCAACTGAAAATTTTAATTCCCGTGGGGACAGCAATGATTATTGTAGCAGAGGTGAAATAAGCTCGGGTATCTACATCTATTCCAACTGTGAATATATGGTGAGCTCAAACTACAAATCCTAAGATACCAATAGCTAATATAGCATAAATTATCCCTAAAGTGCCGAATGATTCTTTTTTTCCTGATTCTTGTCTTACAATATGGGAGATTATACCGAAGGCTGGAAGAATAAGAATATAAACTTCAGGGTGACCGAAGAATCAAAATAAATGTTGATAGAGGACAGGGTCTCC
TTGATTCTTCGGTCACCCTGAAGTTTATATTCTTATTCTTCCAGCCTTCGGTATAATCTCCCATATTGTAAGACAAGAATCAGGAAAAAAAGAATCATTCGGCACTTTAGGGATAATTTATGCTATATTAGCTATTGGTATCTTAGGATTTGTAGTTTGAGCTCACCATATATTCACAGTTGGAATAGATGTAGATACCCGAGCTTATTTCACCTCTGCTACAATAATCATTGCTGTCCCCACGGGAATTAAAATTTTCAGTTGGTTAAGAACCCTTCATGGAACTCAAATTAATTATAGACCTTCCTTAATATGAGCCTTAGGGTTTATTTTTTTATTTACTATAGGTGGATTAACAGGAGTAGTATTAGCCAACTCCTCAATTGATATTATTTTACATGATACATATTATGTTGTAGCCCATTTCCACTACGTTTTATCAATAGGTGCCGTATTTGGTATTTTCGGAGGAATCGCTCACTGATTTCCCCTGTTTACAGGCCTATCATTAAACCCACGATGAATAAAAATTCATTTCTTCACTATATTTGTAGGAGTTAACACAACCTTTTTTCCACAACATTTCCTAGGTTTAAATGGTATGCCACGACGATATTCGGACTACCCA
8) the COI gene standard sequence of special-shaped beans porcelain crab (crustacean door, Ci Xie section) Pisidia dispar (Stimpson, 1858) is as shown in SEQ ID NO.8
GGAATAGTTGGTACATCTCTTAGTTTAATTATTCGAGCTGAACTAGGTCAATTAGGAAGATTAATTGGTGACGATCAAATTTACAATGTTGTAGTCACTGCTCACGCTTTTGTTATAATTTTTTTTATAGTTATACCAATTTTAATTGGAGGATTCGGGAACTGACTAGTTCCTTTAATACTCGGAGCCCCTGATATAGCTTTTCCTCGAATAAATAATATAAGATTTTGACTTTTACCCCCAGCATTGTCACTTCTTTTAATAAGAGGATTAGTTGAAAGAGGAGTGGGAACGGGATGAACCGTTTACCCACCCTTAGCCTCAAATATCGCCCACGCTGGCGCTTCTGTAGACATAGGAATTTTTTCTCTCCACCTTGCAGGAGTTTCCTCAATTTTAGGAGCTGTAAATTTTATTTCTACTGTTATCAACATACGATCAGTTGGAATAACCTTTGACCGTATACCACTTTTTGTTTGATCGGTTTTTATTACAGCAATTTTACTTCTTTTATCTTTACCCGTTTTAGCTGGGGCAATTACCATACTTCTAACAGATCGAAACCTTAATACTTCATTTTTCGACCCAGCAGGGGGAGGAGATCCTATTCTTTATCAACACTTATTTTGATTTTTTGGTCACC
TCCTCCCCCTGCTGGGTCGAAAAATGAAGTATTAAGGTTTCGATCTGTTAGAAGTATGGTAATTGCCCCAGCTAAAACGGGTAAAGATAAAAGAAGTAAAATTGCTGTAATAAAAACCGATCAAACAAAAAGTGGTATACGGTCAAAGGTTATTCCAACTGATCGTATGTTGATAACAGTAGAAATAAAATTTACAGCTCCTAAAATTGAGGAAACTCCTGCAAGGTGGAGAGAAAAAATTCCTATGTCTACAGAAGCGCCAGCGTGGGCGATATTTGAGGCTAAGGGTGGGTAAACGGTTCATCCCGTTCCCACTCCTCTTTCAACTAATCCTCTTATTAAAAGAAGTGACAATGCTGGGGGTAAAAGTCAAAATCTTATATTATTTATTCGAGGAAAAGCTATATCAGGGGCTCCGAGTATTAAAGGAACTAGTCAGTTCCCGAATCCTCCAATTAAAATTGGTATAACTATAAAAAAAATTATAACAAAAGCGTGAGCAGTGACTACAACATTGTAAATTTGATCGTCACCAATTAATCTTCCTAATTGACCTAGTTCAGCTCGAATAATTAAACTAAGAGATGTACCAACTATTCCTGCTCATGCGCCAAAAATAAAATATAATGTTCCAATATCTTTATGT
9) the knurl palm closes the COI gene standard sequence of drum shrimp (crustacean door, Alphaediae) Synalpheus tumiolomanus (Paul'son, 1875) as shown in SEQ ID NO.9
ATTCGAGCCGACTTGGTCAACCTGGTAGCCTAATTGGGAACGACCAAATTTATAACGTAATTGTCACTGCACACGCATTTGTCATAATTTTCTTTATGGTTATACCTATTATAATCGGGGGATTCGGAAACTGACTAGTTCCCCTTATATTAGGAGCTCCAGATATAGCATTTCCCCGAATAAATAACATAAGATTTTGATTACTTCCTCCCTCTCTTACCCTCCTTTTATCTAGAGGAATAGTGGAAAGAGGAGTGGGAACTGGTTGAACAGTCTATCCACCTCTTGCAAGAGGAATTGCCCATGCAGGAGCTTCAGTTGACATAGGAATTTTTTCATTACATCTAGCAGGAGTCTCCTCAATTTTAGGAGCTGTTAATTTTATAACCACTGTAATCAACATACGAGCAGCAGGAATAACAATAGACCAAATACCACTTTTTGTATGAGCAGTTTTTTTAACTGCAATCTTATTATTATTAAGACTTCCAGTTTTAGCTGGAGCTATTACTATACTATTAACAGACCGAAATCTAAAACACATAC
CTAACTGGAGTCTTAATAATAATAAGATTGCAGTTAAAAAAACTGCTCATACAAAAAGTGGTATTTGGTCTATTGTTATTCCTGCTGCTCGTATGTTGATTACAGTGGTTATAAAATTAACAGCTCCTAAAATTGAGGAGACTCCTGCTAGATGTAATGAAAAAATTCCTATGTCAACTGAAGCTCCTGCATGGGCAATTCCTCTTGCAAGAGGTGGATAGACTGTTCAACCAGTTCCCACTCCTCTTTCCACTATTCCTCTAGATAAAAGGAGGGTAAGAGAGGGAGGAAGTAATCAAAATCTTATGTTATTTATTCGGGGAAATGCTATATCTGGAGCTCCTAATATAAGGGGAACTAGTCAGTTTCCGAATCCCCCGATTATAATAGGTATAACCATAAAGAAAATTATGACAAATGCGTGTGCAGTGACAATTACGTTATAAATTTGGTCGTTCCCAATTAGGCTACCAGGTTGACCAAGTTCGGCTCGAATTAAAAGTCTGAGAGAAGTACCAACTATTCCTGCTCACGCTC
10) carefully sting greatly ophiruid (Echinodermata, thorn ophiruid section) Macrophiothrix lorioli A.M.Clark, the COI gene standard sequence of 1968 is as shown in SEQ ID NO.10
TCTCAGCTAGAGTGTTGGGGAAATGTAAGGTTGACTCCAATAAACATTAAAAAGAAGTGGGTAAGTGAGCTGGCCCGATCAATTGTGGCACCGGTAAATAGAGAGAATCAGTGATTAAATCCGGCAAAAATTGCAAATACTGCTCCCATTGACAGTACATAGTGGAAGTGAGCCGTGACGTAATATGTATCGTGGAGGGCTACGTTTAAAGATGAGTTTGATAAAATTATTCCAGTTAAACCCCCTATGGTGAATAAAAATATGAATCCTAGGGCTCAATACAGAGAGGGGTGTGCCTTGTTTATGTGGAAGAATGCTCCCTGGAGTGTTGCAAGTCATCTAAAAACCTTGACCCCTGTCGGAATAGCTATTATCATTGTGGCTGCTGTAAAGTAGGCGCGTGTGTCCACGTCCATGCCTACGGTGAACATGTGGTGGGCTCAGACGATGAAGCCTAGTATTCCAATGGAGATCATTGCGTACATCATACCTAAATACCCGAAGGGGGTAGCCTTCCCTGTTCGGTTAGCCACTACGTGTGAAATAATCCCAAATCCTGGTAAAATTAAGATATAAACTTCGGGGTGCCCAAAAAATCAAAATAGGTGTTGAAATAGGATTGGGTCTCCTCT
GGGCACCCCGAGTTTATATCTTAATTTTACCAGGATTTGGGATTATTTCACACGTAGTGGCTAACCGAACAGGGAAGGCTACCCCCTTCGGGTATTTAGGTATGATGTACGCAATGATCTCCATTGGAATACTAGGCTTCATCGTCTGAGCCCACCACATGTTCACCGTAGGCATGGACGTGGACACACGCGCCTACTTTACAGCAGCCACAATGATAATAGCTATTCCGACAGGGGTCAAGGTTTTTAGATGACTTGCAACACTCCAGGGAGCATTCTTCCACATAAACAAGGCACACCCCTCTCTGTATTGAGCCCTAGGATTCATATTTTTATTCACCATAGGGGGTTTAACTGGAATAATTTTATCAAACTCATCTTTAAACGTAGCCCTCCACGATACATATTACGTCACGGCTCACTTCCACTATGTACTGTCAATGGGAGCAGTATTTGCAATTTTTGCCGGATTTAATCACTGATTCTCTCTATTTACCGGTGCCACAATTGATCGGGCCAGCTCACTTACCCACTTCTTTTTAATGTTTATTGGAGTCAACCTTACATTTTTTCCCCAACACTTCTTAGGCCTTGCAGGAATGCCACGGCGATACTCAGACTACCCAGAG
11) the COI gene standard sequence of spoke ophiruid (Echinodermata, spoke ophiruid section) Ophiactis savignyi (M ü ller & Troschel, 1842) is as shown in SEQ ID NO.11
AGGGTCTCCTCCCCCTGTTGGATCGAAGAAGGTAGTGTTAATTTTTCGGTCTGTGAGGAGCATGGTGATGGCTCCGGCTAGAACAGGTAAGGAGAGCAGGAGGAGGATTGTGGTTAACAAAATGGATCAAACGAAAAGAGGCATACGGTCCATGGTCATTCCAGGGGCCCGCATGTTAAGAATAGTGGTTATAAAGTTTATAGAGGCCATAATAGATGAGGCTCCGGCCAGGTGTAGAGAAAAGATCGCGAGGTCTACACACCCTCCTGCGTGGGCGGTTGGGCCAGATAAGGGCGGGTAAACTGTTCACCCGGTTCCTACTCCCCCTTCTTTGGCAGCCGAAGCTAAGAGTAAAATGAAGGCGGGAGGTACAAGTCAGAATCTCATTTTTTTCATACGTGGGAATGCCATATCTGGAGCTCCTAACATTAAAGGGACCAGTCAGTTTCCAAATCCT
GATATGGCATTCCCACGTATGAAAAAAATGAGATTCTGACTTGTACCTCCCGCCTTCATTTTACTCTTAGCTTCGGCTGCCAAAGAAGGGGGAGTAGGAACCGGGTGAACAGTTTACCCGCCCTTATCTGGCCCAACCGCCCACGCAGGAGGGTGTGTAGACCTCGCGATCTTTTCTCTACACCTGGCCGGAGCCTCATCTATTATGGCCTCTATAAACTTTATAACCACTATTCTTAACATGCGGGCCCCTGGAATGACCATGGACCGTATGCCTCTTTTCGTTTGATCCATTTTGTTAACCACAATCCTCCTCCTGCTCTCCTTACCTGTTCTAGCCGGAGCCATCACCATGCTCCTCACAGACCGAAAAATTAACACTACCTTCTTCGATCCAACAGGGGGAGGAGACCCTATTTTATTTCAACACTTGTTCTGATTTTTTGGTC
12) the COI gene standard sequence of coarse two finger squama worm (Annelita, Shuan Zhilin worm section) Iphione muricata (Lamarck, 1818) is as shown in SEQ ID NO.12
TGATCTGGATTATTGGGCACATCAATAAGTCTTTTAATTCGGGCCGAGCTCGGCCAACCAGGGTCCCTTCTAGGCAGAGACCAACTTTATAATACGATTGTTACGGCTCATGCTTTTCTTATAATCTTCTTTCTGGTAATACCAGTATTAGTAGGAGGATTTGGTAACTGACTTATCCCGTTAATACTAGGGGCTCCGGACATGGCTTTCCCACGTCTGAATAATATAAGATTCTGACTCTTACCCCCTTCACTTATCCTTCTTTTATCATCTAGTGCTGTTGAAAAAGGAGTTGGGACTGGCTGAACTGTCTACCCCCCTCTTGCTAGAAACATTGCTCATGCTGGACCATCTGTTGACCTAGCAATTTTTTCTTTACATATTGCAGGAGTCTCTTCAATTCTAGGTGCTCTAAATTTTATTACTACTGTAATTAATATACGATATAAAGGCCTCCGTCTAGAACGTGTCCCATTGTTTGTATGAGCCGCCAAGATTACTGCCATTCTTCTTTTACTTTCTCTCCCCGTCTTAGCCGGCGCTATTACTATACTTCTCACCGACCGTAATCTAAACACTGCTTTCTTTGACCCTGCTGGTGGTGGTGACCCCGTCCTTTATCAGCATTTATTTTGATTTTTTGGTCAC
CCACCACCAGCAGGGTCAAAGAAAGCAGTGTTTAGATTACGGTCGGTGAGAAGTATAGTAATAGCGCCGGCTAAGACGGGGAGAGAAAGTAAAAGAAGAATGGCAGTAATCTTGGCGGCTCATACAAACAATGGGACACGTTCTAGACGGAGGCCTTTATATCGTATATTAATTACAGTAGTAATAAAATTTAGAGCACCTAGAATTGAAGAGACTCCTGCAATATGTAAAGAAAAAATTGCTAGGTCAACAGATGGTCCAGCATGAGCAATGTTTCTAGCAAGAGGGGGGTAGACAGTTCAGCCAGTCCCAACTCCTTTTTCAACAGCACTAGATGATAAAAGAAGGATAAGTGAAGGGGGTAAGAGTCAGAATCTTATATTATTCAGACGTGGGAAAGCCATGTCCGGAGCCCCTAGTATTAACGGGATAAGTCAGTTACCAAATCCTCCTACTAATACTGGTATTACCAGAAAGAAGATTATAAGAAAAGCATGAGCCGTAACAATCGTATTATAAAGTTGGTCTCTGCCTAGAAGGGACCCTGGTTGGCCGAGCTCGGCCCGAATTAAAAGACTTATTGATGTGCCCAATAATCCAGATCATGTACCAAAAATGAAGTAAAGAGTACCAATATCTTTAT
13) pearl palpus rock clam worm (Annelita, Eunicidae) Leodice antennata Savignyin Lamarck, the COI gene standard sequence of 1818 is as shown in SEQ ID NO.13
GACAGCAGGGGGATAGCAGGGACAGCCATTAGAATGCAAATTCGTTTGGCCTTAGCCCAACCCGGCTCTCTCTTAGGAAATGACCAACTTTATAATACACTAGTAACCGCCCACGCTTTTTTAATAATTTTCTTTTTAGTAATGCCCGTAATAATTGGTGGATTTGGTAATTGACTTCTCCCCCTTATACTAGGGGCTCCAGATATAGCCTTCCCTCGAATAAATAATATAAGCTTCTGACTCTTGCCCCCCTCTCTAATTCTTTTATTAGCCTCTGCAGCAGCAGAAAAAGGAGCCGGGACAGGATGAACGGTTTACCCCCCCTTATCAAGAAATATTGCCCATGCCGGACCCTCTGTAGACCTAGCTATTTTTTCTCTGCACTTAGCAGGAGTATCTTCTATTCTAGGATCAATTAACTTTATTACAACCGCCTTCAACATACGATGAGAAGGAATGCGAGTAGAACGTATACCCTTATTTGCATGGGGGGTAGTTATTACTACTGTCCTTTTACTTTTATCTCTCCCGGTTTTAGCAGGAGCAATCACTATACTACTAACAGACCGAAACCTAAATACCTCATTCTTTGACCCAGCCGGAGGGGGGGACCCTATCTTATTCCAACACTTGTTCTGATTTTTTGGTCAC
CCCCTCCGGCTGGGTCAAAGAATGAGGTATTTAGGTTTCGGTCTGTTAGTAGTATAGTGATTGCTCCTGCTAAAACCGGGAGAGATAAAAGTAAAAGGACAGTAGTAATAACTACCCCCCATGCAAATAAGGGTATACGTTCTACTCGCATTCCTTCTCATCGTATGTTGAAGGCGGTTGTAATAAAGTTAATTGATCCTAGAATAGAAGATACTCCTGCTAAGTGCAGAGAAAAAATAGCTAGGTCTACAGAGGGTCCGGCATGGGCAATATTTCTTGATAAGGGGGGGTAAACCGTTCATCCTGTCCCGGCTCCTTTTTCTGCTGCTGCAGAGGCTAATAAAAGAATTAGAGAGGGGGGCAAGAGTCAGAAGCTTATATTATTTATTCGAGGGAAGGCTATATCTGGAGCCCCTAGTATAAGGGGGAGAAGTCAATTACCAAATCCACCAATTATTACGGGCATTACTAAAAAGAAAATTATTAAAAAAGCGTGGGCGGTTACTAGTGTATTATAAAGTTGGTCATTTCCTAAGAGAGAGCCGGGTTGGGCTAAGGCCAAACGAATTTGCATTCTAATGGCTGTCCCTGCTATTCCCCCTGCTGTTCCGAAGATAAAGTAAAGGGTTCCAATATCTTTAT
14) flap pine worm (Annelita, Eunicidae) Lysidice ninetta Audouin & Mi lne-Edwards, the COI gene standard sequence of 1833 is as shown in SEQ ID NO.14
AATTCTCATCCGCCTTGAATTAAGGCAACCTGGATCATTCCTCGGAAGGGACCACTTATACAACACAATTGTAACCGCACATGCTTTCCTAATAATTTTCTTCATAGTAATACCTATTATAATTGGTGGGTTTGGAAATTGACTCCTCCCCCTAATACTTGGCGCCCCAGACATAACATTCCCCCGAATAAATAATCTCAGATTCTGACTTCTACCACCAGCCTTATTACTCCTAATCTCTTCTGCCACAGTAGAAGGAGGTGCTGGCACAGGATGAACAGTTTACCCCCCCCTATCTAGAAACATTGCCCACAGAGGCCCCTCTGTAGACTTAGCTATCTTTTCCCTTCACTTGGCTGGTGCCTCATCCATTTTAGGATCAATTAATTTTATTGCCACCATAATTAATATACGCAACAAAGAAATACGCTTCGAACAAGTTACCTTATTCGTATGAGCAGCCATCATCACCACCATCCTCCTCCTTCTATCTTTACCCGTCCTAGCCGGCGCAATTACCATGCTCCTAACAGACCGAAATTTCAATACCGCTTTCTTTGATCCCTCTGGGGGCGGGGACCCAATTCTATTCCAACATCTATTTTGATTTTTTGGTCACC
TTGAAATTTCGGTCTGTTAGGAGCATGGTAAATGCGCCGGCTAGGACGGGTAAAGATAGAAGGAGGAGGATGGTGGTGATGATGGCTGCTCATACGAATAAGGTAACTTGTTCGAAGCGTATTTCTTTGTTGCGTATATTAATTATGGTGGCAATAAAATTAATTGATCCTAAAATGGATGAGGCACCAGCCAAGTGAAGGGAAAAGATAGCTAAGTCTACAGAGGGGCCTCTGTGGGCAATGTTTCTAGATAGGGGGGGGTAAACTGTTCATCCTGTGCCAGCACCTCCTTCTACTGTGGCAGAAGAGATTAGGAGTAATAAGGCTGGTGGTAAAAGTCAGAATCTGAGATTATTTATTCGGGGGAATGTTATGTCTGGGGCGCCAAGTATTAGGGGGAGGAGTCAATTTCCAAACCCACCAATTATAATAGGTATTACTATGAAGAAAATTATTAGGAAAGCATGTGCGGTTACAATTGTGTTGTATAAGTGGTCCCTTCCGAGGAATGATCCAGGTTGCCCTAATTCAAGGCGGATGAGAATTCTTATGGCTGTTCCTAAAAGGCCACCCCAAGTTCCAAGGATGAAGTAGAGTGTGCCAATATCTTTATG。
2. a method for Rapid identification karang bottom-dwelling species, is characterized in that:
Extract the genomic dna of each species, with the mitochondrial COI gene sequence of the specific amplification of Auele Specific Primer about 450-650bp;
Primer sequence is:
Forward COI-1490:5 '-GGTCAACAAATCATAAAGATATTGG-3 '
Reverse COI-2198:5 '-TAAACTTCAGGGTGACCAAAAAATCA-3 '
Or,
Forward COI-f:5 '-CCTGCAGGAGGAGGAGAYCC-3 '
Reverse COI-a:5 '-AGTATAAGCGTCTGGGTAGTC-3 ';
Wherein, the Y in COI-f is C or T.
3., by the method for Rapid identification karang bottom-dwelling species according to claim 2, it is characterized in that: the nucleotide sequence of described specific amplification products and the standard C OI sequence alignment of above-mentioned species, difference be less than 3% can be considered same species.
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| CN109234409A (en) * | 2018-11-01 | 2019-01-18 | 浙江海洋大学 | A kind of Portumidae mitochondrial COI gene universal primer and design and amplification method |
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| CN110894526A (en) * | 2019-09-05 | 2020-03-20 | 中国环境科学研究院 | High-throughput sequencing method for benthonic animal COI gene and application thereof |
| CN112359119A (en) * | 2020-11-18 | 2021-02-12 | 中国科学院生态环境研究中心 | Environmental DNA macro-bar code method for large-scale benthonic animal community structure research |
| CN113337621A (en) * | 2021-07-19 | 2021-09-03 | 中国科学院南海海洋研究所 | Molecular monitoring method for starfish larvae in coral reef area |
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