1989 Fiscal Year Final Research Report Summary
Studies on the mutation of mitochondrial DNA in salmonids as a genetic tag for stock management.
| Project/Area Number |
62440016
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| Research Category |
Grant-in-Aid for General Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Research Field |
General fisheries
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| Research Institution | The University of Tokyo |
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Principal Investigator |
NUMACHI Ken-ichi Ocean Res. Inst., Univ. of Tokyo Professor, 海洋研究所, 教授 (30013569)
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| Co-Investigator(Kenkyū-buntansha) |
KOBAYASHI Takanori Ocean Res. Inst., Univ. of Tokyo Research Associate, 海洋研究所, 助手 (70205467)
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| Project Period (FY) |
1987 – 1989
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| Keywords | mitochondrial DNA / salmonids / stock management / restriction fragmental patterns |
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| Research Abstract |
Rapid and simple purification of mt DNA in fish species was established by applying SDS-NaCl precipitation instead of DNase treatment of mitochondria pellets, and by adopting low temperature below 20ﾟ C throughout all the treatments including SDS and RNase treatment. In this procedure, yields of ccDNA were much increased (X 2- 400) compared with the method used so far, and crude mt DNA from 50-60 specimens were prepared within a day. Furthermore, a improved ultrracentrifugal procedure that can separate a larger amount of mtDNA (200mug or more) within 3 hr was developed.
Applying this purification method, 106 specimens of chum salmon from 5 rivers and 86 specimens of 5 natural and artificial sustained populations were analyzed by 10 kinds of 6 base-pair restriction endonuclease. Totally 13 types of mutations were demonstrated in the fragmental patterns produced by digestion of 5 kinds of enzymes, and 9 clonal genotypes of mtDNA were posturated. River populations were consisted of common
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types ( II and III) and some river specific types. The base pair substitutions (delta, Nei and Li) among the clonal genotypes were 0.2-1 %.
In the masu salmon, 22 clonal genotypes in combination of 20 types of fragmental patterns in 7 kinds of enzymes were demonstrated. The base-pair substitutions (delta) among the clonal genotypes were 0.2-2.85 %, and the nucleotide diversity (pi, Nei and Li ) among populations were very high, 0.893-1.531. Each population has their own specific genotypes abundantly. These genotypes were suggested as a useful genetic tags to study population. The Formosan masu salmon consisted of 29 specimens showed only one clonal genotypes, which showed much resemblance with the types in the masu distributed in the northern part of Japan. The base pair substitutions between them were 0.2-1.59 %. We concluded that the Formosan salmon originated from the masu salmon in the Sea of Japan through the Tsushima channel at 100-800 thousands years ago, and the unusual homogeneity observed in the Formosan salmon should be caused by recent contracted stage of population. Less
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