CN109006693A - A method of induction lefteye flounder gene mutation - Google Patents
A method of induction lefteye flounder gene mutation Download PDFInfo
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- CN109006693A CN109006693A CN201810876494.8A CN201810876494A CN109006693A CN 109006693 A CN109006693 A CN 109006693A CN 201810876494 A CN201810876494 A CN 201810876494A CN 109006693 A CN109006693 A CN 109006693A
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- 238000000034 method Methods 0.000 title claims abstract description 44
- 241000694873 Paralichthyidae Species 0.000 title claims abstract description 29
- 206010064571 Gene mutation Diseases 0.000 title claims abstract description 8
- 230000006698 induction Effects 0.000 title description 5
- 235000013601 eggs Nutrition 0.000 claims abstract description 36
- 230000035772 mutation Effects 0.000 claims abstract description 20
- 241000269980 Pleuronectidae Species 0.000 claims abstract description 11
- 230000001939 inductive effect Effects 0.000 claims abstract description 6
- 108010000912 Egg Proteins Proteins 0.000 claims description 32
- 102000002322 Egg Proteins Human genes 0.000 claims description 32
- 210000004681 ovum Anatomy 0.000 claims description 32
- 239000012895 dilution Substances 0.000 claims description 20
- 238000010790 dilution Methods 0.000 claims description 20
- 230000012447 hatching Effects 0.000 claims description 18
- 239000013535 sea water Substances 0.000 claims description 17
- 230000009027 insemination Effects 0.000 claims description 7
- 239000000203 mixture Substances 0.000 claims description 7
- 238000003756 stirring Methods 0.000 claims description 7
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 6
- 230000004720 fertilization Effects 0.000 claims description 5
- 230000004913 activation Effects 0.000 claims description 4
- 238000001514 detection method Methods 0.000 claims description 4
- 241000581364 Clinitrachus argentatus Species 0.000 claims description 3
- 238000011534 incubation Methods 0.000 claims description 3
- 230000000149 penetrating effect Effects 0.000 claims description 2
- 238000009395 breeding Methods 0.000 abstract description 9
- 230000001488 breeding effect Effects 0.000 abstract description 9
- 231100000350 mutagenesis Toxicity 0.000 abstract description 9
- 241000251468 Actinopterygii Species 0.000 abstract description 8
- 238000002703 mutagenesis Methods 0.000 abstract description 8
- 239000000243 solution Substances 0.000 description 10
- 230000002159 abnormal effect Effects 0.000 description 6
- 239000002773 nucleotide Substances 0.000 description 5
- 125000003729 nucleotide group Chemical group 0.000 description 5
- FUSGACRLAFQQRL-UHFFFAOYSA-N N-Ethyl-N-nitrosourea Chemical compound CCN(N=O)C(N)=O FUSGACRLAFQQRL-UHFFFAOYSA-N 0.000 description 4
- 230000007423 decrease Effects 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 230000018109 developmental process Effects 0.000 description 3
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 238000009360 aquaculture Methods 0.000 description 2
- 244000144974 aquaculture Species 0.000 description 2
- 238000012217 deletion Methods 0.000 description 2
- 230000005251 gamma ray Effects 0.000 description 2
- 230000002068 genetic effect Effects 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 description 2
- 238000012165 high-throughput sequencing Methods 0.000 description 2
- 238000003780 insertion Methods 0.000 description 2
- 230000037431 insertion Effects 0.000 description 2
- 230000001678 irradiating effect Effects 0.000 description 2
- 238000009832 plasma treatment Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 230000029936 alkylation Effects 0.000 description 1
- 238000005804 alkylation reaction Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003183 carcinogenic agent Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000002962 chemical mutagen Substances 0.000 description 1
- 238000003851 corona treatment Methods 0.000 description 1
- 238000005034 decoration Methods 0.000 description 1
- 230000037430 deletion Effects 0.000 description 1
- 238000006073 displacement reaction Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- 210000001161 mammalian embryo Anatomy 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 231100000310 mutation rate increase Toxicity 0.000 description 1
- 102000054765 polymorphisms of proteins Human genes 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- UIIMBOGNXHQVGW-UHFFFAOYSA-M sodium bicarbonate Substances [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 description 1
- 229910000162 sodium phosphate Inorganic materials 0.000 description 1
- 239000002023 wood Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/027—New or modified breeds of vertebrates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2227/00—Animals characterised by species
- A01K2227/40—Fish
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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Farming Of Fish And Shellfish (AREA)
Abstract
The present invention relates to a kind of method for inducing lefteye flounder gene mutation, the method uses normal-temperature plasma, is irradiated to halibut spermatozoon or lefteye flounder fertilized eggs.It is preferred that the irradiation power of normal-temperature plasma is 120~200W, irradiation time is 1.5~30min, irradiates spacing≤2mm.The present invention carries out mutagenesis using fertilized eggs or sperm of the normal-temperature plasma to lefteye flounder, can generate a certain proportion of mutated individual.This method is easy to operate, securely and reliably, can be used successfully to the breeding work of lefteye flounder and other fish, the efficiency that mutation rate is significantly increased, promotes New idioplasm resource initiative.
Description
Technical field
The present invention relates to field of biotechnology, and in particular to a method of gene mutation occurs for induction lefteye flounder.
Background technique
The features such as lefteye flounder is the important marine fish in China, has fine and tender taste delicious, and economic value is high, it is deep to be disappeared
Expense person's likes.Breeding is the important foundation guarantee of culture fishery sustainable health development, breeder's breeding in China
Go out multiple lefteye flounder improved Varieties, provides provenance support for the development of lefteye flounder aquaculture.In recent years, with overfishing and
Environmental pollution causes lefteye flounder natural resources to decay, causes for germ plasm resource used in breeding in quantity and genetic diversity
Show a sharp decline.Therefore, for the sustainable development of lefteye flounder fine-variety breeding and aquaculture, it is necessary to be provided to its germplasm
It is efficiently formulated in source.
Mutagenesis is a kind of effective means for carrying out Germplasm enhancement.Method of mutagenesis currently used for fish is predominantly penetrated
Line mutagenesis and chemical mutagenesis, used mutation source mainly include gamma ray, X-ray and N-ethyl-N-nitrosourea (N-
Ethyl-N-nitrosourea, abbreviation ENU).Wherein, gamma ray and X-ray have radiativity, and equipment is expensive, and need special
The protection facility property to ensure safety of door.ENU is a kind of chemical mutagen, it is repaired by the alkylation to genomic DNA base
Decorations, induction DNA occur mispairing in duplication and generate mutation.This substance is 2A class carcinogenic substance, can be caused to the health of human body
Significant damage.Meanwhile the efficiency of inducing mutation of above-mentioned several mutation sources is all lower.Accordingly, it is desirable to provide a kind of safety for lefteye flounder
Reliable and high induced mutation rate method of mutagenesis.
In consideration of it, the present invention is specifically proposed.
Summary of the invention
The purpose of the present invention is to provide a kind of methods for inducing lefteye flounder gene mutation.
To achieve the above object, technical scheme is as follows:
The present invention relates to a kind of methods for inducing lefteye flounder gene mutation, and the method uses normal-temperature plasma, to lefteye flounder
Sperm or lefteye flounder fertilized eggs are irradiated.
Preferably, the irradiation power of the normal-temperature plasma is 120~200W, and irradiation time is 1.5~30min, is shone
Penetrate spacing≤2mm.
Preferably, when being irradiated to fertilized eggs, the irradiation power of the plasma is 120~160W, the photograph
Penetrating the time is 20~25min.
Preferably, when being irradiated to sperm, the irradiation power of the plasma is 160~200W, the irradiation
Time is 2~10min.
Preferably, it when being irradiated to fertilized eggs, the described method comprises the following steps:
1) fresh halibut spermatozoon and ovum are acquired, and is placed in different containers；
2) 6~40 times of dilution is carried out to the sperm with dilution；
3) sperm after dilution is mixed with ovum, is stirring evenly and then adding into seawater, obtained through sea-water activated essence
The smart ovum mixture is hatched 60 minutes in the seawater, obtains fertilized eggs by ovum mixture；
4) fertilized eggs are transferred in culture dish, using plasma is irradiated at normal temperature, then will be described
Fertilized eggs, which are transferred in hatching cylinder, is hatched.
Preferably, the incubation temperature in step 3) and step 4) is 15~20 DEG C, preferably 17 DEG C.
Preferably, it when being irradiated to sperm, the described method comprises the following steps:
1) fresh halibut spermatozoon and ovum are acquired, and is placed in different containers；
2) dilution for carrying out 6~40 times to the sperm with dilution, is then transferred to culture dish for the sperm after dilution
In, using plasma is irradiated at normal temperature；
3) sperm after irradiation is mixed with ovum, is stirring evenly and then adding into seawater, activation ovum complete manually by
Essence obtains fertilized eggs；
4) fertilized eggs are transferred in hatching cylinder and are hatched.
Preferably, above two method further includes the detection of step 4) mutation rate: fry is obtained after incubating oosperm, in March
The fin ray of clip fish when age, the method that sequence is resurveyed by high-throughput full-length genome, obtain each fry individual SNP number and
InDel number obtains mutation rate by the sum of SNP number and InDel number divided by full-length genome base number.
Beneficial effects of the present invention:
The present invention carries out mutagenesis using fertilized eggs or sperm of the normal-temperature plasma to lefteye flounder, can generate a certain proportion of
Mutated individual.This method is easy to operate, securely and reliably, can be used successfully to the breeding work of lefteye flounder and other fish, be significantly increased
Mutation rate, the efficiency for promoting New idioplasm resource initiative.
Specific embodiment
To make the object, technical solutions and advantages of the present invention clearer, technical solution of the present invention will be carried out below
Detailed description.Obviously, described embodiment is only a part of the embodiment of the present invention, instead of all the embodiments.It is based on
Embodiment in the present invention, those of ordinary skill in the art without making creative work it is obtained it is all its
Its embodiment belongs to the range that the present invention is protected.
The present invention relates to a kind of methods for inducing lefteye flounder gene mutation, and method uses normal-temperature plasma, to halibut spermatozoon
Or lefteye flounder fertilized eggs are irradiated.
Normal-temperature plasma (Atmospheric and Room Temperature Plasma, ARTP) is a kind of available
In the novel method of mutation breeding, there is easy to operate, environmental nonpollution and the features such as endangering.ARTP hair used in the present invention
It penetrates source to be produced by clear sky wood Biotechnology Co., Ltd of Luoyang China, model ARTP-P.
In one embodiment of the invention, the irradiation power of normal-temperature plasma is 120~200W, and irradiation time is
1.5~30min irradiates spacing≤2mm.The irradiation power of usual normal-temperature plasma is higher, the abnormal rate of irradiation object and prominent
Variability is bigger.However, the death rate that excessively high irradiation power will lead to irradiation object ramps.Irradiation time is to irradiation object
Also there are above-mentioned affecting laws.Therefore, it is necessary to further limit the relationship of irradiation power and irradiation time.
In a preferred embodiment of the invention, when being irradiated to fertilized eggs, the irradiation power of plasma is
120~160W, irradiation time are 20~25min；When being irradiated to sperm, the irradiation power of plasma is 160~
When 200W, irradiation time is 2~10min.Relatively high abnormal rate and mutation rate, while lefteye flounder fertilized eggs can be obtained in this way
The death rate can also receive.
In one embodiment of the invention, if be irradiated using normal-temperature plasma to fertilized eggs, this method packet
Include following steps:
1) fresh halibut spermatozoon and ovum are acquired, and is placed in different containers.
2) 6~40 times of dilution is carried out to sperm with dilution.
Ringer's solution (Ringer's Solution) is selected to be used as dilution, formula in the present invention are as follows: every 1000ml steams
NaCl 8.22g, KCl 0.39g, CaCl is added in distilled water2·2H2O 0.72, MgCl2·6H2O 0.23g, NaH2PO4·2H2O
0.28g, NaHCO30.20g, glucose 1.00g, and pH value is adjusted to 6.5~7.2.
3) artificial insemination: the sperm after the dilution of above-mentioned ringer's solution is mixed with ovum, is stirring evenly and then adding into sea
Water obtains that smart ovum mixture is hatched 60 in 15~20 DEG C, preferably 17 DEG C of seawater through sea-water activated smart ovum mixture
Minute, obtain fertilized eggs.
4) normal-temperature plasma is handled: fertilized eggs being transferred in culture dish, using plasma is shone at normal temperature
It penetrates, the time of irradiation and power are seen above, and then fertilized eggs are transferred in hatching cylinder, are hatched under discharge condition.
In one embodiment of the invention, if be irradiated using normal-temperature plasma to halibut spermatozoon, this method
The following steps are included:
1) fresh halibut spermatozoon and ovum are acquired, and is placed in different containers.
2) corona treatment: using dilution, and preferably ringer's solution carries out 6~40 times of dilution to sperm, then will be dilute
Sperm after releasing is transferred in culture dish, and using plasma is irradiated at normal temperature.
3) artificial insemination: the sperm after irradiation is mixed with ovum, is stirring evenly and then adding into seawater, and activation ovum is complete
At artificial insemination, fertilized eggs are obtained.
4) fertilized eggs are transferred in hatching cylinder and are hatched.
The present invention also provides the mutation rate detection method after a kind of mutagenesis, this method is carried out at 3 monthly age of fry, specifically
Process includes:
After being irradiated halibut spermatozoon or lefteye flounder fertilized eggs using normal-temperature plasma, hatched to obtain according to conventional
Fry.The clip fin ray at 3 monthly age of fry, the method for resurveying sequence by high-throughput full-length genome obtain each fry individual
SNP number and InDel number.Since SNP number and InDel number can almost represent the type of all variations, by sum of the two divided by complete
Genome base number, obtains mutation rate.
Wherein, high throughput sequencing technologies (High-throughput sequencing), with can be once parallel to hundreds of thousands
It is mark to millions of DNA moleculars progress sequencings and long shorter wait of general reading.Illumina is used in the present invention
Hiseq1500 is sequenced.
Single nucleotide polymorphism (Single Nucleotide Polymorphisms, abbreviation SNP), refers in genome
The genetic marker that the variation of upper single nucleotide acid is formed.Refer to the variation of single nucleotide acid in the genome, including displacement, transversion,
Missing and insertion.There are many its quantity, rich polymorphism.Insertion and deletion marks (insertion-deletion, abbreviation InDel),
Refer to the difference in two kinds of parents in full-length genome.For another opposite parent, in the genome of one of parent
There are a certain number of nucleotides inserteds or missing.
Embodiment 1
Normal-temperature plasma irradiation is carried out to fertilized eggs
1) acquisition of sperm and ovum: fresh lefteye flounder milter sperm and female fish egg are acquired, is kept in dark place；
2) artificial insemination: being diluted with sperm of the ringer's solution to above-mentioned acquisition, by after dilution sperm and ovum into
Row mixing.After mixing evenly, seawater is added thereto to be activated, ocean temperature is 17 DEG C, is obtained through sea-water activated smart ovum
Mixture hatches the smart ovum mixture 60 minutes in 17 DEG C of seawater；
3) normal-temperature plasma treatment with irradiation: after sixty minutes, fertilized eggs are transferred in culture dish for fertilization, and one layer of tiling is simultaneously
A little seawater is stayed, normal-temperature plasma irradiation is carried out, irradiation spacing is 2mm.After the completion of irradiation, fertilized eggs are transferred to hatching cylinder
Interior Lotic hatching；
4) mutation rate detects: at 3 monthly age, the fin ray of 3 tail fish of clip resurveys the side of sequence using high-throughput full-length genome
Method, the SNP number and InDel number for obtaining each individual are dashed forward by the sum of SNP number and InDel number divided by full-length genome base number
Variability.
Wherein, the volume ratio of ringer's solution and sperm, the volume ratio of sperm and ovum, normal-temperature plasma power and irradiation
Time and test result are shown in Table 1.
Table 1
* in table 1, rate of fertilization is the percentage that gastrul stage floating embryo number accounts for induction total ovum number used；
Hatching rate is the percentage hatched prelarva number and account for fertilized eggs number；
Abnormal rate is the percentage that lopsided prelarva number accounts for hatching prelarva number；
Mutation rate is the percentage that SNP number and InDel number account for full-length genome base number.
From table 1 it follows that being fertilized as the irradiation time increases under conditions of plasma irradiating power is constant
Rate and hatching rate gradually decrease, and abnormal rate and mutation rate gradually rise.Hatching rate is reduced to 40% when being 30min between upon irradiation
Hereinafter, being unfavorable for breeding.
Embodiment 2
Normal-temperature plasma irradiation is carried out to halibut spermatozoon
1) acquisition of sperm and ovum: fresh lefteye flounder milter sperm and the female fish egg of lefteye flounder are acquired, is kept in dark place；
2) normal-temperature plasma treatment with irradiation: sperm is diluted with ringer's solution, the sperm after dilution is laid in
Normal-temperature plasma irradiation is carried out in culture dish, irradiation spacing is 2mm；
3) artificial insemination: the sperm after irradiation is kept in dark place, is mixed when needing with ovum, is stirring evenly and then adding into
Seawater, activation ovum complete artificial insemination.Fertilized eggs are transferred to Lotic hatching in hatching cylinder；
4) mutation rate detects: at 3 monthly age, the fin ray of 3 tail fish of clip resurveys the side of sequence using high-throughput full-length genome
Method, the SNP number and InDel number for obtaining each individual are dashed forward by the sum of SNP number and InDel number divided by full-length genome base number
Variability.
Wherein, the volume ratio of ringer's solution and sperm, the volume ratio of sperm and ovum, normal-temperature plasma power and irradiation
Time and test result are shown in Table 2.
Table 2
* in table 2, rate of fertilization, hatching rate, abnormal rate and mutation rate definition are the same as table 1.
From Table 2, it can be seen that being fertilized as the irradiation time increases under conditions of plasma irradiating power is constant
Rate and hatching rate gradually decrease, and abnormal rate and mutation rate gradually rise.Hatching rate is reduced to when being 12min between upon irradiation
24.8% or so, it is unfavorable for breeding.
The above description is merely a specific embodiment, but scope of protection of the present invention is not limited thereto, any
Those familiar with the art in the technical scope disclosed by the present invention, can easily think of the change or the replacement, and should all contain
Lid is within protection scope of the present invention.Therefore, protection scope of the present invention should be based on the protection scope of the described claims.
Claims (10)
1. a kind of method for inducing lefteye flounder gene mutation, which is characterized in that the method uses normal-temperature plasma, to lefteye flounder essence
Son or lefteye flounder fertilized eggs are irradiated.
2. the method according to claim 1, wherein the irradiation power of the normal-temperature plasma be 120~
200W, irradiation time are 1.5~30min, irradiate spacing≤2mm.
3. according to the method described in claim 2, it is characterized in that, when being irradiated to fertilized eggs, the plasma
Irradiation power is 120~160W, and the irradiation time is 20~25min.
4. according to the method described in claim 2, it is characterized in that, when being irradiated to sperm, the photograph of the plasma
Penetrating power is 160~200W, and the irradiation time is 2~10min.
5. according to the method described in claim 3, it is characterized in that, when being irradiated to fertilized eggs, the method includes with
Lower step:
1) fresh halibut spermatozoon and ovum are acquired, and is placed in different containers；
2) 6~40 times of dilution is carried out to the sperm with dilution；
3) sperm after dilution is mixed with ovum, is stirring evenly and then adding into seawater, obtained mixed through sea-water activated smart ovum
Object is closed, the smart ovum mixture is hatched 60 minutes in the seawater, obtains fertilized eggs；
4) fertilized eggs are transferred in culture dish, using plasma is irradiated at normal temperature, then by the fertilization
Ovum, which is transferred in hatching cylinder, is hatched.
6. according to the method described in claim 5, it is characterized in that, the incubation temperature in step 3) and step 4) is 15~20
DEG C, preferably 17 DEG C.
7. according to the method described in claim 5, it is characterized in that, the method also includes the detections of step 4) mutation rate: fertilization
Fry is obtained after egg hatching, the fin ray of clip fish at 3 monthly age, the method for resurveying sequence by high-throughput full-length genome obtains every
The SNP number and InDel number of one fry individual obtain mutation rate by the sum of SNP number and InDel number divided by full-length genome base number.
8. according to the method described in claim 4, it is characterized in that, the method includes following when being irradiated to sperm
Step:
1) fresh halibut spermatozoon and ovum are acquired, and is placed in different containers；
2) sperm after dilution, is then transferred in culture dish by the dilution for carrying out 6~40 times to the sperm with dilution,
Using plasma is irradiated under room temperature；
3) sperm after irradiation is mixed with ovum, is stirring evenly and then adding into seawater, activation ovum is completed artificial insemination, obtained
To fertilized eggs；
4) fertilized eggs are transferred in hatching cylinder and are hatched.
9. according to the method described in claim 8, it is characterized in that, the incubation temperature in step 3) and step 4) is 15~20
DEG C, preferably 17 DEG C.
10. according to the method described in claim 8, it is characterized in that, above two method further includes the detection of step 4) mutation rate:
Fry is obtained after incubating oosperm, the fin ray of clip fish at 3 monthly age, the method for resurveying sequence by high-throughput full-length genome obtains
The SNP number and InDel number for obtaining each fry individual are dashed forward by the sum of SNP number and InDel number divided by full-length genome base number
Variability.
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| CN115104559A (en) * | 2022-07-22 | 2022-09-27 | 福建省水产研究所（福建水产病害防治中心） | Method for mutagenizing black male clown fish body color to become red by ARTP (ARTP) |
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