CN116804225B - Ji Shi tilapia sex chromosome specific molecular marker, detection primer, kit and application thereof - Google Patents
Ji Shi tilapia sex chromosome specific molecular marker, detection primer, kit and application thereof Download PDFInfo
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
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Abstract
The invention discloses a Ji Shi tilapia sex chromosome specific molecular marker, a detection primer, a kit and application thereof, and belongs to the technical field of molecular markers. The Ji Shi tilapia sex chromosome specific molecular marker comprises an X chromosome and a Y chromosome specific molecular marker; the nucleotide sequence of the X chromosome specific molecular marker is shown as SEQ ID NO.1, and the nucleotide sequence of the Y chromosome specific molecular marker is shown as SEQ ID NO. 2. According to the method, specific molecular markers of X and Y chromosomes of Ji Shi tilapia are screened for the first time, a Ji Shi tilapia genetic sex PCR identification method is established, and the method is applied to genetic sex identification in the cultivation process of all-male Ji Shi tilapia. The whole process has the characteristics of high efficiency, accuracy and stability, and has important application value in Ji Shi tilapia sex control and continuous large-scale production of all-male Ji Shiluo non-fish fries.
Description
Technical Field
The invention belongs to the technical field of molecular markers, and particularly relates to a Ji Shi tilapia sex chromosome specific molecular marker, a detection primer, a kit and application thereof.
Background
Research on sex determination, differentiation mechanism and sex-linked molecular marker assisted breeding technology of fish has direct commercial value in aquaculture because many economic traits including growth rate and body color have sex-tographic properties (Mei and guide, 2015). In many aquatic organisms cultivated at present, obvious individual size two states between male and female can be observed. For example, in some species females grow faster than males and are larger in size, including cynoglossus semilaevis (Chen et al, 2007) and carp (Cyprinus carpio) (Wu and Gui, 1999); in contrast, males of nile tilapia (Mair et al, 1995) and Huang (Pelteobagrus fulvidraco) (Wang et al, 2009 b) grow faster and have a larger body size. Thus, screening of parthenocarpic fish can greatly enhance the benefits of aquaculture, and identification of sex-specific markers for these species is an important focus of aquaculture research. Although fish are generally present with sex dichotomy in phenotypic traits, the sex chromosomes of most fish are morphologically very similar and indistinguishable by cytological analysis. Finding genetic markers associated with sex loci is therefore a more scientific and accurate approach (charles worth et al 2005).
Ji Shi Tilapia (Coptodon zillii) belonging to the genus Mylopharyngodon, family Mylopharyngodon, genus Mylopharyngodon, is native to the water system from Africa to about denier river, and introduced into China in 1978 as a breeding variety. The plant has strong environmental adaptability, can resist pollution and low-oxygen environment, has extremely strong reproductive capacity and can grow rapidly, and becomes a common breeding variety. Ji Shi the male tilapia grows faster than the female tilapia, and the fully-androgenic culture can obviously improve the culture efficiency of Ji Shi tilapia (Xie Xiaojun et al, 1994). Meanwhile, the obtaining of the super-male fish and the total-male fish is helpful for basic research. Therefore, the research of the genetic sex identification and sex control technology of Ji Shi tilapia is not only of great scientific significance, but also has wide application prospect.
Ji Shi tilapia chromosome 2n=22 is XY sex chromosome. Due to the plasticity of the sex of the fishes, on one hand, the sex can be treated by a male steroid hormone, an antiestrogen, an aromatase (estrogen synthesis key enzyme) inhibitor and the like, the individuals with the genetic sex XX are induced to develop into pseudo-male fishes (XX still genetically), the XX pseudo-male fishes and normal XX female fishes are bred in a mating way, and finally, the full female offspring is obtained, so that the full female single fingerling cultivation of the Ji Shi tilapia is realized. On the other hand, by treatment with an estrogen and an androgen inhibitor (Sun et al, 2014), development of individuals with genetic sex XY into pseudo-female fish (XY is still genetically) is induced, and the XY pseudo-female fish is bred with normal XY male fish to obtain YY super-male fish, and the YY super-male fish is subjected to female treatment, and the obtained YY pseudo-female fish is bred with normal XY male fish to obtain whole male fish. The traditional fish sex control breeding technology is a method for judging the genotype of a parent through test cross, depends on the growth cycle of fish and consumes time; and based on sex-linked molecular markers, the genetic sex can be simply and rapidly identified, and manpower and material resources are greatly saved. However, there is no report on the development of molecular markers in Ji Shi tilapia.
Disclosure of Invention
Aiming at the defects in the prior art, the invention provides a Ji Shi tilapia sex chromosome specific molecular marker, a detection primer, a kit and application thereof, which can be suitable for identifying the genetic sex of wild Ji Shi tilapia in laboratories and different watersheds. And the method can economically, rapidly and accurately distinguish individuals of normal female fish XX, normal male fish XY, transformed female fish XY and YY, transformed male fish XX and super male fish YY. In addition, the application also provides a method for rapidly and efficiently producing the all-male fish based on the molecular marker, so that the large-scale production of the all-male fish fries is realized, the cultivation yield is obviously improved, the cultivation cost is reduced, and the economic benefit is improved; the fourth aim is to provide a method for rapidly and efficiently producing all-male and super-male fish based on the molecular marker, so as to obtain the unisexual fish fry applied to basic research.
In order to achieve the above purpose, the technical scheme adopted by the invention for solving the technical problems is as follows:
a Ji Shi tilapia sex chromosome specific molecular marker, comprising an X chromosome and a Y chromosome specific molecular marker; the nucleotide sequence of the X chromosome specific molecular marker is shown as SEQ ID NO.1, and the nucleotide sequence of the Y chromosome specific molecular marker is shown as SEQ ID NO. 2.
The primer for detecting the Ji Shi tilapia sex chromosome specific molecular marker has a primer sequence shown as SEQ ID NO.3 and SEQ ID NO.4, and the specific sequence is as follows:
an upstream primer: 5'-AGTTAATGTGTTCCTAGTGTCC-3'; (SEQ ID NO. 3)
A downstream primer: 5'-GCCTCTGACATGCTGAACAT-3'. (SEQ ID NO. 4)
Further, the primers can amplify X chromosome specific molecular markers with the length of 570bp and Y chromosome specific molecular markers with the length of 704 bp.
A kit for detecting the Ji Shi tilapia sex chromosome specific molecular marker, which comprises the primer.
The molecular marker, the primer or the kit is applied to the breeding of Ji Shi tilapia.
A method for identifying the genetic sex of Ji Shi tilapia comprises the steps of amplifying the genome of Ji Shi tilapia to be detected by adopting the primer or the kit, and comparing the amplified fragment with Ji Shi tilapia sex chromosome specific molecular markers to determine the genetic sex.
Further, the genetic sex of the Ji Shi tilapia to be detected is determined based on the length of the amplified fragment, specifically:
when only the X chromosome specific molecular marker with the length of 570bp is amplified from the genome DNA of the Ji Shi tilapia to be detected, judging that the Ji Shi tilapia to be detected is a inherited female fish, and the sex chromosome genotype is XX;
when only a Y chromosome specific molecular marker with the length of 704bp is amplified from the genome DNA of the Ji Shi tilapia to be detected, judging that the Ji Shi tilapia to be detected is a genetic super-male fish, and the sex chromosome genotype is YY;
when the X chromosome specific molecular marker and the Y chromosome specific molecular marker are amplified simultaneously from the genomic DNA of the Ji Shi tilapia to be detected, the Ji Shi tilapia to be detected is judged to be a genetic male fish, and the sex chromosome genotype is XY.
A method for cultivating a total female Ji Shi tilapia, comprising the steps of:
(1) Carrying out maleization treatment on offspring generated after mating of Ji Shi tilapia XX female fish and XY male fish, and screening out a transformed male fish with sex chromosome genotype XX by using the primers;
(2) Mating the XX-transformed male fish with the XX female fish of the normal Ji Shi tilapia to obtain the XX full female fish.
A method for cultivating supermale and Quan Xiongji tilapia, comprising the steps of:
(1) Carrying out feminization treatment on offspring generated after mating of Ji Shi tilapia XX female fish and XY male fish, and screening out transformed female fish with sex chromosome genotype XY by using the primers;
(2) Mating XY transformed female fish with XY male fish of Ji Shi tilapia, and screening supermale fish with sex chromosome genotype YY from offspring by using the primers; in addition, the mating offspring of XY transformed female fish and Ji Shi tilapia XY male fish are taken for female treatment, and the primers are used for screening out the transformed female fish with sex chromosome genotypes of XY and YY;
(3) Mating YY super-male fish with XY or YY converted female fish to obtain YY super-male fish;
(4) And mating the YY super-male fish with the XX normal female fish to obtain the XY full-male fish.
The invention has the beneficial effects that:
according to the method, specific molecular markers of X and Y chromosomes of Ji Shi tilapia are screened for the first time, a Ji Shi tilapia genetic sex PCR identification method is established, and the method is applied to genetic sex identification in the cultivation process of all-male Ji Shi tilapia. The whole process has the characteristics of high efficiency, accuracy and stability, and has important application value in Ji Shi tilapia sex control and continuous large-scale production of all-male Ji Shiluo non-fish fries.
The Ji Shi tilapia X and Y chromosome specific molecular marker provided by the invention can be applied to wild Ji Shi tilapia in laboratories and in different drainage areas; the Ji Shi tilapia genetic sex identification method constructed based on the molecular marker can conveniently, quickly and accurately distinguish normal female fish XX, normal male fish XY, converted female fish XY and YY, converted male fish XX and super male fish YY; based on the genetic sex identification method, the large-scale production of the full female fish fries can be realized by mating XX-transformed male fish with XX-female normal female fish, so that the breeding yield is obviously improved, the breeding cost is reduced, and the economic benefit is improved; through mating normal female fish XX (male) with super-male fish YY (female) to generate XY all-male fish, and converting female fish YY (female) and super-male fish YY (male) to mate to generate YY super-male fish, a large number of single fish fries can be obtained, so that the method is applied to basic research.
Drawings
FIG. 1 is a sequence alignment diagram of specific molecular markers of X and Y chromosomes according to the present invention; in the figure, primer positions are marked with arrows, the black background is the consistent sequence of the X chromosome and Y chromosome DNA fragments, "-" indicates the deleted sequence;
FIG. 2 shows the results of genetic sex identification of female 7-tail Ji Shi tilapia XX and male 7-tail Ji Shi tilapia XY using X and Y chromosome-specific molecular markers according to the present invention; wherein M represents a DNA molecular weight standard DL2000;
FIG. 3 shows the results of genetic sex determination of Ji Shi tilapia (96 total) from different watercourses using X and Y chromosome specific molecular markers according to the present invention; wherein M represents a DNA molecular weight standard DL2000;
FIG. 4 shows a technical route for producing genetic all-female fish (A), all-male fish and super-male fish (B) by using the X and Y chromosome specific molecular markers for auxiliary breeding; FAD, fadrozole, aromatase inhibitors; e2,17β -estradiol (17β -estradiol); tr, trilostane.
Detailed Description
The following description of the embodiments of the present invention is provided to facilitate understanding of the present invention by those skilled in the art, but it should be understood that the present invention is not limited to the scope of the embodiments, and all the inventions which make use of the inventive concept are protected by the spirit and scope of the present invention as defined and defined in the appended claims to those skilled in the art.
The experimental methods of the present invention, in which specific conditions are not specified, are generally carried out according to conventional conditions, for example, conditions described in the guidelines for molecular cloning experiments (third edition, J. Sam Brookfield et al, huang Peitang et al, science Press, 2002) or according to the conditions recommended by the manufacturer.
Example 1 Ji Shi obtaining of Tilapia sex-specific molecular markers
1. Extraction of genomic DNA
(1) Shearing fin of Ji Shi tilapia mossambica in a 1.5mL EP tube, adding 600 mu L of STNE [10mM Tris-HCl (pH8.0) +100mM EDTA (pH8.0) +100mM NaCl+5mg/mL SDS ] buffer and 10 mu L (20 mg/mL) proteinase K, shaking and mixing uniformly;
(2) Placing the mixture in a shaking table at a constant temperature of 55 ℃ to oscillate for 5-8 hours;
(3) According to Tris saturated phenol: chloroform: isoamyl alcohol=25:24:1, adding the mixed solution with equal volume (600 mu L), slowly reversing the mixed solution to the centrifuge tube, and uniformly mixing the solution;
(4) Centrifugation at 12,000rpm at 4℃for 10min, pipetting the supernatant and transferring to a fresh 1.5mL EP tube;
(5) Repeating the steps 3 and 4 twice;
(6) Adding chloroform with the same volume as the supernatant, mixing, centrifuging at 12,000rpm for 10min at 4 ℃ for extraction, sucking the supernatant, and transferring to a new 1.5mL EP tube;
(7) Adding 1/10 volume of 3mol/L NaAc and equal volume of isopropanol, mixing, standing at-20deg.C for 2 hr;
(8) Centrifuging at 12000rpm at 4deg.C for 20min to precipitate DNA, and discarding supernatant;
(9) Adding 1mL of 75% ethanol, centrifuging at 12000rpm at 4 ℃ for 2min, washing the precipitate, and pouring out the ethanol;
(10) Adding 1mL of absolute ethanol, centrifuging at 12000rpm at 4 ℃ for 2min, washing the precipitate, pouring out the ethanol and drying at room temperature;
(11) Dissolving the dried precipitate with DDW;
(12) Detection of the concentration of extracted DNA and OD value (OD) Using a nucleic acid protein quantitative measuring apparatus 260 /OD 280 ＝1.76～1.80)。
2. Screening of sex-specific molecular markers
Based on the data of full genome sequencing and male and female body weight sequencing of the Ji Shi tilapia in the early stage of a laboratory, comparing the obtained clean reads obtained by sequencing and re-sequencing and filtering the XY male fish and XX female fish individuals with Ji Shi tilapia reference genome by software BWA, positioning Ji Shi sex chromosomes and sex determination regions of the tilapia by analyzing and counting the distribution of sex-linked SNP by GWAS, sequencing and indexing the compared bam files, opening by using visual software Integrative Genomics Viewer (IGV), checking, designing primers at the places with obvious structural variation of the male and female individuals, respectively carrying out PCR amplification on XX, XY Ji Shi tilapia genome pools, and screening sex-specific molecular markers.
As a result, it was found that the primer sequences shown in SEQ ID NO.3 and SEQ ID NO.4 have obvious sex specificities, and can amplify an X chromosome specific band of 570bp in XX and XY individuals, while amplifying a Y chromosome specific band of 704bp in XY individuals.
3. Cloning and sequence analysis of sex-specific molecular markers
Sepharose gel containing the specific X chromosome band and specific Y chromosome band is taken, target fragments are recovered, cloned into a pMD-19T vector, competent cells of escherichia coli are transformed, positive clones are screened by a blue colony PCR method, white spot single colonies are selected, positive clones are identified by a PCR method, and sequencing is carried out.
Sequence alignment analysis was performed on the X chromosome specific molecular marker (sequence shown as SEQ ID NO.1, named CzX) and the Y chromosome specific molecular marker (sequence shown as SEQ ID NO.2, named CzY) obtained by sequencing. As a result, as shown in FIG. 1, the sequence differences of the X and Y chromosome specific molecular markers were mainly: compared with the X chromosome specific molecular marker, the Y chromosome specific molecular marker inserts a 130bp sequence at 355 bp.
Example 2 identification of Ji Shi genetic sex of Tilapia
The molecular marker obtained in the example 1 is used for identifying Ji Shi tilapia genetic sex, and the specific process is as follows:
(1) And (3) PCR amplification: shearing fin bars of Ji Shi tilapia to be detected, and extracting genome DNA; using the obtained genome DNA as a template, and adopting an upstream primer F shown in SEQ ID NO.3 and a downstream primer R shown in SEQ ID NO.4 for PCR amplification; the PCR amplification parameters were: pre-denaturation at 95℃for 3 min, then denaturation at 95℃for 30 sec, annealing at 60℃for 30 sec, extension at 72℃for 45 sec for 30 cycles, and finally extension at 72℃for 10 min; the PCR product was subjected to agarose gel electrophoresis at 1.5%;
(2) And (3) judging results: when only an X chromosome specific band with the length of 570bp is amplified from the genome DNA of the Ji Shi tilapia to be detected (namely the X chromosome specific molecular marker CzX), judging that the Ji Shi tilapia to be detected is a genetic female fish, and the sex chromosome genotype is XX; when the X chromosome specific band with the length of 570bp and the Y chromosome specific band with the length of 704bp are amplified simultaneously from the genome DNA of the Ji Shi tilapia to be detected, the Ji Shi tilapia to be detected is judged to be a genetic male fish, and the sex chromosome genotype is XY.
The genetic sex determination is carried out on 7-tail Ji Shi tilapia female fish XX and 7-tail Ji Shi tilapia male fish XY by the method, the result is shown in FIG. 2, only X chromosome specific bands with the length of 570bp are amplified in all XX individuals, and X chromosome specific bands with the length of 570bp and Y chromosome specific bands with the length of 704bp are simultaneously amplified in XY individuals, which are consistent with the theoretical result.
Example 3 genetic sex determination of wild Ji Shi Tilapia mossambica in different watershed based on sex-specific molecular markers
Genetic sex determination was performed on 50-tail Ji Shi tilapia and 144-tail Ji Shi tilapia from the Jiangsu tin-free freshwater fish research center by applying the method constructed in example 2, and the result is shown in FIG. 3.
Only the X chromosome specific band of 570bp in all female Ji Shi tilapia individuals tested was amplified, while the X chromosome specific band of 570bp and the Y chromosome specific band of 704bp were simultaneously amplified in all male Ji Shi tilapia individuals tested, indicating that the 96 to-be-tested Ji Shi tilapia phenotypes were completely identical to genotypes (table 1). From the above results, the sex-specific molecular marker developed in the present experiment was applicable to Ji Shi tilapia of 2 populations, and had universality.
Table 12 genetic sex determination results of different populations of Ji Shi tilapia
Example 4 method for culturing all-male and all-female Ji Shi Tilapia based on specific molecular markers
Genetic sex identification of tilapia Ji Shi is carried out based on the sex-specific molecular markers CzX/CzY obtained in example 1, and tilapia Quan Xiongji is cultivated by the following specific procedures:
1. cultivation of full female Ji Shi tilapia
A. Performing Fadrozole dipping bath on the offspring of the female and male fishes XX and XY mating of Ji Shi tilapia in 5 days after hatching, and screening out a transformed male fish with sex chromosome genotype XX by adopting the molecular marking method for identifying the genetic sex of Ji Shi tilapia;
B. mating XX-transformed male fish with normal Ji Shi tilapia XX female fish to obtain XX full female fish, thereby mass-producing full female commercial fish. (A in FIG. 4)
2. The sex-specific molecular marker also establishes a method for cultivating the super-male and Quan Xiongji tilapia, and the specific process is as follows:
A. carrying out feminization treatment (mixed dipping bath of androgen inhibitor Tr and estrogen E2) on mating offspring of Ji Shi tilapia female fish XX and male fish XY, and screening out a transformed female fish with sex chromosome genotype XY by adopting the molecular marking method for identifying the genetic sex of Ji Shi tilapia;
B. mating XY transformed female fish with XY male fish of Ji Shi tilapia, and screening supermale fish with sex chromosome genotype YY from offspring by adopting the molecular marking method for Ji Shi tilapia genetic sex identification; in addition, the XY transformed female fish and the XY male fish of Ji Shi tilapia are mated to be subjected to female treatment, and the method for molecular marking for identifying the genetic sex of Ji Shi tilapia is adopted to screen the transformed female fish with sex chromosome genotypes of XY and YY;
C. mating YY super-male fish with XY or YY transformed female fish to obtain YY super-male fish;
D. and mating the YY super-male fish with the XX normal female fish to obtain the XY total-male fish. (B in FIG. 4)
Finally, it should be noted that the above-mentioned embodiments are only for illustrating the technical solution of the present invention, and not for limiting the same, and although the present invention has been described in detail with reference to examples, it should be understood by those skilled in the art that modifications and equivalents may be made to the technical solution of the present invention without departing from the spirit and scope of the technical solution of the present invention, and all such modifications and equivalents are intended to be encompassed in the scope of the claims of the present invention.
Claims (9)
1. A primer for detecting Ji Shi tilapia sex chromosome specific molecular markers is characterized in that the primer sequences are shown as SEQ ID NO.3 and SEQ ID NO. 4.
2. The primer of claim 1, wherein the primer amplified Ji Shi tilapia sex chromosome specific molecular markers comprise X chromosome and Y chromosome specific molecular markers; the nucleotide sequence of the X chromosome specific molecular marker is shown as SEQ ID NO.1, and the nucleotide sequence of the Y chromosome specific molecular marker is shown as SEQ ID NO. 2.
3. The primer according to claim 1 or 2, wherein the primer can amplify an X chromosome specific molecular marker of 570bp in length and a Y chromosome specific molecular marker of 704bp in length.
4. A kit for detecting the Ji Shi tilapia sex chromosome specific molecular marker according to claim 1, comprising the primer according to any one of claims 1 to 3.
5. Use of the primer of any one of claims 1-3, the primer-amplified Ji Shi tilapia sex chromosome specific molecular marker of claim 2, or the kit of claim 4 in Ji Shi tilapia breeding.
6. A method for identifying the genetic sex of Ji Shi tilapia, which is characterized in that the primer of any one of claims 1-3 or the kit of claim 4 is adopted to amplify the genome of Ji Shi tilapia to be detected, and then the amplified fragment is compared with Ji Shi tilapia sex chromosome specific molecular marker amplified by the primer of claim 2 to determine the genetic sex.
7. The method according to claim 6, wherein the genetic sex of the Ji Shi tilapia to be tested is determined based on the length of the amplified fragment, in particular:
when only the X chromosome specific molecular marker with the length of 570bp is amplified from the genome DNA of the Ji Shi tilapia to be detected, judging that the Ji Shi tilapia to be detected is a inherited female fish, and the sex chromosome genotype is XX;
when only a Y chromosome specific molecular marker with the length of 704bp is amplified from the genome DNA of the Ji Shi tilapia to be detected, judging that the Ji Shi tilapia to be detected is a genetic super-male fish, and the sex chromosome genotype is YY;
when the X chromosome specific molecular marker and the Y chromosome specific molecular marker are amplified simultaneously from the genomic DNA of the Ji Shi tilapia to be detected, the Ji Shi tilapia to be detected is judged to be a genetic male fish, and the sex chromosome genotype is XY.
8. A method for cultivating a full female Ji Shi tilapia, comprising the steps of:
(1) Carrying out maleation treatment on offspring generated after mating of Ji Shi tilapia XX female fish and XY male fish, and screening out a transformed male fish with sex chromosome genotype XX by using the primer according to any one of claims 1-3;
(2) Mating the XX-transformed male fish with the XX female fish of the normal Ji Shi tilapia to obtain the XX full female fish.
9. A method for cultivating supermale and Quan Xiongji tilapia, comprising the steps of:
(1) Carrying out feminization treatment on offspring generated after mating of Ji Shi tilapia XX female fish and XY male fish, and screening out transformed female fish with sex chromosome genotype XY by using the primer according to any one of claims 1-3;
(2) Mating XY transformed female fish with XY male fish of Ji Shi tilapia, and screening supermale fish with sex chromosome genotype YY from offspring by using the primer of claim 2; in addition, the mating offspring of XY transformed female fish and Ji Shi tilapia XY male fish are taken for female treatment, and the primers in claim 2 are used for screening out the transformed female fish with sex chromosome genotypes of XY and YY;
(3) Mating YY super-male fish with XY or YY converted female fish to obtain YY super-male fish;
(4) And mating the YY super-male fish with the XX normal female fish to obtain the XY full-male fish.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101962641A (en) * | 2010-10-09 | 2011-02-02 | 西南大学 | Specific molecular markers of sex chromosomes of Tilapia nilotica and genetic sex identification method |
| CN103409420A (en) * | 2013-08-31 | 2013-11-27 | 西南大学 | Y-chromosome specific molecular marker of Nile tilapia, and genetic sex determination method and supermale producing method both based on molecular marker |
| CN110938626A (en) * | 2019-12-12 | 2020-03-31 | 西南大学 | Specific molecular marker of sex chromosome of Oreochromis aureus, genetic sex identification based on molecular marker and method for producing unisex fish |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101962641A (en) * | 2010-10-09 | 2011-02-02 | 西南大学 | Specific molecular markers of sex chromosomes of Tilapia nilotica and genetic sex identification method |
| CN103409420A (en) * | 2013-08-31 | 2013-11-27 | 西南大学 | Y-chromosome specific molecular marker of Nile tilapia, and genetic sex determination method and supermale producing method both based on molecular marker |
| CN110938626A (en) * | 2019-12-12 | 2020-03-31 | 西南大学 | Specific molecular marker of sex chromosome of Oreochromis aureus, genetic sex identification based on molecular marker and method for producing unisex fish |
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