CN114731973A - Rapid breeding method for small yellow croakers - Google Patents
Rapid breeding method for small yellow croakers Download PDFInfo
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- CN114731973A CN114731973A CN202210549532.5A CN202210549532A CN114731973A CN 114731973 A CN114731973 A CN 114731973A CN 202210549532 A CN202210549532 A CN 202210549532A CN 114731973 A CN114731973 A CN 114731973A
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- 238000009395 breeding Methods 0.000 title claims abstract description 50
- 241001596950 Larimichthys crocea Species 0.000 title claims abstract description 29
- 241000251468 Actinopterygii Species 0.000 claims abstract description 91
- 230000001488 breeding effect Effects 0.000 claims abstract description 39
- 235000013601 eggs Nutrition 0.000 claims abstract description 22
- 238000000034 method Methods 0.000 claims abstract description 19
- 238000011156 evaluation Methods 0.000 claims abstract description 14
- 230000002068 genetic effect Effects 0.000 claims abstract description 14
- 210000000582 semen Anatomy 0.000 claims abstract description 10
- 230000002196 ecbolic effect Effects 0.000 claims abstract description 7
- 230000032696 parturition Effects 0.000 claims abstract description 6
- 239000011148 porous material Substances 0.000 claims abstract description 6
- 238000011534 incubation Methods 0.000 claims abstract description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 15
- 238000006555 catalytic reaction Methods 0.000 claims description 3
- 210000004392 genitalia Anatomy 0.000 claims description 3
- 230000001568 sexual effect Effects 0.000 claims description 2
- 230000001850 reproductive effect Effects 0.000 abstract description 3
- 230000000052 comparative effect Effects 0.000 description 13
- 229920003023 plastic Polymers 0.000 description 12
- 239000013535 sea water Substances 0.000 description 9
- 210000001015 abdomen Anatomy 0.000 description 8
- 230000000694 effects Effects 0.000 description 6
- 230000006698 induction Effects 0.000 description 6
- 238000011161 development Methods 0.000 description 4
- 230000018109 developmental process Effects 0.000 description 4
- 230000007613 environmental effect Effects 0.000 description 4
- 230000009027 insemination Effects 0.000 description 4
- 210000004681 ovum Anatomy 0.000 description 4
- CPKVUHPKYQGHMW-UHFFFAOYSA-N 1-ethenylpyrrolidin-2-one;molecular iodine Chemical compound II.C=CN1CCCC1=O CPKVUHPKYQGHMW-UHFFFAOYSA-N 0.000 description 3
- 102000002322 Egg Proteins Human genes 0.000 description 3
- 108010000912 Egg Proteins Proteins 0.000 description 3
- 229920000153 Povidone-iodine Polymers 0.000 description 3
- 230000012447 hatching Effects 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 229960001621 povidone-iodine Drugs 0.000 description 3
- 241000356842 Larimichthys polyactis Species 0.000 description 2
- 239000004568 cement Substances 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000001125 extrusion Methods 0.000 description 2
- 230000004720 fertilization Effects 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 108700012941 GNRH1 Proteins 0.000 description 1
- 239000000579 Gonadotropin-Releasing Hormone Substances 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 238000009360 aquaculture Methods 0.000 description 1
- 244000144974 aquaculture Species 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 238000012252 genetic analysis Methods 0.000 description 1
- 210000002149 gonad Anatomy 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000007427 paired t-test Methods 0.000 description 1
- 238000010831 paired-sample T-test Methods 0.000 description 1
- 238000004904 shortening Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 239000010902 straw Substances 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
Classifications
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- 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
- A01K61/00—Culture of aquatic animals
- A01K61/10—Culture of aquatic animals of fish
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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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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Marine Sciences & Fisheries (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Farming Of Fish And Shellfish (AREA)
Abstract
The invention discloses a method for quickly breeding small yellow croakers. The breeding method comprises the following steps: (1) selecting female fish and male fish of 1 age as parents; (2) artificially hastening parturition on the selected parents by using an oxytocic; (3) 1, independently pairing male fishes which are manually and lightly extruded to flow a large amount of seminal fluid with female fishes with round and swollen reproductive pores 0.5-1h before spawning of parent fishes, collecting fertilized eggs for incubation and cultivation, and constructing different families; the female fish do not need to squeeze eggs manually; (4) and (4) performing genetic evaluation on different families by using the body quality data at the age of 8 months, and breeding parents according to the genetic evaluation result. The parent strain of the invention is 1 age individual, the breeding period is shortened, the number of the obtained high-quality fertilized eggs is high, the male and female parent fishes are almost not damaged, and the female and male parent fishes can be recycled through short-time intensive cultivation. The invention has the highest accuracy of genetic evaluation when the breeding is limited to 8 months of age, and can reduce the breeding quantity and effectively reduce the breeding cost.
Description
Technical Field
The invention relates to the field of aquaculture, in particular to a method for quickly breeding small yellow croakers.
Background
The small yellow croaker (Larimichthys polyactis) has very important economic value, and due to long-term fishing pressure and marine environment change, the resource structure of wild small yellow croaker populations is seriously damaged in recent years, so that the small yellow croaker populations show germplasm decline phenomena such as individual miniaturization, advanced sexual maturity and the like. The artificial propagation of the small yellow croakers is successful in 2015, and then the large-scale breeding of the seedlings is broken through, the large-scale cage culture is realized, and a solid foundation is laid for the industrial development of the small yellow croakers.
However, at present, in the process of breeding small yellow croakers, the growth difference between individuals is large, and improved varieties with excellent growth characteristics are seriously deficient, so that the development of improved variety breeding to obtain new small yellow croaker varieties (lines) with excellent characteristics is a necessary prerequisite for the healthy and rapid development of the breeding industry of small yellow croakers.
Patent CN106577382A, a method for constructing and breeding a good pedigree of small yellow croakers, provides a pedigree breeding method based on conventional phenotypic value screening. The breeding method has low accuracy and long time for obtaining new species with excellent characters. Meanwhile, the artificial spawning induction and egg extrusion insemination modes provided by the invention have certain defects, such as inevitable parent fish damage in the artificial egg extrusion and semen collection processes, particularly strong stress of the little yellow croakers, and fatal damage caused by manual operation. Meanwhile, the window period of spawning after spawning promotion of the small yellow croakers is relatively short, the spawning quality at night is optimal, in order to successfully obtain parent fish just about to spawn after gonad maturation, continuous observation is needed, the labor force of personnel is increased, meanwhile, the parent fish can be frightened, and the quality of ova is further influenced.
Disclosure of Invention
The invention aims to provide a method for quickly breeding small yellow croakers, which accelerates the obtaining of improved varieties of the small yellow croakers through 4 aspects of parent fish selection, artificial induced spawning, family construction and individual genetic assessment, and further promotes the quick development of the small yellow croaker breeding industry.
In order to achieve the purpose, the invention provides the following scheme:
the first technical scheme is as follows: a method for quickly breeding small yellow croakers comprises the following steps:
1. a method for rapidly breeding small yellow croakers is characterized by comprising the following steps:
(1) selecting female fish and male fish of 1 age as parents;
(2) artificially hastening parturition on the selected parents by using an oxytocic;
(3) 1, independently pairing male fishes which are manually and lightly extruded to flow a large amount of seminal fluid with female fishes which are round and raised in genital pores 0.5-1h before spawning of parent fishes, collecting fertilized eggs for incubation and cultivation, and constructing different families; the female fish does not need to extrude eggs manually;
(4) and (4) performing genetic evaluation on different families by using the body quality data at the age of 8 months, and breeding parents according to the genetic evaluation result.
Further, the number of the male fish in the parents in the step (2) is 1.2-1.5 times that of the female fish.
Further, the time of the individual pairing in step (3) is: when the water temperature is 16 +/-0.5 ℃, carrying out artificial catalysis for 48-50 h; or 39-40 h after artificial catalysis when the water temperature is 18 +/-0.5 ℃.
Further, the water temperature of the single pair in the step (3) is controlled to be 16-18 ℃.
Further, the genetic evaluation in step (4) uses pedigree information of a plurality of generations to increase the accuracy of evaluation, wherein each generation selects an individual of 1 age of primary maturity as a parent.
The invention discloses the following technical effects:
(1) the invention fully utilizes the characteristic that the yellow croaker is mature for the first time at age 1, and female fish with soft and raised belly and male fish with little semen flowing out when slightly pressing the belly are selected as excellent parents from the breeding basic population at age 1 in the breeding period of the yellow croaker for artificial spawning induction, thereby shortening the breeding period.
(2) The rapid breeding method of the small yellow croaker is 3-5 times higher than the high-quality fertilized eggs obtained by artificial egg-extruding insemination, and meanwhile, the male and female parent fish are almost not damaged and can be recycled through short-time intensive cultivation.
(3) The reliability of the physical quality trait breeding value estimation in 3 growth periods is compared by using a pairing T test method, and the result shows that the reliability of the breeding value estimation in 8-month age is obviously higher than that in 6-month age and 10-month age. The invention can reduce the breeding quantity and effectively reduce the breeding cost by limiting the breeding to 8 months age to carry out genetic evaluation and screening of excellent parents as parent fish breeding. Meanwhile, individual genetic evaluation is carried out by utilizing a plurality of generation pedigrees and phenotype data information, the accuracy of individual genetic evaluation is further increased, and then excellent individuals are accurately screened to serve as parents for cultivation.
Detailed Description
Reference will now be made in detail to various exemplary embodiments of the invention, the detailed description should not be construed as limiting the invention but as a more detailed description of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. Further, for numerical ranges in this disclosure, it is understood that each intervening value, between the upper and lower limit of that range, is also specifically disclosed. Every smaller range between any stated value or intervening value in a stated range and any other stated or intervening value in a stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference herein for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
As used herein, the terms "comprising," "including," "having," "containing," and the like are open-ended terms that mean including, but not limited to.
The oxytocides in examples 1-2 and comparative example 1 were: luteinizing hormone releasing hormone A2(LRH-A2) was formulated into 1ug/mL injection using 9 ‰ saline.
The calculation process of the mortality of the female fish after spawning comprises the following steps: the mortality rate (%) of the female fish after spawning was (number of female fish after spawning/number of female fish of parent fish induced to spawn) × 100%
Example 1
Rapid breeding method for small yellow croakers
(1) Female fish with raised and soft abdomen and male fish with slight semen flow out when slightly pressing the abdomen are selected from the breeding basic group of 1 age as excellent parents for artificial spawning induction;
(2) artificial hastening parturition: the spawning agent is applied to female fish according to the dosage of 0.5mL/100g of the spawning agent, the dosage of male fish is reduced by half, the number of spawned male fish is 1.2 times of that of the female fish, specifically, 15 female fish and 18 male fish are mixed and cultured in a barrel with the volume of 300L;
(3) according to the effect time of an oxytocic, 1h before spawning of parent fishes (when the water temperature is 16 ℃, 50h after artificial spawning induction), selecting female fishes with round and swollen reproductive pores, independently placing the female fishes to spawn in an opaque plastic barrel with the diameter of 80cm, adding clean seawater with the salinity of 25 into the barrel, controlling the water depth in the barrel to be 25cm and each female fish to be one barrel, simultaneously selecting 1 male fish which slightly extrudes a large amount of semen to be matched with the male fishes, controlling the water temperature to be 16 ℃, standing for the female fishes to spawn naturally, observing for 1 time every 1h, waiting for 20min after spawning, collecting eggs by using a 40-mesh net bag, washing by using povidone iodine sterilized seawater, placing in a 2000mL plastic beaker, adding seawater with the salinity of 25, standing for 15min, collecting floating high-quality fertilized eggs, and transferring to 2.5m3And (5) hatching and cultivating in a plastic bucket.
Example 2
Rapid breeding method for small yellow croakers
(1) Female fish with raised and soft abdomen and male fish with slight semen flow out when slightly pressing the abdomen are selected from the breeding basic group of 1 age as excellent parents for artificial spawning induction;
(2) artificial hastening parturition: 0.6mL/100g of oxytocic is used for female fish, the dosage of male fish is halved, the number of the induced spawning male fish is 1.5 times that of the female fish, the specific number of the female fish is 16, the number of the male fish is 24, and the female fish and the male fish are mixed and cultured in a barrel with the volume of 300L;
(3) according to the effect time of the oxytocic, the parent fish is 0.5h before spawning (the water temperature is 18.5 ℃, after artificial oxytocic39h) Selecting female fish with round and raised reproductive pore and about to lay eggs, placing in an opaque plastic barrel with diameter of 80cm, adding clean seawater with salinity of 28, controlling water depth in the barrel at 35cm, placing one barrel for each female fish, selecting 1 male fish lightly squeezing out a large amount of semen, pairing with the male fish, controlling water temperature at 18 deg.C, standing for natural spawning, observing for 1 time every 1h, waiting for 30min after spawning, collecting ovum povidone iodine with 40 mesh net bag, washing with sterile seawater, placing in 2000mL plastic beaker, adding seawater with salinity of 28, standing for 10min, collecting high-quality floated fertilized ovum, transferring to 2.5m fertilized ovum, and transferring to a transparent plastic barrel with diameter of 80cm3And (5) hatching and cultivating in a plastic bucket.
Example 3
The difference between this example and example 1 is that in the artificial induced spawning step of this example, the number of induced spawning male fish is 1.4 times that of female fish, specifically, 15 female fish and 21 male fish. The remaining procedure was the same as in example 1.
Comparative example 1
(1) Female fish with raised and soft abdomen and male fish with slight semen flow out when slightly pressing the abdomen are selected from the breeding basic group of 1 age as excellent parents for artificial spawning induction;
(2) artificial hastening parturition: 0.5mL/100g of the oxytocic is applied to female fish, the dosage of male fish is halved, the number of the induced spawning male fish is 1.2 times of that of the female fish, the specific number of the female fish is 15, and the number of the male fish is 18. Mixed culture is carried out in a barrel with the volume of 300L;
(3) artificial egg-extruding insemination: selecting female fish with round and raised genital pore and about to lay eggs, placing the female fish into an opaque plastic barrel with diameter of 80cm at water temperature of 16 ℃ for 48h after artificial induced spawning, adding clean seawater with salinity of 25 into the barrel, controlling the water depth in the barrel to be 25cm, obtaining eggs by each female fish in an artificial egg squeezing mode, placing the collected eggs into a barrel separately, collecting sperm of male fish by a plastic straw, controlling the water temperature to be 16 ℃, performing artificial dry fertilization, standing for 20min after fertilization, collecting the eggs by a 40-mesh net bag, placing the eggs into a 2000mL plastic beaker after sterilizing seawater by povidone iodine, adding seawater with salinity of 25, standing for 15min, collecting floating high-quality fertilized eggs, transferring the fertilized eggs into a 2.5m plastic beaker3In a plastic bucketAnd (5) hatching and cultivating.
Comparative example 2
The difference between the comparative example and the comparative example 1 is that in the artificial induced spawning step of the comparative example, the number of induced spawning male fish is 1.5 times that of female fish, the specific number of female fish is 16, and the number of male fish is 24. The remaining procedure was the same as in comparative example 1.
Comparative example 3
The difference between the comparative example and the comparative example 1 is that in the step of artificial induced spawning of the comparative example, the number of induced spawning male fish is 1.4 times that of female fish, the number of specific female fish is 15, and the number of male fish is 21. The remaining procedure was the same as in comparative example 1.
Analysis of results
The effects of different ways of inseminating eggs in examples 1 to 3 and comparative examples 1 to 3 were compared and are detailed in table 1.
TABLE 1 comparison of the effects of different insemination modes of spawning
39 families were constructed by the breeding method of example 1, each family was independently bred, and 30 random samples were taken from each family to measure the body mass by 3 months of age. At 6 months of age, each family randomly selects 100 individuals to measure body mass, and after PIT marking, the individuals are mixed and cultured in the same cement pond, and the body mass of each individual is measured at 8 months of age and 10 months of age respectively. The mean values of the body mass of each family at 3 months of age were used as covariates, the growth days of each family were used as fixed effects, the animal models in the ASReml software were used to analyze the measured body mass data at 6, 8 and 10 months of age, respectively, to obtain variance components and calculate the body mass property heritability and common environmental effects, the results are shown in Table 2.
TABLE 2 body Mass Property variance components and heritability estimation results at different growth periods
The genetic force obtained at the age of 8 months is higher than that obtained at the ages of 6 months and 10 months, the common environmental effect is very large at the age of 6 months, and the common environmental effect almost disappears at the ages of 8 months and 10 months, mainly because all families before the age of 6 months are separately cultured in an isolated way, all individuals are cultured in the same cement pond by PIT marking from the age of 6 months, and the common environmental effect is obviously reduced along with the prolonging of the mixed culture time.
The reliability of the estimates of the quality trait breeding values for 3 growth stages was compared using the paired T test, as detailed in table 3. It can be seen that the reliability of the seed value estimation at 8 months of age is significantly higher than at 6 and 10 months of age.
TABLE 3 reliability of estimated breeding values for different growth periods paired sample T test
Genetic analysis was performed on the data of physical quantities of the families of 2 generations of 34 families constructed in 2019 and 39 families constructed in 2020 by using the breeding method of example 1. It was found that the reliability of the obtained breeding values was significantly higher than the reliability of the breeding values estimated using the physical quality data of a single generation in 2020, as detailed in table 4.
TABLE 4 sample T test for reliability of single generation and multi-generation data breeding value
| Pairing | Mean ± standard deviation | t value | Degree of freedom df | Significant sig. |
| Multi-generation-single generation | 0.006±0.045 | 5.567 | 1767 | <0.001 |
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the present disclosure without departing from the scope or spirit of the disclosure. Other embodiments will be apparent to those skilled in the art from consideration of the specification. The specification and examples are exemplary only.
Claims (5)
1. A method for rapidly breeding small yellow croakers is characterized by comprising the following steps:
(1) selecting female fish and male fish of 1 age as parents;
(2) artificially hastening parturition on the selected parents by using an oxytocic;
(3) 1, independently pairing male fishes which are manually and lightly extruded to flow a large amount of seminal fluid with female fishes which are round and raised in genital pores 0.5-1h before spawning of parent fishes, collecting fertilized eggs for incubation and cultivation, and constructing different families; the female fish does not need to extrude eggs manually;
(4) and (4) performing genetic evaluation on different families by using the body quality data at the age of 8 months, and breeding parents according to the genetic evaluation result.
2. The rapid breeding method of yellow croaker according to claim 1, wherein the number of male fish in the parents in step (2) is 1.2-1.5 times that of female fish.
3. The method for rapidly breeding the small yellow croakers according to claim 1, wherein the time for the individual pairing in the step (3) is as follows: when the water temperature is 16 +/-0.5 ℃, carrying out artificial catalysis for 48-50 h; or 39 to 40 hours after artificial spawning when the water temperature is 18 +/-0.5 ℃.
4. The method for rapidly breeding the small yellow croakers according to claim 1, wherein the water temperature of the individual pairs in the step (3) is controlled to be 16-18 ℃.
5. The method for rapid breeding of yellow croaker according to claim 1, wherein the genetic evaluation in step (4) is performed by using pedigree information of multiple generations to increase the accuracy of the evaluation, wherein each generation uses the 1-age individual of primary sexual maturity as a parent.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104904636A (en) * | 2015-06-10 | 2015-09-16 | 浙江省海洋水产研究所 | Method for artificially-induced spawning of little yellow croaker |
| CN106172117A (en) * | 2016-07-14 | 2016-12-07 | 浙江省海洋水产研究所 | Carnis Pseudosciaenae raun and Carnis Pseudosciaenae milter cross breeding method |
| CN112154943A (en) * | 2020-08-27 | 2021-01-01 | 浙江省农业科学院 | Efficient screening method for parent needed for preparing new fast-growing type little yellow croaker line |
-
2022
- 2022-05-20 CN CN202210549532.5A patent/CN114731973A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104904636A (en) * | 2015-06-10 | 2015-09-16 | 浙江省海洋水产研究所 | Method for artificially-induced spawning of little yellow croaker |
| CN106172117A (en) * | 2016-07-14 | 2016-12-07 | 浙江省海洋水产研究所 | Carnis Pseudosciaenae raun and Carnis Pseudosciaenae milter cross breeding method |
| CN112154943A (en) * | 2020-08-27 | 2021-01-01 | 浙江省农业科学院 | Efficient screening method for parent needed for preparing new fast-growing type little yellow croaker line |
Non-Patent Citations (3)
| Title |
|---|
| 刘峰等: "小黄鱼(Pseudosciaena polyactis)形态性状与体质量的相关性及通径分析", 《海洋与湖沼》 * |
| 楼宝等: "小黄鱼全人工繁育技术研究", 《浙江海洋学院学报(自然科学版)》 * |
| 王纪亭 等: "《养鱼手册》", 30 April 2015 * |
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