CN112154943A - Efficient screening method for parent needed for preparing new fast-growing type little yellow croaker line - Google Patents
Efficient screening method for parent needed for preparing new fast-growing type little yellow croaker line Download PDFInfo
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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
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- 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
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Abstract
A high-efficiency screening method for parent needed by preparation of a new fast-growing type small yellow croaker strain belongs to the technical field of small yellow croaker parent screening. The screening method comprises the steps of constructing a yellow croaker family, marking PIT of the yellow croaker with high survival rate, completing recovery and temporary rearing of the yellow croaker after the PIT marking, measuring the physical quality and evaluating the breeding performance, and identifying and selecting parents needed by preparation of a fast-growing new strain. The method can obviously improve the accuracy of selecting excellent parents and lay a foundation for quickly obtaining a new fast-growing type little yellow croaker strain. With the method of the invention, 5.38% relative genetic progress can be achieved per generation. Comparing the breeding value selection and phenotype selection results can find that the breeding value selection accuracy is improved by 94.35% compared with phenotype selection.
Description
Technical Field
The invention belongs to the technical field of small yellow croaker parent screening, and particularly relates to a high-efficiency screening method for parent required by preparation of a new fast-growing small yellow croaker strain.
Background
Yellow croaker (Larimichthys polyactis) Belonging to the genus Pseudosciaenae of the order Perciformes, family Shikoyudae, is a very important economic fish, and is mixed with radix et rhizoma RheiThe fish, hairtail and Sepiella maindroni are called as the traditional four-large marine products in China. Since the 70 s of the last century, the yield of the small yellow croakers is sharply reduced due to over-fishing, environmental pollution, climate environmental changes and the like, and the captured small yellow croakers are in the phenomenon of resource decline such as low age, miniaturization and the like. Under the condition, the artificial propagation and fine breeding of the small yellow croakers are developed, the artificial breeding of the small yellow croakers is promoted to be a necessary trend, the artificial propagation of the small yellow croakers is successful in 2015, the number of the artificially propagated seedlings is more than 200 thousands of seedlings in 2020, the large-scale breeding technology is mature day by day, and an important foundation is laid for the promotion and breeding of the small yellow croakers. The growth speed of the fishes is closely related to the economic income of breeding manufacturers, and the fast-growing new species bring huge economic income, so that the development of the fine breeding of fast-growing small yellow croakers is of great significance to the development of the small yellow croaker breeding industry. Accurate estimation of genetic parameters and breeding values is the basis for implementing breeding plans and directly influences the genetic progress and breeding effect of animal breeding target characters. The conventional phenotype selection has very low accuracy due to the influence of environmental factors in the culture process, so the patent provides a technical method for accurately selecting the new strain parent of the fast-growing type little yellow croaker.
Disclosure of Invention
Aiming at the problems in the prior art, the invention aims to design and provide a high-efficiency screening method for parents needed by preparation of a new fast-growing yellow croaker line. The method is based on the quantitative genetics theory, and utilizes a genetic breeding method to accurately screen new line breeding parents capable of obtaining the fast-growing small yellow croakers.
In order to achieve the purpose, the invention adopts the following technical scheme:
a method for efficiently screening parents required by preparation of a new fast-growing type little yellow croaker strain is characterized by comprising the following steps:
(1) collecting more than 4000 small yellow croakers, culturing in the same net cage to 8 months old, randomly taking 200 small yellow croakers, measuring the body mass of the small yellow croakers, sequencing the former 30% of body mass value, measuring the rest small yellow croakers by taking the method as a standard, selecting individuals with the body mass of the first 30% as a generation parent, feeding living clamworms for intensive culture, regulating and controlling the water temperature to be kept at 15-16 ℃, and promoting the gonad development;
(2) selecting male and female parents with excellent gonad development, constructing more than 30 yellow croaker families by artificial induced spawning, fertilization and incubation according to the mating mode of the 1-male parent and the 1-2 female parent, transferring fertilized eggs into a cultivation bucket for standardized cultivation of the families when the fertilized eggs develop to the intramembranous fry stage, randomly taking 30 fish in each bucket to determine the average body quality of each family when the fish is cultivated to 5 months old, randomly selecting 200 individuals from each family, and marking the individuals by PIT;
(3) and (3) recovering and temporarily culturing the small yellow croakers after the PIT marking is finished: after marking, the breeding density is adjusted to 150 tails/m3And the following, after water is changed by 80%, adding 2ppm of povidone iodine and 5ppm of oxytetracycline hydrochloride, resuming feeding every other day, after the morning is fed with satiation, changing water by 80% and adding 2ppm of povidone iodine and 5ppm of oxytetracycline hydrochloride, after the afternoon is fed with satiation, changing water by 80% and adding 2ppm of vitamin C excitant and 0.5ppm of substrate modifier EM golden dew, resuming normal culture after 7 consecutive days by the method;
(4) mixed culture of recovered small yellow croaker at 45m3Culturing in an indoor cement pond until the age of 13 months, carrying out in-vivo marking scanning on all individuals, measuring the body mass, establishing a unisexual animal model by taking the mean value of the body mass when a family is marked as a covariate and the construction time of the family as a fixed effect, estimating the body mass heritability of the little yellow croaker, and predicting the breeding value of the individuals;
(5) sorting all the small yellow croaker individuals from high to low according to individual breeding values, scanning individual PIT marks one by one, selecting and reserving 30% of the individuals in the top rank, and performing intensive cultivation to obtain the new small yellow croaker line capable of quickly breeding the small yellow croakers.
The efficient screening method for parent strains required by preparation of the new fast-growing type little yellow croaker strain is characterized in that the artificial induced spawning, fertilization and hatching in the step (2) is specifically as follows: hatching the fertilized eggs in a 300L hatching barrel with water temperature controlled at 15-18 deg.c and sea water salinity controlled at 26-28, and oxygenating.
The efficient screening method for preparing the parents required by the new fast-growing type small yellow croaker line is characterized in that the standardized breeding of the families in the step (2) is specifically as follows: each family is independently cultured in a 2500L cultivation barrel for standardized culture, when the family is 2 months old, the first thinning and seedling dividing is carried out, the number of the seedlings is 2000 in each barrel, when the family is 3 months old, the second thinning and seedling dividing is carried out, the number of the seedlings is 800 in each barrel, when the family is 4 months old, the third thinning and seedling dividing is carried out, and the number of the seedlings is 400 in each barrel.
The efficient screening method for parents required by preparation of the new fast-growing type small yellow croaker line is characterized in that the PIT marking method in the step (2) is as follows: placing an individual to be marked into seawater with the concentration of 0.1ppm eugenol for 1min for anesthesia, taking out the individual after tail by tail, injecting a PIT mark into muscle, inserting a needle into superficial muscle from the middle position of the back of the fish at an oblique angle of 15 degrees, quickly injecting the mark, smearing a wound with erythromycin ointment, smearing a liquid wound plaster to completely isolate the wound from the outside, taking out all the anesthetized individuals within 5 min, marking, and transferring the anesthetized individuals into clean seawater for recovery.
The efficient screening method for preparing the parents required by the new fast-growing yellow croaker strain is characterized in that the single-character animal model formula in the step (4) is as follows:y ijk = μ + D i + B j + A k + e ijk whereiny ijk Is shown as k The physical quality of the individual is determined,μthe overall mean value is represented as a function of,D i is shown as i The fixed effect of the individual family construction period,B j is shown asj The mass mean value of the individual family marking time is used as a covariate,A k indicating an additive genetic effect, i.e. an individual breeding value,e ijk indicating residual effects.
The efficient screening method for preparing parents required by the new fast-growing type yellow croaker line is characterized in that the individual breeding value prediction method comprises the following steps: estimation of phenotypic variance using REML
Additive variance
And a residual variance component
According to the formula
And calculating the heritability of the body quality characters, and predicting the individual breeding value by using a BLUP method on the basis of the heritability of the body quality characters.
The method can obviously improve the accuracy of selecting excellent parents and lay a foundation for quickly obtaining a new fast-growing type little yellow croaker strain. With the method of the invention, 5.38% relative genetic progress can be achieved per generation. Comparing the breeding value selection and phenotype selection results can find that the breeding value selection accuracy is improved by 94.35% compared with phenotype selection.
Drawings
FIG. 1 shows the results of the statistics of the physical quality homogeneity and survival rate of each family.
Detailed Description
Example 1:
(1) family construction
Collecting about 4000 small yellow croakers from an artificial culture colony, culturing in the same net cage for 12 months (the colony is about 8 months old), randomly taking 200 small yellow croakers, measuring the body mass of the small yellow croakers, sequencing to determine the body mass value of the former 30% of the sequencing, determining the rest about 3800 small yellow croakers by taking the body mass value as the standard, selecting and reserving individuals with the body mass of the former 30% as a generation parent, feeding living clamworms for parent reinforced culture, regulating and controlling the water temperature to be kept at 15 ℃ and promoting the gonad development.
(2) PIT marker for small yellow croaker with high survival rate
Selecting male and female parents with excellent gonad development at the beginning of 4 months in the next year, constructing 30 little yellow croaker families by artificial induced spawning, fertilization and incubation according to the mating mode of the 1 male parent and the 1-2 female parents, transferring fertilized eggs into a 2500L cultivation barrel for standardized cultivation of the families when the fertilized eggs develop to the intramembranous fry stage, randomly taking 30 eggs in each barrel to determine the average body quality of each family when the eggs are cultivated to 5 months old, randomly selecting 200 individuals in each family, and marking the individuals by using PIT.
Artificial induced spawning insemination: in 10 a.m.: hasten parturition before 00, every other morning 5: 00 starting to check the parent gonad development condition, carrying out artificial egg squeezing fertilization, placing the fertilized eggs in a 300L hatching barrel for hatching, controlling the water temperature at 15-18 ℃, controlling the salinity of seawater at more than 26, and carrying out micro-oxygenation.
Standardized breeding of families: each family is independently cultivated in 2500L cultivation barrels for standardized cultivation, the first thinning and seedling dividing is carried out when the family is 2 months old, 2000 tails of seedlings are reserved in each barrel, 800 tails of seedlings are reserved in each barrel when the family is 3 months old, and 400 tails of seedlings are reserved in each barrel when the family is 4 months old.
Preparing a small yellow croaker anesthetic: dissolving eugenol oil (Henan Fishery Yao biotechnology Co., Ltd.) in 70% ethanol at equal volume, stirring with 18 deg.C seawater to obtain 0.1ppm, and oxygenating for use.
PIT mark type: the length of the syringe is 1.4 mm, the diameter of the syringe is 8mm, the syringe is matched with a matched needle tube, the mark and the syringe are soaked in absolute ethyl alcohol for more than 30min for disinfection, and the syringe is used after being dried.
The marking method comprises the following steps: after the marked individual is put into seawater with the concentration of 0.1ppm eugenol for 1min, the individual starts to fish out and inject PIT marks into muscles one by one, the needle head is inserted into superficial muscles from the middle position of the back of the fish body at an oblique angle of 15 degrees, the mark is quickly injected, then the erythromycin ointment is used for smearing the wound, and the liquid wound plaster is smeared to completely isolate the wound from the outside, so that the wound is prevented from being infected by germs. Within 5 minutes, all anesthetized individuals need to be fished out and marked and transferred into clean seawater for recovery, so that death caused by deep anesthesia is avoided.
(3) Recovery and temporary rearing of small yellow croakers after PIT marking
After marking, the breeding density is adjusted to 150 tails/m3And below, water is changed by 80 percent, and 2ppm of povidone iodine (Beijing Yangguangyo Co., Ltd.) and 5ppm of oxytetracycline hydrochloride (Beijing Yangyo Co., Ltd.) are added. The next day after the injection of the marker, feed was resumed, 7 a.m: 00 feeding, wherein the feed is a Nippon fully-cooked aquatic product compound feed (Nippon feed science and technology Co., Ltd.), the particle diameter is about 3cm, feeding is performed after full eating, 80% of water is changed after half an hour, and povidone iodine 2ppm + oxytetracycline hydrochloride 5ppm are addedFeeding in the same manner at noon, and after changing water by 80% half an hour later, adding 2ppm of vitamin C synephrine (Zhenjiang Yishengtang aquatic organism Co., Ltd.) and 0.5ppm of substrate modifier EM golden dew (Qingdao Zhongren animal medicine Co., Ltd.). The normal culture is resumed after the operation is continued for 7 days.
When the marking operation is carried out, the little yellow croaker can generate more mucus under stress, the fish body is prevented from being anoxic due to excessive mucus in the water body, and the experimental fish culture density after the marking is less than or equal to 150 tails/m3Meanwhile, running water treatment is carried out, and seawater containing more mucus is timely discharged.
By adopting the technical measures, the time for the small yellow croakers to enter the anesthesia is short, the small yellow croakers leave water for a short time to keep a low-stress state by PIT marking operation, the recovery time after the anesthesia is short, and the survival rate after the anesthesia and the marking injection is high.
(4) Mass measurement and breeding performance assessment
Mixed culture of recovered small yellow croaker at 45m3The method comprises the steps of culturing the yellow croaker in an indoor cement pond until the age of 13 months, carrying out in-vivo marking scanning on all individuals, measuring the body mass, establishing a unisexual animal model by taking the mean value of the body mass when a family is marked as a covariate and the construction time of the family as a fixed effect, estimating the body mass heritability of the yellow croaker, and predicting the breeding value of the individuals.
Establishing a parthenocarpic animal model:y ijk = μ + D i + B j + A k + e ijk wherein, the water-soluble polymer is a polymer,y ijk is shown as k The physical size of the individual;μrepresents the overall mean;D i is shown as i Fixed effects of individual family construction period;B j is shown asj The mass mean value of the individual family marking time is used as a covariate;A k indicates additive genetic effect (individual breeding value);e ijk indicating residual effects.
Estimation of phenotypic variance using REML
Additive variance
And a residual variance component
According to the formula
And calculating the heritability of the body quality characters, and predicting the individual breeding value by using a BLUP method on the basis of the heritability of the body quality characters.
(5) Identification and selection of parents needed for preparation of new fast-growing strains
All individual breeding values are sorted from high to low, individual PIT marks are scanned one by one, and 30% of individuals (namely good parents needed for preparing the fast-growing new strains) at the top are selected and reserved for intensive breeding for breeding the fast-growing new strains of little yellow croakers.
Example 2:
the experiment is carried out by the method of the embodiment 1, about 5200 small yellow croakers are collected from an artificial culture colony, the small yellow croakers are cultured in the same net cage for 12 months (the colony is about 8 months old), 200 small yellow croakers are randomly selected, the body mass of the small yellow croakers is measured, the numerical value of the body mass of the first 30% of the sequence is determined by sequencing, the rest 5000 small yellow croakers are measured by taking the numerical value as the standard, individuals with the body mass of the first 30% are selected and remained, the number of the small yellow croakers is 589, the small yellow croakers are finally obtained and used as first generation parents, the living clamworms are fed for parent intensive culture. Artificially induced spawning, fertilization and hatching to obtain 34 families of the yellow croaker, wherein all families are independently cultured in a 2500L barrel, density and seedling division are performed regularly according to the claim, the average body mass of each family is measured by sampling at the age of 5 months, 200 tails of each family are randomly selected to perform PIT marker mixed culture, all marked individuals are scanned at the age of 13 months, the body mass is measured, the statistical analysis result of the body mass is shown in Table 1, and the survival rate and the average value of the body mass of each family are shown in figure 1.
TABLE 1 descriptive statistics of 11-month-old body mass of little yellow croakers
Variance component estimation was performed according to the single-trait animal analysis model described in the previous paragraph and body mass trait heritability was calculated and the results are shown in table 2. The heritability of the little yellow croaker body mass can be found to be 0.17. The genetic variation coefficient of the body mass and the genetic progress obtained for each generation under the condition of 30% seed retention rate were calculated by using the heritability and the additive variance, and the results are shown in table 3, and it can be known that 5.38% of the relative genetic progress can be obtained for each generation by using this selection method. Comparison of the breeding value selection and phenotype selection results revealed that the breeding value selection accuracy was improved by 94.35% over phenotype selection (table 4).
TABLE 2 weight variance component of little yellow croaker estimated using a parthenocarpic animal model
TABLE 3 genetic variation coefficient and genetic progression of little yellow croaker body mass
Note: GCV is the genetic variation coefficient; θ =1.159 is the selection intensity corresponding to 30% seed retention; Δ G is the expected genetic progress; Δ G' is the relative genetic progression.
TABLE 4 comparison of individual selection based on phenotypic and breeding values (at 30% seed retention)
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| CN102047851A (en) * | 2009-10-29 | 2011-05-11 | 上海海洋大学 | Construction and stock breeding method for cultured grass carp families |
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