CN115104557A - method for inducing heterotetraploid carassius pengzhi gynogenesis - Google Patents
method for inducing heterotetraploid carassius pengzhi gynogenesis Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
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Abstract
The invention discloses a method for inducing gynogenesis of allotetraploid carassius auratus pengzi by , which comprises the steps of carrying out continuous two-generation hybridization breeding on eggs of carassius auratus pengzi and sperms of male Xingguo red carps to obtain hybrids, and then selecting excellent male and female parents, carrying out enhanced cultivation on the parents, carrying out artificial induced spawning on the female parents of the allotetraploid carassius auratus pengzi, carrying out artificial dry insemination, carrying out artificial incubation, cultivating gynogenesis seedlings, breeding juvenile fishes and the like, so that finally obtained induced gynogenesis of the allotetraploid carassius auratus pengzi has the advantages of quick growth, large individual, delicious meat, poor appetite, strong stress resistance and strong disease resistance. The method has strong operability and good adaptability, and in the implementation process, the fertilization rate can reach 37.37%, the hatching rate can reach 25.43%, and the emergence rate can reach 16.55%. the induction of the heterotetraploid pengzh crucian gynogenesis filial generation shows larger growth advantage and body type advantage compared with the heterotetraploid pengzh crucian, and is suitable for large-scale popularization and cultivation.
Description
Technical Field
The invention relates to the technical field of fish gynogenesis, in particular to a gynogenesis method for inducing allotetraploid pengzhe crucian carp by .
Background
Carassius auratus var. pengze, also called pengze Carassius auratus, Phragmites crucian, belonging to the order Cyprinales, Cyprinaceae, Cyprinidae, Carassius, originally produced in natural waters such as Taibai lake, Dingjia lake and Fanghu in Pengze county in Jiangxi province. The Pengze crucian carp is bred for 6 continuous generations by the research institute of Water science in Jiangxi province and research personnel of Water science in Jiujiang city, and becomes a freshwater aquaculture variety which is mainly popularized by the Ministry of agriculture in the 90 s of the 20 th century. The pengze crucian carp is a food hybrid and has a wide recipe, and the pengze crucian carp of 1 age and the common carassius auratus of 1 age have a body weight ratio of 3.6:1 under the same culture conditions by eating bacterial clusters, organic debris, animal and plant debris, small aquatic animals and the like in a natural water body, so that the growth advantage is extremely obvious. Carassius pengzensis is a triploid gynogenesis reproductive fish + The sexual maturity is achieved when the age is 3 months to 6 months, the breeding season is from the bottom of the Yangtze river midstream region, the spawning and breeding are started when the water temperature reaches above 17 ℃, and the absolute fecundity of well-developed females is above 2.6 ten thousand grains. The carassius auratus pengze is a carassius auratus variety with high protein content, high meat eating rate, good taste and rich nutrition, and is deeply favored by consumers. In the last decade, the wild pengze crucian carp population is reduced due to the damage of natural environment, and the pengze crucian carp germplasm is degraded due to the close-relative propagation, so that the disease resistance, the growth speed and the breeding benefit of the pengze crucian carp are gradually reduced. Therefore, further artificial breeding is required to cultivate good seeds with fast growth and strong disease resistance. The allotetraploid pengze crucian is a fish species obtained by hybridizing pengze crucian and Xingguo red carp, and the growth speed of the body mass and the body length of the allotetraploid pengze crucian is obviously higher than that of the pengze crucian. The gynogenesis filial generation obtained by inducing carassius auratus pengze ovum by using heterologous sperms is obviously superior to carassius auratus pengze selfing in growth performance. Based on the method, the filial generation obtained by inducing the heterotetraploid carassius auratus pengze ovum through the heterogeneously has more excellent growth performance, which has great significance for the carassius auratus pengze breeding research and the further improvement of the fish species growth performance.
(Elopichths Bambusa), also called as Clarias fuscus and Acronychia, belonging to Cyprinales, Cyprinaceae, Ardisidae, , living in middle and upper layers of rivers and lakes, swimming quickly, moving quickly, and mainly feeding on fish. is a large-scale economic fish with fresh water, which grows fast, is large and has delicious meat taste, and is always regarded as high-grade fresh water fish. the age of the breeding group is 3-4 scales, when the water temperature reaches above 18 deg.C in 4-6 months per year, it can lay eggs in the river torrent, the eggs are floating eggs, and after water absorption and expansion, the development can be completed with water drift. At present, can be caught in the Yangtze river and the lakes directly communicated with the Yangtze river, and most rivers and lakes have difficulty finding tracks.
Since the 90 s in the 20 th century, Chinese scholars have made many attempts to improve the variety of crucian carps, for example, square silver crucian carps (male parent) x diploid red carps (female parent), hibiscus carps (male parent) x red carps (male parent), white carps (female parent) x black dragon carps (male parent), white carps (male parent) x red carps (female parent), white carps (female parent) x red carps (male parent), color carps (male parent) and carp construction (female parent), color carps (female parent) and carp construction carp (male parent), which are all cross breeding. Besides a crossbreeding method, gynogenesis is also an important breeding method, has extremely high application value in aquaculture, and the induction of gynogenesis becomes a common means for breeding fishes. The pengze crucian carp is an important freshwater economic fish in China, researchers carry out a great deal of research and practice on gynogenesis breeding of the pengze crucian carp, and successfully obtain a plurality of excellent breeding varieties, and typical examples include the pengze crucian carp (female parent) multiplied by the white crucian carp (male parent), the pengze crucian carp (female parent) multiplied by the sea carp (male parent), the pengze crucian carp (female parent) multiplied by the tip carp (male parent), the pengze crucian carp (female parent) multiplied by the mouth-tipping culter (male parent), the pengze crucian carp multiplied by the megalobrama (male parent) and the like.
The heterospermia induces gynogenesis to obtain offspring with excellent characters, and research reports show that gynogenesis offspring of Pengze crucian carp (male parent) multiplied by (male parent) and Pengze crucian carp (female parent) multiplied by mouth-uping culter (male parent) are remarkably higher than that of Pengze crucian carp in terms of body quality and body length. The allotetraploid pengze crucian carp and belong to fishes of different subfamilies, have larger difference in morphological structure and growth characteristics, are superior to in the aspects of cultivation adaptability, hypoxia resistance, disease resistance and the like, and are to have more advantages in the aspects of growth speed, body form and fish meat quality, so that the method for exploring the gynogenesis of the allotetraploid pengze crucian carp induced by and cultivating a new fish species with growth characteristics superior to the allotetraploid pengze crucian carp has important application value.
Disclosure of Invention
The invention aims to improve the existing variety of carassius auratus pengze, and provides an induced allopetraploid carassius auratus gynogenesis method.
In order to achieve the purpose, the invention designs a method for inducing the gynogenesis of the allotetraploid carassius auratus pengze by , which comprises the following steps:
1) selection and cultivation of male and female parents:
the female parent is selected from sexually mature Pengze crucian (purchased from the institute of Water-saving and obstetrics in Jiangxi), the male parent is selected from sexually mature Xingguo carp (an aquaculture base of Huazhong university of agriculture), and the two parents are respectively subjected to enhanced cultivation (so that the good development of gonads of the parents is guaranteed) before delivery;
2) artificial induction of spawning of the female parent pengze crucian:
injecting an oxytocic drug into the female parent of Carassius pengzensis by adopting an injection oxytocic method twice, injecting a luteinizing hormone releasing hormone analogue (LRH-A2) into a first needle, and injecting a compound agent into a second needle, wherein the time interval between two injections is 8-12 h;
3) obtaining sperms of male parent Xingguo red carps:
selecting male Kyoho carp with good gonad development before artificial insemination, slowly squeezing the abdomen of the male Kyoho carp, collecting semen at the cloaca, and storing for later use;
4) artificial insemination (dry insemination):
firstly, extruding eggs of carassius auratus pengze into a dry steel basin, then adding sperm of the cyprinus carpio, stirring, adding water, and quickly stirring to ensure that the sperm and the eggs are fully contacted to obtain fertilized eggs;
5) artificial incubation:
putting the fertilized eggs into an incubation barrel for incubation in running water, and incubating first-filial generation fry;
6) and (3) seed cultivation of the first filial generation:
after the first-filial generation of fish fry swims smoothly, transferring the first-filial generation of fish fry to a seedling raising pool for cultivation, feeding zooplankton initially, and feeding powder when the length of the fish fry reaches 2-3 cm;
7) and (3) young fish culture of the first filial generation:
after the surfaces of the fry are covered with scales, transferring the fry into a culture pond, and feeding the first filial generation juvenile fish to adult fish of 1 age according to the method for culturing Pengze crucian;
8) selecting female parents from the first filial generation adult fish and carrying out intensive culture:
selecting a female with large individual and good development in the first filial generation adult fish as a parent, and carrying out intensive culture (tetraploid is not found in the first filial generation);
9) artificial oxytocic of first filial generation female parent Pengze crucian and acquisition of Xingguo red carp sperms:
injecting an oxytocic drug into the female parent of the carassius auratus pekinensis by adopting a two-injection oxytocic method, injecting a luteinizing hormone releasing hormone analogue (LRH-A2) into a first needle, and injecting a compound medicament into a second needle, wherein the time interval of the two injections is 8-12 h; selecting male Kyoho carps with good gonad development before artificial insemination, obtaining semen, and placing in a dark place at 4 ℃ for short-time preservation;
10) artificial insemination, artificial incubation, breeding of second-generation fish fry and young fish culture:
performing dry insemination, putting fertilized eggs into an incubation barrel for running water incubation after fertilization, incubating second-generation young fishes, transferring the second-generation young fishes into a seedling culture pond for cultivation after the second-generation young fishes swim, feeding zooplankton initially, feeding powder when the body length of the second-generation young fishes reaches 2-3cm, transferring the second-generation young fishes into a culture pond after the body surfaces of the second-generation young fishes are covered with scales, and feeding the second-generation young fishes to adult fishes of 1 age according to the method for culturing the Pengze crucian carp;
11) selecting female parents from the second generation adult fish and carrying out intensive culture:
selecting female tetraploid (allotetraploid pengze crucian carp) with large individual and good development in the second-generation adult fish as a female parent, selecting a male parent from (an aquaculture base of university of agriculture in Huazhong) with sexual maturity, and respectively carrying out enhanced cultivation on the two parents before delivery (to ensure good development of gonads of the parents);
12) artificial induced spawning of female tetraploid (allotetraploid pengzh crucian carp) female parent:
injecting an oxytocic drug into a female tetraploid (allotetraploid Pengze crucian) female parent by adopting an injection oxytocic method twice, injecting a luteinizing hormone releasing hormone analogue (LRH-A2) into a first needle, and injecting a compound agent into a second needle, wherein the time interval of the two injections is 8-12 h;
13) acquisition of male parent sperm:
selecting male with good gonad development before artificial insemination, slowly squeezing abdomen, collecting semen at cloaca, and storing at 4 deg.C in dark place for a short time;
14) artificial insemination (dry insemination):
firstly, extruding the ovum of the allotetraploid Pengze crucian carp into a dry steel basin, then adding semen, slightly stirring, adding water and quickly stirring to ensure that the sperm and the ovum are fully contacted to obtain a fertilized ovum;
15) artificial incubation: putting the fertilized eggs into an incubation barrel for incubation in running water, and incubating fish fries;
16) and (3) seedling cultivation: after the fry swims smoothly, transferring the fry to a nursery pond for cultivation, feeding zooplankton initially, and feeding artificial mixed powder when the body length of the fry reaches 2-3 cm;
17) and (3) juvenile fish culture: and after the surfaces of the fry are covered with the scales, transferring the fry into a culture pond, and feeding the young fish to 1 age according to the method for culturing the Pengze crucian carp to obtain induced allopetraploid Pengze crucian gynogenesis filial generation.
Further, in the step 2), the step 9) and the step 12), the injection amount of the luteinizing hormone releasing hormone analogue in the first needle injection is 0.5-2.0 mu g/kg.
Still further, the injection amount of the luteinizing hormone releasing hormone analogue by the first injection is 1 mug/kg.
Still further, in the step 2), the step 9) and the step 12), in the second needle injection, the compound medicament is a combined medicament of luteinizing hormone releasing hormone analogue (LRH-A2) and Delphinone (DOM), and the injection amounts of the compound medicament and the combined medicament are respectively 2-8 mug/kg and 2-8 mg/kg.
Still further, in the second needle injection, the injection amount of the luteinizing hormone releasing hormone analogue is 4 mug/kg, and the injection amount of the diketone is 4 mg/kg.
The invention has the beneficial effects that:
1. the carassius auratus pengze and belong to different subfamilies of fishes, and have large differences in morphological structure and growth characteristics. The pengze crucian carp is superior to in the aspects of cultivation adaptability, hypoxia resistance, disease resistance and the like, is more superior in the aspects of growth speed, body form and fish quality, gynogenesis filial generation of the pengze crucian carp and is still tetraploid through flow cytometry identification, and a chromosome karyotype analysis result shows that the chromosome group of the gynogenesis filial generation of the induced allopetraploid pengze crucian carp consists of 200+ chromosomes (table 1 and figure 1). the growth performance of the heterotetraploid pengze crucian gynogenesis offspring is superior to that of the heterotetraploid pengze crucian inbred offspring.
2. The method for inducing the gynogenesis of the allotetraploid carassius auratus pengze by has strong operability, low cost and good adaptability, and in the implementation process, the hatchability can reach 23 percent and the rate of emergence can reach 16.55 percent. Research shows that the filial generation obtained by inducing the gynogenesis of the allotetraploid carassius auratus pengzhi through is biased to the female parent in morphological characteristics, and the growth speed of the filial generation for inducing the gynogenesis of the allotetraploid carassius auratus pengzhi through is obviously faster than that of the allotetraploid carassius auratus pengzhi selfing filial generation.
3. The invention measures the length and the weight of the gynogenesis progeny population of the allotetraploid pengzhi crucian carp and the inbred population of the allotetraploid pengzhi crucian carp by with the age of 30-180 days, and obtains the absolute growth rate and the absolute weight gain rate of the two zygotic generations through analysis (figure 2 and figure 3). Research results show that induced heterotetraploid pengzhi gynogenesis progeny population is more advantageous in the aspects of body length and weight increase from 60 days old, and the growth speed is obviously faster than that of the heterotetraploid pengzhi crucian inbred population.
Drawings
FIG. 1 is a diagram of induction of nuclear chromosome metaphase of the gynogenesis progeny of allotetraploid Carassius pengzensis;
FIG. 2 is a graph of the absolute growth rate of the gynogenesis progeny of induced allotetraploid carassius pengzensis;
FIG. 3 is a graph of the absolute weight gain rate of induced heterotetraploid Carassius pengzensis gynogenesis progeny;
in fig. 2-3, PX-G is -induced heterotetraploid gynogenesis of pengzh crucian, and PX is hybridization offspring of pengzh crucian and xing country red carp.
Detailed Description
The present invention is described in further detail below with reference to specific examples so as to be understood by those skilled in the art.
Example 1
This example was carried out in the key laboratory of department of education of genetic breeding and reproduction of agricultural animals at the university of agriculture in Huazhong, and the specific implementation of the method for inducing gynogenesis of allotetraploid carassius auratus pengzensis by is as follows
1) Selection and cultivation of male and female parents:
the female parent is selected from sexually mature Pengze crucian (purchased from the institute of Water-saving and obstetrics in Jiangxi), the male parent is selected from sexually mature Xingguo carp (an aquaculture base of Huazhong university of agriculture), and the two parents are respectively subjected to enhanced cultivation (so that the good development of gonads of the parents is guaranteed) before delivery;
2) artificial oxytocic of female parent carassius auratus pengze:
injecting an oxytocic drug into the female parent of the carassius auratus pekinensis by adopting a two-injection oxytocic method, injecting a luteinizing hormone releasing hormone analogue (LRH-A2) into a first needle, and injecting a compound medicament into a second needle, wherein the time interval of the two injections is 8-12 h; wherein in the first needle injection, the injection amount of the luteinizing hormone releasing hormone analogue is 0.5-2.0 mug/kg, and in the second needle injection, the composite medicament is a combined medicament of the luteinizing hormone releasing hormone analogue (LRH-A2) and Dixolone (DOM), and the injection amounts of the composite medicament and the dixolone are 2-8 mug/kg and 2-8 mg/kg respectively;
3) obtaining male parent Xingguan red carp sperms:
selecting male Kyoho carp with good gonad development before artificial insemination, slowly squeezing the abdomen of the male Kyoho carp, collecting semen at the cloaca, and storing for later use;
4) artificial insemination (dry insemination):
firstly, extruding eggs of carassius auratus pengze into a dry steel basin, then adding sperm of the cyprinus carpio, stirring, adding water, and quickly stirring to ensure that the sperm and the eggs are fully contacted to obtain fertilized eggs;
5) artificial incubation:
putting the fertilized eggs into an incubation barrel for incubation in running water, and incubating first-filial generation fry;
6) and (3) seed cultivation of the first filial generation:
after the first-filial generation of fish fry swims smoothly, transferring the first-filial generation of fish fry to a seedling raising pool for cultivation, feeding zooplankton initially, and feeding powder when the length of the fish fry reaches 2-3 cm;
7) and (3) young fish culture of the first filial generation:
after the surfaces of the fry are covered with scales, transferring the fry into a culture pond, and feeding the first filial generation of young fishes to the age of 1 by the method for culturing the Pengze crucian carp;
8) selecting female parents from the first filial generation and intensive culture:
selecting a female with a large individual and good development in the first filial generation of adult fish as a parent, and carrying out intensive cultivation (tetraploid is not found in the first filial generation);
9) artificial oxytocic of first filial generation female parent Pengze crucian and acquisition of Xingguo red carp sperms:
injecting oxytocic drug into female parent of Carassius pengzensis by adopting two-time injection oxytocic method, injecting luteinizing hormone releasing hormone analogue (LRH-A2) with the injection amount of 1 mug/kg at the first needle, injecting compound agent at the second needle, selecting male Xingguo red carp with good gonad development before artificial insemination, obtaining semen, and placing in a dark place at 4 ℃ for short-time preservation; in the first needle injection, the injection amount of the luteinizing hormone releasing hormone analogue is 0.5-2.0 mug/kg, in the second needle injection, the composite medicament is a combined medicament of the luteinizing hormone releasing hormone analogue (LRH-A2) and Dixolone (DOM), the injection amounts are 2-8 mug/kg and 2-8 mg/kg respectively, and the time interval between two injections is 8-12 h;
10) artificial insemination, artificial incubation, second generation offspring seed cultivation and juvenile fish cultivation:
performing dry insemination, putting fertilized eggs into an incubation barrel for running water incubation after fertilization, incubating the second generation of young fish, transferring the young fish to a seedling pool for cultivation after the young fish is flat, feeding zooplankton initially, feeding powder when the body length of the young fish reaches 2-3cm, transferring the young fish to a culture pond after the body surface of the young fish is covered with scales, and feeding the young fish to the second generation of young fish with the age of 1 according to the method for culturing the Pengze crucian carp;
11) selecting female parents from the second generation and intensive culture:
selecting female tetraploid (allotetraploid pengze crucian carp) with large individual and good development in the second-generation adult fish as a female parent, selecting a male parent from (an aquaculture base of university of agriculture in Huazhong) with sexual maturity, and respectively carrying out enhanced cultivation on the two parents before delivery (to ensure good development of gonads of the parents);
12) artificial induction of female parent allotetraploid pengze crucian:
injecting an oxytocic drug into a female tetraploid (allotetraploid Pengze crucian) female parent by adopting a twice-injection oxytocic method, wherein a first needle firstly injects a luteinizing hormone releasing hormone analogue (LRH-A2) at the time of the day 22, and a second needle injects a compound agent, and the time interval of the two injections is 8-12 h; wherein in the first needle injection, the injection amount of the luteinizing hormone releasing hormone analogue is 0.5-2.0 mug/kg, and in the second needle injection, the composite medicament is a combined medicament of the luteinizing hormone releasing hormone analogue (LRH-A2) and Dixolone (DOM), and the injection amounts of the composite medicament and the dixolone are 2-8 mug/kg and 2-8 mg/kg respectively;
13) acquisition of male parent sperm:
selecting male with good gonad development before artificial insemination, slowly squeezing abdomen, collecting semen at cloaca, and storing at 4 deg.C in dark place for a short time;
14) artificial insemination (dry insemination):
firstly, extruding the ovum of the allotetraploid Pengze crucian carp into a dry steel basin, then adding semen, slightly stirring, adding water and quickly stirring to ensure that the sperm and the ovum are fully contacted to obtain a fertilized ovum;
15) artificial incubation: putting the fertilized eggs into an incubation barrel for incubation in running water, and incubating fish fries;
16) and (3) seedling cultivation: after the fry swims smoothly, transferring the fry to a nursery pond for cultivation, feeding zooplankton initially, and feeding artificial mixed powder when the body length of the fry reaches 2-3 cm;
17) and (3) juvenile fish culture: and after the surfaces of the fry are covered with scales, transferring the fry to a culture pond, and feeding the fry to 1 age according to the carassius auratus pengze breeding method to obtain induced heterotetraploid carassius auratus gynogenesis filial generation.
1. Chromosome karyotype analysis of gynogenesis progeny of the -induced heterotetraploid carassius pengzhi
In this example, the gynogenesis individuals obtained by mating the allotetraploid pengze crucian carp with have small difference in quantifiability from the allotetraploid pengze crucian carp, and are difficult to distinguish from each other in appearance, the ploidy identification result shows that the gynogenesis filial generations of the allotetraploid pengze crucian carp and are still tetraploid, and the chromosome karyotype analysis result shows that the chromosome group of the gynogenesis filial generations of the induced allotetraploid pengze crucian carp consists of 200+ chromosomes (table 1 and fig. 1).
TABLE 1 chromosome karyotype analysis of gynogenesis progeny of allotetraploid carassius pengzensis
2. Explore whether the -induced heterotetraploid pengze crucian gynogenesis progeny and the heterotetraploid pengze crucian inbred progeny have difference in growth traits
In this example, two zygotic generations were subjected to growth contrast test, and 150 well-developed offspring were selected immediately after the fry was flat-swimming and transferred to 0.8m 3 The white round barrels are used for feeding, 3 parallel breeding barrels are respectively arranged in each combination, and 50 fish fries are respectively placed in each barrel. In the test process, the fry is subjected to standardized breeding, and is fed regularly, at fixed points and quantitatively every day and is subjected to dirt absorption once a week, so that the breeding conditions are ensured to be as consistent as possible. Randomly taking out 30 fishes from each barrel for measuring the body length and the body quality at the ages of 30 days, 60 days, 90 days and 120 days respectively, wherein the operation is fine in the measuring process, the excessive stress is prevented, and the fishes are placed back after the measurement.
The growth comparison test result shows that the induced heterotetraploid carassius auratus gynogenesis progeny population is more advantageous in the aspects of body length and weight increase from the beginning of 60 days old, and the growth speed is obviously faster than that of the heterotetraploid carassius auratus inbred population (fig. 2 and 3).
3. statistical analysis of fertilization rate, hatchability and emergence rate of gynogenesis filial generation of allotetraploid carassius pengzensis
In this embodiment, the artificial dry insemination of sperm and the ovum of the allotetraploid pengzhe crucian is performed, fertilized eggs are counted when the embryo grows to the late stage of the primitive intestine, the hatchability is counted after all the fry rupture the membrane, and the rate of emergence is counted after all the fries are in the parallel trip. results of fertilization rate, hatchability and emergence rate of gynogenesis offspring of allotetraploid carassius pengzensis induced by the induction are shown in Table 2.
TABLE 2 statistics of fertilization rate, hatchability and emergence rate of gynogenesis progeny of induced heterotetraploid carassius pengzensis
The above studies show that the progeny obtained by inducing the gynogenesis of the allotetraploid carassius auratus pengzi by is biased to the female parent in morphological characteristics, and the growth speed of the progeny for inducing the gynogenesis of the allotetraploid carassius auratus pengzi by is obviously faster than that of the inbred progeny of the allotetraploid carassius auratus pengzi.
Other parts not described in detail are prior art. Although the present invention has been described in detail with reference to the above embodiments, it is only a part of the embodiments of the present invention, not all of the embodiments, and other embodiments can be obtained without inventive step according to the embodiments, and the embodiments are within the scope of the present invention.
Claims (5)
1. The gynogenesis method of induced allotetraploid carassius auratus pengze is characterized in that: the method comprises the following steps:
1) selection and cultivation of male and female parents:
the female parent is selected from sexually mature Pengze crucian carp, the male parent is selected from sexually mature Xingguo carp, and the two parents are respectively subjected to intensive culture before delivery;
2) artificial oxytocic of female parent carassius auratus pengze:
injecting an oxytocic drug into the female parent of the carassius auratus pekinensis by adopting a two-injection oxytocic method, injecting a luteinizing hormone releasing hormone analogue into a first needle, and injecting a compound agent into a second needle, wherein the time interval between two injections is 8-12 h;
3) obtaining male parent Xingguan red carp sperms:
selecting male Xingguo red carps with good gonad development before artificial insemination, slowly squeezing the abdomens of the male Xingguo red carps, collecting semen at cloacae holes, and storing for later use;
4) artificial insemination:
firstly, extruding eggs of carassius auratus pengze into a dry steel basin, then adding sperm of the cyprinus carpio, stirring, adding water, and quickly stirring to ensure that the sperm and the eggs are fully contacted to obtain fertilized eggs;
5) artificial incubation:
putting the fertilized eggs into an incubation barrel for incubation in running water, and incubating first-filial generation fry;
6) and (3) seed cultivation of the first filial generation:
after the first-filial generation of fish fry swims smoothly, transferring the first-filial generation of fish fry to a seedling raising pool for cultivation, feeding zooplankton initially, and feeding powder when the length of the fish fry reaches 2-3 cm;
7) and (3) young fish culture of the first filial generation:
after the surfaces of the fry are covered with scales, transferring the fry into a culture pond, and feeding first-generation juvenile fishes to adult fishes of 1 age according to the method for culturing Pengze crucian carps;
8) selecting female parents from the first filial generation adult fish and carrying out intensive culture:
selecting a female with large individual and good development in the first filial generation adult fish as a parent, and carrying out reinforced cultivation;
9) artificial oxytocic of first filial generation female parent Pengze crucian and acquisition of Xingguo red carp sperms:
injecting an oxytocic drug into the female parent of the carassius auratus pekinensis by adopting a two-injection oxytocic method, injecting a luteinizing hormone releasing hormone analogue into a first needle, and injecting a compound agent into a second needle, wherein the time interval between two injections is 8-12 h; selecting male Kyoho carps with good gonad development before artificial insemination, obtaining semen, and placing in a dark place at 4 ℃ for short-time preservation;
10) artificial insemination, artificial incubation, breeding of second-generation offspring fry and breeding of juvenile fish:
performing dry insemination, putting fertilized eggs into an incubation barrel for running water incubation after fertilization, incubating second-generation young fishes, transferring the second-generation young fishes into a seedling culture pond for cultivation after the second-generation young fishes swim, feeding zooplankton initially, feeding powder when the body length of the second-generation young fishes reaches 2-3cm, transferring the second-generation young fishes into a culture pond after the body surfaces of the second-generation young fishes are covered with scales, and feeding the second-generation young fishes to adult fishes of 1 age according to the method for culturing the Pengze crucian carp;
11) selecting female parents from the second generation adult fish and carrying out intensive culture:
selecting large and well-developed female tetraploids in the second-generation adult fish as female parents, selecting the male parents from with sexual maturity, and respectively carrying out intensive culture on the two parents before delivery;
12) artificial induction of female tetraploid female parent:
injecting an oxytocic drug into a female tetraploid female parent by adopting a two-time injection oxytocic method, injecting a luteinizing hormone releasing hormone analogue into a first needle, and injecting a compound medicament into a second needle, wherein the time interval of the two injections is 8-12 hours;
13) acquisition of male parent sperm:
selecting male with well developed gonad before artificial insemination, slowly squeezing the abdomen, collecting semen at cloaca, and storing in dark place at 4 deg.C for a short time;
14) artificial insemination:
firstly, extruding the ovum of the allotetraploid Pengze crucian carp into a dry steel basin, then adding semen, slightly stirring, adding water and quickly stirring to ensure that the sperm and the ovum are fully contacted to obtain a fertilized ovum;
15) artificial incubation: putting the fertilized eggs into an incubation barrel for incubation in running water, and incubating fish fries;
16) and (3) seedling cultivation: after the fry swims smoothly, transferring the fry to a nursery pond for cultivation, feeding zooplankton initially, and feeding artificial mixed powder when the body length of the fry reaches 2-3 cm;
17) and (3) juvenile fish culture: and after the surfaces of the fry are covered with the scales, transferring the fry into a culture pond, and feeding the young fish to 1 age according to the method for culturing the Pengze crucian carp to obtain induced allopetraploid Pengze crucian gynogenesis filial generation.
2. The method for inducing the gynogenesis of allotetraploid carassius pengzensis as claimed in claim 1, wherein the method comprises the following steps:
in the step 2), the step 9) and the step 12), the injection amount of the luteinizing hormone releasing hormone analogue in the first needle injection is 0.5-2.0 mu g/kg.
3. The method for inducing the gynogenesis of an allotetraploid carassius auratus pengzi according to claim 2, wherein the method comprises the following steps: the injection amount of the luteinizing hormone releasing hormone analogue is 1 mug/kg by the first injection.
4. The method for inducing the gynogenesis of allotetraploid carassius pengzensis as claimed in claim 1, wherein the method comprises the following steps:
in the step 2), the step 9) and the step 12), in the second needle injection, the composite medicament is a combined medicament of a luteinizing hormone releasing hormone analogue and a diketone, and the injection amounts of the composite medicament and the diketone are respectively 2-8 mug/kg and 2-8 mg/kg.
5. The method for inducing the gynogenesis of the allotetraploid carassius pengzensis as claimed in claim 4, wherein: in the second needle injection, the injection amount of the luteinizing hormone releasing hormone analogue is 4 mug/kg, and the injection amount of the diketone is 4 mg/kg.
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| US20080115232A1 (en) * | 2004-09-29 | 2008-05-15 | Hiroshi Onozato | Amphidiploid Aquatic Animal and Method of Breeding the Same |
| CN104161000A (en) * | 2014-08-28 | 2014-11-26 | 沈阳华泰渔业有限公司 | Hybrid carp and pengze crucian carp hybrid breeding method |
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| US20080115232A1 (en) * | 2004-09-29 | 2008-05-15 | Hiroshi Onozato | Amphidiploid Aquatic Animal and Method of Breeding the Same |
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