CN114451335B - Breeding method of ternary hybrid scallop commercial seedlings - Google Patents

Breeding method of ternary hybrid scallop commercial seedlings Download PDF

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CN114451335B
CN114451335B CN202111085297.2A CN202111085297A CN114451335B CN 114451335 B CN114451335 B CN 114451335B CN 202111085297 A CN202111085297 A CN 202111085297A CN 114451335 B CN114451335 B CN 114451335B
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scallops
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CN114451335A (en
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宁军号
王春德
刘桂龙
曹为安
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Yantai Spring Sea Aquaseeds Co ltd
Yantai Institute of Coastal Zone Research of CAS
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Yantai Institute of Coastal Zone Research of CAS
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    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01KANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K61/00Culture of aquatic animals
    • A01K61/50Culture of aquatic animals of shellfish
    • A01K61/54Culture of aquatic animals of shellfish of bivalves, e.g. oysters or mussels
    • YGENERAL 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
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02ATECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/80Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
    • Y02A40/81Aquaculture, e.g. of fish

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  • Marine Sciences & Fisheries (AREA)
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Abstract

The invention relates to the technical field of marine economic shellfish crossbreeding, in particular to a method for cultivating a commercial scallop seedling based on the three-way hybridization of purple scallops, bay scallops and gulf scallops. The bay scallop and the gulf scallop are subjected to subspecies hybridization, then the obtained subspecies hybridization first-generation scallop and the purple scallop are used as seed scallops to be respectively subjected to high selfing purification, and the purified sea ink (or black sea) hybrid scallop and the purple scallop are subjected to interspecific hybridization, so that the large-scale preparation of the sea ink purple (or black sea purple) ternary hybrid scallop commercial seedlings is realized. The invention prepares the commercial fry of the three-element hybrid scallop through scale production, the seashell used in the three-element hybrid scallop fry is the highly self-bred purple scallop strain and the seashell (or the black sea) hybrid scallop strain, thus the heterosis of the first-element hybrid scallop can be obviously increased, and the growth and the adverse resistance advantage are more obvious.

Description

Method for cultivating three-way hybrid scallop commercial seedlings
The technical field is as follows:
the invention relates to the technical field of marine economic shellfish crossbreeding, in particular to a method for cultivating a commercial scallop seedling based on the three-way hybridization of purple scallops, bay scallops and gulf scallops.
Background art:
the northern subspecies of bay scallops (Argopecten irradians irradians, abbreviated as bay scallops) and the southern subspecies of bay scallops (Argopecten irradians concentricus, also known as Mexico bay scallops) are originally produced on the coasts of America, are all fast-growing shellfish, are introduced into China in the eighties and nineties of the last century, and rapidly develop into main scallop breeding varieties in the northern and southern coastal areas of China, and cause a high development tide of aquaculture industry in China (Zhang Fu, 2014; han Jiajun et al, 1998). In 2018, the culture area of Argopecten irradians and Argopecten irradians in the whole country is about 200 ten thousand mu, and the annual output is about 120 ten thousand tons (Chinese fishery statistics yearbook, 2019). However, due to the reasons of small number of introduced species, sterile breeding for many years, inbreeding recession and the like, two types of scallop germplasm are seriously degraded, and the outstanding problems of seedling culture failure, high death rate of offspring seeds, slow growth, small specification of commercial scallops and the like often occur (Zhang national model and the like, 2003). In addition, although bay scallops and mexico bay scallops are wide-temperature shellfish, the water temperature of the surface layer in summer in the south and north sea areas of China exceeds the upper temperature tolerance limit, so that the death rate of the bay scallops cultured in the floating raft is high, the bay scallops grow stagnated and are not fat and thin, and the floating raft culture of the mexico bay scallops can avoid the high-temperature season in summer as far as possible, thereby seriously restricting the healthy and sustainable development of the scallop culture industry in China. Therefore, the scallop breeding industry needs high-quality seedlings with fast growth, strong stress resistance and high identification degree to promote the quality improvement and the efficiency improvement of the scallop breeding industry.
At present, the application of genetic engineering breeding and whole genome selective breeding in the cultivation and seed production of new marine economic shellfish varieties is not mature. Hybridization is an important method for improving the quality of animals and plants, increasing the yield and breeding new varieties, and can effectively utilize the heterosis to combine the excellent characters of parents into a whole or accumulate different micro-effect genes for controlling the same excellent characters in the parents. In order to improve the germplasm of cultured Argopecten irradians, argopecten purpuratus is introduced from Peru by the Wangchund team in 2008, and the Argopecten irradians are respectively hybridized to breed one-generation hybrid scallops of purple sea and purple ink with extremely obvious advantages of growth, survival and stress resistance, and the individual weight average of the Argopecten irradians is improved by 80-100% compared with that of the Argopecten irradians cultured in the same period when the Argopecten irradians and the Argopecten irradians are harvested (Wangchund et al 2009; nanlehong et al 2012). Furthermore, zheng et al (2006) performed subspecial hybridization of bay scallops with bay scallops, and the results indicated that the first filial generation of bay scallops exhibited significant growth and survival advantages. In conclusion, the binary hybrid first-generation scallop seedlings have the advantages of fast growth, high survival rate, strong stress resistance and the like no matter interspecific hybridization or subspecies hybridization.
The ternary hybridization is only popularized and applied in animal husbandry, and researches show that the ternary hybridization performance of animals such as pigs, cows and the like is obviously superior to that of binary hybridization. The early-stage research of a team discovers that the first ternary hybrid generation of the Argopecten irradians, the Argopecten irradians and the Argopecten irradians has the excellent characteristics of remarkable growth advantages, strong stress resistance, high survival rate, high meat yield, beautiful shell color and the like. However, most of the first filial generations of bay scallops and gulf scallops and purple scallops are male sterile and the fertilization rate and the hatchability of eggs thereof are low, which significantly increases the difficulty of large-scale preparation of the ternary hybrid scallop commercial seedlings. Therefore, the application of the patent focuses on solving the technical problem of ternary hybrid seed production between the Argopecten irradians and the Argopecten purpuratus.
The invention content is as follows:
the invention provides a cultivation method of a commercial hybrid scallop seedling based on purple scallop, bay scallop and gulf scallop, aiming at the problems that the scallop cultivation industry lacks high-quality seedlings and the male sterility seed production of hybrid offspring of bay scallop, gulf scallop and purple scallop is difficult.
In order to achieve the purpose, the invention adopts the technical scheme that:
a method for cultivating the three-element hybridized scallop commercial seedlings includes such steps as interspecific hybridization between bay scallop and Argopecten irradians, selfing and purifying to obtain purified hybrid scallop, and interspecific hybridization between purified hybrid scallop and Argopecten irradians.
The self-breeding purification is to take subspecies cross first-generation scallop (sea ink (or sea ink) cross first-generation scallop) and purple scallop as seed scallops, respectively establish more than 100 self-breeding families each year, and continuously purify each obtained self-breeding family for four generations through a self-breeding method in the group, thereby obtaining various purified scallop populations.
Further, the following steps are carried out:
1) Establishing and purifying a first filial generation family among sea ink (or sea ink) subspecies: taking bay scallops and gulf scallops as seed scallops, culturing until the gonad maturity is approximately synchronous until the gonads are mature, drying the eggs after maturation, stimulating by raising the temperature of seawater, inducing spawning and collecting unfertilized eggs and sperms of each scallop, carrying out artificial insemination on the eggs of the bay scallops and the sperms of the gulf scallops or the sperms of the gulf scallops and the eggs of the gulf scallops, respectively establishing more than 100 first-filial generation families of sea ink or sea ink, carrying out mixed culture on all larvae of the first-filial generation families of the sea ink (or the sea) after incubation, and further culturing and culturing the first-filial generation larva groups to obtain the first-filial generation scallops of the sea ink (or the sea); then, using the first filial generation of the fast-growing and large individual Mexico (or Mexico sea) hybrid scallop as the seed scallop, respectively establishing over 100 inbred lines of the Mexico (or Mexico sea) hybrid scallop every year, and continuously purifying for three generations by using a self-propagation method in a group to obtain the inbred line of the Mexico (or Mexico sea) hybrid scallop;
2) Establishing a purple scallop inbred line: carrying out in-vivo self-propagation in a group by taking fast-growing and large-individual purple scallops as seed scallops by a method for establishing an inbred line every year, and continuously purifying for four generations to obtain the inbred line of the purple scallops;
3) The large-scale preparation of the three-element hybrid commercial seedlings of the sepia purple (or the sepia purple): the purified purple scallop inbred line and the sea ink (or sea) hybrid scallop inbred line are bred until the gonad maturity degree tends to be synchronous until the gonad is mature, the purified purple scallop inbred line and the sea ink (or sea) hybrid scallop inbred line are simultaneously dried in shade after maturation, the unfertilized eggs and sperms of each scallop are induced to spawn and collected through heating seawater stimulation, then the eggs and sperms of the same kind of scallops are mixed, artificial insemination is carried out for hatching, and the purpose of obtaining the ternary hybrid scallop seedlings in large scale is achieved.
In the step 1), the tolerance of the Argopecten irradians to low temperature is poorer than that of the Argopecten irradians, so that the initial culture water temperature of the Argopecten irradians is higher than that of the Argopecten irradians, the heating strategy is to preferentially heat the culture seawater of the Argopecten irradians at the speed of 0.5 ℃/day, the temperature is stabilized for 2 days after the continuous heating is carried out for 2 ℃, after the culture seawater of the Argopecten irradians is heated to the culture temperature of the Argopecten irradians, the culture seawater of the two Argopecten irradians is heated at the speed of 0.5 ℃/day, when the culture water is heated to 19 ℃, the heating is stopped and the water temperature is kept constant (19 +/-0.5 ℃) until the gonads of the two Argopecten irradians are completely mature, and the culture conditions such as the water temperature and the bait density are regulated, so that the maturity of the two Argopecten irradians tends to be synchronous, and the culture water temperature is kept below 22 ℃ during fattening period; after gonads of Argopecten irradians and Argopecten irradians are mature, the two kinds of Argopecten irradians are simultaneously dried in the shade for 30 minutes, and each Argopecten irradians is put into seawater with the temperature of 24 ℃ in advance for spawning induction.
The self-reproduction continuous purification is carried out in the establishment of the first filial generation family group of the sea ink or the sea ink to obtain the inbred line, and the specific operation is as follows: the method comprises the steps of taking the first filial generation of the fast-growing individual large sea ink (or sea ink) hybrid scallop as a seed scallop, respectively establishing more than 100 sea ink or sea ink selfing families, respectively establishing more than 100 sea ink or sea ink selfing second-generation families after the seed scallop is bred, and continuously purifying the three generations by using a self-propagation method in the group, thus obtaining the selfing line of the sea ink (or sea ink) hybrid scallop.
The ternary hybrid seedling in the step 3) is obtained by hybridizing the sea ink (or black sea) hybrid scallop self-bred line and the purple scallop self-bred line, wherein the tolerance of the purple scallop self-bred line to low temperature is poorer than that of the sea ink (or black sea) hybrid scallop self-bred line, so that the initial culture water temperature of the sea ink (or black sea) hybrid scallop self-bred line is higher than that of the purple scallop self-bred line, the temperature raising strategy is that the purple scallop culture seawater is preferentially raised at the speed of 0.5 ℃/day and is stabilized for 2 days after 2 ℃ continuous temperature raising, after the purple scallop culture seawater is raised to the culture temperature of the sea ink (or black sea) hybrid scallop self-bred line, the two kinds of scallop culture seawater are raised at the speed of 0.5 ℃/day, when the culture water is raised to 19 ℃, the temperature is stopped to raise, the water temperature is kept constant (19 +/-0.5 ℃) until the scallop gonads are fully mature, and the culture conditions such as the water temperature and the bait density are regulated, so that the gonads of the scallops tend to be synchronous, and the culture water temperature is maintained below 22 ℃ during fattening period; the inbred line of sea scallop and the inbred line of purple scallop with mature gonad development are induced to obtain the ovum and sperm of various scallops respectively to carry out artificial insemination, hatching and larva cultivation, thus realizing the large-scale production of the ternary hybrid scallop seedlings.
More specifically:
1. establishing and purifying first filial generation families of the sea ink (or the sea of the sea): in the middle and late ten days of 1 month in the first year, selecting large and strong-activity bay scallops and mexico bay scallops to be transported back to a breeding workshop for fattening, controlling breeding conditions such as breeding water temperature, bait density and the like during fattening to enable the gonad maturity of the two types of scallops to tend to be synchronous, and maintaining the breeding water temperature of the scallops below 22 ℃ during fattening; after gonads of bay scallops and gulf scallops are mature, the two scallops are simultaneously dried in the shade for 30 minutes, and each scallop is placed in a beaker which is added with seawater at 24 ℃ in advance for spawning promotion; collecting eggs into a clean beaker if the scallops lay eggs firstly, filtering and collecting semen by using a 500-mesh bolting silk if the scallops are discharged, then soaking the scallops and the beaker into a fresh water bucket for 3 seconds to kill the semen, and then adding 24 ℃ seawater into the beaker to hasten parturition until the scallops lay eggs and collecting the eggs into the clean beaker; and performing microscopic examination after 15 minutes of oviposition, and if the ovum is discharged from a polar body, indicating that the ovum is fertilized and discarded. The eggs of the bay scallops and the essence of the gulf scallops or the essence of the gulf scallops and the eggs of the gulf scallops are subjected to artificial insemination, more than 100 sea ink or first-filial generation families are respectively established, the first-filial generation larvae of the sea ink (or the sea ink) are mixed and cultured after incubation, and the larvae are cultured, thrown into an attaching medium, kept in a pond, cultured in the middle of a sea area, cultured and the like, and the cultured eggs can reach the commodity specification after 6-8 months according to the culture process of the common gulf scallops. In the following three years, 2000 individual large sea scallops (or black sea) hybrid scallops with fast growth are selected every year, and are continuously purified for three generations by a method for establishing a selfing line, namely the selfing line of the sea scallop (or black sea) hybrid scallop is obtained. The inbred line of the next generation of hybrid scallops is bred every year by in-population self-breeding.
2. Establishing a purple scallop inbred line: from the first year, 2000 individual and strong-vigor purple scallops are selected in the middle and last ten days of 1 month every year, and the selfing line of the purple scallops is obtained by continuously purifying for four generations through the method for establishing the selfing line. Then, the purple scallop inbred line of the next generation is bred by a self-breeding method in the colony every year.
3. The large-scale preparation of the three-element hybrid commercial seedlings of the sepia purple (or the sepia purple): selecting large-individual and strong-activity scallops from the selfing lines of the purple scallops and the sea ink (or black sea) hybrid scallops in the middle and last ten days of each year from the fifth year, transporting the scallops back to a culture workshop for fattening, enabling the gonad maturity of the scallops to be synchronous by regulating and controlling culture conditions such as culture water temperature, bait density and the like during fattening, and maintaining the culture water temperature of the scallops below 22 ℃ during fattening; after gonads of hybrid scallops of purple scallops and sea scallops (or black scallops) are mature, the purple scallops are dried in shade for 30 minutes, then a plurality of purple scallops are placed in a 2-L beaker which is pre-added with about 500mL of seawater at 24 ℃ for spawning promotion, when the purple scallops are sperminated to cause the seawater in the beaker to be turbid, seminal fluid of the purple scallops is filtered into a 100-L plastic barrel by utilizing a 500-mesh bolting silk for micro-aeration for standby application, and after the seminal fluid of the purple scallops is obtained, artificial insemination is preferably performed within 5 hours; after a large amount of sperms of the purple scallops are discharged, taking the sea ink (or the black sea) hybrid scallops to start drying in the shade, after drying in the shade for 30 minutes, putting the single sea ink (or the black sea) hybrid scallop into a 2-L beaker which is pre-added with seawater of about 500mL and 24 ℃ to hasten parturition, simultaneously pouring 2L of sperms of the purple scallops into a 100-L plastic barrel, adding clean seawater of 24 ℃ to dilute for 5 times, if the sea ink (or the black sea) hybrid scallops lay eggs firstly, pouring the eggs into a barrel which is added with diluted 5 times of purple scallop sperms to fertilize, pouring the scallops and the beaker after the sperms are discharged firstly, soaking the scallops and the beaker into a fresh water barrel for 3 seconds to kill the sperms, then adding about 500mL of seawater of 24 ℃ to hasten parturition until the scallops lay eggs, pouring the eggs into the barrel which contains the purple scallop sperms to fertilize; according to the artificial insemination method, about 200 sea ink (or sea ink) hybrid scallops can lay eggs for 4-5 barrels, after egg liquid is filled in the barrels, fertilization is carried out for 5 minutes, the scallops are poured into a hatching pond with a 25-30 square for fertilization and hatching, the hatching density is 40-50/mL, and the breeding density can be controlled during breeding of D-type larvae after hatching. Because the concentration of the semen of the purple scallop is controlled in advance, eggs do not need to be washed in the large-scale production process, and the hybridization rate of the three-way hybrid scallop seedlings obtained by the method is 60-80 percent. Through simple training, 4 workers can finish the spawning and seedling distribution work of 25-30 square water bodies within 1 hour.
The invention has the advantages that:
the invention realizes the large-scale preparation method of the interspecific ternary hybrid scallop commercial seedlings in the scallops for the first time, thereby solving the problem that the interspecific hybrid male sterility of the scallops is difficult to produce seeds; the ternary hybrid scallop commercial seedling can integrate the excellent properties of the purple scallop, the gulf scallop and the gulf scallop, and improve the excellent seedling supply capacity and the culture yield of the scallop culture industry.
The invention has the following advantages:
1. the invention overcomes the problem of interspecific hybrid male sterility of bay scallops (or gulf scallops) and purple scallops, firstly, subspecies hybrid between the bay scallops and the gulf scallops is carried out, so that the first generation fertility of the sea ink (or ink sea) hybrid is not influenced, and the problems that the bay scallops, the gulf scallops and the purple scallops hybrid first generation is expressed as male sterility, the fertility rate and the hatchability of ova are low and the population purification can not be carried out by a method for establishing an inbred family line can be solved; therefore, the method can establish a sea ink (or sea ink) hybrid scallop inbred line for purification.
2. The hybrid scallop used by the three-way hybrid scallop seedling is a highly selfed purple scallop line and an ink (or ink) hybrid scallop line respectively, and the genetic distance between the two highly selfed lines of the scallops is larger, so that the hybrid vigor of the first-generation three-way hybrid scallop can be obviously increased, and the growth and stress resistance vigor are more obvious.
Detailed Description
The following examples are presented to further illustrate embodiments of the present invention, and it should be understood that the embodiments described herein are for purposes of illustration and explanation only and are not intended to limit the invention.
The technical solution of the invention is as follows: firstly, bay scallops and gulf scallops are used for carrying out subspecies hybridization, and the fertility of offspring generated by the subspecies hybridization of sea ink (or ink sea) is not influenced; then utilizing the first filial generation scallop and purple scallop of the sea ink (or the sea ink) as seed shells, respectively establishing more than 100 selfing families each year, respectively carrying out mixed culture on the bred larvae, and continuously breeding and purifying for more than 3 generations by utilizing the selfing family method; and then interspecific hybridization is carried out on the purified sea ink (or sea ink) hybrid scallop inbred line and the purple scallop inbred line, so that the large-scale preparation of the sea ink purple (or sea ink purple) ternary hybrid commercial offspring seeds is realized. The first three-hybrid scallop seeds can normally develop and have remarkable growth and stress resistance advantages, but the first three-hybrid scallops are male sterile and have low egg fertility rate and hatchability, so that the first three-hybrid scallops are not suitable for being directly used as breeding offspring of breeding scallops and are beneficial to protecting the benefits of breeders.
The strategy for cultivating the ternary hybrid commercial seedling by utilizing the purple scallops, the bay scallops and the gulf scallops comprises the steps of firstly cultivating the first filial generation of the sea ink (or the sea) hybrid and carrying out continuous selfing and purification for the third generation, simultaneously carrying out continuous selfing for the fourth generation to purify the purple scallops, and then carrying out ternary hybridization on the purified sea ink (or the sea) hybrid scallop selfing line and the purple scallop selfing line to obtain the ternary hybrid commercial seedling of the sea ink (or the sea purple).
The embodiment is as follows:
in 1 st ten days of 2019, 200 varieties of bay scallops and gulf scallops with large individuals and strong vitality are selected and put into a nursery room for fattening. The tolerance of the Argopecten irradians to low temperature is poorer than that of the Argopecten irradians, so that the initial culture water temperature of the Argopecten irradians is higher than that of the Argopecten irradians, the temperature rise strategy is to preferentially raise the temperature of the culture seawater of the Argopecten irradians at the speed of 0.5 ℃/day, the culture seawater of the Argopecten irradians is stabilized for 2 days after the temperature rise of the culture seawater of the Argopecten irradians is 2 ℃, the temperature rise of the culture seawater of the two Argopectens is further carried out at the speed of 0.5 ℃/day after the culture seawater of the Argopecten irradians is raised to the culture temperature of the Argopecten irradians, and the temperature rise is stopped and is kept constant (19 +/-0.5 ℃) until the gonads of the two Argopectenes are completely mature. After the gonads mature in 3 middle-month, the scallop is dried in the shade for 30 minutes, and the single scallop is placed in a beaker which is added with seawater at 24 ℃ in advance for spawning induction. If the scallop lays eggs firstly, collecting the eggs into a clean beaker; and if the scallops are subjected to semen discharging, filtering by using a 500-mesh bolting silk to collect semen, then soaking the scallops and the beaker into fresh water for 3 seconds to kill the semen, then adding 24 ℃ seawater into the beaker to hasten parturition until the scallops are found to lay eggs, and collecting the eggs into a clean beaker.
The above scallops lay eggs for 15 minutes, then are microscopically inspected, and are discarded if fertilized. Carrying out artificial insemination on the eggs of the bay scallops and the essence of the gulf scallops or the essence of the gulf scallops and the eggs of the gulf scallops according to a conventional mode, controlling 3-5 sperms around each egg, respectively establishing more than 100 sea ink or first-filial generation families, carrying out mixed culture on the first-filial generation larvae of the sea ink (or the sea ink) after hatching, carrying out culture processes of larva culture, attaching medium feeding, pond seedling protection, middle culture in sea areas, culture formation and the like on the collected larvae according to the culture process of the common gulf scallops, and culturing for 6-8 months to reach the commodity specification, namely obtaining the first-filial generation scallops of the sea ink (or the sea ink) with good fertility. In the culture process, the fertility rate, the hatching rate, the growth rate, the eyepoint time and the larva size of the first filial generation of the sea ink (or the ink sea) hybridization are not obviously different from those of the breeding offspring of the bay scallop or the gulf scallop, but the first filial generation of the sea ink (or the ink sea) hybridization inherits the shell colors of the two scallops and obviously increases the commodity specification (the weight of each individual is increased by 20 to 30 percent, and the pillage rate is improved by 5 to 10 percent).
In 1 st month of 2020, 200 individual and powerful sea scallop (or black sea) hybrid first-generation scallops and purple scallops are selected from the above mixed first-generation, and are put into a nursery room for fattening. The tolerance of the purple scallops to low temperature is poorer than that of the first filial generation scallops of the sea ink (or the ink) so that the initial culture water temperature of the first filial generation scallops of the sea ink (or the ink) is higher than that of the purple scallops, the temperature rise strategy is to preferentially heat the culture seawater of the purple scallops at the rate of 0.5 ℃/day, the temperature is stabilized for 2 days after the temperature of the culture seawater of the purple scallops is raised to the culture temperature of the self-bred strain of the hybrid scallops of the sea ink (or the ink), the culture seawater of the two scallops is heated at the rate of 0.5 ℃/day, and when the temperature of the culture water is raised to 19 ℃, the temperature rise is stopped and the water temperature is kept constant (19 +/-0.5 ℃) until the gonads of the scallops are completely mature. After the gonads mature in 3 middle-month, the scallop is dried in the shade for 30 minutes, and the single scallop is placed in a beaker which is added with seawater at 24 ℃ in advance for spawning induction. If the scallops lay eggs firstly, collecting the eggs into a clean beaker, if the scallops are discharged, filtering and collecting semen by using a 500-mesh bolting silk, then soaking the scallops and the beaker into fresh water for 3 seconds to kill the semen, and then adding 24 ℃ seawater into the beaker to hasten parturition until the scallops lay eggs and collecting the eggs into the clean beaker.
The above scallops lay eggs for 15 minutes, then are examined microscopically, and if fertilized, are discarded. Mixing unfertilized eggs of the first filial generation of sea ink (or ink sea) scallop after microscopic examination, and mixing semen of purple scallop to respectively obtain eggs of the first filial generation of sea ink (or ink sea) scallop and semen of purple scallop; carrying out artificial insemination on eggs of the first filial generation scallop of the sea ink (or the sea of the ink) and the essence of the purple scallop, controlling 8-10 sperms around each egg, and hatching to obtain a first filial generation larva population of the three-element hybridization of the sea ink purple (or the sea ink purple). The collected larvae are cultured according to the culture process of common bay scallops, such as larva culture, attaching medium feeding, pond seedling protection, intermediate culture in sea area, cultivation and the like, and the bred larvae can reach the commodity specification after being cultured for 6-8 months, namely the ternary hybrid first-generation scallop with fast growth, large individual and male sterility is obtained. In the culture process, the fertility rate (< 70%) and the hatchability (< 10%) of the first filial generation of the dark purple (or dark purple) are obviously lower than those of the parents, but the growth speed is high, the eyespot time is 1-2 days later, and the eyespot larvae (210-240 μm) are 30-40 μm larger than the bay scallops (180-200 μm). After the scallop is cultivated for 6 months, the individual weight (58.6 +/-11.4 g) of the ternary hybrid first-generation scallop is improved by 60 to 100 percent compared with the common bay scallop (31.2 +/-5.4 g), and the weight (7.5 +/-1.4 g) of the scallop is improved by 40 to 60 percent compared with the common bay scallop (3.9 +/-1.0 g).
In conclusion, the ternary hybrid scallop commercial seedlings obtained by the seedling cultivation method have high fertilization rate and hatching rate, normal development of floating larvae and obvious growth advantages. The method can solve the current situation of insufficient supply of high-quality seedlings in the scallop breeding industry, and improve the breeding yield and economic benefit of scallop farmers. However, the three-hybrid first-generation scallop shows male sterility, has low egg fertility rate and hatchability, is not suitable for being directly used as a breeding scallop breeding progeny, and is beneficial to protecting the benefit of a breeder.

Claims (6)

1. A method for cultivating a ternary hybrid scallop commercial seedling is characterized by comprising the following steps: carrying out subspecies hybridization on bay scallops and gulf scallops, then using the obtained subspecies hybridization first-generation scallops and purple scallops as seed scallops, respectively carrying out high selfing purification, obtaining purified sea ink or carrying out interspecific hybridization on the sea ink hybrid scallops and the purple scallops, and thus realizing the large-scale preparation of sea ink purple or sea ink purple three-element hybrid scallop commercial seedlings.
2. The method for cultivating the three-way hybrid scallop commercial seedlings according to claim 1, which is characterized in that: the high self-breeding purification is to take the first-generation interspecific hybrid scallop and the purple scallop as seed scallops, respectively establish more than 100 self-breeding families, continuously purify the first-generation interspecific hybrid scallop self-breeding family obtained by the above three generations through a method of self-breeding in the scallop group, and continuously purify the purple scallop self-breeding family through the four generations through the method of self-breeding in the scallop group, thus obtaining various purified scallop populations.
3. The method for cultivating the commercial seedlings of the ternary hybrid scallops according to claim 1 or 2, wherein the method comprises the following steps:
1) Establishing and purifying a first-filial generation family: taking bay scallops and gulf scallops as seed scallops, culturing until the gonad maturity is approximately synchronous until the gonads are mature, drying the eggs after maturation, stimulating by heating seawater, inducing spawning and collecting unfertilized eggs and sperms of each scallop, performing artificial insemination on the eggs of the bay scallops and the sperms of the gulf scallops or the sperms of the gulf scallops and the eggs of the gulf scallops, respectively establishing more than 100 first-filial generation families of sea ink or first-filial generation families of sea, performing mixed culture on all larvae of the first-filial generation families of sea ink or first-filial generation families after incubation, and further culturing and culturing first-filial generation larva groups to obtain first-filial generation scallops of sea ink or first-filial generation scallops of sea; then, respectively establishing more than 100 inbred lines of the sea ink or the black sea hybrid scallop as seed scallops by taking the fast-growing and large-individual sea ink or black sea hybrid first-generation scallops as the seed scallops every year, and continuously purifying for three generations by utilizing a method for establishing the inbred lines to obtain the inbred lines of the sea ink or the black sea hybrid scallops;
2) Establishing a purple scallop inbred line: carrying out in-vivo self-propagation in a group by taking fast-growing and large-individual purple scallops as seed scallops by a method for establishing an inbred line every year, and continuously purifying for four generations to obtain the inbred line of the purple scallops;
3) Preparing commercial sea purple or black sea purple ternary hybrid scallop seedlings in large scale: the purified purple scallop inbred line and the sea ink or black sea hybrid scallop inbred line are cultured until the gonad maturity approaches to synchronization until the gonad is mature, the gonads are simultaneously dried in shade after maturation, the temperature rise seawater is used for stimulation, spawning is promoted, unfertilized eggs and sperms of each scallop are collected, then the eggs and the sperms of the same type of scallop are mixed, artificial insemination is carried out for hatching, and the purpose of obtaining the ternary hybrid scallop seedlings in large scale is achieved.
4. The method for cultivating the commercial hybrid scallop seedlings according to the claim 3, which is characterized in that: in the step 1), the bay scallop culture seawater is heated at a heating rate of 0.5 ℃/day, the bay scallop culture seawater is stabilized for 2 days after being heated continuously for 2 ℃, after the bay scallop culture seawater is heated to the culture temperature of the gulf scallops, the two kinds of scallop culture seawater are heated at a heating rate of 0.5 ℃/day, when the culture water is heated to 19 ℃, the heating is stopped, and the water temperature is kept constant for 19 +/-0.5 ℃ until the gonads of the two kinds of scallops are completely mature; and the water temperature, the bait density and other culture conditions are regulated and controlled, so that the gonad maturity of the two scallops tends to be synchronous, and the water temperature for scallop culture is maintained below 22 ℃ during fattening; after the gonads of the bay scallop and the gulf scallop are mature, the two scallops are simultaneously dried in the shade for 30 minutes, and each scallop is put into seawater with the temperature of 24 ℃ in advance for spawning induction.
5. The method for cultivating the commercial hybrid scallop seedlings according to the claim 3, which is characterized in that: the method comprises the steps of establishing a sea ink or black sea first-filial generation family group, carrying out self-reproduction and continuous purification to obtain an inbred line, taking the fast-growing and large-individual sea ink or black sea first-filial generation scallops as seed scallops, respectively establishing more than 100 sea ink or black sea inbred family systems, culturing, then taking the sea ink or black sea first-filial generation family systems as seed scallops, respectively establishing more than 100 sea ink or black sea inbred second-generation family systems, and continuously purifying three generations by using a method of self-reproduction in the group, namely obtaining the inbred line of the sea ink or black sea hybrid scallops.
6. The method for cultivating three-way hybrid scallop commercial seedlings according to claim 3, which is characterized in that: the three-way hybrid scallop seedling in the step 3) is obtained by hybridizing the sea ink or black sea hybrid scallop self-bred line and the purple scallop self-bred line, the purple scallop cultivation seawater is heated at the speed of 0.5 ℃/day, the temperature is continuously raised for 2 days after each continuous heating, the two kinds of scallop cultivation seawater are heated at the speed of 0.5 ℃/day after the purple scallop cultivation seawater is heated to the cultivation temperature of the sea ink or black sea hybrid scallop self-bred line, when the cultivation water temperature is raised to 19 ℃, the heating is stopped, the water temperature is kept constant at 19 +/-0.5 ℃ until the gonads of the scallops are completely mature, the cultivation conditions such as the water temperature and the bait density are regulated, the gonad maturity of the scallops tend to be synchronous, and the scallop cultivation water temperature is kept below 22 ℃ during fattening; spawning sea ink with mature gonad development or the inbred line of hybrid scallop and the inbred line of purple scallop, respectively obtaining the ovum and sperm of various scallops, and carrying out artificial insemination, hatching and larva cultivation, thus realizing the large-scale production of the ternary hybrid scallop seedlings.
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