WO2016029492A1 - 紫海杂交扇贝三系配套育种体系的构建方法及应用 - Google Patents

紫海杂交扇贝三系配套育种体系的构建方法及应用 Download PDF

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Publication number
WO2016029492A1
WO2016029492A1 PCT/CN2014/085721 CN2014085721W WO2016029492A1 WO 2016029492 A1 WO2016029492 A1 WO 2016029492A1 CN 2014085721 W CN2014085721 W CN 2014085721W WO 2016029492 A1 WO2016029492 A1 WO 2016029492A1
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scallop
line
scallops
hybrid
purple
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PCT/CN2014/085721
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English (en)
French (fr)
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王春德
孙奎府
马斌
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王春德
孙奎府
马斌
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Priority to US15/310,393 priority Critical patent/US10973209B2/en
Publication of WO2016029492A1 publication Critical patent/WO2016029492A1/zh

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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
    • A01K61/50—Culture of aquatic animals of shellfish
    • A01K61/54—Culture of aquatic animals of shellfish of bivalves, e.g. oysters or mussels
    • 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
    • A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
    • A01K67/033—Rearing or breeding invertebrates; New breeds of invertebrates
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
    • Y02A40/81—Aquaculture, e.g. of fish

Definitions

  • the invention belongs to a shellfish breeding technology--the hybrid scallop commodity seed seed breeding method, in particular to a purple sea hybrid scallop including a male sterile line, a maintainer line and a restorer line (a hybrid offspring of the purple scallop egg and the bay scallop sperm)
  • a purple sea hybrid scallop including a male sterile line, a maintainer line and a restorer line (a hybrid offspring of the purple scallop egg and the bay scallop sperm)
  • the Argopecten irradians irradians was native to the Atlantic coast of the United States and was introduced to China in 1982.
  • Bay scallops have a fast growth rate, wide temperature adaptability and strong resistance to stress. They are very suitable for aquaculture in the northern seas of China. At present, the yield and aquaculture area account for more than 50% of scallop culture in China.
  • the scallops in the culture process are seriously degraded during the breeding process.
  • the performance of the shellfish is getting smaller and smaller, and the mortality during the breeding and breeding process is higher. Therefore, the scallop breeding industry urgently needs genetic improvement of the bay scallop.
  • cross breeding is one of the most effective methods for improving germplasm.
  • Purple scallop is a fast-growing medium-sized scallop native to the South Pacific. It belongs to the Argopecten genus and has the same number of chromosomes and is hermaphroditic, but the two are complementary in terms of individual size, growth and temperature adaptability. Therefore, the inventors also cultivated two hybrids of orthogonal (Zifan scallop egg and bay scallop sperm fertilization) and reciprocal (Zi scallop egg and purple scallop sperm fertilization) by interspecific hybridization between purple scallop and bay scallop. The weight of the hybrid generation is about 100% higher than that of the bay scallops cultivated at the same time.
  • the object of the present invention is to achieve the first construction of a three-line hybrid breeding system of purple sea scallops including male sterile lines, maintain lines and restore lines in animal breeding, in particular in hermaphroditic scallops, to overcome the existing Insufficient hybrid scallop breeding technology.
  • Another object of the present invention is to establish a large-scale, low-cost production method for hybrid sea scallop seedlings by using the above three-line breeding system.
  • the above three lines refer to the scallop male sterile line and its maintainer and restorer lines.
  • male sterile lines are male gonads that are abnormally developed and cannot produce functional sperm, but their eggs are normally developed, can receive normal sperm and fertilize, and can inherit their male sterility characteristics to the offspring.
  • the male sterile line is the core strain in the three-line system, and its main function is to provide a large number of unfertilized eggs in the commercial production of hybrid seed, and its breeding depends on the supporting line.
  • the male sterile line is a cross between the male sterility and the ovarian fertile individual who has found the male sterility and the patriarchal backcross to establish the backcross pedigree. When all the individuals in the backcross pedigree are male sterility and the ovum is fertile, the hybrid is obtained. Scallop line.
  • the maintainer line also known as the male sterile line maintainer, can maintain the male sterility characteristics when the male parent is crossed with the male sterile line.
  • the main function of the maintainer line is to provide the sperm necessary for male sterile line breeding.
  • the maintainer line is a hermaphroditic line that can be normally bred, and the next generation maintainer can be bred by selfing.
  • the restorer line also known as the male sterile line restorer line, uses its sperm to fertilize the eggs of the male sterile line, and the resulting female offspring and male fertility return to normal. Therefore, in animal breeding, the role of the restorer line is to provide fertilization of the sperm and the male sterile line to achieve the breeding of excellent commercial hybrid seed.
  • the restorer line is a hermaphroditic line that can be normally bred, and can breed the next generation of restorer lines by selfing.
  • the present invention is based on the following concepts and practices: 1) The inventors found that the purple sea hybrid scallops (purple scallops) There are 5-10% male sterile individuals in the progeny of the egg and the scallop sperm. These individuals do not produce active sperm, but their eggs can be fertilized by the sperm of the bay scallop, and the fertilized eggs obtained can Normal development, that is, the backcrossing scallops; 2) The study further found that about 5-8% of the backseat scallops obtained after maturity are still male sterile and their eggs can be fertilized by bay scallop sperm, so As long as the eggs of the above male sterile individuals and the sperm of the self-crossing progeny of the scallops of the scallops are backcrossed for many generations, the male sterile lines (mothers) in which the offspring are all male sterile and the eggs are fertile can be selected.
  • Bay scallop (parental) combination the scallop line in this combination has the ability to maintain the male sterility characteristics of the corresponding male sterile line, that is, the male sterile line of the purple sea hybrid scallop and the male sterility The maintainer of the line; 3)
  • the study also found that about 90% of the back-spotted scallops cultivated by the backcross of the male sterility of the bay scallop sperm and the purple sea scallop are still hermaphroditic. Therefore, by establishing a plurality of backcrossing families and performing a fitness test, a male sterile line (mother) and a bay scallop (parent) combination in which all offspring are hermaphroditic and have excellent traits can be selected.
  • the scallop strain has the ability to restore male fertility to male fertility, that is, to obtain the restorer line of the hybrid scallop male sterile line in purple sea; 4) the application of purple sea hybrid scallop male sterile line-maintaining line hybrid can mass produce male Infertile and fertile cultivars; by crossing these male sterile cultivars with the corresponding restorer sperm, a convenient, efficient, and large-scale production of hybrid scallop commercial seedlings can be achieved, and the obtained hybrid scallop products are obtained. Seeds have significant advantages in terms of growth, individual size and temperature tolerance.
  • the construction method of the male sterile line, the maintainer line and the restorer line of the purple sea scallop of the present invention is as follows:
  • Zihai hybrid scallop Zifan scallop egg and bay scallop sperm fertilization sub-generation family
  • Zihai hybrid scallop Zifan scallop egg and bay scallop sperm fertilization sub-generation family
  • to be used to select male sterile individuals while fertilizing the eggs of bay scallops with autologous sperm to establish the self-bred family of the bay scallop. Since the proportion of male sterile individuals in the hybrid generation is very low, it is necessary to construct a large number of hybrid generation families to find male sterile individuals for breeding male sterile lines.
  • the fertilized bay scallops are self-fertilized to establish the inbred family;
  • the family and its father-in-law, the scallops, were self-bred, and the individuals who were still male-sterile in the back-crossing family were selected to stimulate the spawning with conventional bay scallop spawning methods.
  • the sperm continues to be returned, and the fertilized bay scallops continue to be self-sufficient to establish self-branched families; afterwards, the breeding methods are continuously backed up in the same way every year until the offspring are all male-sterile and the eggs are fertile, ie Obtained the purple male hybrid scallop male sterile line.
  • the male sterile line is the core strain of the three-line breeding system. It is the premise.
  • the purpose of establishing the purple scallop male sterile line is to provide a large number of male sterile scallop species for the production of hybrid scallop seed, thus achieving convenience. Efficient, large-scale access to a large number of unfertilized eggs.
  • the male sterile line produced by the male sterile line-maintaining line can be selected and the egg can be selected.
  • the large individual scallops are fertilized and fertilized with different scallop sperms with good traits to establish a large number of backcross families.
  • the fertilized bay scallops are self-propagated to establish self-crossing families, and the growth and survival of hybrid progeny are measured.
  • the situation is also tested for cohesion; from the second year onwards, according to the results of the combining ability test and the development of the gonadal gland, the backcross family with good traits and hermaphroditic scallops is selected, and the related male sterile family is continuously and fertilized.
  • the scallop hybrids of the bay scallops are backcrossed to establish the backcross family.
  • the fertilized bay scallop self-bred families are self-propagated and expanded by the conventional bay scallop seedling method, that is, the male is obtained. Restorer of the sterile line.
  • the role of the restorer is to provide fertilization of sperm and male sterile lines to achieve the breeding of good commercial hybrids.
  • the invention not only establishes the male sterile line, the maintainer line and the restorer line of the purple sea scallop, but also establishes the expansion method of each line to achieve continuous use, that is, the series can be continued every year and used for the following Zihai hybrid scallop commodity seed production.
  • the method of producing hybrid sea scallop commercial seedlings by three-line breeding system Firstly, the male sterile line and the restorer type seedlings of purple sea scallops were selected according to the ratio of 5:1, and then cooked according to the conventional method. Stimulate male sterile males to lay eggs and restore the breeding eggs, and then use the 500 mesh sieve to obtain the semen of the restorer, and add the semen of the restorer to the spawning. Stir in the pool, and add the amount of sperm to 5-6 sperm around each egg, and finally incubate after the conventional method That is to obtain the hybrid scallop commodity seed of purple sea.
  • the invention constructs a three-line breeding system including the male sterile line, the maintainer line and the restorer line of the purple sea scallop, and establishes a method for producing the purple sea hybrid scallop product seed by using the three-line breeding system.
  • the specific steps are as follows: :
  • At least 20 such hybrid families were established by the above method, and the fertilized bay scallop sperm eggs were mixed and fertilized, and a corresponding number of bay scallop self-bred families were established. After the larvae cultivation, seedling cultivation and cultivation according to the conventional scallop seedling raising method, For screening of male sterile individuals.
  • Self-communication family establish at least 20 back-crossing families and corresponding bay scallop self-bred families and carry out larval breeding, seedling cultivation and cultivation according to the conventional bay scallop seedling method; in the third year of spring, the back-crossing family and bay scallops will be self-interested.
  • the male sterile individuals in the backcross family with 50 traits are selected, and the fertilized eggs of the normal fertilized and developed fertilized eggs are collected by the self-crossing spermatozoa of the father's bay scallops to continue to establish the backcrossing family.
  • Semen fertilization with different bay scallops establish at least 20 backcross families, and at the same time, the fertilized bay scallops will establish self-crossing families, measure the growth and survival of the backcross progeny and conduct the combining ability test; from the second year According to the results of the combining ability test and the development of the gonadal gland, the backcross pedigrees with good traits and hermaphroditic scallops are selected each year, and the male sterility family of the cultivar and the fertilized bay scallop hybrid family are backcrossed to establish the backcross family.
  • the fertilized bay scallop family is self-propagated and expanded, that is, the restorer line of the purple scallop male sterile line is obtained, which is used for the hybrid scallop commodity seed of the purple sea. Large-scale production.
  • the purple scallops with mature gonads were obtained, and the scallop eggs were obtained after temperature stimulation.
  • the hybrids of purple scallops and bay scallops were obtained and cultivated.
  • the scallops from the first generation of the above-mentioned hybrids were selected to select the scallops that were large and only developed in the female area.
  • the scallops were placed in a container containing 23 °C seawater to stimulate spawning, from multiple bays.
  • the spermatozoon of scallops is inseminated and cultivated to produce a backcross population of Zihai hybrid scallops and bay scallops.
  • the backcross population showed mostly hermaphrodites after spring ripening in 2010, but ⁇ 10% of the individuals still showed male infertility.
  • the male sterile lines (mothers) whose male offspring are all male sterile and the eggs are fertile can be obtained.
  • - Maintaining a lineage (parental) combination to obtain a male sterile line and a maintainer line, respectively.
  • the artificial scallops can be used to induce the production of male sterile scallops to obtain a large number of unfertilized eggs.
  • Fertilization with the sperm of the maintainer line can be used to obtain large-scale production of hybrid scallops for Zihai.
  • Male sterile line of shellfish can be used to obtain large-scale production of hybrid scallops for Zihai.
  • the eggs of the male sterile scallops in the above combination are fertilized with the sperm of different unrelated bay scallops, at least 20 backcross families are established and the combining ability test is carried out, and the offspring containing the hermaphroditic scallops are selected, and the female parent is not male. Breeding species continue to fertilize the spermatozoa of the self-property bay scallops, and continue to breed according to the growth traits and gonad development until the backcross progeny are all hermaphroditic and have good traits, so that the male sterile line is obtained after fertilization. The offspring are hermaphroditic and traits with excellent traits of the scallop family.
  • the eggs of a single male sterile individual are backcrossed with the sperm of a single bay scallop to establish a number of backcross families, and the fertilized bay scallops are selfed to establish a corresponding self-bred family.
  • the gonad development of the family returned to the family was examined. If there were male sterility individuals in the family, the fertilization ability of the eggs was further tested by the sperm of the father's self-interested family, and the family and father were returned.
  • the Bay scallops are self-bred to carry out the next generation of breeding; after one or more generations of breeding, when the backcross progeny are all male sterile and the eggs are fertile, the male sterile line and its corresponding maintainer are obtained. .
  • the eggs of a single male sterile individual are backcrossed with the sperm of a single bay scallop to establish a number of backcross families, and the fertilized bay scallops are selfed to establish a corresponding self-bred family.
  • the gonad development of the family returned to the family was examined.
  • the male sterile line-recovery line combination was obtained only by one generation; if only the part of the returned family was hermaphrodite, then The male sterile scallops continue to establish a backcrossing family with the spermatozoa of the corresponding bay scallops, and continue to establish the self-branched family of the scallops of the scallops for the next generation of breeding; after one or more generations of breeding, When the backcross progeny are all hermaphroditic, the restorer line of the male sterile line is obtained.
  • the method for constructing the male sterile line, the maintainer line and the restorative line established by the invention is also applied to the breeding of the sea purple hybrid scallop (the hybrid offspring of the bay scallop egg and the purple scallop sperm), and is applied to establish the sea purple hybrid scallop commodity seed. Large-scale production methods.
  • the bay scallops and purple scallops are selected to promote ripening and stimulate simultaneous spawning and ejaculation.
  • the eggs of bay scallops and the sperm and eggs of purple scallops are collected separately.
  • the eggs of bay scallops and the sperm of purple scallops are fertilized to establish the generation of sea purple hybrids.
  • the family is used to select male sterile individuals.
  • the sperm and eggs of the purple scallops are mixed and fertilized to establish a self-bred family of purple scallops.
  • male sterile individuals are selected from the family of Haizi hybrids, and the purple scallops are used.
  • the high-yield, convenient and large-scale production of sea-breeding scallop male sterile cultivars can be achieved by using the above-mentioned hybrids of the sea-sterile hybrid scallops with the sperm of the maintainer line; the sea-brown hybrid scallop male sterile seedlings and restorer lines
  • the sperm hybridization can achieve high-efficiency, convenient and large-scale production of sea purple hybrid scallop seed.
  • the method for constructing the male sterile line, the maintainer line and the restorative line established by the invention is also applied to the breeding of the purple-stained scallop (the hybrid offspring of the purple scallop egg and the scallop sperm of the Gulf of Mexico), and is applied to establish the purple scallop hybrid scallop commodity seed.
  • the scale of production methods are also applied to the breeding of the purple-stained scallop (the hybrid offspring of the purple scallop egg and the scallop sperm of the Gulf of Mexico), and is applied to establish the purple scallop hybrid scallop commodity seed.
  • the purple scallops and the scallops of the Gulf of Mexico were selected to promote ripening and stimulate simultaneous spawning and ejaculation.
  • the eggs of the purple scallops and the sperm and eggs of the scallops of the Gulf of Mexico were collected respectively, and the eggs of the purple scallops and the sperm of the scallops of the Gulf of Mexico were fertilized to establish purple ink.
  • a family of hybrids used to select male sterile individuals.
  • the sperm and eggs of the scallops of the Gulf of Mexico were mixed and fertilized to establish a self-crossing family of scallops in the Gulf of Mexico.
  • male sterile individuals were selected from the family of purple-violet hybrids.
  • the male sterile line-recovery line combination if only the part of the returning family is hermaphrodite, the male sterile scallop continues to establish a backcross family with the sperm back of the corresponding Gulf Bay scallop self-family, while continuing to establish the male parent
  • the self-branched family of scallops in the Gulf of Mexico carries out the next generation breeding; after one or more generations of breeding, when the backcross progeny are all hermaphroditic, the recovery line of the purple-soil hybrid scallop male sterile line is obtained.
  • the high-quality, convenient and large-scale production of purple-breeding scallop male sterile cultivars can be achieved by hybridizing the male sterile line of the purple-stained scallops with the sperm of the maintainer; the purple-breeding scallop male sterile shell and restorer line
  • the hybridization of sperm can achieve high-efficiency, convenient and large-scale production of hybrid scallop seedlings of purple ink.
  • the method for constructing the male sterile line, the maintainer line and the restorative line established by the invention is also applied to the breeding practice of the ink purple hybrid scallop (the hybrid offspring of the scallop egg and the purple scallop sperm in the Gulf of Mexico), and is applied to establish the purple purple hybrid scallop commercial seedling.
  • a large-scale production method is also applied to the breeding practice of the ink purple hybrid scallop (the hybrid offspring of the scallop egg and the purple scallop sperm in the Gulf of Mexico), and is applied to establish the purple purple hybrid scallop commercial seedling.
  • the purple scallops and the scallops of the Gulf of Mexico were selected to promote ripening and stimulate simultaneous spawning and ejaculation.
  • the eggs of the scallops of the Gulf of Mexico and the sperm and eggs of the purple scallops were collected respectively.
  • the eggs of the scallops of the Gulf of Mexico and the sperm of the purple scallops were fertilized to establish purple
  • a family of hybrids is used to select male sterile individuals.
  • the sperm and eggs of the purple scallops are mixed and fertilized to establish a self-crossing family of purple scallops.
  • male sterile individuals are selected from the family of the purple-violet hybrids.
  • the sperm of the purple scallop is fertilized, if the fertilized egg develops normally Establish a back-crossing family to continue to breed male infertile families, and at the same time, self-fertilization of the male scallops to establish a self-respecting family; thus, after one generation to multiple generations of choice, when the backcross progeny all manifested as male infertility, they obtained The male sterile line of the purple-violet scallop and its corresponding maintainer.
  • the high-efficiency, convenient and large-scale production of the purple-stained hybrid scallop male sterile seedling can be realized; the hybrid purple scallop male sterile seedling and the restorer line
  • the sperm hybridization can realize the efficient, convenient and large-scale production of the hybrid purple scallop seed.
  • the method for constructing the three-line breeding system of the invention is also applied to the sea purple hybrid scallop (the hybrid offspring of the bay scallop egg and the purple scallop sperm), the purple ink hybrid scallop (the hybrid scallop of the purple scallop egg and the scallop sperm of the Gulf of Mexico) and the purple-hybrid hybrid.
  • the breeding practice of scallops (the hybrid offspring of scallop egg and scallop sperm in the Gulf of Mexico), such as the Argopecten hybrid scallop, is applied to the establishment of large-scale production methods for these hybrid scallop commercial seed.
  • the method of the present invention is also applied to interspecific cross breeding of other hermaphroditic scallops, such as interspecific hybridization of scallops of the genus Pecten.
  • the basic method of the present invention is also applicable to interspecific cross breeding of other hermaphroditic invertebrates.
  • the purple male hybrid scallop male sterile line, maintainer line and restorer line constructed by the invention are the first to establish a three-line breeding breeding system of scallop in the animal kingdom, which should be a major breakthrough in animal breeding technology.
  • the invention effectively overcomes the obstacle of self-fertilization in the process of hybridization of the androgynous scallop, and realizes the low-cost and large-scale obtaining complete hybrid progeny, so that the hybrid advantage can be fully utilized, and the cultivated purple sea hybrid scallop commodity seed has the same
  • the characteristics of bay scallops and purple scallops have significant advantages over bay scallops in terms of growth, individual size and temperature tolerance, so they can be successfully cultured in the northern seas such as Qingdao and Dalian, and the annual yield can be increased by more than 40% compared with bay scallops. .

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Abstract

一种紫海杂交扇贝三系配套育种体系的构建方法及应用，所述三系包括紫海杂交扇贝的雄性不育系、保持系和恢复系。紫海杂交扇贝是紫扇贝卵子与海湾扇贝精子的杂交后代。首先构建大量的紫海杂交子一代家系，从中找到雄性不育的个体用于雄性不育系的选育；然后用海湾扇贝的精子连续回交，分别选育后代全部为雄性不育个体的雄性不育系-保持系组合，和后代全部为雌雄同体个体的雄性不育系-恢复系组合；最后用雄性不育系与保持系杂交生产雄性不育系亲贝，用雄性不育系与恢复系杂交生产紫海杂交扇贝商品苗种。上述紫海杂交扇贝三系配套育种体系的构建方法及应用在动物种建立了扇贝三系配套育种体系，有效克服了雌雄同体扇贝杂交种自体受精的障碍，且当年产量比海湾扇贝提高40％以上。

Description

紫海杂交扇贝三系配套育种体系的构建方法及应用 技术领域
本发明属于贝类育种技术--杂交扇贝商品苗种培育方法，尤其是涉及一种包括雄性不育系、保持系和恢复系的紫海杂交扇贝(紫扇贝卵子与海湾扇贝精子的杂交后代)三系配套育种体系的构建方法及应用。
背景技术
海湾扇贝(Argopecten irradians irradians)原产于美国大西洋沿岸，于1982年引入我国。海湾扇贝生长速度快，温度适应范围广，抗逆性强，非常适合我国北方海域的养殖，目前产量和养殖面积占我国扇贝养殖的50％以上。但是由于引进后长期近交，海湾扇贝在养殖过程中种质退化严重，表现为商品贝规格越来越小，育苗养殖过程中死亡率较高，因此扇贝养殖业迫切需要对海湾扇贝进行遗传改良，其中杂交育种是最有效的种质改良方法之一。
为此，发明人自2007年首次从秘鲁引进了紫扇贝(Argopecten purpuratus)。紫扇贝是原产于南太平洋的一种速生型中型扇贝，与海湾扇贝都属于Argopecten属，染色体数相同，且都是雌雄同体，但二者在个体大小、生长和温度适应性等方面性状互补。因此发明人又通过将紫扇贝与海湾扇贝进行种间杂交，培养出正交(紫扇贝卵子和海湾扇贝精子受精)和反交(海湾扇贝卵子和紫扇贝精子受精)两种杂交子一代。杂交子一代的个体重比同期培育的海湾扇贝提高约100％，当年可长到7-8公分，而且第二年可继续养殖达到更大规格，个体重达到200克以上，其抗逆性增强，特别是在山东地区能够顺利度夏和越冬，显示出巨大的产业潜力(王春德等，2009.紫扇贝与海湾扇贝种间杂交的研究。海洋科学，33(10)：84-87；Chunde Wang等，2011.Introduction of the Peruvian scallop and its hybridization with the bay scallop in China，Aquaculture,2011,310:380-387)。
上述杂交子一代虽然可以产生较大的杂种优势，但是要大面积推广仍存在着较大的困难，这是因为海湾扇贝和紫扇贝都是雌雄同体的扇贝，繁殖时同时排放精子和卵子，排放的卵子容易受到同种精子的污染，难以大量获得未受精卵用于种间杂交，从而导致所获得的商品苗种中真正的杂交后代比例大大降低，因而使其产量的提高幅度大大受限。
研究发现，上述两种Argopecten扇贝的雌雄同体特性与许多农作物的雌雄同株特性相似。在雌雄同株的农作物育种中也存在着同花授粉的问题，需要人 工去雄，这一点与紫扇贝与海湾扇贝杂交过程中遇到的自体受精问题相似。农业上解决这个问题的关键依赖于雄性不育植株的发现，利用雄性不育株与其父本连续回交，就可以选育出雄性不育系、保持系和恢复系，以此来构建农作物育种中的三系配套育种体系，实现杂交种的大规模生产，从而解决杂交制种难题。迄今为止，国内外已经在众多的农作物中建立了三系配套育种体系，并广泛应用于杂交种的生产中，成为大幅度提高农作物产量的主要途径。
但是在动物育种中，国内外迄今为止尚无发现雄性不育且雌性可育个体的报道，因而也无法建立类似于农作物的雄性不育系、保持系和恢复系，用于杂交育种。显然，如果在动物育种中能够构建起类似的包括雄性不育系、保持系和恢复系的三系配套育种体系，也就可以实现雌雄同体型动物的低成本、规模化种间杂交，提高杂交效率，提高育种动物的抗逆性和产量，从而实现动物育种技术的重大突破。
发明内容
本发明的目的是实现在动物育种中，具体地说是在雌雄同体型扇贝中首次构建包括雄性不育系、保持系和恢复系等品系的紫海杂交扇贝三系配套育种体系，以克服现有杂交扇贝育种技术的不足。
本发明的另一目的是利用上述三系配套育种体系建立大规模、低成本生产紫海杂交扇贝苗种的应用方法。
上述的三系是指扇贝雄性不育系及其保持系和恢复系。其中的雄性不育系是雄性性腺发育不正常，不能产生有功能的精子，但其卵子发育正常，能接受正常精子而受精，并能将其雄性不育特性遗传给后代的品系。显然雄性不育系是三系配套体系中的核心品系，其主要作用是在杂交苗种商业化生产中提供大量的未受精卵子，同时其繁育依赖于与其配套的保持系。雄性不育系是利用偶然发现的雄性不育且卵子可育的个体与其父本连续回交建立回交家系，当回交家系的所有个体均为雄性不育且卵子可育时所获得的杂交扇贝品系。
保持系又称雄性不育系保持系，当作为父本与雄性不育系杂交时，能使子一代保持雄性不育特性，保持系的主要作用是提供雄性不育系繁育所必须的精子。保持系是能够正常繁育的雌雄同体品系，可以通过自交繁育下一代保持系。
恢复系又称雄性不育系恢复系，用它的精子给雄性不育系的卵子受精后，所产生的杂交后代雌性和雄性育性均恢复正常。因此，在动物育种中，恢复系的作用是提供精子与雄性不育系的卵子受精，以实现优良商品杂交苗种的培育。恢复系是能够正常繁育的雌雄同体品系，可以通过自交繁育下一代恢复系。
本发明基于以下构思和实践：1)发明人研究发现了紫海杂交扇贝(紫扇贝 卵子与海湾扇贝精子的杂交后代)子一代中存在5-10％的雄性不育个体，这些个体不产生有活性的精子，但其卵子可以被海湾扇贝的精子受精，且所获得的受精卵能够正常发育，即获得了回交扇贝；2)研究进一步发现，由此获得的回交扇贝性腺成熟后约有5-8％的个体仍为雄性不育且其卵子可被海湾扇贝精子受精，因此只要将上述雄性不育个体的卵子与父本海湾扇贝自交后代的精子连续多代回交，就能选育出后代全部为雄性不育且卵子可育的雄性不育系(母本)-海湾扇贝(父本)组合，该组合中的海湾扇贝品系具有使相应的雄性不育系的后代仍保持雄性不育特性的能力，即获得紫海杂交扇贝的雄性不育系和该雄性不育系的保持系；3)研究还发现，用海湾扇贝精子与紫海杂交扇贝中雄性不育个体的卵子回交所培育的回交扇贝养成后约90％个体仍表现为雌雄同体，因此可以通过建立多个回交家系并进行配合力测验的方法，选育出其后代全部为雌雄同体且性状优良的雄性不育系(母本)-海湾扇贝(父本)组合，这个组合中的海湾扇贝品系具有使雄性不育系恢复雄性育性的能力，即获得紫海杂交扇贝雄性不育系的恢复系；4)应用紫海杂交扇贝雄性不育系-保持系杂交可以大规模生产雄性不育且卵子可育的种贝；通过将这些雄性不育种贝与相应的恢复系的精子杂交，能实现方便、高效、规模化生产紫海杂交扇贝商品苗种，且所获得的杂交扇贝商品苗种在生长、个体大小和温度耐受性方面具有显著的优势。
本发明的紫海杂交扇贝雄性不育系、保持系和恢复系的构建方法如下：
1)紫海杂交扇贝雄性不育个体的筛选及其父本的繁育：选择优良的紫扇贝与海湾扇贝个体同步促熟后，用常规的海湾扇贝阴干和温度刺激方法诱导紫扇贝和海湾扇贝产卵，并分别收集紫扇贝和海湾扇贝的卵子，同时诱导海湾扇贝排精，并用500目筛绢过滤获得海湾扇贝精子，然后向紫扇贝的卵子中加入适量的海湾扇贝精子，受精卵正常发育后即获得紫海杂交扇贝(紫扇贝卵子与海湾扇贝精子受精)子一代家系，待用于选择雄性不育个体，同时将海湾扇贝的卵子用自体的精子受精，建立该海湾扇贝的自交家系。由于杂交子一代中雄性不育个体存在的比例很低，因此需要构建大量的杂交子一代家系，从而找到雄性不育的个体用于雄性不育系的选育。
2)紫海杂交扇贝雄性不育系的选育：选择上述紫海杂交扇贝子一代家系中的雄性不育个体，与父本海湾扇贝自交家系中的优良个体同步促熟，性腺成熟后用常规的海湾扇贝阴干和温度刺激的方法诱导上述杂交子一代扇贝排卵并收集卵子，同时诱导海湾扇贝排精，并用500目筛绢过滤获得海湾扇贝精子，将上述获得的杂交子一代的卵子与海湾扇贝精子受精，收集受精且发育正常的受精卵建立回交家系，同时将供精的海湾扇贝自体受精建立自交家系；翌年将回 交家系及其父本海湾扇贝自交家系促熟，选择回交家系中仍为雄性不育的个体用常规的海湾扇贝产卵诱导方法刺激产卵后，用其父本海湾扇贝自交后代的精子继续回交，同时仍将供精的海湾扇贝继续自交建立自交家系；以后每年按照同样的方法连续回交选育，直到回交后代全部为雄性不育且卵子可育的个体，即获得紫海杂交扇贝雄性不育系。雄性不育系是三系配套育种体系的核心品系，是前提，建立紫海杂交扇贝雄性不育系的目的是为杂交扇贝苗种的生产提供大量的雄性不育扇贝种贝，从而实现方便、高效、规模化获得大量的未受精卵。
3)紫海杂交扇贝保持系的选育：将上述能使雄性不育系后代继续保持雄性不育特性的海湾扇贝自交家系用常规的海湾扇贝育苗方法自交扩繁，即获得该雄性不育系的保持系。保持系的主要作用是提供雄性不育系繁育所必须的精子，它是雄性不育系传代所必需的父本品系。
4)紫海杂交扇贝恢复系的选育：获得上述雄性不育系(母本)-保持系(父本)组合后，选择用该雄性不育系-保持系生产的雄性不育且卵子可育的大个体扇贝，促熟产卵后分别用性状优良的不同海湾扇贝精子授精，建立大量的回交家系，同时将供精的海湾扇贝自繁建立自交家系，测量杂交后代的生长和存活情况并进行配合力测验；从第二年起每年根据配合力测验结果和性腺发育情况，选择其中性状优良且含有雌雄同体扇贝的回交家系，将与其相关的雄性不育家系种贝连续与供精的海湾扇贝杂交家系精子回交建立回交家系，直到回交家系全部为雌雄同体且性状优良，则将供精的海湾扇贝自交家系用常规的海湾扇贝育苗方法自交扩繁，即获得该雄性不育系的恢复系。恢复系的作用是提供精子与雄性不育系的卵子受精，以实现优良商品杂交苗种的培育。
5)紫海杂交扇贝雄性不育系、保持系和恢复系的扩繁：本发明通过紫海杂交扇贝雄性不育系与保持系的杂交繁育下一代紫海杂交扇贝雄性不育系扇贝，从而实现紫海杂交扇贝雄性不育系的扩繁；保持系和恢复系均为雌雄同体且正常发育的海湾扇贝品系，因此通过常规的海湾扇贝群体自交方法分别繁育下一代保持系和恢复系。本发明不但建立了紫海杂交扇贝的雄性不育系、保持系和恢复系，而且建立了各品系的扩繁方法以达到持续利用，即保证了各品系能够每年延续下去，并用于下述的紫海杂交扇贝商品苗种生产。
6)用三系配套育种体系生产紫海杂交扇贝商品苗种的方法：首先按照5:1的比例选择紫海杂交扇贝的雄性不育系和恢复系种贝，并按常规方法促熟后分别刺激雄性不育系种贝产卵和恢复系种贝产卵排精，再将该恢复系的精卵混合液用500目筛绢过滤即获得恢复系的精液，将恢复系的精液加入产卵池中并搅拌，且加入精子的量以每个卵子周围有5-6个精子为宜，最后按常规方法孵化后 即获紫海杂交扇贝商品苗种。
具体实施方式
本发明构建了紫海杂交扇贝的包括雄性不育系、保持系和恢复系的三系配套育种体系，并建立了用三系配套育种体系生产紫海杂交扇贝商品苗种的方法，具体步骤如下：
实施例1
1)紫海杂交扇贝雄性不育个体的筛选及其父本的繁育：第一年选择50个性腺发育成熟的紫扇贝和50个海湾扇贝，阴干刺激30分钟后将每个扇贝放在预先加入23℃海水的单独的容器中使其产卵、排精，单独收集每个紫扇贝的卵子和每个海湾扇贝的精子和卵子，向每个紫扇贝的卵子悬浮液中加入少量来自单个海湾扇贝的精液，搅拌使精卵混合均匀，显微镜下观察使每个卵子周围有5-6个精子，并将受精卵置于黑暗环境下孵化，即获得紫海杂交子一代家系。用上述方法建立至少20个这样的杂交家系，同时将供精的海湾扇贝精卵混合受精，建立相应数量的海湾扇贝自交家系，按照常规的扇贝育苗方法进行幼虫培育、保苗和养成后，用于雄性不育个体的筛选。
2)紫海杂交扇贝雄性不育系的选育：第二年春季将紫海杂交子一代家系促熟后，选择50个性状优良的雄性不育个体，用常规的海湾扇贝产卵诱导方法刺激产卵，分别用其父本海湾扇贝的精子受精，收集正常受精、发育的受精卵建立回交家系，收集正常受精、发育的受精卵建立回交家系，同时将提供精子的海湾扇贝自繁建立自交家系，建立至少20个回交家系和相应的海湾扇贝自交家系并按照常规的海湾扇贝育苗方法进行幼虫培育、保苗、养成；第三年春季，将回交家系和海湾扇贝自交家系促熟后，选择50个性状优良的回交家系中的雄性不育个体，用其父本海湾扇贝的自交后代精子回交，收集正常受精、发育的受精卵继续建立回交家系，同时继续繁育海湾扇贝的自交家系，选择建立20个以上性状优良的回交家系及其相应的海湾扇贝自交家系；以后每年按照同样的方法选择性状优良的雄性不育扇贝与其父本海湾扇贝的自交家系回交，直到回交后代的所有个体均表现为雄性不育且卵子可育，即获得紫海杂交扇贝雄性不育系。
3)紫海杂交扇贝保持系的选育：将上述能使雄性不育系个体的回交后代所有个体均表现为雄性不育且卵子可育的父本海湾扇贝按照常规的海湾扇贝育苗 方法大量自交繁育，即获得该紫海杂交扇贝雄性不育系的保持系。
4)紫海杂交扇贝恢复系的选育：选择用上述选育的紫海杂交扇贝雄性不育系和保持系生产的雄性不育且卵子可育的大个体扇贝50个，促熟产卵后分别用不同海湾扇贝的精子受精，建立至少20个回交家系，同时将供精的海湾扇贝自繁建立自交家系，测量回交后代的生长和存活情况并进行配合力测验；从第二年起每年根据配合力测验结果和性腺发育情况选择其中性状优良且含有雌雄同体扇贝的回交家系，将与其相关的雄性不育家系种贝连续与供精的海湾扇贝杂交家系精子回交建立回交家系，直到回交家系全部为雌雄同体且性状优良，则将供精的海湾扇贝家系自交扩繁，即获得该紫海杂交扇贝雄性不育系的恢复系，用于紫海杂交扇贝商品苗种的规模化生产。
5)用三系配套育种体系生产紫海杂交扇贝商品苗种的方法：每年用紫海杂交扇贝雄性不育系和保持系种贝杂交扩繁紫海杂交扇贝雄性不育系扇贝，同时按常规海湾扇贝育苗方法自交扩繁恢复系；春季育苗时按照5:1的比例选择紫海杂交扇贝雄性不育系和恢复系种贝促熟后，在产卵池中刺激雄性不育系种贝产卵，在水桶中刺激恢复系种贝排精，并用500目筛绢过滤获得不含卵子的恢复系海湾扇贝精液；将恢复系的精液均匀泼洒到产卵池中并立即搅拌，随时取样在显微镜下观察受精情况，精液加入量以每个卵子周围有5-6个精子为宜；受精卵按照常规方法孵化，发育后即获得紫海杂交扇贝商业苗种。
实施例2
2008年3月底，取性腺发育成熟的紫扇贝，经温度刺激后获得紫扇贝卵子，用海湾扇贝的精子授精后获得紫扇贝与海湾扇贝杂交子一代苗种并养成。2009年春天，将上述杂交子一代扇贝促熟后从中选择个体大且仅雌区发育的扇贝，阴干30分钟后将扇贝置于盛有23℃海水的容器中刺激产卵，以来自多个海湾扇贝的精子混合授精，培育出子一代紫海杂交扇贝与海湾扇贝的回交群体。该回交群体在2010年春季促熟后大部分表现为雌雄同体，但还有<10％的个体仍表现为雄性不育。
继续将上述雄性不育的个体用其父本海湾扇贝自交后代的精子回交，经过一代至多代选育就可获得后代全部为雄性不育且卵子可育的雄性不育系(母本)-保持系(父本)组合，从而分别获得雄性不育系和保持系。性腺促熟后用常规海湾扇贝产卵诱导方法催产雄性不育系扇贝可获得大量未受精的卵子，将其用保持系的精子受精，就可大规模获得用于紫海杂交扇贝商品苗种生产的雄性不育系种贝。
将上述组合中的雄性不育系扇贝的卵子用不同的非相关海湾扇贝的精子受精，建立至少20个回交家系并进行配合力测验，选择后代含有雌雄同体扇贝的家系，将其母本雄性不育系种贝继续用供精的海湾扇贝自交家系的精子受精，并根据生长性状和性腺发育情况连续选育，直到回交后代全部为雌雄同体且性状优良，就获得受精后使雄性不育系后代均为雌雄同体且性状优良的恢复系海湾扇贝家系。
最后将上述获得的紫海杂交扇贝的雄性不育系种贝性腺促熟后，用常规的海湾扇贝催产方法获得大量未受精的卵子，将其用恢复系的精子受精，就可大规模生产紫海杂交扇贝商品苗种。
实施例3
2010年春天，选择2009年建立的紫扇贝卵子与海湾扇贝精子杂交子一代中雄性不育的个体，经促熟后刺激产卵，用海湾扇贝的精子受精，建立了12个回交家系并养成。2011年春天促熟后发现，其中的11个家系大部分个体表现为雌雄同体，而其中的1个家系的大部分个体仅雌区发育，少量个体性腺不发育。
依此方法，将其中的单个雄性不育个体的卵子与单个海湾扇贝的精子回交建立若干回交家系，并将供精海湾扇贝自交建立相应的自交家系。第二年检查回交家系的性腺发育情况，若回交家系存在有雄性不育的个体，则进一步用其父本自交家系的精子检测其卵子的受精能力，并继续建立回交家系和父本海湾扇贝自交家系，进行下一代选育；经一代或多代选育后，当回交后代全部为雄性不育且卵子可育时，就获得了雄性不育系及其相应的保持系。
同样，将其中的单个雄性不育个体的卵子与单个海湾扇贝的精子回交建立若干回交家系，并将供精的海湾扇贝自交建立相应的自交家系。第二年检查回交家系的性腺发育情况，若回交家系全部为雌雄同体的扇贝，则仅需选择一代就获得了雄性不育系-恢复系组合；若回交家系仅有部分为雌雄同体，则用雄性不育扇贝继续与相应的海湾扇贝自交家系的精子回交建立回交家系，同时继续建立父本海湾扇贝的自交家系，进行下一代选育；经一代或多代选育后，当回交后代全部为雌雄同体时，就获得了该雄性不育系的恢复系。
实施例4
2012年用紫海杂交子一代家系中仅卵区发育的雄性不育个体，经刺激后其中有11个雄性不育个体产卵，将这些卵子与单个海湾扇贝的精子回交，大部分个体的卵子不能正常受精，少数虽能够受精但大部分发育不正常，仅仅发育出带纤毛的畸形胚胎。但产卵的11个个体中有1个个体的卵子能够正常发育，从而培育出回交家系并养殖到成体阶段。在2012年11月中旬收获时，与同期的 海湾扇贝比较，回交家系的壳高、壳长、壳宽和体重分别提高34.1％、29.8％、23.0％和111.5％，杂种优势极其显著。2013年春季将回交扇贝促熟后，发现后代全部为雌雄同体，证明通过用海湾扇贝精子与雄性不育个体回交完全能成功选育出紫海杂交扇贝雄性不育系的恢复系。
实施例5
2013年和2014年，利用引进的紫扇贝与海湾扇贝共建立了紫扇贝卵子与海湾扇贝精子的杂交子一代家系18个，并建立了供精海湾扇贝的自交家系，按照实施例1的方法，可在1-3年内选育出紫海杂交扇贝的雄性不育家系及其相应的保持系和恢复系，实现紫海杂交扇贝苗种的商业化生产。
实施例6
本发明用于海紫杂交扇贝雄性不育系、保持系和恢复系的构建及应用
本发明建立的雄性不育系、保持系和恢复系的构建方法也应用于海紫杂交扇贝(海湾扇贝卵子与紫扇贝精子的杂交后代)的育种，并且应用于建立海紫杂交扇贝商品苗种的规模化生产方法。
首先选择海湾扇贝和紫扇贝同步促熟并刺激同时产卵、排精，分别收集海湾扇贝的卵子以及紫扇贝的精子和卵子，将海湾扇贝的卵子与紫扇贝的精子受精建立海紫杂交子一代家系，用于选择雄性不育个体，同时将紫扇贝的精子和卵子混合受精，建立紫扇贝自交家系；第二年从海紫杂交子一代家系中选择雄性不育的个体，用紫扇贝的精子与其受精，若受精卵正常发育则建立回交家系，用于继续选育雄性不育系，同时将父本紫扇贝自体受精建立自交家系；这样经过一代至多代选择，当回交后代全部表现为雄性不育时，就获得了海紫杂交扇贝的雄性不育系及相应的保持系。
选择上述海紫杂交扇贝雄性不育系中的若干大个体扇贝，刺激产卵后用多个非相关紫扇贝的精子受精，若回交家系全部为雌雄同体的扇贝，则仅需选择一代就获得了雄性不育系-恢复系组合；若回交家系仅有部分为雌雄同体，则用雄性不育扇贝继续与相应的紫扇贝自交家系的精子回交建立回交家系，同时继续建立父本紫扇贝的自交家系，进行下一代选育；经一代或多代选育后，当回交后代全部为雌雄同体时，就获得了海紫杂交扇贝雄性不育系的恢复系。
用上述海紫杂交扇贝的雄性不育系卵子与保持系的精子杂交，即可实现高效、方便、规模化生产海紫杂交扇贝雄性不育种贝；用海紫杂交扇贝雄性不育种贝与恢复系的精子杂交，即可实现高效、方便、规模化生产海紫杂交扇贝苗种。
实施例7
本发明用于紫墨杂交扇贝雄性不育系、保持系和恢复系的构建及应用
本发明建立的雄性不育系、保持系和恢复系的构建方法也应用于紫墨杂交扇贝(紫扇贝卵子与墨西哥湾扇贝精子的杂交后代)育种，并且应用于建立紫墨杂交扇贝商品苗种的规模化生产方法。
首先选择紫扇贝和墨西哥湾扇贝同步促熟并刺激同时产卵、排精，分别收集紫扇贝的卵子以及墨西哥湾扇贝的精子和卵子，将紫扇贝的卵子与墨西哥湾扇贝的精子受精建立紫墨杂交子一代家系，用于选择雄性不育个体，同时将墨西哥湾扇贝的精子和卵子混合受精，建立墨西哥湾扇贝自交家系；第二年从紫墨杂交子一代家系中选择雄性不育的个体，用墨西哥湾扇贝的精子与其受精，若受精卵正常发育则建立回交家系，用于继续选育雄性不育家系，同时将父本墨西哥湾扇贝自体受精建立自交家系；这样经过一代至多代选择，当回交后代全部表现为雄性不育时，就获得了紫墨杂交扇贝的雄性不育系及相应的保持系。
选择上述紫墨杂交扇贝雄性不育系中的若干大个体扇贝，刺激产卵后用多个非相关墨西哥湾扇贝的精子受精，若回交家系全部为雌雄同体的扇贝，则仅需选择一代就获得了雄性不育系-恢复系组合；若回交家系仅有部分为雌雄同体，则用雄性不育扇贝继续与相应的墨西哥湾扇贝自交家系的精子回交建立回交家系，同时继续建立父本墨西哥湾扇贝的自交家系，进行下一代选育；经一代或多代选育后，当回交后代全部为雌雄同体时，就获得了紫墨杂交扇贝雄性不育系的恢复系。
用上述紫墨杂交扇贝的雄性不育系卵子与保持系的精子杂交，即可实现高效、方便、规模化生产紫墨杂交扇贝雄性不育种贝；用紫墨杂交扇贝雄性不育种贝与恢复系的精子杂交，即可实现高效、方便、规模化生产紫墨杂交扇贝苗种。
实施例8
本发明用于墨紫杂交扇贝雄性不育系、保持系和恢复系的构建及应用
本发明建立的雄性不育系、保持系和恢复系的构建方法也应用于墨紫杂交扇贝(墨西哥湾扇贝卵子与紫扇贝精子的杂交后代)育种实践，并且应用于建立墨紫杂交扇贝商品苗种的规模化生产方法。
首先选择紫扇贝和墨西哥湾扇贝同步促熟并刺激同时产卵、排精，分别收集墨西哥湾扇贝的卵子以及紫扇贝的精子和卵子，将墨西哥湾扇贝的卵子与紫扇贝的精子受精建立墨紫杂交子一代家系，用于选择雄性不育个体，同时将紫扇贝的精子和卵子混合受精，建立紫扇贝自交家系；第二年从墨紫杂交子一代家系中选择雄性不育的个体，用紫扇贝的精子与其受精，若受精卵正常发育则 建立回交家系，用于继续选育雄性不育家系，同时将父本紫扇贝自体受精建立自交家系；这样经过一代至多代选择，当回交后代全部表现为雄性不育时，就获得了墨紫杂交扇贝的雄性不育系及其相应的保持系。
选择上述墨紫杂交扇贝雄性不育系中的若干大个体扇贝，刺激产卵后用多个非相关紫扇贝的精子受精，若回交家系全部为雌雄同体的扇贝，则仅需选择一代就获得了雄性不育系-恢复系组合；若回交家系仅有部分为雌雄同体，则用雄性不育扇贝继续与相应的紫扇贝自交家系的精子回交建立回交家系，同时继续建立父本紫扇贝的自交家系，进行下一代选育；经一代或多代选育后，当回交后代全部为雌雄同体时，就获得了墨紫杂交扇贝雄性不育系的恢复系。
用上述墨紫杂交扇贝的雄性不育系卵子与保持系的精子杂交，即可实现高效、方便、规模化生产墨紫杂交扇贝雄性不育种贝；用墨紫杂交扇贝雄性不育种贝与恢复系的精子杂交，即可实现高效、方便、规模化生产墨紫杂交扇贝苗种。
工业实用性
本发明的三系配套育种体系构建方法也应用于海紫杂交扇贝(海湾扇贝卵子与紫扇贝精子的杂交后代)、紫墨杂交扇贝(紫扇贝卵子与墨西哥湾扇贝精子的杂交后代)和墨紫杂交扇贝(墨西哥湾扇贝卵子与紫扇贝精子的杂交后代)等Argopecten属杂交扇贝的育种实践，并且应用于建立这些杂交扇贝商品苗种的规模化生产方法。
本发明的方法也应用于其他雌雄同体型扇贝的种间杂交育种，例如Pecten属扇贝的种间杂交，本发明的基本方法也适用于其他雌雄同体型无脊椎动物的种间杂交育种。
本发明构建的紫海杂交扇贝雄性不育系、保持系和恢复系是首次在动物界中建立了扇贝的三系配套育种体系，应属于动物育种技术中的重大突破。本发明有效克服了雌雄同体扇贝杂交过程中自体受精的障碍，实现了低成本和规模化获得完全的杂交后代，因此可充分利用杂种优势，达到了使所培育的紫海杂交扇贝商品苗种同时具有海湾扇贝和紫扇贝的特性，在生长、个体大小和温度耐受性方面比海湾扇贝具有显著优势，因此能在北方海区如青岛、大连等海域成功养殖，且当年产量可比海湾扇贝提高40％以上。

Claims (10)

  1. 一种紫海杂交扇贝三系配套育种体系，其特征在于包括紫海杂交扇贝的雄性不育系、保持系和恢复系，其中紫海杂交扇贝是紫扇贝卵子与海湾扇贝精子的杂交后代。
  2. 权利要求1的紫海杂交扇贝三系配套育种体系的构建方法，其特征在于：
    1)紫海杂交扇贝雄性不育个体的筛选及其父本的繁育：首先构建20个以上紫海杂交扇贝子一代家系，同时建立供精的海湾扇贝自交家系，用于下述雄性不育系的选育；
    2)紫海杂交扇贝雄性不育系的选育：第二年从上述紫海杂交扇贝子一代家系中选择50个雄性不育个体，促熟后刺激其排卵，用该雄性不育个体的父本海湾扇贝自交家系中优良个体的精子受精，收集正常发育的受精卵建立回交家系20个以上，同时将供精的海湾扇贝自体受精建立自交家系；翌年选择回交家系中仍为雄性不育的个体50个促熟并刺激产卵后，用该雄性不育个体的父本海湾扇贝自交后代的精子继续回交，建立20个以上回交家系，同时仍将供精的海湾扇贝继续自交建立自交家系；以后每年按照同样的方法连续回交选育，直到回交后代全部为雄性不育且卵子可育的个体，即获得紫海杂交扇贝雄性不育系；
    3)紫海杂交扇贝保持系的选育：将上述能使雄性不育系后代继续保持雄性不育特性的海湾扇贝自交家系用常规的扇贝育苗方法自交扩繁，即获得该雄性不育系的保持系。
    4)紫海杂交扇贝恢复系的选育：选择用上述雄性不育系-保持系生产的雄性不育且卵子可育的大个体扇贝，促熟产卵后分别用来自不同个体的海湾扇贝精子授精，建立20个以上回交家系，同时将供精的海湾扇贝自繁建立自交家系，测量杂交后代的生长和存活情况并进行配合力测验；从第二年起每年根据配合力测验结果和性腺发育情况，选择其中性状优良且含有雌雄同体扇贝的回交家系，将与回交家系相关的雄性不育家系种贝连续与供精的海湾扇贝杂交家系精子回交，建立回交家系，直到回交家系全部为雌雄同体且性状优良，则将供精的海湾扇贝自交家系用常规的扇贝育苗方法自交扩繁，即获得该 雄性不育系的恢复系。
    5)紫海杂交扇贝雄性不育系、保持系和恢复系的扩繁：通过上述雄性不育系与保持系的杂交繁育下一代雄性不育系扇贝，从而实现雄性不育系的扩繁；通过常规的扇贝群体自交方法分别繁育下一代保持系和恢复系。
  3. 上述权利要求1和权利要求2的紫海杂交扇贝三系配套育种体系应用于规模化生产紫海杂交扇贝雄性不育系种贝，即用紫海杂交扇贝雄性不育系与相应的保持系杂交，以规模化繁育雄性不育且卵子可育的下一代紫海杂交扇贝雄性不育系种贝。
  4. 上述权利要求1和权利要求2的紫海杂交扇贝三系配套育种体系应用于生产紫海杂交扇贝商品苗种，即用紫海杂交扇贝雄性不育系的卵子与相应的恢复系的精子杂交，获得紫海杂交扇贝商品苗种。
  5. 权利要求2建立的紫海杂交扇贝三系配套育种体系的构建方法也应用于以下的各杂交扇贝的育种：
    (1)用海湾扇贝卵子与紫扇贝精子受精培育的海紫杂交扇贝；
    (2)用紫扇贝卵子与墨西哥湾扇贝精子受精培育的紫墨杂交扇
    贝；
    (3)用墨西哥湾扇贝卵子与紫扇贝精子受精培育的墨紫杂交扇
    贝。
  6. 权利要求3和权利要求4建立的紫海杂交扇贝三系配套育种体系的应用方法也应用于以下的各杂交扇贝的育种：
    (1)用海湾扇贝卵子与紫扇贝精子受精培育的海紫杂交扇贝；
    (2)用紫扇贝卵子与墨西哥湾扇贝精子受精培育的紫墨杂交扇
    贝；
    (3)用墨西哥湾扇贝卵子与紫扇贝精子受精培育的墨紫杂交扇
    贝。
  7. 权利要求2建立的紫海杂交扇贝三系配套育种体系的构建方法也应用于其他雌雄同体型扇贝，例如Argopecten属和Pecten属扇贝的种间杂交育种。
  8. 权利要求3和权利要求4建立的紫海杂交扇贝三系配套育种体系应用方法也应用于其他雌雄同体型扇贝，例如Argopecten属和Pecten属扇贝的种间杂交育种。
  9. 权利要求2建立的紫海杂交扇贝三系配套育种体系的基本构建方法也应用于其他雌雄同体型无脊椎动物的种间杂交育种。
  10. 权利要求3和权利要求4建立的紫海杂交扇贝三系配套育种体系的应用方法也应用于其他雌雄同体型无脊椎动物的种间杂交育种。
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