CN103004648B - Breeding method for improving DHA (Docosahexaenioc Acid) content in oyster body - Google Patents
Breeding method for improving DHA (Docosahexaenioc Acid) content in oyster body Download PDFInfo
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- CN103004648B CN103004648B CN201110283181.XA CN201110283181A CN103004648B CN 103004648 B CN103004648 B CN 103004648B CN 201110283181 A CN201110283181 A CN 201110283181A CN 103004648 B CN103004648 B CN 103004648B
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Abstract
The invention provides a breeding method for improving DHA (Docosahexaenioc Acid) content in an oyster body, wherein purebred oysters from various subspecies or populations of the same species are used as parents; and next, hybrid progenies with high DHA content can be generated through homologous outbreeding between the subspecies or populations of the same species. With the adoption of the breeding method for improving the DHA content in the oyster body, the DHA content in the oyster body can be improved by more than 20% and the efficiency is high; in addition, techniques such as transgenosis and medication are not required by the breeding method for improving the DHA content in the oyster body; therefore, the operation is simple and convenient and the practicability is strong; the breeding method for improving the DHA content in the oyster body can enable nutritional ingredients in the oyster body to be improved remarkably, so that the problem that the nutritional quality of the current bred oysters is low can be effectively suppressed; and therefore, a new route is provided for the cultivation of a high-valued oyster variety and the market prospect is wide.
Description
Technical field
The present invention relates to a kind of breeding method that improves nutrient component in Oysters, is a kind of breeding method that improves DHA content in Oysters specifically.
Background technology
DHA(DHA, 22:6n-3), be commonly called as " docosapentaenoic acid ", it is a kind of long-chain polyvalent unsaturated fatty acid of needed by human, be the important composition composition of brain cell film, participate in formation and the growth of brain cell, to the important function that is formed with of the extension of nerve cell aixs cylinder and new projection, the normal physiological activity of nerve cell be can maintain, human thinking and memory forming process participated in.The abundantest long-chain polyvalent unsaturated fatty acid in DHA or Retinal acceptor, is amphiblestroid important composition composition, to promoting the growth of retinal tissue, promotes the development of visual function, improves depending on keen grade and has important function.The same vitamin of DHA, mineral matter is the same is also the essential nutriment of human body, and in human brain cortex, in human brain cortex, content is up to 20% up to 20% for content, and in eye retina, proportion maximum, accounts for 50%.Therefore, most important to tire infant intelligence and visual acuity, also can effectively prevent senile dementia, the blurring of image etc. simultaneously.Research in recent years shows, DHA can also reduce cholesterol, suppress platelet aggregation, thus angiocardiopathy preventing.In addition, DHA also has very strong anti-oxidation function, can improve the hyperplasia of immunity of organisms, prevention tumour cell.But human body can not oneself synthesize DHA, must obtain from food.Therefore the DHA content, how improving in food more and more causes people's attention.
Oyster is commonly called as " oyster ", " oyster " etc., delicious flavour, nutritious, and protein content is high, have the title of " ocean milk ".In oyster, also contain higher unsaturated fatty acid, its content is conventionally at 4.74%-19.96%.Traditional Chinese medicine thinks, oyster is transquilization with heavy material, checking exuberance of yang tonifying yin, and softening and resolving hard mass, restrains astringent or styptic treatment for spontaneous sweating; Effective in cure for diseases such as palpitation with fear insomnia, a tax on one's mind impairment of the spleen, impotence, pain in the back, uterine bleedings.Because of kind and regional difference, the nutrient component of oyster also has certain difference.
Oyster is also the highest kind of world aquaculture output.According to the World Food Programme's statistics, the whole world only Pacific oyster cultured output just reached 4,400,000 tons in 2003.China is shellfish culture big country, the big country of Ye Shi oyster culture simultaneously, and 2009, oyster culture output reached 350.38 ten thousand tons, accounted for 32.55% of cultivated shellfish gross yield.Jin10Nian Lai China oyster output is always in rising trend.At present, the oyster of China's cultivation mostly is huge oyster and belongs to (Crassostrea) kind, as Pacific oyster C.gigas, Crassostrea rivularis C.ariakensis, the huge oyster C.hongkongensis in Hong Kong, Portuguese oyster C.angulata etc.But due to the expansion of cultivation scale, " water oyster " day by day increases, and causes the nutritional quality of oyster constantly to decline, and seriously affected output and the benefit of oyster.Therefore, must seek a kind of method and improve the nutrient component of oyster, solve oyster culture nutritional quality decline problem.
" homology outbreeding " refers to individuality between the different subspecies (population) that stem from same species or the mating of population level.As everyone knows, hybridization is the important means of animals and plants genetic improvement, and its object is exactly in order to obtain hybrid vigour.Hybrid vigour refers to that two different parents of genetic background hybridize the hybrid F of generation
1on growth potential, vitality, fecundity, resistance, yield and quality than parent's a side or the superior phenomenon of parents.The advantage but not all hybridization can both hybridize.In the time that the phenotype of hybrid surmounts the mean value of parents' phenotype, such hybridization has produced hybrid vigour, and when the phenotypic value of hybrid is during lower than the mean value of parents' phenotype, such hybridization has just produced breakthrough.Research shows, between the kind of oyster, the distant hybridization overwhelming majority is unsuccessful.But, utilize the homology outbreeding method improvement oyster varietal character of kind of interior different subspecies or different geographic populations but to obtain gratifying progress, mostly can produce positive hybrid vigour.But the breeding method of utilizing up to now, homology outbreeding improves DHA content in Oysters but never to be reported.
Summary of the invention
Object of the present invention, for a kind of easy, with clearly defined objective breeding method that improves oyster nutrient component is provided, solves the problem that China's oyster culture nutritional quality declines, thereby promotes the development of China's oyster culture industry.
Technical scheme of the present invention is: plant that shellfish is selected, foundation, seed rearing and the juvenile mollusk of purebred qualification and gonad maturity accelerating, hybrid and purebred combination are formed, DHA assay and the process such as relatively, can make the DHA content of hybrid significantly improve, its operating procedure is:
1, plant that shellfish is selected, purebred qualification and gonad maturity accelerating: plant shellfish and must stem from the purebred of the different subspecies of same oyster or population, adopt morphology and molecular biology method to carry out purebred qualification to parent, by synchronously maturation of manual method regulation and control kind of shellfish sexual gland.
2, the foundation of hybrid and purebred combination: taking the kind shellfish of gonad maturity as material, dissect and obtain respectively sperm and ovum.Then the outbreeding group that hybridizes between subspecies or population in adopting kind.Meanwhile, adopt mating in subspecies or in population to produce to be derived from purebred group of same parent, as the control group of hybrid group.
3, larvae cultivation and juvenile mollusk are formed: the cultivation density, bait, water temperature, the salinity that comprise larva; Juvenile mollusk is in order to reduce environmental effect, and hybrid group and purebred group, from larva, juvenile mollusk, cultivate under identical environmental condition during becoming shellfish always.
4, the mensuration of DHA content and comparison: adopt the DHA content of gas Chromatographic Determination hybrid and purebred oyster, then hybrid group compares with purebred group.
Advantage of the present invention and beneficial effect are as follows:
1, just can make the DHA content in Oysters improve significantly more than 20% by the hybridization of 1 generation, efficiency is high.
2, improve first the nutritional quality of cultivated shellfish from improving the angle of content of polyunsaturated fatty acid, opened up the new way of shellfish breed of variety, for exemplary role has been played in the cultivation of other shellfish new varieties.
3, in view of the special role of DHA to aspects such as medical treatment, health cares, the raising of DHA content can also can improve oyster value-added content of product, and economic worth is improved, and increases breeding enterprise and raiser's income, and market prospects are wide.
4, this method does not need the technology such as transgenosis, drug treating, easy and simple to handle, practical.
The present invention is a kind of method that improves oyster nutrient component, can significantly improve the DHA content in Oysters by this technology.The oyster that this method is cultivated, owing to being rich in DHA, has special value at aspects such as medical treatment and health cares.Can improve nutrition kind and the economic worth of oyster by the present invention, effectively contain the problems such as current aquaculture product price is low-priced, the output value is low, for the cultivation of high-valued oyster kind provides a new way.The present invention does not need by the complex technology such as transgenosis, drug treating, and therefore this product does not affect biological and Environmental security.
Embodiment
1, plant that shellfish is selected, purebred qualification and gonad maturity accelerating: in June, 2010, from Shantou and two places, Qingdao each select 100 health, undamaged 1 age oyster be kind of a shellfish.First respectively get 10, by morphology and 16S order-checking, kind of shellfish is carried out to purebred qualification, result shows that the oyster of Qingdao cultivation is that the oyster that Crassostrea gigas gigas, Shantou cultivate is Crassostrea gigas angulata, the two all belongs to Pacific oyster and plants, is purebred, but belongs to different subspecies.From the oyster of Qingdao cultivation be called for short Qingdao oyster, from Shantou cultivation oyster be called for short Shantou oyster.Kind of the shellfish two weeks that first naturally accelerates the ripening in sea area, and then move on in indoor, to be placed on 4 500L polyethylene drum sexual gland is carried out to artificial ripening.Change water every day 2-3 time, mix with the heart-shaped flat algae+Dicrateria inornata in Asia+yeast throw something and feed, uninterruptedly inflation.About approximately 10 days, planting shellfish sexual gland can be synchronously ripe.
2, the foundation of hybrid and purebred combination: taking the Qingdao oyster of gonad maturity and Shantou oyster as kind of a shellfish, dissect and obtain respectively sperm and ovum separately.The male and female kind shellfish of each subspecies is used 20, then the ovum of same hybrid is mixed and the sperm that is divided into two parts, same subspecies also mixes.Then get appropriate Qingdao oyster sperm and Shantou oyster ovum fertilization and carry out inter-subspecies hybrid, set up homology hybrid group; Get appropriate Qingdao oyster sperm and Qingdao oyster ovum fertilization simultaneously, get appropriate Shantou oyster sperm and Shantou oyster ovum fertilization and carry out inbreeding in subspecies, set up purebred group of two homologies, as the control group of hybrid group.Under 27 DEG C-28 DEG C, salinity 31-32 condition, through 24h, development of fertilized ova is to D type larva.
3, larvae cultivation and juvenile mollusk are formed: larvae cultivation carries out indoor, mainly comprise and cultivate density, bait, water temperature, salinity; Juvenile mollusk is formed at sea and is carried out.Put into the polyethylene drum inflation of 500L cultivates with 300 mesh sieve tulles collection D type larvas.Approximately 2/ml of the initial density of larvae cultivation, each group arrange 3 repetitions.The bait of larva is taking equilateral chrysophyceae and flat algae as main, bait throwing in every day 3 times, first 3 days each 10,000 alga cells/ml of daily ration, feeding quantity, next 7 days each 50,000 alga cells/ml, 100,000 the alga cells/ml that at every turn throw something and feed afterwards; Change water every day 1 time, shell top phase and larva being changed to while going out eyespot in the polyethylene drum of other clean 500L cultivates.Larva culture density was carried out quantitatively in every two days, and cultivate water body so that the larval density in each cultivation bucket is basically identical by adjusting.Added with antibiotic not during larvae cultivation.After 15 days, larva eyespot rate exceedes 30 ﹪, throws in the adherances such as oyster shell or scallop shell, adheres to 8-10 juvenile mollusk throw in by each adherance.Throw in adherance after approximately 1 week, larva all metamorphosis is juvenile mollusk.Juvenile mollusk continues cultivation 1 week in the polyethylene drum of indoor 500L, by stringing and carry out mark with rope with oyster shell or the scallop shell of juvenile mollusk, goes out pond in fine afternoon, is suspended to sea and carries out juvenile mollusk and form; Afterwards, make regular check on growth and the survival condition of juvenile mollusk.After 6 months, can grow into 5cm-6cm.In order to reduce the impact of environment, from larva, juvenile mollusk to forming stage, hybrid group remains consistent with the breeding environment of purebred group.
4, the mensuration of DHA content and comparison: respectively getting 15 adult oysters from hybrid group and purebred group is material, and vivisection obtains fresh soft tissue.The soft tissue of same first stirs evenly, vacuum cooling drying is pulverized again.First use chloroform-methanol mixed liquor extracting fat, then with NaOH methanol solution, fat is carried out to esterification, finally analyze fatty acid component with gas chromatograph, determine the DHA content in hybrid group and purebred group of sample.The DHA content of hybrid oyster reaches 16.24%, the DHA content mean value of purebred oyster is 12.54%, then hybrid and purebred DHA content is compared, and finds that the DHA content in hybrid Oysters has exceeded 29.51% than the DHA content in purebred Oysters.Therefore the DHA content that, homology outbreeding can improve in Oysters reaches more than 20%.
Obviously the present invention can improve the DHA content in Oysters significantly, thereby can effectively contain the low problem of current oyster culture nutritional quality, for the cultivation of high-valued oyster kind provides a new way.
Claims (1)
1. a breeding method that improves DHA content in Oysters, comprises the following steps:
(1) plant that shellfish is selected, purebred qualification and gonad maturity accelerating: plant different subspecies purebred that shellfish must stem from same oyster, adopt morphology and molecular biology method to carry out purebred qualification to parent, by synchronously maturation of manual method regulation and control kind of shellfish sexual gland;
(2) foundation of hybrid and purebred combination: taking the kind shellfish of gonad maturity as material, dissect and obtain respectively sperm and ovum, at least 20 of the male and female individual number of every all shellfishes, then the subspecies outbreeding group that hybridizes in adopting kind, simultaneously, adopt sub-intrabreeding to produce to be derived from purebred group of same parent, as the control group of hybrid group;
(3) larvae cultivation and juvenile mollusk are formed: larvae cultivation carries out indoor, mainly comprise the selection to cultivating density, bait, water temperature, salinity, juvenile mollusk is formed at sea and is carried out, and the initial density of larvae cultivation is 2/ml, and entirely change water once every day, and inflation is cultivated, bait is Isochrysis galbana and sub-heart-shaped flat algae, day bait throwing in 3 times, and daily ration, feeding quantity first 3 days is 10,000 alga cells/ml, next within 7 days, is increased to 50,000 alga cells/ml, is increased to afterwards 100,000 alga cells/ml, larva culture density was carried out quantitatively in every two days, and cultivate water body so that the larval density in each cultivation bucket is basically identical by adjusting, added with antibiotic not during larvae cultivation, in the time there is eyespot in 20%～30% larva, the oyster shell that input is handled well, scallop shell collector is collected seedling, adhering to 8-10 juvenile mollusks by each adherance throws in, when after the whole settlement and metamorphosis of the floating larvae in water body, continue indoor cultivation about a week, then oyster shell or scallop shell with juvenile mollusk are strung and carry out mark with rope, go out pond in fine afternoon, being suspended to sea carries out juvenile mollusk and forms, afterwards, make regular check on growth and the survival condition of juvenile mollusk, after 6 months, can grow into 5cm-6cm, in order to reduce environmental effect, hybrid group and purebred group, from larva, juvenile mollusk, cultivate under identical environmental condition during becoming shellfish always,
(4) mensuration of DHA content and comparison: be respectively taken to from hybrid group and purebred group that to lack 15 adult oysters be material, vivisection obtains fresh soft tissue, organizing first of same stirs evenly, vacuum cooling drying is pulverized again, first use chloroform-methanol mixed liquor extracting fat, then with NaOH methanol solution, fat is carried out to esterification, finally analyze fatty acid component with gas chromatograph, determine the DHA content in hybrid group and purebred group of sample and compare.
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| CN103798166B (en) * | 2014-01-24 | 2015-07-29 | 中国科学院南海海洋研究所 | The indoor extensive artificial breeding method of a kind of the South China coastal Hong Kong oyster |
| CN103814810B (en) * | 2014-02-28 | 2015-05-20 | 中国科学院南海海洋研究所 | Method of obtaining oyster distant intergeneric hybrid |
| CN104115773B (en) * | 2014-07-29 | 2016-05-18 | 大连海洋大学 | The method of DHA content in indoor raising Ruditapes philippinarum |
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