39 results about "Haliotis discus" patented technology

The invention discloses a non-invasive extraction method of Haliotis discus hannai ino genome DNA, comprising the steps of using 75% of alcohol to sterilize and irritate pallium of Haliotis discus hannai ino; scraping to obtain a mixture of mucus and epidermal tissue fragments; treating the mixture, centrifugally separating the mixture to obtain precipitate; adding a cell lysate and protease K to the precipitate; after digesting and oscillating, cooling the mixture for a certain time; adding NH4Ac to the sample; centrifugally separating to collect supernate; adding isopropanol to the supernate; after oscillating and mixing uniformly, and then centrifugally separating to obtain precipitate; using ethanol to wash the precipitate; drying the precipitate at room temperature for 5 minutes to obtain the Haliotis discus hannai ino genome DNA. The non-invasive extraction method of the Haliotis discus hannai ino genome DNA is simple, efficient and convenient, does not damage the Haliotis discus hannai ino and does not influence survival ability and production performance.

The invention discloses a use of Shewanella as a feed additive in the abalone culture. Shewanella (comprising WA64 and WA65 strains) has strong a capability of antagonism of abalone pathogenic bacteria and thus Shewanella can be used as a feed additive for abalone culture. The Shewanella as feed additive for abalone culture has the advantages that Shewanella promotes that juvenile abalones vie with pathogenic bacteria for adhesion sites, maintains a normal microecological balance of the intestinal canal, can colonize in the intestinal canal of an abalone, promotes nutrient absorption, improves immunity thereby improving a growth speed and disease resistance of a Haliotis discus hannai, can replace a chemical drug for disease treatment, and does not damage the ecological environment.

The invention discloses a method for inducing triploid of Haliotis discus Hannai through medicament, which comprises the following steps, preparing 6-dimethoxy aminopurine solution, obtaining abalone sperms and ova through the conventional method, obtaining oosperm through artificial insemination, placing oosperm of seed abalone into sea water, finally inhibiting the second polar body of the Haliotis discus Hannai with 6-dimethoxy aminopurine solution.

The breeding method of tangerine shell colour Haliotis discus hannai Ino includes the following steps: using the mutant of it, planting it in sea-water, adding natural food, applying natural air to make its sex gland be developed and matured, adopting the processes of drying in the shade, heating and irradiating sea-water by UV-ray to make stimutation to hasten parturition to obtain female gameteand male gamete, using single cross or group cross mode to make oritificial insemination, and utilizing conventional hatching and post-cultivation method so as to obtain the invented new variety.

The invention discloses a parent haliotis discus hannai mature-promoting method capable of realizing early breeding, which can improve the fatness, egg laying amount, fertility rate, egg rate and fry survival rate of parent haliotis discus hannai, and comprises the following steps: selecting grown haliotis discus hannai, which is characterized by shells of at least 9 centimeters, gland maturity of 30 to 40 percent, G/L3 of more than 0.145, high activity and sorptive ability and no diseases as parent haliotis discus hannai, wherein the ratio of the number of male haliotis discus hannai to the number of female haliotis discus hannai is 80-100:500-1,000; and culturing the parent haliotis discus hannai in an early period, a middle period and a later period, wherein in the early period, the culture time is 40 to 50 days, the water temperature is 15 to 18 DEG C, and the parent haliotis discus hannai are fed with saline kelp; in the middle period, the culture time is 40 to 50 days, the watertemperature is 17 to 22 DEG C and the partent haliotis discus hannai are fed with fresh kelp; and in the later period, the culture time is 20 days, the water temperature is 9 to 22 DEG C, and the parent haliotis discus hannai are fed with dried nori.

The invention discloses a haliotis discus hannai microsatellite molecular marker which can analyze the genetic diversity of haliotis discus hannai wild population and identify a genetic relationship between cultured stocks and a preparation method thereof. According to the steps of establishing a haliotis discus hannai microsatellite (AC)n library, screening microsatellite sequence positive clone, designing a microsatellite primer, performing polymerase chain reaction (PCR) amplification and screening, ten highly polymorphic microsatellite molecular markers of Hd1, Hd2, Hd3, Hd4, Hd5, Hd6, Hd7, Hd8, Hd9 and Hd10 are obtained; and the polymorphic detection proves that the microsatellite molecular marker has the characteristic of high repeatability, is an effective and reliable molecular marker, can establish a haliotis discus hannai microsatellite DNA marker technology system and is used for evaluating the genetic diversity of haliotis discus hannai, analyzing the genetic structure and identifying the genetic relationship between the cultured stocks.

The invention relates to a haliotis discus hannnai and haliotis iris interspecific hybrid production method with a higher fertility rate and a higher hatching rate. According to the interspecific hybrid production method, Chinese haliotis discus hannnai is taken as female parent abalone, and Zelanian haliotis iris is taken as father parent abalone; the female parent abalone and the father parent abalone are put into different parent abalone cultivation ponds for cultivation; the female haliotis discus hannnai with the gonad fully developed is subjected to spawning induction with a method combining shady drying and ultraviolet irradiation seawater stimulation until the female haliotis discus hannnai lays eggs, and the male Zelanian haliotis iris with the gonad fully developed is subjected to spawning induction with a method combining shady drying and hydrogen peroxide seawater soaking until the male Zelanian haliotis iris releases sperms; fertilized eggs of haliotis discus hannnai and haliotis iris hybrids are obtained in the following steps: the eggs of the haliotis discus hannnai and the sperms of the haliotis iris are taken respectively and mixed, the temperature of seawater where fertilization is performed is adjusted, and the eggs are washed after the fertilized eggs are split to two cell stages; the haliotis discus hannnai and haliotis iris hybrid fries can be obtained with conventional hatching and later cultivation methods.

The invention aims to provide an SNP (single nucleotide polymorphism) marker for distinguishing Chinese population and Japan population of haliotis discus hannai. The SNP marker is SNP loci for distinguishing Chinese population and Japan population of haliotis discus hannai. The invention further provides a primer group and a probe for detecting the SNP loci. The SNP marker is used for identifying the Chinese population and the Japanese population of haliotis discus hannai, so that breeding parent identification at a group level is simpler, more convenient and more feasible.

The invention belongs to the technical field of marine organisms and particularly relates to a method for inducing and collecting high-quality sperms of haliotis discus hannai. The method comprises the steps of selection of high-quality male parent abalones, drying stimulation of the parent abalones in shade, preparation of ozone seawater, sperm collection, sperm motility detection and determination of the fertilization rate and the hatchability. According to the method, 3-4-year plump male parent abalones with gonads mature are selected, drying stimulation in shade is performed, the parent abalones are placed in seawater under the ultraviolet irradiation and arranged in the quiet and dark environment so as to release the sperms, and the high-concentration sperms are collected by centrifugation. The collected abalone sperms are subjected to artificial insemination, and the fertilization rate is over 95%. Practice proves that the abalone sperms collected in the abalone collection mode can be applied to production time and research. The method for inducing and collecting the high-quality sperms of the haliotis discus hannai is of great significance for artificial breeding, germplasm conservation and genetic diversity protection and the like of the haliotis discus hannai.

The invention relates to a rapid primary culture method of haliotis discus hannai gill cells. The rapid primary culture method comprises the following steps of performing disinfection treatment on experimental abalones; taking out the gills of the disinfected experimental abalones by using a sterilized dissecting tool, and putting the experimental abalones into sterile seawater for cleaning; taking the cleaned gills, cutting the gills into small sections, and adding the gills in a culture dish containing 0.1% of type II collagenase for digestion for 30 minutes; adding the digested gills into aserum-containing culture medium to terminate digestion, performing grinding, performing sieving with a 100 [mu]m cell sieve, and collecting the sieved liquid; centrifuging the sieved liquid, discarding supernatant, performing resuspending with a complete medium, and collecting the supernatant; and inoculating the resuspended and collected supernatant into a T25 culture flask, and performing culturing in an incubator at 22 DEG C;. The method can meet the requirements of obtaining gill cells of the abalones in a short time to perform in vitro experiments, solves the problem of bacterial pollution in gill tissues of the abalones, and solves the problem of gill cell viability reduction caused by enzymolysis of the gill tissues by using a trypsin enzymolysis method. The invention further discloses an application of the method.

The present invention relates to the field of aquaculture, and particularly relates to a shallow sea ecological culture method for kelp, gracilaria, haliotis discus hanai, strongylocentrotus intermedius and apostichopus japonicus. The method comprises the specific steps of selecting a proper culture sea region; dividing the sea region into culture regions each with the area of 13 ha, wherein the two adjacent culture regions are an alga culture region and an abalone culture region respectively; and putting regular tetrahedron-shaped artificial fishing reefs at the bottoms of the alga culture region and the abalone culture region. The culture method effectively improves the system internal material and energy utilization efficiency, the annual production value per unit area is improved by 50% or above, the influence of culture production on the environment is effectively reduced, and ecological, efficient and low-carbon culture is realized.

The invention provides a strain for promoting attachment and metamorphosis of shellfish larvae and application. The strain is classified and named as Pseudoalteromonas sp., the strain is preserved in China General Microbiological Culture Collection Center (CGMCC) on June 8, 2020, the preservation address is Beijing, China, and the preservation number is CGMCC NO.20040. The invention further relates to application of the strain in the attachment, metamorphosis and development process of shellfish seedlings. The strain is adopted for induction, so that the attachment and metamorphosis rate of haliotis discus hannai seedlings can be increased. There is no toxic or side effect, and no influence is caused on haliotis discus hannai. The strain can be matched with other active substances for use, so that the attachment rate of the haliotis discus hannai seedlings is increased. The strain is high in operability, convenient, simple and high in practical applicability.

The invention discloses a molecular marker for identifying genetic sex of haliotis discus hannai and an application of the molecular marker. The molecular marker is shown as insertion/deletion lengthpolymorphism of a nucleotide sequence shown as SEQ ID NO: 1; an individual with the nucleotide sequence as shown in SEQ ID NO: 1 is expressed as genetic male haliotis discus hannai. During detection,SEQ ID NO: 2 and SEQ ID NO: 3 are adopted as primers for PCR amplification, and when the amplified fragments are 252bp and 240bp, the haliotis discus hannai to-be-detected lacks the gene of the nucleotide sequence shown in SEQ ID NO: 1 and is genetic male haliotis discus hannai; when only 252bp amplified fragments exist, the haliotis discus hannai to-be-detected has a nucleotide sequence shown asSEQ ID NO: 1 and is genetic female haliotis discus hannai. The molecular marker disclosed by the invention can be used for simply, conveniently, quickly and stably identifying the genetic sex of the haliotis discus hannai, is beneficial to developing a unisexual breeding technology of the haliotis discus hannai and developing haliotis discus hannai breeding, further improves the breeding benefit and increases the economic benefit of haliotis discus hannai breeding.

The invention discloses an abalone hybridization seedling culture method for increasing utilization rate of bred abalone. The method is characterized by including the steps: preparing male abalone offirst filial generation of green abalone, wrinkle haliotis discus discus female abalone and wrinkle haliotis discus discus male abalone according to the ratio of 5:50:2 to serve as the bred abalone; maturing and spawning induction to obtain sperms and eggs; incubating fertilized eggs; selecting optimization products. The seedling culture method is simple to operate, can effectively utilize spermsand eggs of the abalone, the utilization rate of the bred abalone is increased, and artificial breeding benefits are improved.

The invention relates to a haliotis discus hannai superoxide dismutase, and a preparation method and application thereof. The invention discloses a haliotis discus hannai superoxide dismutase gene which contains a base sequence described in SEQ ID No.1 or a gene with a code shown by protein (a) or (b) as follows: (a) protein consisting of an amino acid sequence shown in SEQ ID No.1; and (b) protein derived by (a), which is formed by replacing, losing or adding one or a plurality of amino acids in the amino acid sequence restricted by (a) and has superoxide dismutase activity. Meanwhile, the invention discloses protein encoded by the gene and a recombinant vector as well as the preparation method of the superoxide dismutase. By the preparation method, the superoxide dismutase is obtained with the help of genetic engineering technology, and the protein has very strong activities in removal of superoxide anion free radicals and hydroxyl radicals, safety, damage repair and the like, and can give play to application value in the fields such as food additives, drug preparation, cosmetics and the like.

The invention provides a method for detecting and identifying haliotis discus hannai ino through fluorogenic quantitative PCR, and belongs to the technical field of species resource identification. The method comprises the step of identifying molecular characteristics of the haliotis discus hannai ino by applying a specific primer and a probe. Through the adoption of the method, the haliotis discus hannai ino can be quickly and accurately detected and identified, the method has the advantages of high specificity, high sensitivity, short consumed time, and accurate and convenient determinationof results, PCR aftertreatment is not needed, and false positive and cross contamination can be effectively avoided. A new technological means is provided for quickly and accurately identifying the haliotis discus hannai ino in entry and exit species resource inspection, fishery production, consumption and trade.

The invention relates to an artificial breeding method for abalone, and the method is capable of resisting viruses and diseases and improving the specifications of abalone fries. The method comprises the steps of maturity promotion of parent abalone, spawning induction, incubation, fry collection, breeding with diatom, stripping, advection breeding, application of fish-bait techniques and obtainment of commercial abalone fries. The method has the following characteristics: hybridization is carried out with New Zealand Black Gold as a female parent and Chinese Haliotis discus Hannai as a male parent in the stage of maturity promotion of parent abalone; abalone fry collection time is advanced, abalone fry collection is lowered down, the area of abalone fry collection is halved, the time for breeding with diatom is prolonged and the scale of stripping is enlarged. The artificial breeding method for abalone provided by the invention greatly improves the survival rate and the scale of the artificial breeding of abalone and enhances economic benefits.

The invention discloses a method for centralized spermatogenesis of male haliotis fulgens of a first filial generation of haliotis fulgens. The method comprises the following steps: (1) placing a spermatogenesis box into a water groove, wherein the spermatogenesis box comprises a plurality of independent small chambers, and a through hole is arranged on the wall of each small chamber, and placingone shade-dry stimulated male haliotis fulgens of the first filial generation of the haliotis fulgens into each small chamber; (2) putting disinfected seawater into the water groove until the spermatogenesis box is submerged, and continuously inflating air into the water groove; (3) putting an ovum-free egg liquid of female haliotis discus hannai into the water groove, stirring a water body by inflating, and allowing the ovum-free egg liquid of the female haliotis discus hannai to stimulate the male haliotis fulgens to produce sperms; and (4) after any one male haliotis fulgens of the first filial generation of the haliotis fulgens produces sperms, and allowing the sperms to flow to the small chambers through the through holes arranged on the walls of the small cambers to stimulate the male haliotis fulgens to produce sperms quickly. The method provided by the invention solves the problem of slow and decentralized spermatogenesis of the male haliotis fulgens of the first filial generation of the haliotis fulgens in the conventional hybridization process, improves the spermatogenesis rate of the male haliotis fulgens of the first filial generation of the haliotis fulgens, and guarantees large-scale production effect of hybrid haliotis fulgens juveniles.

The invention relates to a polyculture method of gracilaria asiatica and hybrid abalone. The density of the hybrid abalone is 400 per pond, the density of the gracilaria asiatica is 38.13g/ m<3>, and the specification of a culture pond is 180cm x 150cm x 130cm. The hybrid abalone is a hybrid offspring of haliotis discus hannai and haliotis discus discus, the hybrid abalone is cultured by using a net cage, the specification of the net cage is 43.5cm x 33.5cm x 14 cm, 40 hybrid abalones are cultured in each cage, every five culture net cages are bundled together to form a string, and 200 hybrid abalones are contained in each string. After the culture net cages are placed into an alga cultivation pond, and two strings of abalone cages are arranged in each pond. After the hybrid abalones are cultured for one week, the algae are introduced for polyculture and suspended in the center of the pond surface by using a net cover, a certain distance is kept between the algae and the abalone cage, and the hybrid abalones are prevented from eating the cultured algae. According to the invention, the polyculture method of the common large-scale algae gracilaria asiatica in China and the hybrid abalone of the haliotis discus hannai and the haliotis discus discus is studied, the quality of water which is purified by the algae in the hybrid abalone culture pond and the influence of the growth of the hybrid abalone are researched, and a necessary scientific basis is provided for further selecting the large-scale algae to establish a novel effective abalone healthy breeding mode.

The invention relates to a polyculture method of ulva lactuca and hybrid abalone. The density of the hybrid abalone is 400 per pond, the density of the ulva lactuca is 40.21g/m<3>, and the specification of a culture pond is 180cm x 150cm x 130cm. The hybrid abalone is a hybrid offspring of haliotis discus hannai and haliotis discus discus, the hybrid abalone is cultured by using a net cage, the specification of the net cage is 43.5cm x 33.5cm x 14 cm, 40 hybrid abalones are cultured in each cage, every five culture net cages are bundled together to form a string, and 200 hybrid abalones are contained in each string. After the culture net cages are placed into an alga cultivation pond, and two strings of abalone net cages are arranged in each pond. After the hybrid abalones are cultured for one week, the algae are introduced for polyculture and suspended in the center of the pond surface by using a net cover, a certain distance is kept between the algae and the abalone cage, and the hybrid abalones are prevented from eating the algae. According to the invention, the polyculture method of the common large-scale algae ulva lactuca in China and the hybrid abalone of the haliotis discus hannai and the haliotis discus discus is studied, the quality of water which is purified by the algae in the hybrid abalone culture pond and the influence on the growth of the hybrid abalone are researched, and a necessary scientific basis is provided for further selecting the large-scale algae to establish a novel effective abalone healthy breeding mode.

The invention relates to application of the polypeptide HDP-1 of Haliotis discus hannai to preparation of drugs, health products or dietary supplements used for preventing and treating tumors and diseases related to oxidation. HDP-1 has strong free radical removing capability and can inhibit free radical-mediated DNA oxidation damage. HDP-1 can also inhibit the migration of HT1080 cells and the expression of MMP-2 and MMP-9, and the inhibitory intensity of HDP-1 is dose-dependent; that is, HDP-1 has antioxidant and antitumor activities.

The invention relates to a gynogenesis inducing method of haliotis discus hannai. According to the method, an electrostimulation method is adopted; ova are transferred into an electrostimulation buffer solution for balance for 2min before electrostimulation; parameters of an electric pulse instrument is regulated; 1ml of ova are sucked with a sucker and put into an electrostimulation cup, and then electrostimulation treatment can be carried out; the electrostimulation buffer solution is a Zimmerman buffer solution which specifically comprises 0.1 mmol/L of Ca(C2H3O2)2.H2O, 1 mmol/L of K2HPO4, 0.5 mmol/L of Mg(C2H3O2)2.4H2O, 0.1 mmol/L of reduced glutathione, 1% of BSA, and 0.30 mmol/L of sucrose; the parameters of the electric pulse instrument include the field intensity of 0.88 kv/cm, the capacitance of 10[mu]F, and 3 times of electric pulses. The density of ova is 7.50x10<4> /ml during electrostimulation. The gynogenesis inducing method is simple and effective. After electrostimulation induction, the cleavage rate of the haliotis discus hannai can reach 28% or above. The embryo survival rate can reach 98% or above.

The invention relates to a haliotis discus hannaiinothioredoxinperoxidaseas well as a preparation method and an application thereof. The invention discloses a haliotis discus hannaiinothioredoxin peroxidase gene containing a base sequence shown by SEQ ID NO.1 or containing a gene of a protein (a) or (b) coded as follows: (a) a protein composed of amino acid sequences shown by SEQ ID NO.2; and (b) a protein derived from (a) by substituting, deleting, or adding one or more amino acids in the amino acid sequences limited by (a) and having thioredoxin peroxidase activity. The invention also discloses a protein and a recombinant vector coded by the gene, and a preparation method of the thioredoxin peroxidase. By virtue of a gene engineering technology, the thioredoxin peroxidase is obtained, has very strong activities of eliminating hydroxy radicals, achieving safety and repairing damages, and will give play to application values in the fields of food additives, drug preparation, cosmetics and the like.