35 results about "Acipenser dabryanus" patented technology

The invention belongs to the field of microsatellite molecular markers and particularly relates to an acipenser dabryanus microsatellite marker as well as a screening method and application of the microsatellite molecular marker, wherein the acipenser dabryanus microsatellite marker has the nucleotide sequence of any one or more of SEQ ID No.1-8. In the invention, the development of microsatellite molecular markers using the high-throughput sequencing technology is faster and costs less than a traditional method; and moreover, the functional traits of corresponding EST sequence can be found while the microsatellite sites are obtained, and the gene relevance is relatively strong. In the invention, through the developed microsatellite sites, basic data is provided for the genetic diversity of acipenser dabryanus, and the study fields such as genetic diversity analysis, genetic map establishment, gene location, variety identification, germplasm conservation, quantitative character gene analysis and parent-child relationship identification can be carried out. According to actual needs, any one or more markers disclosed by the invention can be adopted, or any one or more pairs of specific primers corresponding to related markers can be analyzed and studied.

The invention relates to microsatellite marker primers for identification of an Acipenser dabryanus family. Through a transcriptome sequencing technology, Unigenes of the screened Acipenser dabryanus are screened, microsatellite loci in the Unigenes sequence are detected, according to flanking sequences at two ends of each one of the microsatellite molecular marker loci, specific primers of the molecular marker are labeled, the specific primers are subjected to PCR amplification, the stability and polymorphism of the amplification result are detected, and 18 effective microsatellite markers are acquired. Through the microsatellite marker primers, according to the polymorphism of the microsatellite marker primers, a parentage identification platform of Acipenser dabryanus is built and the basis is provided for Acipenser dabryanus selection and breeding matching.

The invention discloses acipenser dabryanus sperm preserving fluid liquid and a preparing method and application. The acipenser dabryanus sperm preserving fluid liquid comprises saccharose, mycose, trihydroxy aminomethane, potassium chloride, reduced glutathione and carbinol. The acipenser dabryanus sperm preserving fluid liquid has simple components and is low in cryopreservation cost. The added reduced glutathione plays a good role in antioxygenation, well protects sperm plasma membranes and reduces damage to DNA molecules. The added mycose has the effects of preventing protein denaturation and effectively protecting sperms. By means of the acipenser dabryanus sperm preserving fluid liquid, motility of unfreezed sperms is higher than 70%, and the fertility rate reaches 45%.

The invention provides a wild-training method for improving the feeding ability of released Acipenser dabryanus. The wild-training method comprises the following steps that fish bait is produced; feeding domestication is conducted, wherein feeding domestication starts when are hatched, feeding domestication is conducted on the Acipenser dabranus which are fed with artificial feed for 2-4 months and feeding domestication is conducted on the Acipenser dabranus which are fed with the artificial feed for over 5 months; Through the wild-training method for improving the feeding ability of the released Acipenser dabrayanus, after training is over, both the feeding capability and the predation capability of the Acipenser dabryanus with different specificications are obviously enhanced and improved. After analog wild-release is conducted, the trained Acipenser dabryanus is released to the wild or an experimental pond, the capability of preying on wild life live baits is obviously improved, and the growth rate and the survival rate are obviously higher than the released Acipenser dabryanus which is not trained.

The invention discloses a real-time quantitative PCR detection primer set and a real-time quantitative PCR detection method of a differentially expressed acipenser dabryanus gonad gene. The real-timequantitative PCR detection primer set comprises an SOX9 primer pair, an FST primer pair, a DF primer pair or a beta-actin primer pair. The real-time quantitative PCR detection method comprises the following steps: extracting RNA from an acipenser dabryanus sample to be detected, using the total RNA as a template, and synthesizing cDNA of the acipenser dabryanus sample to be detected; using the cDNA of the acipenser dabryanus sample to be detected as a template, adding the real-time quantitative PCR detection primer set of the differentially expressed acipenser dabryanus gonad gene, performingreal-time fluorescent quantitative PCR detection, and detecting an amplification product according to an agarose gel electrophoresis result and an amplification product dissolution curve. The primersand the quantitative PCR detection methods of a real-time quantitative PCR target gene and an internal reference gene are suitable for acipenser dabryanus as well as other sturgeons with a similar kinship, and a basis is provided for identifying genders of the sturgeons and researching a gender determining gene and a gender determining mechanism in the future.

The invention discloses an acipenser dabryanus gonad differential expression gene expression gene GSDF and a screening method thereof, and belongs to the technical field of biology. A high-flux sequencing technique is used for performing male and female transcriptome comparative sequencing on acipenser dabryanus; transcriptome data is subjected to assembling, annotation and differential gene expression analysis. An FPKM method is used for analyzing the gene expression level in male and female gonad. EBSeq is used for performing gene differential expression analysis, so that a differential expression gene set between the male and female samples is obtained. In the screening process, FDR is selected as a key index of the differential expression gene screening; the conditions that the FDR issmaller than 0.05 and the differential multiple is greater than or equal to 2 are used as screening standards. Then, the differential gene is subjected to function annotation; the sex determination-related gene is screened; the basis is provided for the acipenser dabryanus sex determination and the sex determination gene and sex determination mechanism study in future.

The invention relates to the technical field of aquaculture, in particular to an artificial breeding method for acipenser dabryanus. Oxytocin is injected into female acipenser dabryanus two times, wherein the interval between two times of injection is 12 hours, and the injection amount of the first time accounts for 1/3 to 1/2 of the total dose. The one-time injection method is adopted for male acipenser dabryanus, wherein injection of the male acipenser dabryanus is synchronous with the first time of injection of the female acipenser dabryanus, and the dose of the male acipenser dabryanus is half that of the female acipenser dabryanus. Fertilization is conducted according to a half-dry method, water, eggs and seminal fluid are mixed according to the proportion of 75:50:1, and the mixture is stirred for 2 minutes to 3 minutes. A complete acipenser dabryanus artificial breeding technique process is provided, and technical support is provided for large-scale artificial breeding; due to the adoption of the method, the hatchability of acipenser dabryanus can reach 85% or higher at most, and restrict factors in the acipenser dabryanus artificial breeding process are eliminated.

The invention discloses a torrent fish domesticating method. After first filial generation of acipenser dabryanus of 1-5 ages is raised in a culture pond, and then the torrent fishes to domesticated are added to the pond for raising. According to the domesticating method, the acipenser dabryanus and the torrent fishes are raised together, so that the stress response of wild fishes can be reduced, and the ichthyophthirius multifiliis infection resistance and the large individual characteristics of the acipenser dabryanus are utilized, and torrent is created by fishtailing to drive the activity of the wild fishes; the large cluster of the wild fishes is reduced and the probability of reciprocal infection of the ichthyophthirius multifiliis is reduced; as a result, the ichthyophthirius multifiliis can be effectively prevented, the survival rate is increased, the drug use is reduced and the pollution to the water environment is reduced.

The invention provides acipenser dabryanus testis cell cryopreservation liquid as well as a testis cell cryopreservation method and application. The cryopreservation liquid comprises bovine serum albumin, trehalose, ethylene glycol, lecithin and fetal calf serum. Meanwhile, the invention further provides an acipenser dabryanus testis cell cryopreservation method. By using the method, damages of ice crystals on cells can be reduced to the greatest extent by using a relatively low cooling manner under the condition that liquid-nitrogen cooling is not used, and the cells have high survival rate and reproduction rate after recovery, so that warm cryopreservation is possible to realize, and long-period stable cell supply is provided for transplanting acipenser dabryanus spermatogonia.

The invention belongs to fish feed and particularly discloses feed of adult acipenser dabryanus. The feed is prepared from fish meal, soybean protein concentrate, soybean meal, flour, soybean oil, soyabean lecithin, fish oil, calcium biphosphate, a vitamin complex, complex mineral salts, choline, betaine, a mould inhibiter, immune polysaccharides, bentonite and ethoxyquin. The feed provided by the invention can meet the requirement on nutrients of adult acipenser dabryanus, and can be used for enhancing the growth rate of the adult acipenser dabryanus and lowering the death rate of the adult acipenser dabryanus.

The invention belongs to the field of aquatic animal germplasm identification in the field of aquaculture and discloses a microsatellite primer applied to Chinese sturgeon and acipenser dabryanus species identification and application thereof. According to the microsatellite primer disclosed by the invention, a pair of fluorescent labeled microsatellite primers are successfully screened and synthesized, and the primers can be applied to establishing a Chinese sturgeon and acipenser dabryanus species identification technology; after the primers are applied to PCR amplification, allelic genes are measured to perform Chinese sturgeon and acipenser dabryanus species identification and analysis. The identification method disclosed by the invention has the characteristics of simpleness and convenience in operation, low cost, high efficiency, economical performance, convenience and practicability; the identification method can be popularized and applied to Chinese sturgeon and acipenser dabryanus species identification, germplasm identification and genetic management; furthermore, the identification method can evaluate an artificial fecundation drift effect.

The invention provides a method for constructing a spleen tissue cell line of acipenser dabryanus. The method mainly comprises the following steps: (1), an MEM (minimum essential medium) culture solution containing fetal calf serum, human epidermal growth factors, human fibroblast-like cell growth factors and acipenser dabryanus serum and having the pH of 7.2-7.4 is prepared and stored in a refrigerator at the temperature of 4 DEG C for standby application; (2), spleen tissue is separated from the acipenser dabryanus body and subjected to primary culture with a tissue block culture method; (3), when the cell growth forms a single layer and the bottom coverage rate is higher than 70%, 0.25% trypsin digestion is adopted for subculture; (4), vigorously growing spleen cells with uniform forms are subjected to liquid nitrogen cryopreservation and resuscitaion culture. The spleen tissue cell line constructed with the method is fast to reproduce, has multiple forms, is fibroblast-like, and can realize continuous passage and cryopreservation resuscitation. The construction method is simple in technology, easy to operate, can be popularized to culture of spleen cells of other sturgeons and can be expected to be used for acipenser dabryanus species resource conservation as well as separation and identification of sturgeon diseases especially virus pathogens.

The invention provides a method for hatching eggs of Sturgeon dabryi, comprising: using 150-250ppm concentration of formaldehyde solution to statically soak the fertilized eggs for 20-40 minutes during the hatching period of fertilized eggs of Dabry's sturgeon in running water to the stage of rhubarb suppository; Rinse the fertilized eggs with clean water, and then continue to incubate the fertilized eggs with running water until the fish come out of the membrane. The method provided by the invention can prevent saprolegniasis from occurring in a large area in a short period of time without affecting the normal development of fish eggs, and the method is simple and feasible. Experimental results show that the method provided by the invention can greatly improve the survival rate of the dabry's sturgeon.

The invention relates to a sturgeon egg preservation balance solution, wherein the balance solution comprises: 1.0 g/L of glucose, 0.95 g/L of HEPES buffer salt, 0.8-1.2 g/L of bovine serum albumin BSA, 0.41g/L of NaH2PO4, 0.38g/L of KCl, 0.23g/L of MgSO4, 7.25g/L of NaCl, 0.147g/L of CaCl2, 200,000 IU/L of penicillin, and 0. 2g/L of streptomycin sulfate, wherein the pH value is adjusted to be thesame as the body cavity fluid. The balance solution of the invention can realize the short-term preservation of the egg of Chinese sturgeon, the egg of acipenser schrenckii, and the egg of acipenserdabryanus, the storage temperature is 16 DEG C, after 6 hours of storage, the fertilization rate of Chinese sturgeon egg is 77.35%, the hatching rate is 80. 06%, the abnormal rate is 0, the fertilization rate of acipenser schrenckii egg is 89.97%, the hatching rate is 91.24%, the abnormal rate is 0, the fertilization rate of acipenser dabryanus egg is 90.03%, the hatching rate is 89.45%, and the abnormal rate is 0. 47%, so that the solution can be used to solve the technical problems caused by the unsynchronized spawning induction effect time of artificial breeding of male and female sturgeon.

The invention relates to the technical field of artificial breeding, in particular to a secondary debonding method for acipenser dabryanus. The method includes the steps of placing fertilized eggs in a container after the fertilized eggs of acipenser dabryanus are hatched to the gap-shaped blastopore period-wide neural plate period and are hatched for 46 hours to 54 hours at the temperature of 18 DEG C, adding talcum powder with the mass concentration of 8% to 12% or milk powder with the mass concentration of 13% to 17%, stirring the mixture in the same direction for 15 minutes to 30 minutes, and placing the fertilized eggs in a sturgeon hatching device directly or after washing the fertilized eggs to continue to be hatched. By means of the method, stability of water flow in the hatching process is ensured, man-made stimulation of the critical period of fertilized egg hatching is reduced, the probability of fertilized egg blocking is greatly lowered, the probability of blocky water molds is lowered, the loss ratio of live eggs is decreased, and the hatchability is improved.

The invention discloses a GFP-Addnd 3'UTR recombinant expression vector. The vector is an expression vector pCS<2+>, 3'UTR segment of an Acipenser dabryanus dnd gene and a green fluorescent protein coding region gene are connected to the expression vector pCS<2+>. The GFP-Addnd 3'UTR recombinant expression vector can be used for marking an Acipenser dabryanus original germ cell. The specific marking method comprises the steps of synthesizing mRNA of the GFP-Addnd 3'UTR recombinant expression vector in vitro, injecting the mRNA into an embryo in the embryo development 1-cell period of Acipenser dabryanus in a micro-injection mode, then conducting incubated cultivation, and observing expression conditions of the green fluorescent protein in different development periods in real time. Because the Addnd is a maternal gene, and is only expressed in the ovary and the spermary, after the marking method is adopted, expression of the green fluorescent protein can be detected in the development process of the zygote, and thus tracking migration development of the original germ cell and further deep scientific study are promoted.

The invention discloses oligopeptide and an application in the fishery field. At present, diluents and anti-freezing agents are generally used during cryopreservation of acipenser dabryanus sperm, thediluents adopt 0.7% sodium chloride, 0.02% potassium chloride and 3.5% glucose, the anti-freezing agents adopt dimethyl sulfoxide and the like, but these anti-freezing agents have different toxic effects on acipenser dabryanus sperm, and the sperm life is shortened after preservation. Study results prove that oligopeptide A, B or C is significantly superior to the prior art as the anti-freezing agent and can significantly prolong the life of sperm recovered from cryopreservation. The oligopeptide A, B or C can be used as the anti-freezing agent to improve the anti-freezing capacity of the acipenser dabryanus sperm.

The invention discloses oligopeptide and an application thereof. At present, diluents and cryoprotectants are usually used for sperm cryopreservation of acipenser dabryanus, 0.7% sodium chloride, 0.02% potassium chloride and 3.5% glucose are used as the diluent, and dimethyl sulfoxide and the like are used as the cryoprotectants. However, the cryoprotectants have different toxic effects on spermsof acipenser dabryanus, and the sperm life is shortened after cryopreservation. Research results show that oligopeptide A, B or C as the cryoprotectant is significantly better than the cryoprotectantsin the prior art, and can significantly prolong the service life of the sperms after cryopreservation and recovery. The oligopeptide A, B or C can be used as the cryoprotectant to improve the antifreeze capacity of the sperms of acipenser dabryanus.

The invention discloses an accurate and highly efficient transcriptome-based eukaryotic gene identification method. The method includes the following steps that three internal reference genes of acipenser dabryanus are obtained through PCR amplification reaction of a high-fidelity enzyme, real-time fluorescence quantitative primers of the three internal reference genes of the acipenser dabryanus are developed, the quality of the primers of the three internal reference genes of the acipenser dabryanus are evaluated based on a standard curve and a melting curve, and the stability of the three internal reference genes of the acipenser dabryanus in tissue expression is evaluated. According to the accurate and highly efficient transcriptome-based eukaryotic gene identification method, by providing full-length coding regions of the three internal reference genes (ACT, GAPDH and EF1-alpha genes), more internal reference gene choices are provided for fluorescence quantitative research of the acipenser dabryanus; by developing the fluorescence quantitative primers based on nucleotide sequences of ACT, GAPDH and EF1-alpha, the reliability and reproducibility of fluorescence quantitative research are greatly improved; the stability of ACT, GAPDH and EF1-alpha in tissue expression is evaluated, and stable and suitable internal reference genes are provided for the fluorescence quantitativeresearch of the acipenser dabryanus.