21 results about "Crassostrea" patented technology

The invention discloses a tetraploid crassostrea gigas parent-child relationship identification method based on SNP sites, and belongs to the technical field of tetraploid crassostrea gigas molecular assisted breeding. The method for identifying the parent-child relationship of tetraploid crassostrea gigas comprises the following steps: (1) sequencing adductor muscles of tetraploid crassostrea gigas to be detected; (2) comparing the reference genome of the crassostrea gigas to obtain the genotype of the specified SNP site of the tetraploid crassostrea gigas; (3) calculating IBS and LOD of the to-be-detected parents and the to-be-detected offspring according to an SNP typing result; and (4) identifying the parent-child relationship between the to-be-detected parent and the filial generation by combining IBS and LOD. The SNP loci have the advantages that 1000 SNP loci for identifying the parent-child relationship of the tetraploid crassostrea gigas are disclosed for the first time, data support is provided for identifying the parent-child relationship of the tetraploid crassostrea gigas, a foundation is laid for developing a liquid phase chip for identifying the parent-child relationship of the tetraploid crassostrea gigas, and the SNP loci have good application prospects.

The application belongs to the field of genetic engineering and genetic breeding, and particularly relates to a high-temperature response gene HSP70 expression regulation SNP marker of Crassostrea gigas and application of the SNP marker in identifying high-temperature resistant oyster individuals. The SNP marker is the 251th base in the sequence shown in SEQ ID NO: 2, and the mutation type is C / G (trans-regulation site Marker13973). The application provides a SNP marker which is significantly related to the expression of the HSP70 gene, and the advantage of the application is that the genotype of parent oysters can be identified before seed breeding, and the tolerance of offspring to high-temperature environment is improved. The SNP marker obtained in the research has high reliability and stable results.

The invention relates to an InDel marker primer for identifying crassostrea gigas' Haida No.3 'and application of the InDel marker primer, and belongs to the field of molecular biology. The invention provides a method for identifying the specific InDel marker of the Haida No.3 group by using the primer, DNA of a crassostrea gigas sample is extracted by adopting an SDS method, the marker primer is used for carrying out PCR amplification on the sample to be detected, and the specific InDel marker site of the Haida No.3 group is detected; the determination standard is as follows: the amplified fragment length of the 'Haida No.3' crassostrea gigas is 236bp, and the amplified fragment length of the crassostrea gigas of other strains and wild populations is 274bp. Technical support is provided for paternity identification and germplasm resource protection of crassostrea gigas' Haida No.3 '.

The invention relates to the technical field of aquaculture molecular breeding, in particular to an SNP (Single Nucleotide Polymorphism) molecular marker remarkably related to a crassostrea gigas shell shape and application thereof. The SNP (Single Nucleotide Polymorphism) molecular marker obtained by screening disclosed by the invention is as follows: (1) Cg05009-24308790Cgt; t is located at the 24308790th site of the reference genome; (2), Cg05270 to 28618069Agt, Cg05270 to T is located at the 28618069th site of the reference genome; (3) Cg < 18426-31151905 > A < gt >; t is located at the 31151905th site of the reference genome. The SNP molecular markers obtained by screening provide a powerful molecular tool for developing a new oyster variety with a nearly circular shell, so that a targeted breeding plan is supported.

The invention discloses a standardized evaluation method for shell hardness of crassostrea gigas, and belongs to the field of aquatic shellfish mechanical phenotype determination and biomineralization material performance detection. The method comprises the following steps: carrying out cleaning pretreatment on double shells of crassostrea gigas; respectively determining the longest axis and the widest axis in the contours of the left shell and the right shell, constructing an orthogonal reference cross axis, axially arranging three standardized measurement sites of the left shell and the right shell at positions about 5mm away from the inner sides of the edges of the shells, and marking the standardized measurement sites; cutting the marked area to obtain a flat shell sample; applying a load of about 10 gf to each measurement site by using a Vickers microhardness tester, maintaining the pressure for about 10 seconds, and carrying out multiple indentation tests to obtain an original hardness value; carrying out abnormal value elimination and mean value calculation on the data to obtain representative hardness values of all sites; and further comprehensively calculating representative hardness values of six sites of the left shell and the right shell, and constructing an individual comprehensive shell hardness index. Compared with an existing single-point or random sampling measurement method, the method has the advantages that the problems of irregular shell shapes, asymmetry of left shells and right shells and the like of oysters are considered, and the accuracy and repeatability of oyster hardness phenotype measurement are remarkably improved through standardization of measurement sites and multi-site high-repetition measurement. The method can be widely applied to the fields of oyster stress resistance evaluation, marine acidification, environmental monitoring and the like, provides possibility for accurate phenotype determination in GWAS and QTL positioning in breeding, and has a huge application prospect.

The invention discloses application of crassostrea gigas miR-182-5p in negative regulation of melanin synthesis, and belongs to the technical field of aquatic genetic breeding and biology. The miR-182-5p in the crassostrea gigas is interfered by the miR-182-5p inhibitor, synthesis of melanin of the crassostrea gigas is remarkably enhanced after the miR-182-5p is interfered, and the color of the crassostrea gigas is black. In addition, the invention also synthesizes an exclusive miR-182-5p stimulant to treat the crassostrea gigas. Results show that compared with wild crassostrea gigas, the content of melanin in the crassostrea gigas injected with the simulant is lower, and the shell color is white. Therefore, the miR-182-5p is in negative correlation with the synthesis of the melanin of the crassostrea gigas, so that the miR-182-5p not only can be used for selecting or predicting the shell color character of the crassostrea gigas, but also can be used for assisting in screening or culturing varieties with high melanin content by silencing or knocking out the miR-182-5p in the crassostrea gigas. The important effect of miR-182-5p in oyster melanin synthesis is clarified for the first time, the molecular mechanism of mollusk melanin formation can be deeply understood, and the miR-182-5p has important significance in promoting practical application of crassostrea gigas quality breeding.

The invention provides a reef building shellfish bottom sowing and field planting method. The method comprises the following steps of bottom sowing sea area selection, fry selection and treatment, fry transportation and bottom sowing and field planting. The bottom sowing sea area is a leeward area with the annual average water temperature ranging from 20 DEG C to 30 DEG C, the salinity ranging from 29 per mill to 34 per mill, the dissolved oxygen content ranging from 4 mg / L to 7 mg / L and the water flow speed ranging from 0.1 m / s to 0.3 m / s, the area is required to be located in the area suitable for reef building shellfish to grow and distribute, and the sea area substrate is mainly corallite or reef. The reef-building shellfish is ostrea gigas, ostrea marginata or giant clam, parents are collected from a natural sea area to be domesticated, and after gonads of the parents are well developed, offspring seeds are obtained by adopting an artificial breeding method. According to the method, the native species obtained in situ are adopted, and artificial breeding acquisition is carried out; manual bottom sowing and field planting are carried out in the large tide, the tide water law is fully utilized, and shellfish survival is facilitated.

The invention belongs to the field of molecular biology and genetic breeding, and relates to screening of a group of SNP (Single Nucleotide Polymorphism) sites related to gonad development traits of crassostrea gigas and application of the SNP sites in evaluating gonad development level. Four SNP molecular marker loci significantly related to gonad development traits are disclosed by performing whole genome association analysis on a large-scale group of crassostrea gigas, the four SNP molecular marker loci are respectively located in Region1 (BP10, embryo protease 10) and Region2 (GALNT5, polypeptide N-acetylgalactosamine transferase 5) gene regions, and dominant genotypes and dominant genotype combinations of the SNP loci are determined. The SNP molecular marker combination is used for prediction and auxiliary breeding of the gonad development level of the crassostrea gigas, early-stage, lossless and high-throughput screening of individuals with the high gonad development level can be achieved, and the breeding efficiency of breeding traits of the crassostrea gigas is remarkably improved.

The application belongs to the technical field of aquaculture, and particularly relates to a method for improving the attachment rate and disease resistance of long oyster (Crassostrea gigas) nauplii and application thereof. The method is that early nauplii of the long oyster are soaked in filtered seawater containing a certain dose of glutathione. Compared with a control group without adding glutathione, the growth and development speed of the long oyster nauplii soaked in the glutathione is accelerated, the attachment rate is significantly improved, the lysozyme activity is enhanced, and the ability to resist vibrio is significantly increased. The application has a significant effect on reducing the mortality of the long oyster nauplii in the attachment metamorphosis process and improving the attachment metamorphosis rate, and can be used as an immune enhancer for the attachment metamorphosis stage of the long oyster and applied to seed production.

The application discloses a method for obtaining and preserving tetraploid male Crassostrea gigas, and belongs to the technical field of genetic breeding of aquatic breeding varieties. The method comprises the following steps: accelerating the maturity of tetraploid Crassostrea gigas by regulating water temperature and bait, identifying the male and female of the tetraploid Crassostrea gigas by nuclear magnetic resonance technology, further accelerating the maturity of the tetraploid male Crassostrea gigas by regulating water temperature and bait, and preserving the tetraploid male Crassostrea gigas by well storage seawater. The method has the advantages of simple operation, no influence of external air temperature and seawater temperature, no damage to the tetraploid Crassostrea gigas, and can provide the tetraploid male Crassostrea gigas stably throughout the year, thereby laying a solid foundation for the stable cultivation of triploid Crassostrea gigas throughout the year.

The invention belongs to the field of gene chips, and discloses a tetraploid crassostrea gigas 40K breeding liquid phase chip as well as a preparation method and application thereof. The breeding liquid phase chip disclosed by the invention comprises a probe for detecting 41270 SNP (Single Nucleotide Polymorphism) sites; the position information of the 41270 SNP loci is determined by performing sequence alignment on the basis of a reference version of a pacific oyster genome, and the reference version of the pacific oyster genome is GCA011032805.1. The gene chip can be applied to genotyping, genome selection or whole genome association analysis of tetraploid crassostrea gigas. The breeding liquid chip can effectively reduce the genotyping and breeding cost of tetraploid crassostrea gigas, improve the breeding accuracy and efficiency, and promote the development of tetraploid crassostrea gigas genome breeding.

The invention discloses an ascorbic acid-based oyster umami peptide browning targeted inhibition and nutritional ingredient synergistic preservation method, and relates to the technical field of aquatic product processing, and the method comprises the following steps: selecting fresh crassostrea gigas adductor muscle, washing with sterile water, carrying out standing pretreatment, and crushing and pulping by adopting a grinding technology to obtain oyster primary pulp; adding an ascorbic acid solution into the oyster primary pulp, stirring and mixing under a dark condition, and balancing for a preset time, so as to construct a browning substrate targeted binding system, and the like. According to the invention, through targeted combination of ascorbic acid and a browning substrate, phenolic substances, free amino acids and other components which are easy to cause browning are locked in advance; an ascorbic acid-composite antioxidant synergistic effect mechanism is formed by combining with a composite antioxidant composed of a green tea extract and citric acid, the problems that a traditional single anti-browning means is limited in effect and prone to being affected by the environment and losing efficacy are effectively solved, the browning risk of the whole processing flow of the oyster flavor peptide is reduced, and the natural color and luster of the product are reserved to the maximum extent.

The application discloses an active peptide agonist of an insulin-like peptide receptor of Crassostrea gigas and a method for identifying and applying the same, and belongs to the field of bioactive peptides. The amino acid sequence of the active peptide is selected from RWFYVNRLGRF, FELQQYCGFR and APWKSVLKRMMV. The method for identifying the active peptide of the Crassostrea gigas comprises the following steps: protein three-dimensional structure modeling, determination of a secreted peptide fragment, molecular dynamics simulation, construction of a binding free energy landscape, and screening of a candidate active peptide. The active peptide agonist significantly activates the insulin-like peptide receptor of the Crassostrea gigas, enhances the expression of a downstream channel mediated by the insulin-like peptide receptor, and promotes the metabolism and growth of the Crassostrea gigas.

The invention belongs to the technical field of molecular breeding of aquatic products, and relates to a group of SNP (Single Nucleotide Polymorphism) molecular markers remarkably associated with egg diameter and brood quantity characters of crassostrea gigas and application. According to the invention, through high-throughput whole genome re-sequencing and whole genome association analysis (GWAS), a group of SNP loci closely associated with the breeding traits of crassostrea gigas are mined. Wherein the marker associated with the egg diameter character is located at the G / T variation at the 44, 274 and 019 bp of the NC047563.1 chromosome and the A / G variation at the 44, 274 and 322 bp of the NC047563.1 chromosome, and the marker associated with the egg diameter character is located at the T / A variation at the 9, 556 and 289 bp of the NC047568.1 chromosome; the marker associated with the brood quantity character is located at 25, 410 and 510 bp of the NC047564.1 chromosome C / G variation, in addition, the dominant genotypes and combination of the sites are determined, early prediction of the breeding potential of individuals and parent screening can be achieved through genotype detection in the crassostrea gigas seedling stage or non-breeding season, and the breeding potential of the individuals can be predicted. The breeding efficiency of the new high-fecundity crassostrea gigas variety is obviously improved.

The invention discloses a method for accurately evaluating the germplasm purity of crassostrea gigas, and relates to the technical field of aquatic genetic breeding biology, the core of the method is to construct an index based on an NCBI (National Center of Biotechnology Information) crassostrea gigas genome sequence by virtue of a whole genome re-sequencing technology, collect 150 crassostrea gigas and 150 Fujian oysters in a natural population, extract mantle DNA (Deoxyribose Nucleic Acid) for sequencing, screen out 635 specific SNP (Single Nucleotide Polymorphism) sites, and obtain the purity of the crassostrea gigas. The method comprises the following steps: constructing a pure variety feature sequence, sequencing a to-be-detected sample, comparing two detection sequences of 635 sites with the feature sequence, and calculating a consistency average value according to the same base number / sequence length * 100%, namely the germplasm purity value. The method fills the blank that quantitative evaluation cannot be carried out in the prior art, is high in precision and reliability, and can provide key technical support for crassostrea gigas breeding improvement, germplasm protection, industrial standardization and the like.

The application belongs to the technical field of aquaculture, and particularly relates to a method for improving the quality of gametes of Crassostrea gigas. Gonad undeveloped parent oysters of Crassostrea gigas are selected, and the parent oysters are treated with lipopolysaccharide, and then the gametes generated by the parent oysters are used to cultivate offspring after temperature promotion. The diameter of the gametes generated by the parent oysters of Crassostrea gigas treated with lipopolysaccharide is increased, the total antioxidant capacity is improved, and the growth speed and the attachment metamorphosis capacity of the offspring larvae are significantly improved. The application opens up a new way for solving the bottleneck problem of high mortality of larvae in the process of artificial breeding of Crassostrea gigas.

The invention belongs to the technical field of aquatic organisms, and relates to application of a pacific oyster anti-disease gene SNP marker in identification of an anti-bacterial-disease pacific oyster individual, the SNP marker is located at the 1275th base, the 1281th base, the 1285th base, the 1386th base, the 1623th base and the 1701th base from an initiation codon of a hypoxia inducible factor CgHIF-1alpha gene as shown in SEQ ID No.1, and the mutation types are respectively + 1275G / A, + 1281A / G, + 1285C / T, + 1356A / T, + 1386C / T, + 1623C / T and + 1701C / G. The polymorphic sites of the hypoxia-inducible factor CgHIF-1alpha gene in the crassostrea gigas are screened, the SNP marker related to the crassostrea gigas bacterial disease resistance character is developed, and the SNP marker can be used for identification of crassostrea gigas bacterial disease resistance individuals and parent screening in bacterial disease resistance strain breeding.

This invention relates to the field of aquaculture technology, and particularly to a method and application of menthol in enhancing the embryonic development of *Crassostrea gigas* oysters. The invention employs two methods: treating parent oysters or sperm and eggs with menthol. Both methods significantly enhance the embryonic development of *Crassostrea gigas* oysters. When treating parent oysters with menthol, at a final concentration of 20 mg / L and a seawater salinity of 12‰, the hatching rate increased from 16.25% in the control group to 67.75%, an increase of 3.17 times. When treating gametes with menthol, the sperm and eggs of *Crassostrea gigas* were mixed and soaked in a 20-30 mg / L menthol-infused seawater solution with a seawater salinity of 12‰. The hatching rate increased from 40.85% in the control group to a maximum of 77.06%, an increase of 88.64%.

The invention discloses a crassostrea gigas offspring seed cultivation method capable of improving immunity of D-type larvae, which comprises the following steps: in the crassostrea gigas offspring seed cultivation process, when crassostrea gigas fertilized eggs develop to the initial stage of the D-type larvae, culturing the D-type larvae for 24 hours by using a culture water body containing ammonium chloride with the final concentration of 0.053 g / L, and then transferring the D-type larvae into a culture water body without ammonium chloride to be continuously cultured. According to the invention, the autophagy level of the D-type larvae can be obviously improved, the proliferation of blood lymphocytes of the D-type larvae is promoted, and the immune response capability of the D-type larvae to viruses is enhanced. In addition, ammonium chloride can also induce expression of inflammation-related genes of the D-type larvae, so that the immunity of the D-type larvae of the crassostrea gigas is comprehensively improved, the death rate of the larvae is effectively reduced, and the quality of crassostrea gigas fries is improved. Meanwhile, the method has the advantages of simplicity in operation, low cost and the like.

This invention discloses a detection method and application for *Crassostrea gigas* medicinal material or its processed slices, standard decoctions, and traditional Chinese medicine granules, belonging to the field of analytical detection technology. The detection method includes: enzymatic extraction of *Crassostrea gigas* medicinal material or its processed slices, standard decoctions, and traditional Chinese medicine granules to obtain a test solution; enzymatic extraction and enzymatic digestion of *Crassostrea gigas* reference material to obtain a reference solution; and detection of the test solution and the reference solution using ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS / MS). The detected ion pairs are 364.21→307.19, 364.21→417.25, 369.71→512.25, and 369.71→448.26. The detection method of this invention has strong specificity and can effectively identify different oyster origins and adulterants through the organic matter of *Crassostrea gigas*, providing a reliable basis for the quality control and evaluation of oysters and their products.

The invention discloses a method for improving Crassostrea gigas genome selection predictability based on generic transcriptome information and application, and belongs to the field of molecular breeding. Expression frequency information of Crassostrea gigas population level transcripts is systematically integrated, and the Crassostrea gigas genome selection predictability is improved according to conservative degrees of different transcripts. When genome selection is carried out, functional layering and weighting are carried out on a gene functional unit and a regulation and control region thereof, so that the prediction robustness and genetic analysis capability of the model are enhanced. The markers of the transcripts of different conservative types defined by the method can be commonly used among main strains of the crassostrea gigas, multi-group data can be conveniently integrated, and the generalization ability of the model is enhanced.