36 results about "Sexual maturity" patented technology

The invention relates to the technical field of gene engineering, in particular to a transgenic zebrafish system pTo12-efla-tjp1a-P2A-mCherry based on a Tol2 transposition subsystem and a construction method of the transgenic zebrafish system pTo12-efla-tjp1a-P2A-mCherry. The preparation method comprises the following steps: S1, constructing a Tol2 transposon expression vector pTol2-ef1a-tjp1a-P2A-mCherry which contains an ef1a promoter, a tjp1a gene, a P2A peptide sequence and an mCherry reporter gene; s2, carrying out in-vitro transcription to prepare Tol2 transposase mRNA; s3, mixing the expression vector with Tol2 transposase mRNA, and microinjecting the mixture into the single-cell stage embryo of the zebra fish; s4, performing fluorescence screening on the F0 generation embryos surviving after injection to obtain the Founder fish with positive transgenosis; s5, after the F0-generation positive fish is bred to be sexually mature, an F1 generation is obtained through mating, transgenic positive individuals with stable inheritance are screened out after identification, and a transgenic zebrafish strain is established. According to the transgenic zebrafish system pTo12-efla-tjp1a-P2A-mCherry based on the Tol2 transposon system and the construction method of the transgenic zebrafish system pTo12-efla-tjp1a-P2A-mCherry based on the Tol2 transposon system, efficient integration of exogenous genes is achieved by utilizing the Tol2 transposon system, and the transgenic zebrafish line capable of being stably inherited to the F1 generation is successfully obtained.

This invention discloses a method for large-scale breeding of all-male salmon and trout, belonging to the field of fish sex regulation breeding technology. The method includes: collecting fertilized eggs from different families of the same species of salmon and trout, artificially incubating them to obtain fry, and inducing sex reversal in the fry by using 17β-estradiol when they begin feeding; subsequently, raising the fry to sexual maturity, selecting individuals with a female phenotype, and then using sdY as a sex identification gene, selecting pseudo-females with genotype XY by quantitative real-time PCR; pairing the sexually mature pseudo-females with genotype XY with normal males with genotype XY, and obtaining super-males with genotype YY through artificial insemination; pairing the sexually mature super-males with females with genotype XX, and producing all-male salmon and trout with genotype XY on a large scale through artificial insemination. This invention overcomes the problem that existing technologies cannot achieve large-scale breeding of all-male salmon and trout.

The invention discloses a method for promoting ovulation of sexual maturity zebrafish by using Gap27, and relates to the technical field of artificial breeding of fish, a gap connecting protein inhibitor Gap27 is applied to ovulation induction of zebrafish, an intercellular communication network in follicles is accurately intervened, the problem of asynchronous ovulation is solved, and in the final stage of maturation of oocytes, ovulation of sexual maturity zebrafish is promoted. Close substance and information exchange is carried out among follicular granule cells through a gap connecting channel formed by connecting protein Cx43, and Gap27 is used as a specific peptide fragment, can simulate an extracellular ring structure of Cx43, and competitively binds with Cx43 on an adjacent cell membrane, so that formation and functions of the gap connecting channel are physically blocked, and the follicular granule cells are separated from each other. When ovarian tissues of sexually mature female fish are exposed in a Gap27 solution, the transmission of inhibitory signals from granular cells to oocytes can be effectively interrupted, the rupture of nuclear membranes of the cells is promoted, and the final maturation process is completed.

The invention provides a method for calculating a migratory fish sexual maturity cycle through a PIT mark monitoring technology, and relates to the technical field of PIT mark monitoring. Comprising the steps of determining starting time and ending time according to electronic tag endurance attributes; based on the starting time and the ending time, installing an electronic tag monitoring device and an environment monitoring sensor at a preset position of the target monitoring area to obtain environment monitoring data and an initial tag monitoring data set of each year in the target monitoring area; determining an initial sexual maturity cycle data set according to the initial label monitoring data set; and correcting the initial sexual maturity cycle data set according to the environmental monitoring data in each year to obtain a final sexual maturity cycle data set. According to the invention, the problem of low fish sexual cycle monitoring efficiency and precision in the prior art is solved.

The invention discloses a method, a system and equipment for splitting an allopolyploid based on a perl language and a storage medium, and particularly relates to the technical field of bioinformatics analys.The method comprises the steps that a diploid species close to a to-be-split allopolyploid species is selected as a diploid reference species; calculating a distance SD value between chromosomes to form a matrix; a homologous chromosome group HG1. HGN is formed; according to the homologous chromosome group HG1... HGN, an allopolyploid to be split is split into AN subgenomes. The subgenome is split based on the mutation accumulation difference of the sexual maturity cycle, and the method has high flexibility; the method disclosed by the invention has wide applicability; the splitting method disclosed by the invention has the advantages of high accuracy, good visualization effect, simplicity and convenience in operation, reliable data support, capability of promoting genome research and the like; according to the invention, how different mutations are accumulated by different species under the difference of sexual maturity cycles can be well revealed, so that deep understanding of the evolutionary mechanism of the species is promoted.

The application provides a method for on-site rapid artificial insemination of wild Pseudosciaena crocea of Lutjanidae, and belongs to the technical field of aquatic animal breeding. Wild parent fish of Pseudosciaena crocea of Lutjanidae are captured in the breeding season; sperm is collected from the selected sexually mature male parent fish on the capture site; after artificial sperm collection, sexually mature female parent fish are selected, and fish eggs are collected; high-quality fertilized eggs are obtained through artificial insemination; the artificial insemination comprises the following steps: adding sperm into fish eggs, adding natural seawater for stirring, and finally washing with natural seawater. Through comprehensive regulation of key links in the whole process of artificial insemination, the purpose of obtaining high-quality fertilized eggs by using wild parent fish of Pseudosciaena crocea of Lutjanidae and carrying out artificial fry breeding is achieved.

The application belongs to the technical field of fish variety improvement, and discloses a method for creating gynogenetic grass carp with rapid growth. The method comprises the following steps: screening sexually mature male mirror carp and sexually mature female grass carp with obvious growth advantages and stress resistance, collecting sperm of the mirror carp, diluting the sperm with Hank's solution, inactivating the sperm by ultraviolet irradiation, mixing the obtained inactivated sperm with middle-stage eggs of the grass carp, and then performing cold shock doubling treatment, and adding normal temperature water in batches for hatching to obtain gynogenetic grass carp. The gynogenetic grass carp induced by the mirror carp sperm performs excellently in growth speed and disease resistance, which provides a new method and idea for inheritance improvement of the grass carp, provides more possibilities for screening of excellent varieties, and brings greater economic benefits to the grass carp breeding industry.

The application belongs to the technical field of animal model construction, and relates to a construction method and application of a zebrafish aging model. The construction method comprises the following steps: 1, sgRNA, Cas9 protein and a homologous recombination knock-in plasmid containing fluorescent protein A and a knock-in gene are transferred into zebrafish embryos, development is cultured to sexual maturity, F0-I is obtained, and a knock-in strain I is obtained through screening; 2, sgRNA, Cas9 protein and a homologous recombination knock-in plasmid containing fluorescent protein B and a knock-in gene are transferred into zebrafish embryos, development is cultured to sexual maturity, F0-II is obtained, and a knock-in strain II is obtained through screening; 3, the knock-in strain I and the knock-in strain II are mated, zebrafish with fluorescent protein A and fluorescent protein B simultaneously expressed in the heart are screened from the offspring, and a wrn gene knockout homozygote, that is, a zebrafish aging model, is obtained. The construction method is more convenient to breed, significantly reduces breeding costs and the workload of screening and breeding, and has good application value.

The present invention provides a method for accurately locating the spawning grounds of schizothoracine fish, comprising: step 1, determining the study area; step 2, determining the sexual maturity stage based on the egg diameter and the stage of gonad development, and then determining whether spawning has occurred based on whether the eggs are emptied and whether the parent fish's anal fin is broken, thereby determining the spawning time; step 3, based on the study area determined in step 1, identifying the river section of the spawning ground of the genus Gymnosoma schizothoracine fish during the spawning period of the genus Gymnosoma schizothoracine fish determined in step 2; step 4, based on the spawning ground section determined in step 3, accurately locating the spawning ground: determining the specific time of spawning within the spawning period obtained in step 2; after determining the specific spawning time, accurately locating the spawning ground of the genus Gymnosoma schizothoracine fish based on the bottom soil differences of the river section of the spawning ground of the genus Gymnosoma schizothoracine fish. The present invention accurately locates the spawning grounds of schizothoracine fish, fills a gap, and provides scientific support for the protection of fish habitats in the source river of the Yangtze River.

The invention relates to the technical field of fish cultivation, in particular to a method for cultivating all-male tetraploid crucian carp, which comprises the following steps of: performing artificial spawning induction by taking gynogenesis homologous diploid crucian carp as a female parent and taking androgenesis homologous diploid crucian carp as a male parent; wherein the sex chromosome type of the gynogenesis homologous diploid crucian carp is XX and generates non-reduced diploid ovum, and the sex chromosome type of a male individual in the androgenesis homologous diploid crucian carp is XY or YY and generates non-reduced diploid sperm; artificial insemination is carried out, obtained fertilized eggs are hatched and cultured to sexual maturity, and survival offspring is the all-male tetraploid crucian carp population. Based on the characteristic that gynogenesis homologous diploid carassius auratus and androgenesis homologous diploid carassius auratus both generate non-reducing gametes, the gynogenesis homologous diploid carassius auratus and the androgenesis homologous diploid carassius auratus are subjected to artificial insemination, and all-male tetraploid The all-male tetraploid crucian carp can reach a sexual maturity state after being only 120 days old, diploid sperms can be stably generated, and the reproductive performance is improved.

The invention belongs to the field of beekeeping, and relates to a method for controlling mating of queen apis cerana by using a queen bee fixing device, and by designing the specially-made queen bee fixing device, the device can mildly and stably limit the activity range of the queen bee, and meanwhile, the physiological safety and health of the queen bee are ensured. In the sexual maturity stage of the queen bee, the queen bee is placed in the device and placed in a specific mating site, the range and intensity of pheromone released by the queen bee are controlled through the device, drones are attracted to come to mate, and the problems that in natural mating, the queen bee flies away, bad weather influences exist, and non-target drone interference exists are effectively solved. By accurately regulating and controlling the mating environment and time, the mating success rate and the offspring quality are improved. The method is easy and convenient to operate, small in bee colony interference and beneficial to improvement of Chinese bee breeding benefits and population genetic diversity management, and a new way is provided for scientific breeding and breed conservation and breeding of Chinese bees.

The application discloses application of an IRS1 gene promoter region SNP site as a pig sexual maturity molecular genetic marker. The present application takes the IRS1 gene promoter region as a research object, and studies the correlation between the IRS1 gene promoter region and sow sexual maturity by using molecular and cell biology methods: it is found that g.128307611C>A and g.128307308C>T are significantly correlated with the age of sexual maturity of Duhei pig populations; then, the influence of different genotypes of the above sites on the activity of the IRS1 gene promoter is studied by transfecting different vectors. Further, the sow ovarian granulosa cells are taken as experimental materials, and experimental techniques such as transcriptome sequencing, total iron colorimetry and malondialdehyde colorimetry are used to study the influence of the IRS1 gene on the transcription level and ferroptosis of the sow ovarian granulosa cells, and it is found that the IRS1 gene influences the different genes of the cell transcriptome, and is mainly enriched in signal pathways such as iron ion homeostasis, and inhibits the ferroptosis of the cells.

The present application relates to fish breeding technology field, especially to a kind of all-male tetraploid crucian carp breeding method, comprising the following steps: with gynogenesis homologous diploid crucian carp as female parent, and androgenesis homologous diploid crucian carp as male parent, artificial delivery is carried out;Wherein, the gynogenesis homologous diploid crucian carp sex chromosome type is XX, produces non-reduction diploid egg, and the androgenesis homologous diploid crucian carp male individual sex chromosome type is XY or YY, produces non-reduction diploid sperm;Artificial insemination is carried out, and the obtained zygote is hatched, and cultured to sexual maturity, and the surviving offspring is all-male tetraploid crucian carp population.The present application is based on the characteristics that gynogenesis homologous diploid crucian carp and androgenesis homologous diploid crucian carp both produce non-reduction gametes, artificial insemination is carried out, and all-male tetraploid crucian carp can be prepared on a large scale.The all-male tetraploid crucian carp only needs 120 days of age to reach sexual maturity, can stably produce diploid sperm, and the reproductive performance is improved.

The invention belongs to the technical field of fish variety improvement, and discloses a method for creating rapidly growing gynogenetic grass carps, which comprises the following steps of: screening sexual maturity male mirror carps and sexual maturity female grass carps with relatively obvious growth advantages and stress resistance; the method comprises the following steps: collecting mirror carp sperms, diluting the mirror carp sperms with Hank's liquid, inactivating the sperms by ultraviolet radiation, mixing the obtained inactivated sperms with middle-section ova of grass carps, then carrying out cold shock doubling treatment, and then adding normal-temperature water in batches for hatching to obtain gynogenetic grass carps. The gynogenesis grass carps induced by the mirror carp sperms are excellent in growth speed and disease resistance, a new method and thought are provided for heritage improvement of the grass carps, more possibilities are provided for screening good varieties, and greater economic benefits are brought to the breeding industry of the grass carps.

The invention provides a method for promoting gonad maturation of plectropomus leopardus by applying a spectrum, which is characterized in that breeding systems under different illumination conditions, including full-spectrum, green-spectrum and blue-spectrum illumination conditions, are constructed, and are combined with a factory-like circulating water breeding system to breed the plectropomus leopardus for 60 days. Experimental results show that the blue light and the green light can significantly accelerate the development of male and female gonads, especially the blue light can promote the development of oocytes, and the green light can promote the development of male germ cells. The method effectively shortens the sexual maturity period of the plectropomus leopardus by accurately regulating and controlling the illumination condition, improves the culture efficiency, and provides a theoretical basis and technical support for regulating and controlling the gonad development environment in aquaculture.

The invention relates to the technical field of biological feed processing and application, and discloses an artificial feed for monochamus alternatus adult beetles as well as a preparation method and application of the artificial feed. The artificial feed provided by the invention can ensure that the adult monochamus alternatus hope has longer survival time and higher survival rate under the condition of not using a large number of fresh pine branches; meanwhile, by adding the female moth powder and the goose egg yolk powder, lecithin and other components can be introduced, the sexual maturity time of the adult monochamus alternatus can be remarkably shortened, the egg laying amount of the adult monochamus alternatus can be increased, and the breeding efficiency of the adult monochamus alternatus can be remarkably improved; in addition, the components in the artificial feed are reasonably matched, and the problem of feed hardening caused by addition of high wood chips is effectively avoided.

The invention provides a gene influencing sexual precocity of deep-county pigs and a screening method thereof, and belongs to the field of research on sexual maturity of pigs, the gene influencing sexual precocity of deep-county pigs comprises an SRD5A1 gene and / or a CYP11B2 gene, testicular tissues of deep-county pigs before and after sexual maturity are subjected to transcriptome sequencing, differential expression gene functions are analyzed, and the sexual precocity of deep-county pigs before and after sexual maturity is obtained. And screening out candidate genes related to the sexual maturity of the deep county pigs. And then performing transcriptome sequencing on testis tissues of the sexually mature deep county pig and the hybrid pig of the sexually mature deep county pig and the large white pig, analyzing functions of differential expression genes, and screening specific expression genes of the sexual maturity of the deep county pig. Integrating the results of the two times of transcriptome sequencing, analyzing the functions of the coincident genes, and screening candidate genes SRD5A1 and CYP11B2 which influence the sexual precocity of the pigs in the deep county according to the gene functions; and a foundation is laid for subsequent research on precocious puberty of pigs in deep county.

The invention belongs to the technical field of culture, and discloses a grass carp culture method capable of shortening sexual maturity time, which comprises the following steps: 1, parent screening; feeding with feed; managing water temperature. By systematically analyzing the synergistic effect of antioxidant regulation of tocopherol (vitamin E), feed nutrition optimization and water temperature accurate management, the sexual maturity period of large-scale cultured grass carp is successfully and stably controlled within 3 years. The breakthrough has important practical significance on reduction of breeding cost and guarantee of offspring seed supply.

The invention relates to the technical field of fish breeding. The invention provides an artificial breeding method for paraphyllous hemsleyanus, which comprises the following steps: (1) selecting parent paraphyllous hemsleyanus which is healthy, free of injury and disease, good in gonad development and more than 18 months old, breeding male and female parent fishes in different ponds, and controlling the ratio of male fishes to female fishes to be 1: (2-3); (2) selectively injecting a first oxytocic into the mature male fish, and injecting a second oxytocic into the sexually mature female fish to obtain induced spawning parent fish; (3) collecting seminal fluid of the induced spawning male fish and fish egg cells of the induced spawning female fish, carrying out artificial insemination, and hatching to obtain fry. By adopting the breeding method, generation of parent fish egg cells or sperms can be effectively promoted, so that the quality of fertilized eggs and the breeding success rate are improved; by utilizing the compound spawning induction substance, the spawning induction effect is enhanced, meanwhile, the use of chemical substances can be reduced, and drug residues and environmental pollution are reduced. The method is of great significance to artificial breeding and large-scale breeding of the paralifola speculatus.

The invention discloses a method for creating fluorescent zebra fish based on an enhancer of a muscle specific gene. The invention provides a method for creating fluorescent zebrafish, which comprises the following steps: by taking an upstream promoter and an enhancer of a fmyhc2.2 gene as target sequences, amplifying the upstream promoter and the enhancer and seamlessly connecting the upstream promoter and the enhancer with a fluorescent protein carrier to form a recombinant plasmid, carrying out microinjection on a zebrafish embryo, selecting F0 generation individuals capable of driving fluorescent protein expression, and culturing until sexual maturity, thereby obtaining the fluorescent zebrafish. Selecting male and female individuals for selfing to obtain F1 generation, observing and selecting individuals with strong fluorescence from F1 generation embryos through a fluorescence microscope, and culturing the individuals with strong fluorescence to sexual maturity to obtain the transgenic zebrafish with remarkably discolored skin. The fluorescent zebra fish is created by utilizing the enhancer of the muscle specific gene, an innovative method is provided for creating a zebra fish ornamental strain, a support is provided for researching a regulation mechanism of zebra fish muscle related genes, and the fluorescent zebra fish has important scientific value and industrial application value.

The invention discloses a method for producing all-male largemouth micropterus salmoides based on soaking induction of sex reversal of largemouth micropterus salmoides, and particularly relates to the field of artificial propagation of all-male largemouth micropterus salmoides, the method comprises the following steps: S01, taking fairy shrimps as initial feed when largemouth micropterus salmoides fries obtained by incubation swim flatly; s02, transferring the largemouth micropterus salmoides fry into a water body in which a sexual reversal induction reagent is dissolved, keeping feeding three times every day during the period, and soaking and breeding the fry for 60 days; and S03, after soaking and culturing for 60 days, transferring the largemouth bass fry into a conventional culture water body for culturing, and after sexual maturity of the largemouth bass fry, obtaining a male population capable of generating seminal fluid, namely the all-male largemouth bass. According to the method, largemouth bass fries are soaked in a sex reversal induction reagent with a specific concentration, sex reversal fishes with a high survival rate and a low malformation rate can be directly obtained, and then large-scale cultivation of all-male largemouth bass is achieved.

This invention provides a method for rapid propagation of superior grass carp families, belonging to the field of aquaculture and fish genetic breeding technology. The method includes: screening for disease-resistant superior families from established grass carp families; isolating and enriching reproductive stem cells from these superior families; transplanting the grass carp family reproductive stem cells into the abdominal cavity or genital ridge region of recipient fish with short sexual maturity cycles; cultivating the recipient fish to sexual maturity, producing sperm, and then fertilizing the sperm with grass carp eggs to obtain F1 generation offspring; performing molecular identification on the offspring to confirm that their genetic material originates from the donor family. This invention can shorten the propagation cycle of superior grass carp families from 4-5 years to approximately 1.5 years, achieving precise and efficient transfer of superior traits, and providing an effective technical means to shorten the grass carp breeding cycle and cultivate superior grass carp varieties.

The present invention discloses an induction method for efficiently producing triploid male Nibea albiflora, belonging to the technical field of triploid induction of male Nibea albiflora, and specifically relates to nutritional regulation and photothermal regulation of sexually mature pseudo-male Nibea albiflora and female Nibea albiflora, sperm and eggs collection; artificial insemination to obtain fertilized eggs, which are immersed in seawater for treatment and then incubated, and finally triploid male Nibea albiflora is induced and cultivated; when the fertilized eggs are immersed in seawater during artificial insemination and incubation, purine derivatives can be added to the seawater to treat the fertilized eggs, thereby improving the success rate of triploid induction of Nibea albiflora. The method of the present invention has a high success rate of triploid induction of Nibea albiflora, and triploid male Nibea albiflora is successfully obtained under subsequent induction.

The application discloses a high-body cobia broodstock feed containing Chinese herbal medicines and a high-body cobia breeding method using the feed, and by matching different raw materials, the high-body cobia broodstock feed can better provide the high-body cobia broodstock with nutrient components and active substances. Before spawning, the high-body cobia broodstock feed can enhance the physical fitness and immunity of the broodstock, promote the development of the broodstock, accelerate sexual maturation, improve the spawning rate, the egg production, the fertilization rate and the hatching rate, and after spawning, the high-body cobia broodstock feed can promote the healing of the broodstock, restore the physical fitness, and guarantee the survival rate and quality of the broodstock after spawning.

The application relates to the field of aquatic organism measuring equipment. A device for rapidly identifying Euphausia superba and measuring biological characteristic parameters comprises a main box body, the top of the main box body is a measuring plate, at least three hollowed-out areas are arranged on the measuring plate in sequence, and a scale unit is arranged on the hollowed-out areas; the scale unit comprises a base plate, and a reference scale line is arranged on the upper side of the base plate; a slide rail with a left-right direction as a guide direction is arranged at the rear of the main box body; the device further comprises a sliding support which is slidably connected with the slide rail, and a camera and a tablet computer are arranged on the sliding support and connected with the tablet computer. The camera can take photos of the Euphausia superba, and then biological characteristics such as the body length, gender, sexual maturity, body color and feeding intensity of the Euphausia superba are recorded. The sliding support facilitates the sequential recording of the Euphausia superba on different scale units.

This invention discloses a method for constructing a zebrafish model of fatty liver based on glucokinase overexpression and its application. The method includes the following steps: using cDNA as a template, a zebrafish GCK gene fragment is obtained by PCR amplification; the vector fragment and the GCK gene fragment are homologously recombined using a homologous recombinase to obtain a recombinant plasmid; the recombinant plasmid is transformed into competent cells, and an endotoxin-free plasmid is extracted as a zebrafish GCK expression vector; a microinjection system is used to inject the plasmid into one-cell-stage fertilized eggs; after the fertilized eggs hatch into juveniles, fluorescent juveniles are selected for further culture until sexual maturity; sexually mature males and females are paired to spawn, and a stably heritable GCK-overexpressing zebrafish strain is selected from the offspring. This invention can obtain a zebrafish strain with stable high GCK expression, which can be used as a zebrafish model of fatty liver based on GCK overexpression.

The invention provides a method for constructing an anti-subtype J avian leukosis gene edited chicken based on adenine editor adenovirus and application of the anti-subtype J avian leukosis gene edited chicken. The method comprises the following steps: taking c.98Tgt of a chicken NHE1 gene; c, c, 100 Tgt; c, c.112 Tgt, c. 112 Tgt; c, c, 116Agt, c, 116Agt; the method comprises the following steps of: designing an adenine editor by taking a nucleotide sequence A and a nucleotide sequence G as targets, constructing plasmids of the adenine editor capable of simultaneously editing all the sites, further packaging an adenovirus containing the sequence, and injecting the adenovirus into an HH15-18 stage chicken embryo through a microinjection method to realize the NHE1 gene c.98Tgt in the chicken embryo; c, c, 100 Tgt; c, c.112 Tgt, c. 112 Tgt; c, c, 116Agt, c, 116Agt; the method comprises the following steps: carrying out site-directed mutagenesis on one or more of G.98Tgt and G. A chick embryo is hatched and developed to sexual maturity, and carrying out hybridization and breeding to obtain an NHE1 gene C, c, 100 Tgt; c, c.112 Tgt, c. 112 Tgt; c, c, 116Agt, c, 116Agt; g, gene edited chicken with one or more point mutations, wherein the gene edited chicken is proved to have the J subtype avian leukosis resisting character. The method has great application value in the fields of J subtype avian leukosis prevention and purification, gene editing chicken disease resistance model research and the like.

The invention provides an RNA (Ribonucleic Acid) molecular marker for judging early gonad differentiation of young river snails in China and an identification method. Dmrt1 and Foxl2 genes are important genes related to gonad differentiation of river snails. The method comprises the following steps: designing and verifying primers SEQ ID NO: 1-2 according to conserved regions of Dmrt1 and Foxl2 genes; the method comprises the following steps: collecting gonad tissue samples in a young snail stage and a sexual maturity stage, wherein n is greater than or equal to 120, extracting total RNA (Ribonucleic Acid) of a field snail tissue, carrying out reverse transcription to obtain cDNA (Complementary Deoxyribonucleic Acid), detecting a double-gene expression quantity through qRT-PCR (Quantitative Reverse Transcription-Polymerase Chain Reaction), calculating a standardized expression value by taking beta-actin as an internal reference, and establishing a discrimination function D = log2 (Dmrt1 / Foxl2); 1, judging that the juvenile snails of the field snails start gonad differentiation; according to the invention, the problems of deficiency of specific molecular markers for sex differentiation of the young escargots in China and indefinite gonad differentiation time point are solved in a breakthrough manner, accurate locking of the gonad differentiation time of the young escargots is realized, the gonad differentiation time is 15-20 days earlier than morphological differentiation, and verification of more than one hundred of samples shows that the identification accuracy reaches 100%.

The present invention discloses SNP sites in the promoter region of the MMP2 gene and their application in pig breeding. The present invention takes the promoter region of the MMP2 gene as the research object, and uses molecular and cell biological methods to study its correlation with the sexual maturity of sows: it is found that the g.30088284G>A site and the g.30088004C>T site are significantly correlated with the age of sexual maturity of the Douro Black pig group; then, by transfecting different vectors, the different genotypes of the above sites are studied to affect the activity of the IRS1 gene promoter. Further, using sow ovarian granulosa cells as experimental materials, transcriptome sequencing, total iron colorimetry, malondialdehyde colorimetry and other experimental techniques are used to study the effect of the MMP2 gene on the transcription level and ferroptosis of sow ovarian granulosa cells. It is found that the differential genes in the cell transcriptome affected by the MMP2 gene are mainly enriched in signal pathways such as cell differentiation, promoting cell ferroptosis.

The invention relates to the technical field of frog breeding, in particular to a method for artificially obtaining fertilized eggs of Pleurotus nigromaculatus. According to the method, high-protein feed and tocopherol are fed to female breeding frogs and male breeding frogs of the rana nigromaculata, and the female breeding frogs and the male breeding frogs are paired after sexual maturity. Practical breeding and oviposition experiment verification shows that when 0.6-0.8 g of tocopherol is added into each 1g of feed for mixed feeding of the paphiophyllum nigromaculatum in the sexual development stage, the sexual maturity time can be remarkably shortened by 14.3% compared with that of a group without addition, the sexual maturity of the paphiophyllum nigromaculatum is promoted by adding the tocopherol preparation, and the sexual maturity rate of the paphiophyllum nigromaculatum is improved. According to the method, the hormone mixed liquid is injected into the abdominal cavity to induce spawning, the spawning success rate is increased, fertilized eggs are obtained, the method is effective and feasible, and compared with a traditional breeding spawning method, the method has remarkable advantages and can be applied to actual production research.