53 results about "Sexual maturity" patented technology

The invention provides a low-temperature-resistant trachinotus ovatus fry breeding method which comprises the following steps: (1) parent selection and breeding: taking trachinotus ovatus as a female parent, and carrying out intensive culture before insemination; then low-temperature stress culture is carried out under the condition of 18-20 DEG C, and the survival and normal ingestion parent fishes are used for the subsequent breeding step; (2) cross breeding: screening out sexually mature parents for induced spawning to complete artificial insemination; when the larvae are hatched out of membranes, the larvae are transferred into a breeding pond for fry breeding; (3) feeding rotifer and copepod larvae to the newly hatched larvae; when the larvae are transformed into juvenile fishes, feed containing drosera burmannii is fed, and when the juvenile fishes are transformed into juvenile fishes, feed containing eucommia ulmoides vines is fed for at least 15 days. According to the method, the low-temperature resistance of the trachinotus ovatus fries is effectively improved, and the method is easy to operate, low in cost and easy to apply and popularize.

The invention discloses an MMP2 gene promoter region SNP (Single Nucleotide Polymorphism) site and application thereof in pig breeding. According to the invention, an MMP2 gene promoter region is taken as a research object, and the relevance between the MMP2 gene promoter region and sow sexual maturity is researched by adopting a molecular and cell biology method, so that the situation that g.30088284G > A site and g.30088004C > T site are obviously related to the group maturity day age of Dumet black pigs is found; then different vectors are transfected, and different influences of different genotypes of the sites on the activity of the IRS1 gene promoter are studied. Further, sow ovarian granular cells are used as experimental materials, experimental technologies such as transcriptome sequencing, a total iron colorimetric method and a malondialdehyde colorimetric method are adopted, the influence of the MMP2 gene on the transcriptome level and ferroptosis of the sow ovarian granular cells is researched, and it is found that cell transcriptome differential genes influenced by the MMP2 gene are mainly enriched to signal channels such as cell differentiation; and ferroptosis of cells is promoted.

The invention discloses a porcine circular RNA, a porcine circLDHC, an overexpression vector and a construction method and application thereof, and relates to the technical field of livestock molecular biology. The porcine circular RNA is formed by cyclizing 3rd to 6th exons of an LDHC gene, and the nucleotide sequence of the porcine circular RNA is as shown in SEQ ID NO: 1. The porcine circLDHC is obtained by carrying out reverse transcription on porcine circular RNA (Ribonucleic Acid). According to the invention, a specific amplification primer is designed for circLDHC, and the real existence of circLDHC is verified through PCR (Polymerase Chain Reaction) amplification, sequencing and RNase R enzyme digestion. Fluorescent quantitative PCR analysis finds that circLDHC is highly expressed in sexual mature testis tissues of pigs, the expression level of circLDHC is related to testis maturation, and meanwhile, the expression quantity of circLDHC in testis of 75-day-old Meishan boars is remarkably higher than that of circLDHC in testis of same-day-old Duroc boars (plt; 0.01) of the substrate. The invention further finds that the over-expressed circLDHC inhibits the proliferation of the porcine sertoli cells, promotes the maturation of the porcine sertoli cells, lays a foundation for further researching the start of the puberty period of the boars, and provides a new thought for analyzing the sexual precocious puberty mechanism of the boars and improving the reproductive performance.

Provided is a method for promoting sexual maturity in fish, comprising a step for inhibiting functional expression of at least one of a leptin receptor and leptin in fish. The present invention addresses the problem of providing a method for promoting sexual maturity of a fish in order to obtain an individual fish capable of ovulating or spermising at an age of 1 more than under natural or conventional culture conditions.

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.

The present invention relates to the technical field of genetic breeding of marine economic shellfish, and in particular to a breeding method for a new Vibrio-resistant variety of scorpion clams. Specifically, the method comprises the following steps: using a semi-lethal concentration of Vibrio parahaemolyticus to stress adult scorpion clams, recovering 15% of the surviving individuals, and selecting scorpion clams with an 8% seed retention rate to obtain Vibrio-resistant scorpion clams, fattening the scorpion clams in an indoor workshop to mature the gonads, and inducing the scorpion clams to produce heritable immune factors by inactivating Vibrio parahaemolyticus before inducing spawning; then inducing the scorpion clams to spawn, obtaining the Vibrio-resistant strain of the F1 generation of scorpion clams, and then rapidly cultivating the scorpion clams to sexual maturity by adopting the breeding and culture technology of "breeding in the north and raising in the south" for the F1 generation; thereafter, selecting scorpion clams by stress, enhancing the immunity of the offspring, and continuously breeding by the method of "breeding in the north and raising in the south" every year to obtain a new Vibrio-resistant variety of scorpion clams with stable genetics and excellent traits. The method of the invention can significantly improve the anti-vibrio ability and the aquaculture survival rate of razor clams, and improve the good variety preservation and supply capacity of the razor clams aquaculture industry.

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 discloses an application of an IRS1 gene promoter region SNP (Single Nucleotide Polymorphism) site as a porcine sexual maturity molecular genetic marker. According to the invention, an IRS1 gene promoter region is taken as a research object, and the relevance between the IRS1 gene promoter region and the sexual maturity of the sow is researched by adopting molecular and cell biological methods: g.128307611Cgt is found; a and g.128307308Cgt, and g.128307308C T is obviously related to group mature day age of Dupin black pigs; then different vectors are transfected, and different influences of different genotypes of the sites on the activity of the IRS1 gene promoter are studied. Furthermore, sow ovarian granular cells are used as experimental materials, experimental technologies such as transcriptome sequencing, a total iron colorimetric method and a malondialdehyde colorimetric method are adopted to research the influence of the IRS1 gene on the transcriptome level and ferroptosis of the sow ovarian granular cells, and the result shows that the IRS1 gene influences the signal paths that cell transcriptome differential genes are mainly enriched to iron ion homeostasis and the like; the ferroptosis of the cells is inhibited.

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 discloses a method for prolonging the spawning period of epinephelus moara, which comprises the following steps: (1) selecting juvenile epinephelus moara with the body weight of 100-150g as a parent fish rearing object, with the dissolved oxygen of 5-9mg / L, the salinity of 25-33, the pH of 7.8-8.2, the water flow exchange capacity of 2-3m < 3 > / h, the water temperature of 25-28 DEG C, the sunlight illumination time of 13.5 h and the rearing density of 6-8kg / m < 3 >; the feed is fed once a day, and the traditional Chinese medicine composition is taken for 10d / month; (2) when the parent fish reaches the sexual maturity age, the water temperature is reduced to 20 DEG C, the sunlight illumination time is 12 h, and the breeding density is 6-8 kg / m < 3 >; the feed is fed twice and three times every day, and the traditional Chinese medicine composition is taken for 10d / month; after one month, the water temperature is gradually increased to 25 DEG C, and the daily illumination time is gradually increased to 13.5 h; (3) when the gonad of the parent fish develops to 3-4 stages, injecting chorionic gonad promoting hormone for spawning induction; (4) after the parent fishes lay eggs for 30 days, the water temperature is gradually reduced to 20 DEG C, and meanwhile, the illumination time is gradually reduced to 12 hours; after 30 days, the water temperature is gradually increased to 25 DEG C, and the illumination time is gradually increased to 13.5 h. According to the method, the breeding period of the parent fish of the epinephelus moara is prolonged.

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 provides a method for artificial insemination and fertilized egg incubation of wild schizothorax parvus, which comprises the following steps: on the basis of ascertaining the biological characteristics of the wild schizothorax parvus through tracking and monitoring for many years, capturing sexually mature male and female parent fishes, carrying out artificial insemination after sperm and egg treatment, obtaining fertilized eggs, and carrying out micro-running water incubation, so that the fertilization rate reaches 85%, and the incubation rate reaches 70%. The artificial insemination of the rare and endangered indigenous fish Schizothorax parvum in the Luagu Lake and the trial culture of the first filial generation of fish fries are successfully carried out for the first time in China, the blank in the field is filled, the artificial domestication, breeding and culture of the Schizothorax parvum are completely feasible and are preliminarily successful, and the specific fish population of the Luagu Lake is recovered, developed and cultured, so that the economic benefits of the Luagu Lake and the Schizothorax parvum are improved. High-quality offspring seeds are provided, and the method has important significance on lake enhancement and release and improvement of fishery economic benefits.

The invention discloses an abalone gender differential expression miRNA and a mining method of target mRNA of the abalone gender differential expression miRNA, and belongs to the technical field of molecular biology. According to the method, transcriptome and sRNA sequencing is carried out on gonad tissues of sexually mature female and male abalones, so that the expression difference of mRNA and miRNA in the female and male abalones is compared, multiple differential expression miRNAs related to the sex and reproduction of the abalones and target mRNA thereof are positioned through sequencing, and a regulatory network of part of miRNA and mRNA is disclosed; according to the method, the sequencing result is accurate and reliable, and the sequenced gene is expected to serve as a molecular marker, is applied to evaluation of sex maturity and sex determination of abalones, and has important practical significance and practical application value for perfecting transcriptomes and miRNA gene banks of ovaries and testis tissues of abalones.

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 rapidly obtaining genotype homozygous intermuscular thorn-free grass carps, which comprises the following steps: carrying out gene editing on wild grass carps by taking runx2b genes as target genes, screening successfully mutated F0-generation individuals, culturing to sexual maturity, then screening mutated F0-generation male and female individuals with the same mutation type, and mating to obtain F1-generation grass carps; the individuals with homozygous runx2b alleles in the F1 generation are obtained through genotype detection, and the individuals are the F1 generation intermuscular thorn-free grass carp new germplasm. The sexual maturity of the grass carp needs 4-5 years, and the period of breeding the first generation is long, so that the invention aims to establish a method for quickly obtaining the grass carp without the intermuscular thorns by utilizing a gene editing technology, the period of obtaining homozygotes is shortened, and a method is provided for genetic improvement of economic characters of economic fishes such as the grass carp and the like. The method provides support for molecular mechanism research of fish intermuscular spines, and has important scientific research and industrial application values.

The invention provides a method for accelerating directional breeding of bumblebees, which specifically comprises the following steps: selecting worker bees with excellent characters as female parent worker bees for directional breeding, laying eggs and hatching the worker bees into male bees in a dark environment with the temperature of 27-29 DEG C and the humidity of 50%-60%, obtaining sperms of the male bees after sexual maturity, and then putting the sperms into a bumblebee breeding box to breed the bumblebees. And inseminating female parent worker bees by using the obtained sperms, establishing a queen bee breeding group by using the inseminated female parent worker bees and more than 30 worker bees which just emerge and come out of the house to produce offspring queen bees, and mating the offspring queen bees with the male bees to obtain an offspring queen bee colony. According to the method, the queen bee colony can be stably and rapidly obtained only through a single female parent worker bee, all genes come from the female parent worker bee, rapid propagation of bumblebees and breeding of excellent characters are facilitated, and the method has important application prospects in natural resource protection and agricultural production.