72 results about "Female gametes" patented technology

The invention provides a method for effectively reducing environmental risk caused by pollen propagation of transgene plant. In the method, two tightly linked gene expression cassettes of 1) a pollen lethal gene ZM-AA1 driven by a pollen growth anaphase singular promoter PG47 and 2) a fluorescence screening mark gene FP driven by a callus / seed coat singular promoter END 2 are transferred to a Zhonghua 11 wild type material. During selfing fructification of a single plant carrying a single copy transgene, a pollen grain without carrying the transgene can be fertilized with a female gamete normally, but a pollen grain carrying the transgene is abortive in pollen growth anaphase and can not be fertilized with a female gamete, so as to reduce risk of transgene drift in the environment.

The invention relates to an undariapinnatifida cultivation method, in particular to an undariapinnatifida seedling cultivation method through parthenogenesis. The method comprises the steps that induction culture is conducted on female gametophytes until juvenile sporophytes reach more than 1mm, aerated feeding culture is conducted on the female gametophytes, then the life history of parthenogenesis sporophytes is completed, and mature sporophytes are obtained; sporophyls of the sporophytes are placed into sea water to be released to obtain planospores, after the color of the sea water where the planospores are obtained through releasing is changed into yellowish-brown, the planospores are attached to a seedling curtain, and then the seedling curtain is put into fresh sea water; after all the germinal planospores develop into the female gametophytes and are sufficiently grown, light intensity is lowered, the development of the gametophytes is delayed to enable the gametophytes to pull through the high-water-temperature summer; the male gametophytes and the female gametophytes attached to the seedling curtain are simultaneously cultured, and the development and fertilization processes are completed; after reaching about 200 micrometers, the juvenile sporophytes are moved to sea to be cultivated, and then undariapinnatifida cultivation is achieved. According to the undariapinnatifida seedling cultivation method through parthenogenesis, female gametophyte clonal update and enlarged cultivation can be achieved through the parthenogernesis life history, and it is of great significance in clonal preservation and crossbreeding.

The invention belongs to the technology field of the gene engineering, in particular relates to a specificity molecule marker which can distinguish laminaria female and male gametophyte. The specificity molecule marker is obtained through the following method: two pairs of primers of SEQ ID No1and SEQ ID No2 are designed according to 5'end flanking sequence of the laminaria female and female gametophyte gene groups, polymerase chain reaction amplification is performed to the deoxyribonucleic acid of the laminaria female and female gametophyte gene groups, and the deoxyribonucleic acid of specificity amplification fragment are reclaimed; large intestine rod competent strain JM109 is cloned through pMD19-T vector to obtain a laminaria female gametophyte specificity molecule marker Rf-763 and a laminaria male gametophyte specificity molecule marker Rm-239, and the sequences thereof are SEQ ID No3 and SEQ ID No4. The genders of the laminaria gametophyte, parthenogenesis laminaria sporophyte parental source and the genders of the offspring thereof can identified through utilizing the specificity molecule marker, and the laminaria gender differentiation in basis of the cell and molecule level can be studied.

A method of producing male or female sterile plants comprising the steps of transforming plant material with a polynucleotide which encodes at least one enzyme which reacts with a non-phytotoxic substance to produce a phytotoxic one, and regenerating the thus transformed material into a plant, wherein the said non-phytotoxic substance is applied to the plant up to the time of male or female gamete formation and / or maturation, so that the non-phytotoxic substance provides for the production of a phytotoxic one which selectively prevents the formation of or otherwise renders the said gametes non-functional, wherein the enzyme is expressed preferentially in either male or female reproductive structures, characterised in that (i) the non-phytotoxic substance is selected from the group consisting of ester derivatives of non-phosphonate herbicides which herbicides are directly phytotoxic to non-green tissue, D-alpha amino acids, peptide derivatives of non-protein D-alpha amino acids, S-enantiomers of aryloxyphenoxypropionates and S-enantiomers of ester derivatives of aryloxyphcnoxypropionatcs and (ii) the enzyme is selected from the group consisting of carboxylesterases, D-amino acid oxidases, D-amino acid dehydrogenases, D-amino acid racemases, 2-arylpropionyl-CoA epimerases, alpha-methylacyl-CoA racemases, thioesterases and acyl-CoA synthetases.

The invention discloses a method for rapidly obtaining a tripod of siraitia grosvenorii. The method comprises the following steps: soaking a female flower at a reduction mitosis stage by using a novel chromosome doubling agent Oryzalin with high induction efficiency and low toxicity according to the floral traits of female flower megaspore mother cells in reduction mitosis to inhibit normal separation of chromosomes in reduction mitosis and induce the production of unreduced female gametes; and after the female flower to be treated blooms, pollinating by using diploid male plant pollen to obtain a tripod descendant. By adopting the method, the technical problems of long cultivation time, the need of screening unreduced gametes during cultivation of the siraitia grosvenorii tripod by manual induction of the unreduced female gametes and the like in the conventional tripod cultivation method of siraitia grosvenorii are solved effectively, the tripod can be successfully obtained conveniently and easily in one growing period, and technical support is provided for the creation of abundant new siraitia grosvenorii tripod germplasms as well as the cultivation of new seedless tripod varieties.

Compositions and methods for producing a plant population lacking sexually derived embryos are provided. Compositions include suppression cassettes encoding polynucleotides and promoters resulting in parthenogenesis. Further provided are parthenogenesis genetic elements used to prevent sexual reproduction in self-reproducing plants.Methods include: utilizing maternal embryo defective recessive mutations which are maintained as a sterile inbred maintenance system, allowing generation of populations that are homozygous for recessive mutant alleles, but transgenically complemented. Methods include utilizing a toxin genes expressed via egg-cell specific promoters, creating a dominant, embryo-less phenotypes, non-transmittable through female gametes. Resultant hemizygous plants are transformed with egg-cell promoters driving the antidote, a pollen ablation PTU and a seed color marker for identification of transgenic seed. The generation of a plants 50% female fertile, having seed which when grown in the next generation will yield plants with 50% viable transgenic seed, and 50% non-viable embryo-less seed.

The invention discloses a pinus massoniana embryogenic callus proliferation and maintenance culture method. The method includes: placing a filamentous embryogenic callus cell cluster obtained by induction of 3-4 weeks of a conventional somatic embryo and an embryogenic callus obtained by separation of a female gamete into a petri dish with a culture medium I; removing dead cells and unnecessary tissues in embryogenic calli in 4-6 weeks; transferring into a centrifugal tube, adding a culture medium II, mixing and well shaking; adding an embryogenic culture obtained after well shaking into a petri dish with a culture medium III; after 6-8 weeks, directly transferring an appropriate size of the embryogenic calli into a culture medium IV for normal subculture in a subculture period of 10-14d to finally obtain a pinus massoniana embryogenic cell line which is fast in growth, vigorous and available for long-term subculture. Evident browning and water soaking of the embryogenic calli in long-term subculture are avoided, the embryogenic calli grow fast, proliferation effects are stably high, the proliferation coefficient reaches 25 or above, and high economic benefits and social benefits are achieved.

The invention discloses a reproductive technique of an intelligent rice sterile line. The reproductive technique is adopted to achieve the effect that reproduced seeds of the intelligent rice sterile line are not required to be subjected to fluorescent sorting. In the method, the intelligent rice sterile line is used as a maintainer line and is hybridized with the corresponding rice sterile line; pollen grains carrying transgenes of the intelligent rice sterile line are aborted and cannot be fertilized, pollen grains without the transgenes are normally fertilized with female gametes of the sterile line and are fruited, and therefore, the fruited seeds of the sterile line are all genetically-modified sterile line seeds and can be used for producing hybrid rice seeds. In the method, the fluorescent sorting procedure is not required in the intelligent rice sterile line reproductive process, and only the reproduction of the maintainer line requires the fluorescent sorting, and therefore, the reproductive process of the sterile line is simplified, the reproductive efficiency of the sterile line is improved and the seed industry benefit is increased.

The invention relates to a building body of mediating plant fertility and application thereof. The building body comprises (a) a first nucleotide sequence, wherein male fertility of a plant is restored when the first nucleotide sequence is expressed; (b) a second nucleotide sequence, wherein formation or function of fertile male gametes in the plant is inhibited when the second nucleotide sequence is expressed, (3) a third nucleotide sequence, wherein inhibition or neutralization on a herbicide is formed when the third nucleotide sequence is expressed. The building body of mediating plant fertility and application thereof disclosed by the invention are applied to a method for keeping a homozygous recessive state of a male sterile plant, so that the building body is good in benefit, and simple to operate; the generated male sterile descendant is non-transgenic; the building body is convenient to identify, high in sorting efficiency, good in effect and especially applicable to the situation that the contained building body is inconveniently identified in the seed stage when the building body is applied to the method for producing seeds from the plant with female gametes and male gametes.

The invention relates to an oryza sativa male sterility gene OsFIGNL1 (oryza sativa Fidgetin-like 1) and an application thereof. The nucleotide sequence of the gene OsFIGNL1 is shown as SEQ ID NO:1. One function lost mutant of OsFIGNL1 is obtained through <60>Co-gamma mutagenesis, and the mutant in a field planting environment shows the following characteristics: vegetative growth of plants are normal, male plants are sterile, pollen is 100% typically abortive, and development of female gametes is not affected. Meiosis squashing operation shows that chromosome behaviors are severely affected by function loss of OsFIGNL1 during meiosis, and developmental defects of microspores are caused finally. The mutant can be applied to breeding of new dominant varieties of oryza sativa hybrids. The gene OsFIGNL1 can serve as a functional gene for regulating meiosis of oryza sativa to be applied to oryza sativa heterosis creation work in future and has huge development and utilization value and broad market application prospect.

The present invention relates to methods for increasing the developmental competence of at least one mammalian germ cell, gamete, zygote, early embryo, implanted blastocyst and / or embryo by administrating a compound which is capable of inhibiting the de novo biosynthesis of sterols and thereby establishing cellular conditions that improve their development and survival. The invention also relates to methods for increasing the sterol efflux prior to fertilisation from at least one mammalian ovary, oocyte, female gamete, or ovary derived cell surrounding an oocyte by administrating a compound which is capable of promoting the sterol efflux and thereby reducing the phopholipid / sterol ratio of said cells.

The invention relates to an establishing method of a Cnobilis inbred line, which can quickly purify the genome of the Cnobilis, so as to accelerate variety breeding. Individuals containing female and male germ cells and appearing as female, namely hermaphrodites, are selected through micrographic examination, an egg is fertilized by sperm in the same individual through induced parturition hastening or dissection fertilization, the amount of the sperms are increased or reduced artificially by taking measures to artificially interfere and adjust the relative proportion of the male and the female gametes in the water, and hatching, breeding and cultivating are carried out according to the ordinary technology of the species, so as to establish the inbred line. The establishing method of the Cnobilis inbred line is simple to operate, requires no special chemical reagent, and is easy to popularize and apply.

The invention belongs to the technical field of seaweed cultivation, and discloses a method for producing caulerpa lentillifera seedlings by utilizing artificial sexual breeding. The method comprisesthe following steps: selecting caulerpa lentillifera which grows for more than two months and is good in growth vigor, cleaning miscellaneous algae on the surfaces of the caulerpa lentillifera, and pre-culturing the caulerpa lentillifera to serve as seed caulerpa lentillifera; inducing the seed caulerpa lentillifera at a low temperature (15-20 DEG C), culturing the seed caulerpa lentillifera underconventional conditions, and changing water at regular intervals until sexual maturity is observed; and transferring the seed caulerpa lentillifera into a disinfected nursery pond laid with an adherance, giving certain illumination stimulation, culturing and releasing male and female gametes, then transferring the seed caulerpa lentillifera out of the nursery pond, and culturing fertilized eggs for 1-2 months after field planting to obtain artificial seedlings. According to the method disclosed by the invention, the caulerpa lentillifera seedlings can be produced by utilizing sexual breeding,the method is suitable for large-scale breeding of the caulerpa lentillifera seedlings, the breeding efficiency is high, and seedling production is not limited by seasons.

The invention belongs to the field of seaweed molecular genetics, and relates to an enteromorpha gender specific molecular marker and application thereof. The enteromorpha gender specific molecular marker is characterized in that one enteromorpha female gametophyte DNA specific molecular marker and one enteromorpha male gametophyte DNA specific molecular marker are provided, wherein the nucleotidesequences of the enteromorpha female gametophyte DNA specific molecular marker and the enteromorpha male gametophyte DNA specific molecular marker are respectively SEQ ID NO. 3 and SEQ ID NO. 4, theSEQ ID NO. 3 is obtained by amplifying a primer sequence SEQ ID NO. 1, and the SEQ ID NO. 4 is obtained by amplifying a primer sequence SEQ ID NO. 2. The specific molecular marker is completed by detecting the specific fragment through PCR (polymerase chain reaction), and the process comprises the following steps of extracting to-be-detected enteromorpha genomic DNA, performing PCR amplification,and performing agarose gel electrophoresis detection on an amplification product. The specific primer is obtained by referring to existing high-throughput data result in a laboratory, combining genomeDNA extraction, target fragment amplification and recovery, cloning by using pGEM-T vector and escherichia coli competent strain TOP10 and the like. By means of the specific molecular marker, ploidyof a large number of enteromorpha algae and gender of gametophytes can be rapidly, simply and accurately identified, and an important molecular tool is provided for enteromorpha life history researchand variety breeding.

The invention provides a novel method for breeding kelp seedlings with a gametophyte cloning method, comprising the following steps of: respectively separating a female cell on a female gametophyte and a male cell on a male gametophyte under a microscope by using a capillary tube separation technology; then respectively cloning the female cell and the male cell by using a cloning vegetative propagation technology to respectively grow the female cell and the male cell into female and male gametophytes; continually breeding the gametophytes; when the gametophytes are bred to a fixed amount, damping a formed cell cluster by a cell stamping machine; mixing the female and male gametophytes and respectively releasing ova and sperms by the female and male gametophytes to spontaneously fertilize;putting fertilized zygotes into nutritive salt to be cultured; and after 10 weeks, obtaining seedlings with the height of about 1.5 cm. According to the invention, a fertilization process and the control of temperature and illumination are realized by manual operation, so that the growth period of offspring seeds is greatly shortened; meanwhile, the female cell and the male cell are separated from the gametophytes by using the capillary tube separation technology to clone and culture, so that the quality and the yield of the seedlings are improved, the cultivation of the kelp can be satisfied; and by utilizing the novel method, the cost for breeding the kelp seedlings is low.

The invention discloses an application of a rice gene OsDES1. The invention discloses for the first time that the rice gene OsDES1 has a biological function of regulating and controlling plant fertility. The function of the gene is verified in rice based on a gene knockout experiment and an overexpression experiment, a CRISPR / Cas9 gene knockout vector is constructed to transform Nipponbare of rice, female gametes of transgenic plants grow abnormally, and fertility is reduced; after the pCUBi1390-OSDES1 overexpression vector is used for converting a mutant des1, the fertility of the female gamete of the transgenic plant is recovered, namely, the female plant is partially aborted due to OSDES1 gene function loss type mutation, and the fertility is recovered to be normal through the overexpression of the gene. The rice OSDES1 gene is not completely aborted after the function is lost, part of seeds can be harvested, the rice OSDES1 gene can form favorable supplementation with other femalesterile genes, and a new gene resource is provided for rice intelligent restorer line breeding.

The invention relates to a recoverable intra-uterine system comprises a housing (1) capable of containing one or a plurality of elements selected from among the group comprising an embryo, male and / or female gametes, a fertilized oocyte, an unfertilized ovum and a combination of these elements, the housing (10) having along an axis (X) a distal end (12) and a proximal end (13), and a device (20) for holding the recoverable intra-uterine device in the uterus. The holding device (20) is arranged at the proximal end (13) of the housing (10) and includes at least one holding arm (23) in the uterine cavity capable of taking at least two positions: - one free position in which at least one holding arm (23) is separated from the axis (X); and - a retracted position in which at least one holding arm (23) is substantially parallel to the axis (X). Use in medically assisted reproduction techniques.

The invention relates to the technical field of plant gene engineering and especially relates to isolation and cloning and functional identification of a paddy rice S5-site hybrid embryo sac fertility gene ORF3 and a use of the paddy rice S5-site hybrid embryo sac fertility gene ORF3 in paddy rice breed improvement. The gene ORF3 codes an Hsp70 albuminoid and has functions of folding and transferring cell proteins and transducing stress signals. Different alleles of the gene ORF3 have insertion / deletion variation of 13bp in a coding zone, and the length of the dominant allele ORF3+ is 13bp more than that of the recessive allele ORF3-. A nucleotide sequence of the recessive allele ORF3- is shown in the formula of SEQ ID NO: 1 and a nucleotide sequence of the dominant allele ORF3+ is shown in the formula of SEQ ID NO: 3. In a S5 oryza sativa L. ssp. Indica / oryza sativa L. ssp. Japonica hybrid, the dominant allele ORF3+ can protect female gametes so that a maturing rate of the S5 oryza sativa L. ssp. Indica / oryza sativa L. ssp. Japonica hybrid is improved. The paddy rice hybrid embryo sac fertility gene has an important meaning for using oryza sativa L. ssp. Indica / oryza sativa L. ssp. Japonica hybrid vigor.

The invention discloses a method for directly identifying homologous recombination of higher plant DNA, which comprises the following steps: inducing chromosome doubling of embryo sac by taking a plant to be identified as a female parent to obtain 2n female gametes; obtaining a hybrid triploid by the hybridization of the 2n female gametes and male gametes of a paternal plant; screening to obtain a codominant molecular marker primer; conducting PCR (Polymerase Chain Reaction) amplification with the screened codominant molecular marker primer; conducting capillary electrophoresis analysis to the amplification product: if only one of codominant molecular markers of the female parent occurs in the amplification product of the triploid, homologous recombination does not occur at relevant locusof the molecular marker, and if all genetic information of the female parent occur in the amplification product of the triploid, homologous recombination occurs to the pistillate parent at the relevant locus of the molecular marker and heteroduplex DNA is formed. The method has important theoretical and application values on plant genetic analysis, discrimination of molecule marker technical method and guidance on triploid breeding and the like.

The invention discloses an artificial insemination method for Filicinae, and belongs to the technical field of plant artificial insemination. In order to solve the problem of low seedling rate of pteridophytes, the invention provides an artificial insemination method of Filicinae, which comprises the following steps: inoculating spores into a culture medium, obtaining female gametophytes and malegametophytes of the Filicinae by using a conventional spore culture method, after a sexual organ is mature, sucking mature sperms and a prepared induction liquid, sequentially dropwise adding the mature sperms and the prepared induction liquid to a mature archegonium of the female gametophyte, and enabling the sperms to enter the archegonium to be combined with ova under the guidance of the induction liquid, so as to complete artificial insemination. According to the sperm taking and artificial directional insemination technology provided by the invention, the fertilization rate of the gametophyte can reach 30%-50% and is 3-5 times that under conventional culture conditions, the phenomenon that a single gametophyte has two embryonic developments often occurs, and the hybridization technology of pteridophytes is simpler, more convenient and easier to implement.

The invention relates to the technical field of marine product production, and discloses a method for increasing the artificial hatching rate of tegillarca granosa, male and female gametes of the tegillarca granosa are artificially obtained and are artificially fertilized in a seawater environment added with calcium and magnesium ions, the pH value of a water body is 8.0-8.2, the concentration of the added calcium ions is 200-300 mg / L, the concentration of the magnesium ions is 150-200 mg / L, and the sperm-egg ratio is controlled to be 100: 1-200: 1; the method is easy to operate, low in cost and short in operation time, and development and growth of the tegillarca granosa larvae are not affected.

A breeding method for solving a color difference problem of colored fresh-eating corn grains is characterized in that conventional colchicine or herbicide induction and other chemical induction methods are adopted, a female gamete chromosomal of an amphiphilic selfing line is induced for doubling to make genotypes including a genotype controlling grain colors highly homozygous; followed by, according to color standards of varieties, grain color and depth phenotypes are subjected to genotype identification and selection; an amphiphilic selfing line with pure and uniform colors and consistent genotypes and phenotypes is obtained, finally, pure colors and uniform color depth of hybrid grains in a fresh-eating period and a hybrid mature period are achieved, and the color ratio is consistent to a theoretical value; and the breeding method has the technical effects that the color genotypes are highly homozygous, the grain colors in the fresh-eating period are pure and uniform, and the color phenotype ratio is consistent to the theoretical value. No matter compared with conventional techniques or molecular techniques, the method has the advantages of good color effect, accurate breeding result, high breeding efficiency, fast homozygous speed, low economic cost, simple techniques, easy operation, and convenience for application.

The invention relates to a chromosomal localization method for FSML (female-specific marker of Laminaria japonica Aresch)-1488. The method comprises the steps of preparation of DNA (deoxyribonucleic acid) with target fragment FSML-1488 plasmids, preparation of gametophyte chromosomes of the Laminaria japonica Aresch, preparation of probes, and fluorescence in situ hybridization (FISH). The method has the advantages of preparing the high-quality chromosomes on the basis of obtaining protoplast of antiphyte and gametophyte of the Laminaria japonica Aresch by the optimal coordination of various tool enzymes such as macerozyme, abalone enzyme, cellulose and pectinase, breaking through the technology bottleneck of the FISH technology in the research of algology, and successfully locating the FSML-1488 with the length being 1488bp on the chromosomes.

The present invention features immunogenic compositions based on pre-fertilization or post-fertilization antigens expressed in the circulating gametocytes in the peripheral blood of infected persons or on the malaria parastes' stages of develop-ment in the mosquito midgut including extracellular male and female gametes, fertilized zygote and ookinete. The invention also features methods to prevent the transmission of malaria using the immunogenic compositions of the invention.

The invention provides a method for facilitating ovulation of kelp female gametophytes. The method includes the steps: resuscitating kelp germplasm filaments in seawater, crushing kelp germplasms, andfiltering crushed liquid to obtain filament fragments with the lengths of 3-6 cells; performing aerated suspension culture on the filament fragments under the conditions of the illumination intensityof 6000+ / -300lx, the illumination cycle of L:D=12h:12h and the temperature of 15+ / -1 DEG C to collect eggs; culturing the collected eggs to obtain young kelp sporophytes, culturing the young kelp sporophytes, and performing culture by running water at the stage of young sporophytes of two rows of cells. According to the method, female kelp germplasms can ovulate in 11-13 days, ovulation time is shortened, ovulation rate is high, ovulation synchronization is good, ovulation rate within two days can reach 65-70%, and egg development rate can reach 70-80%. According to the built culture method of young sporophytes, the aberration rate of the young sporophytes can be reduced to 50-60%.

The present invention relates to methods for increasing the developmental competence of at least one mammalian germ cell, gamete, zygote, early embryo, implanted blastocyst and / or embryo by administering a compound which is capable of inhibiting the de novo biosynthesis of sterols and thereby establishing cellular conditions that improve their development and survival. The invention also relates to methods for increasing the sterol efflux prior to fertilisation from at least one mammalian ovary, oocyte, female gamete, or ovary derived cell surrounding an oocyte by administering a compound which is capable of promoting the sterol efflux and thereby reducing the phospholipid / sterol ratio of said cells.

The invention relates to a cultivation method and development and application of a new variety of eucommia ulmoides triploid. The cultivation method comprises the following steps: a, inducing 2n female gametes: treating female flower buds on eucommia ulmoides female plant bodies by adopting 0.05-0.5 wt% colchicine solution and treating at the high temperature of 40-50 DEG C; b, artificial pollination; c, collecting seeds; d, sowing and seedling raising; e, ploidy detection; f, grafting, wherein buds with the diameter of 0.8-1 cm on the triploid plants are grafted to the diploid plants. The method has the advantage of improving the yield of the eucommia triploid.

The invention discloses a molecular marker for identifying fertility of hybrid progeny of common wild rice and cultivated rice and an application of the molecular marker. The rice molecular marker isnucleotide, corresponding to the 2099th position in sequence 1 of a sequence table, in a rice genome, namely, T or C. Experiments prove that the rice molecular marker is related to fertility of distant hybrid progeny of rice, especially fertility of female gametes; in the hybrid progeny of the common wild rice and the cultivated rice, fertility of TT genotype rice with the rice fertility molecularmarkers in two chromosomes being both T and fertility of TC genotype rice with the rice fertility molecular markers being T in one chromosome and C in the other chromosome are lower than the fertility of CC genotype rice with the rice fertility molecular markers in two chromosomes being both C; the female gametes of the TT genotype rice and the TC genotype rice develop abnormally, while female gametes and male gametes of the CC genotype rice develop normally. Therefore, the molecular marker can be used for detecting the fertility of the distance hybrid progeny of the rice, especially the fertility of the female gametes.