48 results about "Microspore" patented technology

The invention relates to the technical field of plant tissue culture, and provides a method for improving the embryo emergence rate of free microspores of non-heading Chinese cabbage by using 5-AzaC, and in the microspore culture stage, 5-AzaC with the concentration of 0.005-0.020 mg is added into a microspore culture medium. The invention designs a method for increasing the embryogenesis rate of free microspores of the non-heading Chinese cabbage by adding 5-AzaC into a microspore culture medium of the non-heading Chinese cabbage. According to the method, the embryogenesis rate of the free microspores of the non-heading Chinese cabbage can be remarkably improved, a non-heading Chinese cabbage microspore culture system is more easily established, the haploid breeding efficiency is improved, and the breeding process is accelerated.

The invention relates to a haploid induction method based on a brassica napus phospholipase gene BnaPLA2-a and application of the haploid induction method, the BnaPLA2-a gene in brassica napus is knocked out through a gene editing technology, a mutant material capable of inducing haploid generation is obtained, and the application blank of a phospholipase pathway in dicotyledon haploid induction is filled. Compared with a traditional microspore culture technology, the invention provides a brand-new haploid induction path which is derived from the rape and is used for the rape. The invention provides a brand new technical tool and germplasm resources for genetic breeding of brassica napus. Meanwhile, the invention discloses a gene, a mutant, a creation method and application in breeding.

The invention relates to the technical field of bioengineering, in particular to a method for extracting exosomes from microspore ganoderma lucidum immune protein.The method comprises the steps that microspore ganoderma lucidum immune protein extraction comprises pretreatment, extraction, centrifugation, salting-out, dialysis and chromatographic purification; exosome is extracted from cell culture supernate by using the immune protein through an immunoaffinity capture method, and the method comprises the steps of incubation combination, centrifugal separation and elution purification. The microspore ganoderma lucidum immune protein is specifically combined with the exosome for the first time, the extraction method is efficient, mild and low in cost, and the extracted exosome is high in purity and good in integrity, has double activities of the exosome and the ganoderma lucidum immune protein, and has wide application prospects in the fields of disease diagnosis, drug delivery, immune regulation and the like.

The present invention relates to the technical field of plant cultivation, and provides a method for rapidly screening high-embryo-rate radish materials. The method screens high-embryo-rate materials by analyzing the expression levels of the ENHANCER OF SHOOT gene in different radishes. There is a negative correlation between the expression level of the ENHANCER OF SHOOT gene in the radish and the embryo formation rate of the corresponding microspores. When the expression level of the ENHANCER OF SHOOT gene in the measured radish is the lowest, the corresponding microspore embryo formation rate is the highest, and this material is the screened high-embryo-rate radish material. By counting the embryo formation rates of microspores of different radish varieties, the present invention finds that there is a negative correlation between the embryo formation rate of radish microspores and the expression level of the ENHANCER OF SHOOT gene. Based on this, radish varieties with high embryo formation rates can be rapidly screened, effectively saving the manpower and time for screening high-embryo-rate materials.

Plant cell fate and development is altered by treating cells with cellular reprogramming factors. Embryogenesis inducing embryogenesis factor genes and / or morphogenic developmental genes are used as cellular reprogramming factors, specifically comprising polypeptides or polynucleotides encoding gene products for generating doubled haploids or haploid plants from gametes. Maize microspores treated by contacting the isolated cells with an exogenous purified, recombinant embryogenesis inducing embryogenesis factor gene products and / or morphogenic developmental gene polypeptide results in embryogenesis. The gametes of a maize plant develop into embryoids when transformed with a genetic construct including regulatory elements and structural genes capable of acting in a cascading fashion to alter cellular fate of plant cells. Embryogenesis factor proteins and / or developmental morphogenic proteins expressed from a genetic construct are used for ex situ treatment methods and for in planta cellular reprogramming.

The present invention provides a method for detecting single-cell gene sequence variation in rice and its application, which belongs to the field of agricultural technology. The method of the present invention mainly includes three aspects: sampling, sequencing and bioinformatics analysis, wherein the sampling is to obtain a single rice microspore sex cell. The present invention successfully isolates a single rice microspore sex cell, and improves the defects of the existing rice single-cell variation detection technology process, such as weak integrity and detection consistency that need to be improved. In addition, the authenticity of the mutation site is identified and confirmed by heavy ion mutagenesis technology. The method of the present invention broadens the source range of rice single-cell mutagenic materials, and provides new ideas and methods for creating more diverse breeding materials and conducting more efficient mutagenic breeding. At the same time, microspore sex cells serve as male gametes, and the mutations carried in the genome can be passed into the offspring through hybridization, which can realize the inheritance and fixation of mutations, and also provide a way to quickly utilize excellent mutant genes.

This disclosure provides automated, high-throughput methods and systems for generating microspore-derived doubled haploid structures, plantlets, and crop plants, and progeny thereof. Methods of integrating computer vision and artificial intelligence into a variety of steps including microspore selection, culture, sorting, and genotyping are disclosed. Multiplexed genome-editing of microspores through cargo delivery, non-destructive sampling, and genotyping are also described.

The invention discloses a method for creating a new bolting-resistant radish germplasm by using a microspore culture technology, and belongs to the technical field of radish breeding. The method comprises the following steps: hybridizing a radish material R-16 and a bolting-resistant radish material LR-75, carrying out microspore culture, embryoid induction and regeneration on hybrid offspring plants to obtain double haplobionts, carrying out selfing, and carrying out character evaluation and screening on selfing offspring plants to obtain bolting-resistant double haplobionts with stable characters. Experimental results show that by selecting the parent material and optimizing the hormone concentration of the induction culture medium, the embryo emergence rate of the radish material is remarkably increased, and the homozygous bolting-resistant radish germplasm can be efficiently and quickly obtained. According to the method, a rich material foundation is laid for heterosis breeding of the bolting-resistant radishes, the breeding procedure is simplified, the breeding cost is reduced, the breeding process of new varieties of the bolting-resistant radishes is accelerated, and the method has great economic significance and wide application prospects.

The present application relates to the technical field of antibacterial technology, and particularly relates to a natural antibacterial agent and application thereof. The present application provides a method for preparing a composite antibacterial preparation by using Chinese medicine pomegranate peel and Bacillus subtilis AC-1 as raw materials. The composite antibacterial agent has the characteristics of strong resistance to gram-negative bacteria Escherichia coli, gram-negative bacteria Staphylococcus aureus, Staphylococcus pseudintermedius, pathogenic fungus Gymnoascus ruber, Microsporum canis and Microsporum gypseum.

To provide a medicinal drug or a food product produced from small-spored Ganoderma lucidum-derived GMI.SOLUTION: Provided is a medicinal drug or a food product produced from small-spored Ganoderma lucidum-derived GMI (Ganoderma immunoregulatory protein) . The medicinal drug or the food product is produced by using small-spored Ganoderma lucidum-derived GMI (Ganoderma immunoregulatory protein) fermentatively produced by genetically modified Pichia pastoris Ey72 strain, GMI has an amino acid sequence shown by SEQ ID NO:1, and is used for treatment, prevention or amelioration of pulmonary fibrosis caused by a factor other than a virus.SELECTED DRAWING: Figure 1

The invention belongs to the technical field of crop variety breeding in modern agriculture, and discloses a method for creating homozygous tetraploid rice by using haploid anther, which makes full use of the characteristic that the number of chromosomes in cells subjected to meiosis is halved, and microspores formed by meiosis are subjected to anther culture at a proper development stage, so that the homozygous tetraploid rice is created. The method comprises the following steps of: a, selecting a diploid rice line with excellent characters as a parent material, and carrying out continuous secondary induction by colchicine to form a callus, and differentiating to emerge seedlings to form a stable homozygous tetraploid rice line; b, selecting anther with microspores in the late uninucleate stage during observation; c, inducing anther calluses; e, differentiating the anther calluses to emerge seedlings; f, seedling callus differentiation and rooting; and g, determining the authenticity of a homozygous tetraploid plant formed by anther culture through morphological and cytological technologies (flow cytometry). The method is simple, low in cost and easy to operate.

A method for efficiently inducing genomic variation in rice microspore sexual cells, relating to the technical field of plant cell engineering. In the method, a rice male gamete (early uninucleate microspore) is used as a mutagenic material, it is attempted, for the first time, to irradiate rice microspore sexual cells with heavy ions (4 Gy) and γ rays (10 Gy), experimental technologies such as fluorescence microscope observation, FDA-PI dyeing, and immunofluorescence detection are taken into account to determine three indicators comprising a target period for irradiation sample sampling, an appropriate heavy ion / γ ray radiation dose, and an appropriate sampling time after the end of radiation, and a relatively complete rice microspore sexual cell mutagenesis system is established. The method involves clear indicators, strong operability, can support batch mutagenesis operations, and can provide a technical system support for the efficient batch production of rice sexual cell mutants.

The invention discloses a multiple qPCR (quantitative polymerase chain reaction) detection reagent for detecting pathogens of a superficial fungal disease. The multiple qPCR detection reagent comprises specific primers and probes for detecting trichophyton rubrum, epidermophyton flocculent, microsporum canine, trichophyton beard, candida parapsilosis, candida albicans, malassezia furfur and gliocladium breviculare, wherein the specific primers and the probes are used for detecting trichophyton rubrum, epidermophyton flocculent, microsporum canine, trichophyton beard, candida parapsilosis; according to the invention, different fluorescently-labeled probes are utilized to add specific primers and probes of multiple pathogens into a reaction system to detect multiple targets at the same time, and the method has the advantages of good specificity, high sensitivity, high detection speed, capability of detecting multiple pathogens at the same time, rapidness, simplicity, convenience and economy; a convenient method is provided for rapid detection of superficial fungal disease pathogens, and the kit has important significance in epidemiological research, clinical early molecular diagnosis and the like.

A switch to haploid embryogenesis is controlled by the activity of histone deacetylases (HDACs). Blocking HDAC activity with HDAC inhibitors (HDACi), e.g., trichostatin A (TSA), in Brassica napus, B. rapa, Arabidopsis thaliana, and Capsicum annuum male gametophytes leads to a large increase in the proportion of cells that undergo embryogenic growth. In B. napus, treatment with one specific HDACi (SAHA) improves the conversion (i.e., germination) of these embryos into seedlings. Existing methods of culturing microspores of angiosperm plants following stress to produce haploid embryos, haploid plants, and double haploid plants can be improved by adding HDACi to the culture medium. Advantageously, species hitherto recalcitrant to haploid embryogenesis via microspore culture are rendered useful when using HDACi. Haploid and double haploid plants are of industrial application in the plant breeding programmes.

The invention discloses a method for preparing a microspore embryo by using a microspore specific expression rice OsBBM1 gene and application of the method in preparation of the microspore embryo, and belongs to the field of haploid breeding. The rice microspore specific expression transcription factor OsBBM1 is directly cultured through a culture medium, microspore embryogenesis can be carried out without stress treatment, and microspore embryos are obtained. According to the method, the dependency of traditional microspore embryogenesis on species genotypes and culture conditions is overcome, a new thought is provided for the biological effect of the BBM gene in the haploid induction process, and the preparation efficiency of the rice microspore embryo can be improved.

The invention discloses a method for identifying meiosis microspore mother cells and chromosomes of cotton at normal and high temperatures, and belongs to the technical field of plant cell genetics. The method comprises the following steps: determining the length of a flower bud in a tetrad period of a microspore mother cell, selecting the flower buds in different periods before the tetrad period of the microspore mother cell, establishing a corresponding relation between the flower buds in different length ranges and a meiosis process, and observing the chromosome morphology of residual anther in the flower buds of which the meiosis process is determined; and obtaining a chromosome behavior characteristic graph in each period of cotton anther meiosis. Particularly, by establishing the corresponding relation between the flower buds in different length ranges and the meiosis process, more systematic observation of the cotton meiosis microspore mother cells is achieved, and more excellent cotton microspore mother cell chromosome flakes can be prepared. The method is simple to operate, does not need special equipment, and has important practical application significance.

The invention provides an anther liquid culture method of hybrid highland barley, and belongs to the technical field of plant tissue culture. The method comprises the following steps: (1) refrigerating young ears of hybrid highland barley, disinfecting, and collecting anther; (2) inoculating the collected anther into a barley pre-culture solution for culture to obtain a pre-cultured anther; (3) inoculating the pre-cultured anther into a barley induction culture medium for culturing to obtain a callus; (4) inoculating the callus into a differential culture medium for culturing to obtain a green regenerated plant; and (5) inoculating the green regenerated plant into a strong seedling culture medium for culturing to obtain the highland barley seedling. According to the method, single ears are cultured in dishes, the damage and cross contamination probability of a mechanical dissociation mode to microspores is reduced, and the method is a small-scale culture mode suitable for being limited by material drawing. The method solves the problems that the number of materials of hybrid highland barley is limited and the activity of anther free microspores is relatively weak, and has a wide application prospect.

The invention relates to the technical field of agricultural cultivation, and particularly discloses a spring rape variety breeding method suitable for large-density planting, which comprises the following steps: collecting and establishing a germplasm resource library, performing preliminary screening according to local conditions, evaluating the growth speed, close planting resistance, disease resistance and quality in detail, and screening out excellent germplasm resources; selecting parents according to germplasm resource characters, designing a hybridization combination scheme, and performing artificial hybridization operation to obtain filial generations; filial generation F1-generation microspores are extracted for in-vitro culture, haplobionts are obtained and doubled into a homozygous DH line, and plants meeting the breeding target are screened out; designing a high-density planting test scheme, carrying out a planting test, regularly observing and recording plant characters, and screening out a spring rape variety which is excellent in performance and meets a breeding target; the breeding steps of the system comprise germplasm resource screening, hybrid combination construction, microspore culture, DH line breeding and high-density planting test and screening.

The present application relates to an anti-browning method in the process of coconut microspore embryo culture, and belongs to the field of coconut tissue culture technology. The anti-browning method in the process of coconut microspore embryo culture of the present application comprises the following steps: S1, anther collection and pretreatment; S2, anther observation, disinfection and separation; S3, anther culture and embryo culture. The anti-browning method in the process of coconut microspore embryo culture provided in the present application adds PVP to the culture medium to counteract the browning phenomenon during the induction of coconut microspore embryos; and when cutting the explant, dip it in citric acid solution to reduce the browning caused by physical damage; ascorbic acid (vitamin C) is also added to the culture medium to adjust the pH and counteract the browning phenomenon; in addition, low concentration TDZ (thiadiazole) is used as an induction hormone to improve the efficiency of embryo induction.

The present invention is directed to methods of chemical reprogramming of a microspore plant cell using tyrosine kinase inhibitors. The tyrosine kinase inhibitors described facilitate microspore embryoid regeneration in maize, including maize varieties previously recalcitrant to microspore embryoid regeneration, and in a variety of other plant species.

The invention provides application of a glutathione solution in increasing the yield of highland barley microspore calluses, and belongs to the technical field of plant breeding. According to the method disclosed by the invention, the yield of the highland barley microspore calluses can be obviously improved by soaking highland barley young ears in a glutathione (GSH) aqueous solution or adding glutathione (GSH) into a highland barley microspore callus induction culture medium.

The application discloses a kind of multiple qPCR detection primer combinations of superficial mycosis pathogen, it includes the specific primer and probe of detecting Trichophyton rubrum, Trichophyton mentagrophytes, Epidermophyton floccosum, Microsporum canis, Microsporum audouinii, Microsporum gypseum, Candida albicans, Malassezia furfur;The application detects pathogen target gene using real-time fluorescent quantitative PCR technology;The method has the advantages of short time consumption, high specificity, low cost, and can detect multiple pathogens at a time, provides a convenient method for the detection of superficial mycosis pathogen, and has important significance for the early molecular diagnosis of superficial fungal infection patients.

The methods have application to the tissue culture of plant cells, ovules and microspores. Automated imaging, automated cell tracking and predictive modeling are used to develop methods that predict the likelihood that a plant cell will develop into a desired phenotype and / or which cellular reprogramming factors will assist in this development. The methods taught herein can also be used to evaluate the toxicity effects of compounds on plant cells, predict genotypic responses to tissue culture and cellular reprogramming factors, determine cell ploidy status, and predict other types of cell phenotype development.

The invention relates to the technical field of rice microspore culture, in particular to an improved rice multi-variety free microspore efficient culture method based on a callus induction culture medium, which comprises the following steps: step 1, sampling and pretreatment, collecting young panicles of which floret microspores in the middle of the rice young panicles develop from a mononuclear middle-late stage to a binuclear early stage in the morning; 2, preparing an extracting solution and a culture medium: preparing the extracting solution, an induction culture medium, a differential culture medium and a seedling strengthening and rooting culture medium; step 3, separating microspores and inducing calluses, preparing microspore suspension by utilizing the induction culture medium and the microspores, and placing the microspore suspension in an incubator with the temperature of 26 DEG C and the relative humidity of 60-75% for dark culture; 4, plant regeneration, seedling strengthening and rooting; 5, field transplanting, through the culture method, the problem that free microspores are prone to pollution in the culture process can be effectively solved, and the induction effect of the induction culture medium is good.

The invention provides an asparagus microspore induction culture medium and a culture method, and relates to the technical field of crop cultivation. The asparagus microspore induction culture medium comprises the following components, 50 to 1500 mg / L of KNO3, 300 to 340 mg / L of NH4H2PO4, 155 to 195 mg / L of CaCl2. 2H2O, 18 to 224 mg / L of MgSO4. 7H2O, 60 to 700 mg / L of K2SO4, 7.3 to 9.5 mg / L of MnSO4. 4H2O, 2.0 to 4.0 mg / L of ZnSO47H2O, 2.0 to 4.0 mg / L of H3BO3, 0.5 to 1.0 mg / L of KI, 2.0 to 2.4 mg / L of acetylcholine, 20 to 30 mg / L of spermidine, 5.0 to 6.0 mg / L of proline, 1.5 to 2.5 mg / L of glycine, 20 to 240 mg / L of glutathione, 80 to 100 mg / L of inositol, 2.4 to 3.6 mg / L of acetosyringone-220, 2 to 4 mg / L of The addition of acetylcholine, spermidine, proline, glutathione, acetosyringone, epicatechin, hydrocortisone and other additives can promote the induction of the microspore callus.

The present disclosure provides novel methods and compositions for the maturation and regeneration of plantlets from microspore-derived embryos. The compositions provided herein include a liquid maturation composition comprising a first plant hormone and about 430 mM to about 880 mM of a monosaccharide solute, a disaccharide solute, or a polysaccharide solute. The methods provided herein include the steps of contacting a microspore-derived embryo with a liquid maturation medium for a time period sufficient to produce a matured microspore-derived embryo, transferring the matured microspore-derived embryo to a substrate, and regenerating the plantlet from the matured microspore-derived embryo.

The invention discloses a rape high oleic acid Pol restorer line breeding method and application thereof, and relates to the technical field of rape breeding, the breeding method comprises the following steps: hybridizing a high oleic acid content line with a common oleic acid content Pol restorer line fully fertile plant, and combining a rape microspore technology, molecular marker identification and gas chromatography to breed a high oleic acid Pol restorer line material; the molecular marker comprises an oleic acid KASP marker, and primer sequences of the oleic acid KASP marker are shown in SEQ ID NO: 1-SEQ ID NO: 3. According to the molecular marker and the breeding method thereof, single plants can be effectively selected according to genotypes at any period in the breeding process, the selection efficiency is improved, the breeding period is greatly shortened through the rape microspore technology, the breeding efficiency is improved, and the efficient breeding method is provided for breeding of the high-oleic-acid rape Pol restoring line.

The present invention relates to a novel simultaneous multiple all-in-one LOP molecular diagnostic platform structure and a simultaneous multiple molecular diagnostic kit for diagnosing shrimp white spot virus, acute hepatopancreatic necrosis, and microspore infection using same. According to the present invention, shrimp white spot virus, acute hepatopancreatic necrosis, and microspore infection that necessitated PCR analysis can now be easily and simultaneously detected on-site via simultaneous multiple detection.

The present disclosure provides novel methods and compositions for maturation and regeneration of plantlets from embryos of microspore origin. The compositions provided herein include a liquid maturation composition comprising a first plant hormone and from about 430 mM to about 880 mM of a monosaccharide solute, a disaccharide solute, or a polysaccharide solute. The methods provided herein comprise the steps of contacting a microspore-derived embryo with a liquid maturation medium for a period of time sufficient to produce a mature microspore-derived embryo, transferring the mature microspore-derived embryo to a matrix, and regenerating the plantlet from the mature microspore-derived embryo.

The invention discloses a method for preparing a plant haploid by using a microspore specific expression NtBBM1 or NtBBM2 gene and application of the method in preparation of angiosperm haploid, and belongs to the field of haploid breeding. Tobacco microspore specific expression transcription factors NtBBM1 or NtBBM2 are directly cultured through a culture medium, microspore embryogenesis can be carried out without stress treatment, plants are cultured, and haploids are obtained through phenotypic screening. According to the method, the dependency of traditional microspore embryogenesis on species genotypes and culture conditions is overcome, a new thought is provided for the biological effect of the BBM gene in the haploid induction process, and the method can improve the crop haploid breeding efficiency.