26 results about "Microspore" patented technology

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.

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 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

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 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 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 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 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 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 the maturation and regeneration of plantlets from microspore-derived embryos. The compositions provided 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 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 provides an application of N-Methylalline in preparation of a fungus inhibitor, and belongs to the technical field of antifungal drugs. According to the invention, the N-Methylalanine is a natural source and has small toxicity, and the IC50 of the N-Methylalanine to human normal pulmonary epithelial cells is greater than 40 [mu] M; the antibacterial effect is obvious, and under the concentration of 100 [mu] M, the inhibition rates on epidermophyton flocculent, trichophyton rubrum and microsporum gypsum reach 99.385 + / -0.8%, 102.17 + / -1.956% and 100 + / -0.115% respectively. The invention provides a new way for treatment of human and animal epidermis fungal infection.

The invention belongs to the field of plant tissue culture, and particularly discloses a potato microspore culture method and application thereof.The culture method comprises the steps that 4-5 mm single-core late-stage flower buds are selected and pretreated at the temperature of 4 DEG C for 3-4 days, then microspores are separated through a high-speed disperser at the speed of 12000-14000 r / min, and after heat shock treatment, the microspores are obtained; the microspores are suspended and separated in an extracting solution composed of mannitol, calcium chloride (CaCl2) and morpholinoethanesulfonic acid (MES), the microspores are cultured in an MS culture solution, and differentiation culture is performed after callus tissue is induced in the dark. According to the method, the corresponding relation between the potato flower bud morphology and the development period of the microspores is established, by optimizing separation, heat shock and culture systems, the survival rate of the microspores reaches 90% or above, the callus induction rate is increased to 8.9 pieces / bud, 90% of regenerated plants are diploids, the diploids can be naturally doubled, and the survival rate of the microspores is increased. The technical problem of homozygous material creation in potato tetraploid breeding is solved.

The present invention discloses a nucleic acid aptamer and a detection kit for detecting Trichophyton rubrum and Microsporum canis, wherein the nucleic acid aptamer includes a nucleic acid aptamer Seq1-LR, and the nucleotide sequence of the nucleic acid aptamer Seq1-LR is a DNA fragment shown in SEQ ID NO.1. The detection kit is a kit comprising nucleic acid aptamers for detecting Trichophyton rubrum and Microsporum canis. The nucleic acid aptamer and the detection kit of the present invention can sensitively, simply, efficiently, specifically and broadly identify and detect Trichophyton rubrum and Microsporum canis, and the nucleic acid aptamer has the advantages of higher affinity and specificity than protein antibodies, no immunogenicity, chemical synthesis, small molecular weight, stability, easy storage and labeling. The nucleic acid aptamer provides a new idea and method for the detection of skin fungal infections, and is expected to achieve rapid and accurate detection of skin fungal infections in the original site clinically.

The invention provides a breeding method for obtaining haplobionts through apple anther culture, and belongs to the technical field of plant breeding. Comprising the following steps: selecting flower buds with microspores in the late uninucleate stage, carrying out low-temperature pretreatment and heat shock treatment, disinfecting and sterilizing, stripping anther, sequentially selecting corresponding culture mediums, and carrying out induction culture, differentiation culture, subculture multiplication and rooting culture to obtain apple test-tube plantlets, acclimatizing and transplanting to obtain apple haplobionts. According to the method, low-temperature pretreatment cooperates with short-time heat shock treatment, the culture medium in each culture stage is optimized, the efficient, stable and repeatable method for obtaining the haplobiont through apple anther culture is established, the induction rate and the differentiation rate can be increased, and the malformation rate can be reduced.

The invention discloses a method for improving the inductivity of an MDE (Microspore Derived Embryo) of an autumn radish, which specifically comprises the following steps: (1) soaking radish seeds for 2 hours (25 DEG C), and transferring the radish seeds into a refrigerator at 4-6 DEG C to be treated for 21-25 days if the radish seeds are white; (2) selecting 2-3.5 mm flower buds, and spraying 1.5% mannitol one day ahead of time; (3) after the flower buds are disinfected and cleaned, extracting and purifying microspores by using a B5 liquid culture medium (pH 5.8) added with 1.5% mannitol; (4) suspending the purified microspores in an improved NLN-13 culture medium with the pH value of 5.8-6.0, performing heat shock treatment at the temperature of 32.5 DEG C, and performing standing dark culture for 18-25 days at the temperature of 25 DEG C; (5) after the MDE grows out, transferring the MDE to an MS culture medium and culturing for 4-8 days; transferring the hypocotyls to a strong bud culture medium after the hypocotyls extend, and then transferring to a rooting culture medium; and (6) acclimatizing and transplanting after the root system is developed. According to the method, the seasonal limitation of a radish free microspore culture technology can be broken through, the MDE induction rate in the autumn radish free microspore culture process is increased, the radish genotype is rapidly purified, and the radish variety breeding process is accelerated.

The invention relates to the technical field of callus induction, in particular to a method capable of remarkably improving rice in-vitro microspore embryogenesis, which comprises the following steps: step 1, material selection: selecting rice young ears with extracted flag leaves; 2, low-temperature treatment, wherein the rice young ears are placed in a clean container and subjected to low-temperature treatment; 3, microspore dissociation: collecting anther from the rice young ears, carrying out anther pretreatment, and after pretreatment, carrying out high-speed rotary cutting, dissociating and collecting microspores; and 4, callus induction pretreatment: putting the collected microspores in sterile water, and carrying out callus induction pretreatment. According to the method for improving the yield of the free microspore calluses of the rice, the free microspores of the rice are put in the sterile water for 2 days during pretreatment, so that the yield of the microspore calluses can be remarkably improved, and the yield of the free microspore calluses is improved. The microspore culture efficiency is improved, and the production benefit is improved.

The invention provides a method for establishing an ATAC-seq library by using quinoa microspores, and belongs to the technical field of bioengineering. By improving a microspore sample pretreatment process and adjusting parameters of a transposase reaction system, key technical bottlenecks in the prior art are solved, efficient construction of the quinoa microspore ATAC-seq library is realized, and reliable technical support is provided for analyzing a chromatin regulatory network developed by quinoa male gametophytes and excavating and regulating key genes.

The present invention relates to the field of barley planting technology, and in particular to a method for improving the green shoot differentiation rate of barley microspore callus after Agrobacterium infection and its application. The method for improving the green shoot differentiation rate of barley microspore callus after Agrobacterium infection of the present invention comprises the following steps: (1) inoculating barley microspores into an induction medium to obtain microspore callus; (2) inoculating the microspore callus into a solid induction medium for subculture to obtain subculture microspore callus; (3) mixing the subculture microspore callus with an Agrobacterium suspension, transferring the subculture to a co-culture medium for co-culture, and obtaining infected callus; (4) treating the infected callus with negative pressure, transferring the infected callus to a differentiation medium for differentiation culture, and obtaining barley microspore callus green shoots. The method of the present invention can significantly improve the green shoot differentiation rate of barley microspore callus after Agrobacterium infection.