280 results about "Polyploid" patented technology

Methods are provided for the synthesis and secretion of recombinant proteins preferably large mammalian proteins or hetero-multimeric proteins at high levels and for prolonged time in polyploid, preferably diploid yeast. These methods use various mating competent yeast, including Pichia. In a preferred embodiment, a first expression vector is transformed into a first haploid cell; and a second expression vector is transformed into a second haploid cell. The transformed haploid cells, each individually synthesizing a non-identical polypeptide, are identified and then genetically crossed or fused. The resulting diploid strains are utilized to produce and secrete fully assembled and biologically functional hetero-multimeric protein.

A method for producing polyploid plants of orchids includes the steps of: providing a protocorm or protocom-like body (PLB) of an orchid, the protocorm or PLB having an upper portion with a growing point and a lower portion without any growing point; cutting the protocorm or PLB approximately at a point of half height to separate the upper portion; subculturing the lower portion of the protocorm or PLB in an inducing medium, and putting a cut surface of the lower portion of the protocorm or PLB to face upward so that one or more next-generation PLBs grow from the cut surface of the lower portion. The method characterized in using no antimicrotubule agent can simplify the entire process of orchid polyploidy breeding, and can be used in mass-production of the stable polyploid plants.

The present invention provides polyploid rice selectively breeding and identifying method with polyploid meiosis stability (PMeS) gene. The rice selectively breeding process includes: selecting parents; hybridization or complex hybridization; back crossing; doubling chromosome; identifying polyploid; selecting bearing property; observing meiosis behavior; self-crossing to form stable line; and identifying high bearing property. The identifying indexes include: polyploid line of Asia cultivated indica type rice or japonica rice; bearing rate not lower than 65 %; normal meiosis behavior; normal pollen shape and physiological activity and normal dyed pollution with 0.1 5 I-KI not lower than 95 %; bearing rate of hybrid of PMeS polyploid rice and other polyploid rice variety over 70 % and that of PMeS polyploid rice and wild rice variety over 65 %. The present invention solves the problem of low bearing rate of polyploid rice and may be used in breeding polyploid rice variety.

The invention provides a method of breeding fourfold paulownia tomentosa in the combination of isolated culture by using placentas as explants and colchicine treatment, comprising the following twelve steps: a. choice of parent; b. using placentas as best explants; c. placenta callus inducement; d. double treatment of placenta callus; e. callus recovery cultivation after treatment; f. callus division cultivation; g. sprout rooting cultivation; h. transplanting of seedling; i. morphological authentication of paulownia tomentosa plant polyploidy; j. chromosome number authentication of paulownia tomentosa plant polyploidy; k. flow cytometry analysis authentication of paulownia tomentosa plant polyploidy; l. authentication of paulownia tomentosa plant polyploidy in flowering period, etc. Compared with adult plants of other multiploid paulownia tomentosa, the fourfold paulownia tomentosa is characterized by big flower, thick leaf, no burliness, and the like, obviously different from twofold paulownia tomentosa, and has perfect ornamental value and growing vantage. The technology not only can be directly applied to breeding new types of paulownia tomentosa but also builds a fine foundation for polyploid breeding of other trees.

The invention discloses a method for rapid identification of the ploidy of a salicaceae plant. The method comprises the steps of cell nucleus extraction buffer solution preparation, cell nucleus dyeing buffer solution preparation, cell nucleus suspension preparation, cell nucleus DNA specific dyeing, flow cytometry detection and ploidy judgment. The method has the advantages of being wide in application range, low in sample consumption and simple in preparing method when used for determining the ploidy of the salicaceae plant. Extracted cell nucleus DNA is high in average flourescence intensity and small in variable coefficient. The determination result can be obtained quickly and is accurate, and resolution ratio is high. The method provides a reliable technical means for salicaceae plant polyploid breeding and has broad application prospects.

The invention discloses an inter-simple sequence repeat ISSR-SCAR marker specific to E-group chromosomes of agropyron elongatum. The invention provides a kit for detecting the E-group chromosomes of agropyron elongatum. The kit comprises the following primer pair: one primer in the primer pair is nucleotide shown in a sequence 3 of a sequence table, and the other primer in the primer pair is nucleotide shown in a sequence 4 of the sequence table. The E-group chromosomes of thinopyrum elongatum are the basic chromosome group forming the polyploid species of elytrigia and carry the genes beneficial to the genetic breeding of wheat, the detection of exogenous E chromatins in wheat is the key point for identifying recombination lines and introgression lines of the wheat, namely the agropyron elongatum, the work of screening and identifying a large number of filial generations is quite complicated, and tests prove that the ISSR-SCAR marker developed by the invention can lay a foundation for quickly and accurately identifying the E-group chromatins or chromosomes of exogenous agropyron elongatum under the genetic background of wheat.

The invention relates to a polyploid inducement method of Fritillaria pallidiflora Schrenk in vitro culture, which uses fresh bulb to be induced to produce adventitious buds. Inducement treatment is applied through cultivating in MS liquid medium with different density of colchicine to produce Fritillaria pallidiflora Schrenk polyploid plant. Bulb yield and medicinal components content of the plant are improved by changing chromosome ploidy to produce plant with good characteristics. Therefore, the problem of variety degeneration of Fritillaria pallidiflora Schrenk in a long-term course of Fritillaria pallidiflora Schrenk planting is overcomed. The method has the advantages that Fritillaria pallidiflora Schrenk polyploid can be obtained quickly with effectively improved content of active components and a shorter planting cycle.

The invention provides an excised mutagenesis tetraploid method of water melon and ploidy early stage certification technique, wherein dinitro toluene herbicide (DNH) is employed to substitute the conventional colchicines as inducer, whose function is to suppress the Mitosis in the metaphase of cell division through the mechanism of interfering spindle, so as to double the tissue cell chromosome. The method has the advantages of increased inducement success rate and substantially shortened time required for inducement.

Exemplary embodiments provide methods and systems for string graph assembly of polyploid genomes. Aspects of the exemplary embodiment include receiving a string graph generated from sequence reads of at least 0.5 kb in length; identifying unitigs in the string graph and generating a unitig graph; identifying string bundles in the unitig graph; determining a primary contig from each of the string bundles; and determining associated contigs that contain structural variations compared to the primary contig.

The invention priovidese a new Broussonetia papyrifera mulberry tree hybrid distant hybridization and polyploidization breeding method. The method is characterized in that an inductive hybrid fruit callus doubling distant hybridization polyploid hybrid line is established by an embryo engineering and tissue culture technique of distant hybrids through a reproduction engineering kind improvement technology. The method comprises following steps: 1, obtaining distant hybrid fruits; 2, carrying out callus induction of the hybrid fruits; 3, carrying out colchicine treatment of the callus of the hybrid fruits; 4, carrying out recovery culture of treated calluses; 5, differentiating the calluses into bud seedlings; 6, differentiating the bud seedlings into roots; 7, transplanting tissue culture plants; 8, carrying out comparative observation and selection of survival plants; and 9, carrying out asexual propagation of the selected plants to form a stable kind (line). The method solves the problems comprising slow growth,weak adaptability, unclear trunk and bad material quality of present Broussonetia papyrifera, and allows the new polyploid Broussonetia papyrifera mulberry tree hybrid having the advantages of obvious trunk, strong adaptability and good material quality to be cultivated.

The invention discloses a method for cultivating triploid dendrobium huoshanense by a crossbreeding method of tetraploid plants and diploid plants. The method comprises the following steps: (1) dendrobium huoshanense species which are from different producing areas and have characteristics of good stress resistance, rapid growth, thick stalk, long stem, many tillers, low fiber content and high active ingredient content are preferably selected as polyploidy cultivated original plant parents; (2) seeds of the different species of original plants undergo aseptic sowing in a solid medium so as to obtain diploid protocorms; (3) protoplasts of the different species of diploid protocorms are prepared and cell fusion is carried out; (4) detected tetraploid plants undergo asexual tissue culture so as to obtain tetraploid plant seedlings; (5) the tetraploid plant seedlings undergo hardening-seedling acclimatization and cultivation to flowering; (6) the tetraploid plants and diploid plants undergo artificial pollination and crossbreeding, and fructification is completed to obtain triploid dendrobium huoshanense seeds; and (7) the triploid seeds undergo aseptic sowing and culture so as to obtain a lot of triploid seedlings.

The invention discloses a method for cultivating a corn allopolyploid by using an unreduced gamete characteristic of tripsacum dactyloides, belonging to the field of corn distance hybridization. The method comprises the steps of: hybridizing an MTF-1 (metal-responsive transcription factor) as a female parent with corn or tetraploid perennation corn as a male parent, and then selecting a filial generation plant which is capable of overwintering, namely a new corn allopolyploid, wherein the tiller number of the filial generation plant is more than 15 and the chromosome number of the filial generation plant is a sum of the chromosome number of a female parent and the chromosome number of the male parent. According to the method provided by the invention, the dysgenesis of corn affinis species is overcome and a good inheritance basis is provided for breeding ground-breaking corn species by using the bred corn allopolyploid as a bridge material transfer character. In addition, the method provided by the invention provides a model to breed allopolyploids of other species. The corn allopolyploid bred by the method provided by the invention is perennial, adopts vegetative propagation, and provides a material to corn allopolyploid origin and evolution research and allopolyploid breeding. The method provided by the invention is simple, is short in time, high in efficiency and small in workload.

The invention discloses a technology for cultivating triploid momordica grosvenoris which is high in mogroside content and whole momordiaca grosvenori utilization rate. The technology comprises the steps of selecting field fresh momordica grosvenoris with good appearances, disinfecting the whole momordica grosvenoris, inducing young embryos to bud and preparing the budded embryos into virus-free seedlings; performing polyploid induction and cell flow analysis on female plants analyzed and identified from the virus-free seedlings through restricted fragment length polymorphism (RFLP); hybridizing stable tetraploid female plants with diploid male plants with good commodity natures; and harvesting fruits in autumn and sowing seeds in the fruits in a nutrition pot, planting virus-free seedlings which are analyzed to be triploid female plants through a flow cytometry and the RFLP, and hybridizing virus-free seedlings with diploid male plants with good commodity natures to obtain the triploid high-mogroside content momordica grosvenoris, namely, the born fruits.

The invention belongs to the field of plant cell engineering, and relates to a method for pear blade inducing adventitious buds to regenerate plant. The young blade of a test-tube plantlet is used as an explant and is placed on an ultra-clean work bench, 2-4 wounds are crosscut perpendicular to the blade midrib with a sterile blade, then the blade is inoculated on an adventitious bud induction medium, after being dark cultured for about 3 weeks, the blade is placed under light for culturing, and then is transferred to an enrichment medium after growing adventitious buds so as to promote the growth of buds; and after the adventitious buds grow to about 2cm, the blade is transferred to a rooting medium for culturing to obtain complete plants. The method is simple in operation, low in cost and good in result, and with the adoption of the method, a stable and efficient blade renewable system can be quickly established, the technical problems of pear blade adventitious bud regeneration induction can be effectively solved, so that the precondition can be provided for improving fruit tree variety through the two ways of genetic transformation of disease resistant gene and genome doubled polyploid cultivating.

The present invention relates to an ALS inhibitor herbicide tolerant polyploid plants, such as B. napus or B. juncea plants, progeny and parts thereof comprising a mutation of all acetolactase genes.

Exemplary embodiments provide methods and systems for string graph assembly of polyploid genomes. Aspects of the exemplary embodiment include receiving a string graph generated from sequence reads of at least 0.5 kb in length; identifying unitigs in the string graph and generating a unitig graph; and identifying string bundles in the unitig graph by: determining a primary contig from each of the string bundles; and determining associated contigs that contain structural variations compared to the primary contig.

The invention belongs to the field of crop breeding and discloses a sexual polyploidization breeding method of tetraploid Brassica campestris ssp.chinensis Makino. The method comprises the following steps of: treating flower buds by using solution of colchicine in the bolting period of the Brassica campestris ssp.chinensis Makino to induce the flower buds to generate 2n male gametes; and performing manual sexual polyploidization cross breeding on a tetraploid male sterile line serving as a female parent and 2n male gamete-containing diploid obtained by the induction of the solution of colchicine serving as a male parent in a blooming period to obtain the tetraploid Brassica campestris ssp.chinensis Makino. In the method, a sexual polyploidization breeding approach of hybridizing the male sterile line of the tetraploid Brassica campestris ssp.chinensis Makino with the diploid Brassica campestris ssp.chinensis Makino which generates 2n gametes after induction is used for the first time to obtain a new species of the tetraploid Brassica campestris ssp.chinensis Makino which has wider genetic diversity and higher seed setting rate compared with the autotetraploid of asexual polyploidization. The new tetraploid of the invention has the characteristics of polyploid such as largeness, disease resistance and cold resistance, high quality, and the properties of a diploid male parent.

The invention relates to a cultivation method of genuine salvia, which comprises the following steps of: establishment of a salvia germplasm resource garden, selection of representative genuine salvia germplasm resources, identification of genuine salvia genome karyotype structure, determination of genuine salvia DNA bar codes, artificial mutation to obtain autotetraploid, sexual hybridization to obtain triploid salvia permanent hybrids, detoxification treatment, tissue culture seedling cloning and proliferation, bottle seedling transplanting, transplanted seedling management, new species comparison and testing, determination of the medicinal material yield and the main drug effective components, and genuine salvia new species determination and species identification. The genuine salvia obtained in the invention is, so far, the only population that has two great natural advantages: the polyploid advantage and the heterosis, and the advantages are giant and irreplaceable. Therefore, the method of the invention is an effective method for cultivating genuine traditional Chinese medicine with strong biotic stress and abiotic stress capability by using traditional Chinese medicine genuine functional genomes and feature genes and based on inherent genes of genuine traditional Chinese medicine.

The invention relates to a process of producing autotetraploid by inducing Primula forbesii with colchicine. It comprises following steps: immersing the seeds of Primula forbesii for 24 hours; then immersing the seeds with colchicine of 0.5-1.5g / l for 36-48 hours in the dark, controlling temperature within 20+-2deg.C; identifying ploidy with morphology and chromosome of cytology. The method is characterized in that tetraploid Primula forbesii is induced with colchicine, the success rate is high, and the effect is good; the genetic stability of tetraploid Primula forbesii F1 generation can be maintained; when compared to diploid Primula forbesii, the tetraploid shows good characters of bigger flowers, much more odorous and thick scape; and it also improves the ornamental value.

The invention discloses a subfamily distant hybridization for German mirror carp and megalobrama amblycephala. The method comprises the following steps of artificial dry-process fertilization on the ova of the pubescent German mirror carp and sperms of the megalobrama amblycephala by taking the German mirror carp as a female parent and the megalobrama amblycephala as a male parent, hatching fertilized ova at the water temperature of 19-20 DEG C, and breeding fries in a pond; then, detecting the DNA (deoxyribonucleic acid) content, the chromosome number and the blood cell sizes of the fries by a flow cytometry DNA content determination method, a chromosome ploidy detection method of peripheral blood cell culture and a blood smear method, screening, and then preparing to obtain triploid hybrid fishes and tetraploid hybrid fishes. A tetraploid fish strain which is genetically stable and has four sets of crucian carp chromosomes can be formed by subsequent self-cross selective breeding of the tetraploid hybrid fishes, and the tetraploid fish strain can be used as a resource for breeding the infertile triploid fishes. The method disclosed by the invention has the advantages that not only are the characters of hybrid offspring good but also a good foundation can be laid for variety breeding, and a precious resource is provided for subsequent breeding of new polyploid fishes.

The invention discloses a method for performing in-vivo induction on a vaccinium australe tetraploid. Vaccinium australe has the advantages of strong stress resistance, high growth speed and high yield; a tetraploid is taken as one of plant evolution routes and generally has the characteristics of hugeness, low fertility, strong growth, big fruits, thick and strong stems as well as strong stress resistance; and moreover, compared with conventional cross breeding, polyploid breeding is relatively short in cycle, and a new plant variety can be obtained relatively rapidly. According to the method disclosed by the invention, tissue culture seedlings and rooting seedlings are taken as materials, and a vaccinium australe plant with strong stress resistance, high quality, high yield and big single fruits can be obtained by performing tetraploid induction on the vaccinium australe tissue culture seedlings by using an in-vivo induction method so as to lay a foundation for cultivating new big fruit type cowberry blueberry varieties which are suitable for tropical and subtropical zones.

A breeding method of a polyploid rice photo-thermo-sensitive genetic male sterile line includes determining a diploid rice line with photo-thermo-sensitive genetic male sterility or PMeS characteristic as a parent; carrying out hybridization on a diploid photo-thermo-sensitive genetic male sterile line and a diploid PMeS gene line, carrying out doubling culture on a young ear of a hybrid plant into a hybrid tetraploid; back-crossing the hybrid tetraploid with a tetraploid photo-thermo-sensitive genetic male sterile line; selecting a tetraploid male sterile plants from the back-crossed progeny, self-crossing during a low-temperature and short-day fertile period, and then carrying out composite hybridization with another tetraploid rice line having PMeS gene; selecting tetraploid male sterile plants, and detecting the stability of tetraploid male sterile plants after multiple generations of continuous self-crossing; and determining the stable and consistent tetraploid rice sterile line as the polyploid rice photo-thermo-sensitive genetic male sterile line, named as PSXXX.

The invention belongs to the field of vegetable breeding, and in particular relates to a method for breeding a new variety of leaf achillea. The method comprises the following steps of: accelerating germination of hybrid offspring seeds through temperature change; soaking in colchicine solution; screening a polyploidy line from single plants; and performing trait screening and clonal propagation. The ratio of small leaves of the bred new variety in the product is higher, the area of a single leaf is large and round, leaf margin teeth are a few and blunt, the yield is higher, the quality is better, the small leaves can be eaten together with the achillea, and the leaf achillea has higher commercial value and greatly contributes to income increase of farmers and scientific diet of residents.

The invention relates to a method for rapid induction of homogenous polyploids of Chinese jujube, that is the method can double and further develop chromosomes of a single-cell embryo into homogenous tetraploids by simultaneously carrying out induction with colchicine during the somatic embryogenesis process of in vitro leaf blades of the Chinese jujube. The method comprises the steps of somatic embryo induction, doubling of the chromosomes, development of embryoids till nature and detection and screening of the homogenous tetraploids. The specific steps are as follows: selecting tissue culture seedling leaf blades of the Chinese jujube with robust growth and seedling age of 20-30d, and using a pair of scissors after sterilization to shear 5-8 times by being vertical to nervures; then inoculating the leaf blades in an induction culture medium MS plus 15-25g / L of maltose plus 3.0-5.0g / L of agar plus 5.0-10.0mg / L of TDZ plus 1.0-2.0mg / L of AgNO3 by leading the front surfaces to face upwards, and simultaneously adding 15-20mg / L of the colchicine into the culture medium, and carrying out dark culture for 40-50 days; transferring embryoids differentiated from calli of the leaf blades of the Chinese jujube into a hormone-free culture medium of the MS plus 40g / L of sucrose plus 500mg / L of caseinhydrolysate for lighting culture and further developing the embryoids into somatic embryo seedlings; and screening out the homogenous tetraploids from the somatic embryo seedlings through chromosome detection or the detection by a flow cytometer, wherein the pH of the culture medium is 5.0-6.0, and the culture temperature is 25-30 DEG C. The method can avoid the appearance of chimeras, omit the steps of separation and purification, rapidly obtain the homogenous polyploids and have important application value for accelerating the breeding process of the polyploids of the Chinese jujube.

The invention discloses an artificial spawning and insemination method for hyriopsis cumingii, which includes the steps: selecting parents 4-6 years old and identifying sex; isolating female parents and male parents for synchronous cultivation in a culture pond with micro-flow water; sorting out female clams to be pregnant; dividing the female clams to be pregnant into a natural insemination group and an artificial insemination group; injecting male clams with oxytocin; and performing artificial insemination to the male clams. By the method, artificial regulation of insemination of hyriopsis cumingii is realized, artificial insemination rate of hyriopsis cumingii can reach 70%-90% in 10-30 minutes, and basic conditions for developing researches on hyriopsis cumingii, such as polyploid breeding, family selection and the like are created.

A breeding method of a polyploid rice two-line restorer line includes determining hybrid parents for breeding the restorer line; breeding a restorer line of an indica sterile / japonica restorer type, or breeding a restorer line of a japonica sterile / indica restorer type, and carrying out backcrossing or composite hybridization after hybridization of the parents; selecting a single plant that meets the breeding goal, carrying out composite hybridization again and carrying out preliminary molecular marker detection; comparing, and selecting a single plant with good traits for continuous selfing until the line is basically stable; selecting a stable line with good traits and detection molecular markers; carrying out test-crossing on the preferred line as a male parent with different types of polyploid sterile lines; and selecting a good hybrid combination and a restorer line thereof.

The invention provides a method for inducing polyploid through wild jujube adventitious buds, relates to the biotechnical field, and the problem that homozygous tetraploid is hard to obtain through existing methods is solved. The method provided by the invention comprises the following steps: starting of cultivation, induction culture, elongating cultivation, strong seedling cultivation and detection on young leaves. The method can be used for cultivating wild jujube homozygous polyploid so as to support the genetic improvement of jujube trees.

The invention belongs to the technical field of polyploid breeding, and discloses a method for inducing phalaenopsis protocorm to generate polyploid by colchicine. The method comprises the following steps of transferring the protocorm into a 1 / 2MS+1mg / l 6-BA + 0.2 mg / l NAA+5g / l agar+20g / l sucrose solid culture medium containing 0.05% of colchicine, and culturing for 15 days for induction treatment; washing the protocorm for 3-4 times by using sterile water after treatment, absorbing moisture by using filter paper, transferring the protocorm into a 1 / 2MS+5g / l agar+20g / l sucrose culture medium,and carrying out subculture for 2-3 times; performing polyploid identification, namely respectively carrying out morphological observation, stomatal identification and ploidy identification; and performing data statistics. According to the method, colchicines with different concentrations and different treatment time are used for doubling the phalaenopsis protocorm by a co-culture method, so thata foundation is laid for polyploid induction culture of a new phalaenopsis variety with large flower size, bright color and high ornamental value.

The invention relates to an artificial cultivation and seedling industrial production method of a dendrobium nobile polyploid compound, which comprises the following steps of: carrying out artificial pollination in a flowering period of common dendrobium nobile, inoculating harvested seeds in a 1 / 2MS liquid nutrient medium for nonsymbiotic germination cultivation, transferring protocorm of seeds in a proembryo stage into a 1 / 2MS liquid nutrient medium containing 0.01% of colchicin for mutagenesis cultivation, then transferring into 1 / 2MS solid nutrient medium for cultivation till seedlings ofdendrobium nobile are formed, carrying out flowering cultivation on tetraplont seedlings with chromosome numbers of 2n=76, taking tetraplont as a female parent and diploid as a male parent or taking diploid as a female parent and tetraplont as a male parent for hybridization to obtain the triploid; transferring triploid hybridized seeds in a formation early state into the 1 / 2MS liquid nutrient medium containing 0.01% of colchicin for carrying out artificial mutation hexaploid treatment for 150-180h, and then transferring into the 1 / 2MS solid nutrient medium till the seedlings are formed, selecting seedlings with chromosome numbers of 2n=114 as tetraplont dendrobium nobile for carrying out bottle seedling transplanting, and finally cultivating and managing according to a conventional method till flowering. The invention lays the foundation for the popularization of new varieties of the dendrobium nobile.