146 results about "Polyploid Cells" 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.

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.

A method for selectively culturing the irregular allopolyploid rice includes such steps as choosing parents, hybridizing while treating with hormone, embryonic retrieval culturing, reduplicating chromosome, reduplicating calli or fasciculate embryos, growing to become seedlings, rooting culture, transplanting, culturing and discriminating. The resultant rice may be irregular allohexaploid rice or irregular allooctaploid rice.

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 propagating hemerocallis hybridus. The method for propagating the hemerocallis hybridus comprises the following steps: (1) selecting an explant and sterilizing; (2) inoculating the sterilized explant onto a bud induction medium, and carrying out bud induction culture in a culture room; (3) transferring a differentiated explant onto a subculture medium and carrying out subculture, so as to obtain a differentiated seedling; (4) inoculating the differentiated seedling onto a rooting culture medium and carrying out rooting culture; (5) hardening the differentiated seedling, moving the differentiated seedling out of the culture room, and transplanting, wherein the hemerocallis hybridus is a diploid or multiploid. The method for propagating the hemerocallis hybridus is obviously better than other rapid plant tissue culture propagation methods in the aspects of sterilization time, sterilization effect, operation method, regeneration efficiency, production cost and the like, so that an established large-scale commodity production system is stable and the large-scale rapid propagation production can be realized; the method for propagating the hemerocallis hybridus is mainly applied to rapid propagation of new hemerocallis hybridus varieties and is an effective approach for enabling the new hemerocallis hybridus varieties to be rapidly put into the market, so that the current situation that new varieties are pushed over-slowly in China can be changed.

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 invention discloses a selecting and breeding method of a polyploidy novel mulberry tree breed. The breeding method comprises the following steps: implementing artificial hybridization and chromosome induction of polyploidy and other modes by selecting mulberry tree diploid hybridized mulberry seeds or hybridized seed mulberry sapling disease-free plants as materials; selecting out bud variation and branches the shape of which are similar to the polyploidy or plants by excellent plant, single plant selection and system selection; carrying out simple bud grafting on branch break bud variation the shape of which is similar to the polyploidy or plants in a primary selection garden; carrying out microscopical examination on the selected polyploidy single plants; establishing a system selection garden after determining that the single plants are polyploidy; and breeding a mulberry tree polyploidy novel breed by establishing product test gardens and parent gardens. The novel breed has extremely-high quality, ultrahigh yield and wide applicability; and in the actual production, a mulberry tree breed with ultrahigh yield matched with early growth, middle growth and late growth mulberry can be developed and bred, adapts to the requirement of pluralized and multiple silkworm breeding production all the year around on mulberry breeds and has obviously-improved mulberry yield, mulberry quality and other economic characters.

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 relates to a method for cultivating the novel variety of tetraploid dendrobium by use of the seed protocorm (PLBs). After the artificial pollination, the immature dendrobium seed in the development stage is carried out non-symbiotic sproution cultivation for 60 days in the 1 / 2MS liquid culture medium, which is then transferred into the 1 / 2MS liquid culture medium containing 0.02 percent Oryzalin for process and then transferred into the 1 / 2MS solid culture medium for continuous cultivation; after that, till the dendrobium seedling is shaped, the dendrobium is not planted, transplanted and managed by a conventional method until the dendrobium blooms. The invention overcomes the defect of generally low mutagenesis frequency of the polyploid and has better appreciation; moreover, the method is easy and practical, which increases the mutagenesis frequency (by over 60 percent) of the polyploidy in a large scale and provides enough polyploidy (over hundreds of plants) obtained one time to be used for the seed selection with high application value. Besides, the invention is applicable to the large-scale production with the advantages of small investment and high output.

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 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.

The invention discloses a woad triploid breeding method with obvious rhizome weight increase.The method comprises the following steps of germinating seeds; conducting gravity field mutagenizing; conducting directional subculture breeding; collecting obtained woad seeds to be subjected to seed germination, obtaining woad buds and dividing the woad buds into two groups A and B, conducting gamma ray source irradiation treatment on the buds in the group A, using a colchicine solution for performing chromosome doubling treatment on the buds in the group A, and screening out tetraploid buds; obtaining diploid buds in the group B; conducting transplanting treatment on the buds in the group A and the buds in the group B and then obtaining group A tetraploid plants and group B diploid plants respectively; adopting the group A tetraploid plants as a female parent, adopting the group B diploid plants as a male parent, conducting sexual hybridization, and obtaining woad triploid plants.The obtained triploid plants have the dual advantages of polyploids and hybrids and are highly sterile, the hybrid vigor is fixed, the genetic characters can be continuously inherited through a maintainer line, and the problem of posterity segregation is avoided.

The invention relates to a plant variety cultivating method, especially a method of cultivating lily polyploidy. 1. according to routine lily cultivating technique, selecting seed balls and jarovizing at low temperature to break dormancy to complete the jarovizing; 2. cultivating in a greenhouse or making field cultivation and management according to routine cultivating lily technique; 3. cultivating for 30-40days, then dripping 0.02-0.2% colchicines at 20 deg.C-25 deg.C; 4. according to the routine cultivating technique, processing the plants so that they will come into flower in 50-60 days, selecting the plants with extremely large stigmas and huge direct microscopic pollen particles to eliminate stamens so as to obtain 2n-oogamete containing stamens; 5. gathering normal pollens to hybridize with the obtained stamens, then according to routine cultivating technique, cultivating so that the fruits will swell fully in 45-60 days, thus able to gather them; 6. adopting routine triploid embryo in vitro culture method to be able to obtain the lily triploids.

The invention relates to a breeding method for inducing parthenogenesis of corn by propyzamide, belonging to the field of polyploid breeding of plants. The breeding method specifically comprises the following steps of: preparing an inducer with the content of 5 mu mol / L-200mu mol / L by the propyzamide and water; performing induction treatment on filaments, wherein the inducer is sprayed or smeared onto the filaments which grow to about 2.5cm long after emerging, and mature seeds are harvested to get genotype homozygous diploid seeds; and selecting and identifying the seeds according to a conventional method, and selecting a head progeny row which conforms to a breeding goal, thus finally getting a homozygous corn inbred line. The breeding method disclosed by the invention has the positive effects of omitting artificial pollination, being simple, being easy to implement, low in cost, high in efficiency and suitable for large-scale use.

The invention discloses a method for breeding mulberry polyploids through chemical mutagenesis. A mixed solution of 0.4% of colchicine and 3% of DMSO is employed as a mutagenic agent, new shoots of mature mulberries more than two years old, in vigorous growth, with plump growing points and without insect diseases are subjected to chemical mutagenesis processing, and new mulberry strains with genetic stability and with excellent or special biological characters and economic characters are obtained through generation selection. Through the method, high-frequency polyploid mutant plants can be obtained, the variation spectrum is wide, excellent new mulberry strains with genetic stability can be obtained, which is helpful for improvement of mulberry species. In addition, orientation selection of parents can be carried out according to actual demands of silkworm production, furthermore excellent new species with excellent biological characters and economic characters are obtained, demands of diversification and all-year silkworm breeding production for multiple times to mulberry species are met, and the economic benefit of the silkworm industry is raised.

The invention relates to an allopolyploid induction growth method of hybridized Chinese pennisetum. The method comprises the following steps: immersion of allotriploid hybridized Chinese pennisetum F1 seeds in a mutagen containing colchicine and dimethylsulfoxide, initial culture, polyploid identification, proliferation culture, expanded production and transplanting. The method allows a large number of mutagenic seedlings to be obtained in a short time, greatly improves the culture efficiency and saves the production cost.

The invention relates to a polyploid induction method for Chinese gooseberry Hort 16A offspring fine strain tissue culture seedlings. The polyploid induction method comprises the following steps: soaking leaves of the Hort 16A offspring strain tissue culture seedlings in a colchicine water solution to induce polyploidy, washing the leaves, and inoculating in callus differentiation enrichment culture medium and subculture culture medium for light culture according to the steps. The polyploid induction method is simple in operation, good in induction directionality, high in induction success rate, low in cost, shorter in culture period and higher in induction rate, an efficient and feasible new method is provided for polyploidy breeding of Chinese gooseberry, the polyploid induction method has important application values, through the popularization of the polyploid breeding method, economic benefits can be brought for fruit farmers, and demands of more consumers are met.

The invention belongs to the technical field of rape molecular breeding, in particular relates to a preparation method of a codominant SNP (Single Nucleotide Polymorphism) molecular marker closely linked with the linolenic acid content of a cabbage type rape and application thereof as the marker in the auxiliary selection of the low linolenic acid cabbage type rape. The low linolenic acid cabbage type rape strain A254 and high linolenic acid cabbage type rape strain A177 genomes DNA are amplified by using primers to respectively obtain two DNA amplified fragments, then cloning, sequencing and nucleotide sequences comparison are carried out on the four DNA amplified fragments; according to the difference of the sequences, the second bases at 3'-end of a forward primer and a reverse primer are mispaired, primer pairs of YQ-fad31-1, YQ-fad31-2, YQ-fad32-1 and YQ-fad32-2 are designed, the PCR (Polymerase Chain Reaction) amplification is carried out on the primer pairs of YQ-fad31-1, YQ-fad31-2, YQ-fad32-1 and YQ-fad32-2, and the PCR amplification and group detection effects are analyzed to obtain the codominant SNP molecular marker linked with low linolenic acid genes of the cabbage type rape. The invention provides a new marker for the rape molecular breeding. In the invention, a mispairing strategy of the 3'-end of the primer is firstly applied to the development and the detection of the SNP polymorphic marker of the cabbage type rape, and a means of synchronously mispairing the forward primer and the reverse primer is firstly adopted to develop the SNP marker based on the PCR amplification and an agarose gel electrophoresis in the cabbage type rape of an allopolyploid crop. The invention synchronously discloses a method for preparing the molecular marker and the application thereof.

The invention discloses a method of utilizing methods of molecular biology and gene engineering to construct a polyploid tandem gene pBluescriptIIKS-nPFWM, a Pichia pastoris tandem expression vector pPICZ alpha A-pBluescriptIIKS-nPFWM and a Pichia pastoris engineering strain GS115-pPICZ alpha A-pBluescriptIIKS-nPFWM1, wherein n is 1, 2, 4, 6 or 12. A foundation is laid for subsequent efficient and stable expression of antioxidant peptide, industrialized large-scale production and application in the field of cosmetics and healthcare food.

The invention discloses a method and application method for obtaining a novel germ plasm of brassica A genome vegetable. In the method, free microspore culture is carried out on crossbreeds among vegetables of Chinese cabbage subspecies*pakchoi cabbage subspecies or variety in pakchoi cabbage subspecies*variety of the brassica A genome to obtain a regeneration plant and further to create the novel germ plasm which is used for breeding. The method mainly comprises the following steps of: hybridizing the vegetables of Chinese cabbage subspecies*pakchoi cabbage subspecies or variety in pakchoi cabbage subspecies*variety; inducing free microspore culture embryoids; germinating the embryoids to obtain the regeneration plant; carrying out ploidy identification on the regeneration plant obtainedby microspore culture; reserving seeds of double haploids and polyploids; planting an offspring of the regeneration plant to carry out morphological identification and nutritive index identification;and preparing, hybridizing and combining. The method has the advantages of short cycle and high efficiency, provides a material for efficiently breeding Chinese cabbage vegetables and has broad application prospect.