138 results about "Polyploid" patented technology

The invention provides application of a bacteriostatic agent in open type tissue culture and rapid propagation of polyploidy bamboo reeds, and relates to the technical field of tissue culture and rapid propagation of plants, the bacteriostatic agent is applied to the open type tissue culture and rapid propagation process of the polyploidy bamboo reeds, and the use method is that the bacteriostatic agent is added into a culture medium for use or sprayed or oscillated and flushed for use, and the bacteriostatic agent is applied to the open type tissue culture and rapid propagation process of the polyploidy bamboo reeds. The bacteriostatic agent is formed by mixing isothiazolinone and sodium dehydroacetate, the mass addition ratio of the isothiazolinone to the sodium dehydroacetate is (8-5): (2-5), and the method comprises the following steps: S1, primary culture of explants; s2, multiplication culture of seedlings; s3, rooting culture, wherein each liter of the culture medium contains 0.05-1g of the bacteriostatic agent in the above steps. The method has the beneficial effects that under the condition that the method does not need to be controlled in a sterile environment and does not depend on the quality of an inoculator, a large amount of propagation of the polyploidy bamboo reed tissue culture seedlings is realized by adopting an open tissue culture technical mode, the productivity speed of a tissue culture center is improved, and the construction threshold and cost of the tissue culture center are reduced.

The invention discloses a method for differentiating induced pluripotent stem cells into megakaryocytes and platelets, and relates to the technical field of induced pluripotent stem cell directional differentiation, and the method comprises the following steps: inoculating an iPSC single cell into a culture vessel, and carrying out adherent culture through a culture solution; after the cell adheres to the wall and before cloning is not formed, the liquid is changed into a first induction culture liquid, and the cell is induced and differentiated into a mesoderm cell; changing the solution into a second induction culture solution to induce and differentiate the mesoderm cells into hematopoietic endothelial cells; collecting the suspension cells, culturing the suspension cells in an amplification culture solution, and enriching megakaryocytes; inducing megakaryocyte polyploidy through a third induction culture solution; and culturing the polyploidy megakaryocyte through a fourth induction culture solution to promote platelet release. The method for inducing the pluripotent stem cells to differentiate into megakaryocytes and platelets is convenient and rapid, the megakaryocytes and platelets can be obtained through iPSC in a short time, and feeder cells, serum and additional gene editing are not needed; meanwhile, the yield is high.

The invention belongs to the field of gene detection, and provides an STR primer group, a kit and a method for detecting polyploid, UPD and maternal pollution in a sample, the primer group comprises a first primer group, the first primer group comprises 178 pairs of primers, and the nucleotide sequences of the 178 pairs of primers are shown as SEQ ID NO.1-SEQ ID NO.356. The kit comprises an amplification reaction solution, dNTP, DMSO, nuclease-free water and the STR primer group. By adopting the primer group and the kit disclosed by the invention, the polyploidy of a sample, the monoparental diploid of chromosome level and whole genome level and the condition of maternal pollution can be detected by the method at the same time. And Cnv-seq or PGT-A can be configured for use, so that the problem that polyploidy and single-parent diploid maternal pollution cannot be detected by the two technologies at present can be effectively solved.

The invention discloses a k-mer marker-based correlation analysis method, which comprises the following steps of: obtaining re-sequencing data of each sample in a to-be-detected species population, performing quality control, constructing a k-mer abundance matrix based on a k-mer sequence in each sample, performing effective k-mer screening, and performing correlation analysis on the k-mer abundance matrix. And further converting into a continuous dosage genotype matrix, and then carrying out whole genome association analysis. According to the method, accurate GWAS analysis of a species group to be detected can be realized, and particularly in GWAS analysis of polyploidy species, the method can efficiently complete genotype identification and correlation analysis of the polyploidy species by utilizing k-mer abundance; the problems of high homologous region comparison errors and inaccurate allele dose estimation in polyploidy genotype identification of a traditional method are avoided, so that accurate detection and analysis are realized.

The invention discloses a polyploidy breeding method for wheat plant genome doubling, which belongs to the technical field of plant breeding, and comprises the following steps: treating germinated wheat for 30 days under the conditions of low temperature of 2-7 DEG C and 24-hour illumination, then transplanting into soil, culturing for 45-55 days under the conditions that the temperature is 20-25 DEG C, the light-dark culture period is 20-22 hours, and the light-dark culture period is 2-4 hours, the illumination intensity is 10000-20000 lx, and the culture time is 20-30 days. Wherein the ratio of the red light to the blue light is 3: 2, obtaining a large number of chromosome group doubled seeds until the wheat grains are mature, and realizing wheat plant chromosome group doubling. Experiments prove that the method is easy to operate, toxic reagents are avoided, the period is short, and the efficiency of obtaining wheat with doubled chromosome groups is high.

The invention discloses a haplotype analysis and assembly method and device for a polyploid genome. The invention develops a haplotype analysis support construction method which integrates HiC data and an assembly graph to improve accuracy, integrity and continuity. Key limitations of HiC are effectively overcome by utilizing graph connectivity derived from long read length overlap. The method is applied to complex polyploidy genomes, and compared with an existing tool, the method has the advantages that the sequence deletion rate is reduced by dozens of times, the sequence error rate is reduced by one order of magnitude, and the continuity of gapless sequences is improved by several times.

The invention relates to a method for inducing doubling cultivation of polyploidy by girdling calluses of camellia oleifera branches, the influence on tree vigor is reduced through girdling, the efficiency of regeneration buds is improved through thorough removal of cambium, the bottleneck in the prior art is overcome, chimera-free polyploidy material induction can be realized without in-vitro tissue culture, and the method is suitable for large-scale popularization and application. The method is simple and convenient to operate and low in cost, can effectively avoid the problems of rooting difficulty of tissue culture materials, transplanting loss of tissue culture seedlings and the like, provides a new technical approach for woody plant polyploid breeding, and has important popularization and application values.

The current invention pertains to a reliable method for determining the relative frequency of a sequence variant of interest in a nucleic acid sample derived from at least one polyploid cell, wherein the method uses a UMI to correct for any amplification biases. The invention further pertains to the use of a UMI for accurately determining the relative frequency of a sequence variant of interest in a nucleic acid sample derived from at least one polyploid cell.

The utility model belongs to the technical field of citrus cultivation, and particularly relates to a citrus polyploidy efficient induction device which comprises a machine body. A sealing door is rotationally arranged on the side wall of the machine body; the sealing doors are symmetrically arranged on the side wall of the machine body; an induction processor is mounted on the side wall of the machine body; the output end of the induction processor is connected with a built-in discharger; the built-in discharger is arranged on the inner side wall of the machine body; a spraying opening is formed in the side wall of the built-in discharger; the spraying openings are formed in the side wall of the built-in discharger in a linear array mode. A storage rack is fixedly connected to the interior of the machine body; the storage racks are symmetrically arranged in the machine body; an embedding groove is formed in the side wall of the storage rack; by means of the structure, tissue culture is not needed by adopting a living body induction technology, multi-time growth can be achieved from seed germination to regeneration, compared with a traditional in-vitro induction method, the method is more convenient, and multi-time cultivation of citrus can be achieved in a sealed state.

The invention discloses a method for carrying out nine-fold breeding by utilizing female gametes, and relates to the technical field of biological breeding. Comprising the following steps: step 1, pollinating hexaploid persimmon pollen to a'rattan royal house 'containing 2n eggs; 2, collecting young fruits which are born for 65-70 days after pollination, stripping seeds, putting the seeds into a sterilized triangular flask, treating the seeds for 30 seconds in an ultra-clean workbench by using 70% ethanol, pouring out the seeds, adding a 1% NaClO solution, soaking the seeds for 10 minutes, and shaking the seeds once every 3-4 minutes. According to the method for carrying out nine-fold breeding by utilizing female gametes, in persimmon breeding, compared with a 2n pollen path, the 2n egg path is higher in feasibility, 2n eggs and complete male germplasm with good pollen traits are hybridized, pollination populations are expanded, and polyploidy seedless materials can be obtained.

The application provides a method for inducing poly-podophylli rhizoma by using clump buds, comprising the following steps: S1, cutting stem segments from a podophylli rhizoma mother plant, soaking and cleaning to obtain explants; S2, disinfecting and cleaning the explants, inoculating into a clump bud induction medium to obtain podophylli rhizoma clump buds; S3, inoculating the clump buds into doubling liquid I, washing with a liquid clump bud induction medium after culture, and then transferring into doubling liquid II; S4, taking out the clump buds, cleaning, inoculating into a clump bud induction medium, and then inoculating into a bud proliferation medium; S5, waiting until 1-2 cm, screening out potential mutant strains through phenotypes, inoculating into the bud proliferation medium for subculture, and obtaining mutant plants; and S6, rooting culture of the mutant plants, and after root systems grow, planting and raising seedlings, and then poly-podophylli rhizoma plants are obtained. In the research, suitable concentration of colchicine is used for induction treatment twice, and through stage concentration switching and intermittent recovery strategies, the poly-ploid induction efficiency and explant survival are effectively considered.

The invention discloses a method, a system and equipment for splitting an allopolyploid based on a perl language and a storage medium, and particularly relates to the technical field of bioinformatics analys.The method comprises the steps that a diploid species close to a to-be-split allopolyploid species is selected as a diploid reference species; calculating a distance SD value between chromosomes to form a matrix; a homologous chromosome group HG1. HGN is formed; according to the homologous chromosome group HG1... HGN, an allopolyploid to be split is split into AN subgenomes. The subgenome is split based on the mutation accumulation difference of the sexual maturity cycle, and the method has high flexibility; the method disclosed by the invention has wide applicability; the splitting method disclosed by the invention has the advantages of high accuracy, good visualization effect, simplicity and convenience in operation, reliable data support, capability of promoting genome research and the like; according to the invention, how different mutations are accumulated by different species under the difference of sexual maturity cycles can be well revealed, so that deep understanding of the evolutionary mechanism of the species is promoted.

The present inventions relate to a breeding system for the production of polyploid maize seeds, maize plants, or maize plant parts where cycles of meiosis, syngamy, and selection are used for interpopulation improvement of progenitor lines, and sexual polyploidization occurs during hybrid production by inducing clonal gamete formation in the parents that are to be crossed. Reciprocal recurrent selection can be used to inform selection of candidate maize lines that are either advanced to a gene editing or genetic modification system or crossed and selected to induce clonal gamete formation by arresting meiotic recombination and chromosome reduction. Crosses of parent maize plants bearing clonal gametes are planned and executed based upon predicted heterotic performance at the polyploid level. The final product is a homogeneous population of hybrid polyploid maize seed, or derivative thereof, bearing both parents' complete nuclear genomes.

This invention discloses a method for efficiently inducing allopolyploid fish fry, belonging to the field of fish breeding technology. The method of this invention includes the following technical steps: (1) Induction of spawning in distant hybridization parents: Selecting female and male parent fish from different subfamilies of Cyprinidae, and collecting mature eggs and semen respectively; (2) Obtaining hybrid fertilized eggs: Mixing the collected eggs with semen, and performing dry artificial insemination to obtain hybrid fertilized eggs; (3) Inducing allopolyploidy: Placing the fertilized eggs in a water bath for heat shock treatment to inhibit the expulsion of the second polar body or inhibit the first cleavage, and then quickly transferring the treated fertilized eggs to aquaculture water to continue hatching fry. The method of this invention combines the method of distant hybridization between subfamilies with the method of heat shock induction. After induction using this method, the multiplication rate of hatched fry can reach 100%, which significantly improves the yield of allopolyploid fish fry from distant hybridization.

The present invention provides a method for rapidly propagating excellent seedlings from the fruit pods of Bletilla ochracea. The fruit pods of Bletilla ochracea are wiped and sterilized with alcohol and then subjected to a temperature-changing treatment. The seeds after the temperature-changing treatment are inoculated into a starting medium and subjected to light culture until the seeds germinate. The germinated seedlings are inoculated into a protocorm induction and proliferation medium for induction and proliferation culture. The seedlings after the induction and proliferation culture are cultured in a polyploid induction medium. The seedlings after polyploid induction are further cultured on a protocorm-induced bud medium to obtain seedlings. The polyploids obtained by the technical scheme of the present invention can effectively improve the self-crossing ability, thereby increasing the seed setting rate. Thus, a breeding system for rapid propagation of the fruit pods of Bletilla ochracea and selection of excellent varieties with relatively uniform quality is formed.

The application discloses a chrysanthemum constitutive high-expression promoter and application thereof in gene editing. The nucleotide sequence of the promoter CmUbi is shown as SEQ ID NO. 4. Through double luciferase reporter experiments and stable genetic transformation experiments, it is proved that the expression activity of the CmUbi promoter in chrysanthemum protoplast and callus is significantly higher than that of commonly used CaMV 35S, AtRPS5a and corn ZmUbi promoters. A CRISPR / Cas9 gene editing vector is constructed by using the promoter, and a key gene of branch of hexaploid chrysanthemum is edited BRC1b , and a plurality of allele simultaneous mutation complete editing strains are successfully obtained, and the number of branches of the mutants is significantly increased. The endogenous high-activity promoter of the chrysanthemum provided by the application effectively solves the problems of low expression activity of an existing heterologous promoter in the chrysanthemum and poor gene editing efficiency of a polyploid, and provides an efficient and adaptive expression regulation element for chrysanthemum molecular breeding and functional genomics research.

A method and composition for preparing a DNA library for replicating a target nucleic acid sequence are provided. A target DNA template containing the target sequence is circularized via a terminal adapter to form a circular construct, which is extended bidirectionally by polymerase-mediated elongation initiated at a nick site on the terminal adapter. Following polymerase-mediated elongation, a diploid DNA template is formed containing two copies of the target DNA template (and thus two copies of the target sequence). Each strand of the diploid DNA template contains a parent polynucleotide chain ligated to a newly synthesized daughter strand copy of the parent polynucleotide chain. Predetermined sequences, such as primer sequences, unique molecular identifiers, and sequence indices, can be included in the diploid DNA template. Sequencing of the diploid DNA template can reveal genetic / epigenetic information related to the target sequence. A method for constructing asymmetric and multiploid DNA template constructs is also provided.

The present application relates to the technical field of bioinformatics, and particularly relates to a genome selection method and application thereof in breeding of homologous polyploids. The method comprises the following steps: constructing an additive genomic kinship matrix; constructing a genomic selection model based on the additive genomic kinship matrix; and estimating breeding values of individuals of a species according to the genomic selection model. The genotype matrix used in the additive genomic kinship matrix of the homologous polyploids is a polyploid genotype matrix, and the value of the genotype in the polyploid genotype matrix is determined by the copy number of any one allele. The present application proposes a genome selection method based on the homologous polyploid genomic kinship matrix for the species with the characteristics of homologous polyploids, which can accurately predict the genomic breeding values of individuals of the homologous polyploid species and improve the accuracy of genome selection, and has important significance in the field of genome selection breeding.

The invention discloses a centella asiatica polyploidy seedling factory production method based on micro-organ cutting-water culture combination, which comprises the following steps: (1) selecting tetraploid centella asiatica aseptic seedlings as materials, transplanting rooting seedlings of the tetraploid centella asiatica aseptic seedlings into nutrient soil for culture, and cutting growing stolons into stem segments with axillary buds; (2) micro-organ cutting culture: cutting the stem sections treated in the step (1) on an improved MS solid culture medium for culture; simultaneously germinating new buds and new roots; transferring the seedlings into an improved MS liquid culture medium for water culture, transferring the obtained seedlings to outdoor for seedling hardening, and then performing field transplanting and field planting. The invention provides an efficient, economic and large-scale propagation scheme for new germplasm after genetic improvement of centella asiatica, so that large-scale industrialized production can be realized in a short time, and the method has a wide industrial application prospect.

The application discloses a method for creating polyploid fish, which comprises the following steps of: knocking out a spo11 gene by using a CRISPR / Cas9 gene knockout technology, obtaining a spo11 homozygous knockout individual through genotyping and breeding, utilizing the characteristic that a spo11 homozygous knockout diploid female can produce part of unmeiotic oocytes to obtain a triploid, and further screening a spo11 homozygous knockout triploid 3N spo11 ‑ / ‑ / ‑ ; utilizing the characteristic that a 3N spo11 ‑ / ‑ / ‑ female can produce part of unmeiotic oocytes to further screen a heterozygous knockout tetraploid 4N spo11 + / ‑ / ‑ / ‑ ; utilizing the characteristic that a 4N spo11 + / ‑ / ‑ / ‑ can produce a large number of diploid gametes to further prepare a large number of triploid offspring. Compared with other polyploid breeding methods, the application has universality and genetic controllability, and provides important technical support for artificially inducing fish polyploidy.

The invention relates to the field of phytomining and, particularly, to a method for collecting metal from an induced polyploid or mixoploid plant.

The present invention provides a method for inducing Arundo donax polyploids. Through the induction method of the present invention, on the basis of rapid propagation of Arundo donax by tissue culture, the culture medium can be scientifically formulated, the culture conditions can be reasonably adjusted, and Arundo donax polyploids with excellent traits can be obtained quickly and efficiently, effectively reducing the incidence of chimeras. The present invention uses the tillering tissue mass of clustered buds as the induction material, which enriches the existing Arundo donax polyploid induction materials compared with the traditional Arundo donax polyploid induction materials. By using colchicine at a specific concentration to induce the tillering tissue mass, the induction rate of Arundo donax polyploids is increased, the incidence of chimeras is effectively reduced, time and costs are saved, and a new way is provided for the breeding of new Arundo donax varieties.

The invention relates to the technical field of eucommia ulmoides breeding. The invention provides a breeding method and an identification method of triploid eucommia ulmoides, which comprises the following steps: taking eucommia ulmoides branches, cutting, treating with colchicine solution, and culturing to obtain adventitious buds; cultivating the adventitious buds to obtain seedlings, performing ploidy identification on the seedlings, and screening out tetraploid plants; taking the tetraploid plant as a female parent, hybridizing with the diploid male parent, and harvesting triploid eucommia ulmoides seeds. The invention provides the method for efficiently and stably obtaining the homozygous tetraploid eucommia ulmoides clone, the problems of common chimeras and unstable materials in chemical induction are solved, and reliable parents are provided for cross breeding. A set of complete technical system for directionally creating the triploid eucommia ulmoides by taking tetraploid as a female parent and diploid as a male parent (reverse crossing) is established, and the technical blank from creation of a polyploid parent to breeding of a new triploid variety is filled.

The invention discloses a method for separating and purifying haploid nucleuses and polyploid nucleuses of polyploid hepatocytes, and belongs to the field of molecular biology, biological medicine and medical health care, and the method comprises the following steps: S1, performing perfusion by using an improved two-step collagenase digestion method to obtain primary hepatocytes; s2, dyeing the primary hepatocytes and carrying out first flow sorting; s3, respectively grinding the diploid hepatocytes, the tetraploid hepatocytes and the octaploid hepatocytes after the first flow sorting to obtain hepatocyte nucleuses; s4, the hepatocyte nucleuses obtained after grinding are subjected to second-time flow type sorting; therefore, the application of the invention can break through the limitation that the polyploidy hepatocyte nucleus cannot be distinguished in the traditional hepatocyte separation, so that the separation of the hepatocyte nucleus is more accurate, the bottleneck of polyploidy karyotype separation of the hepatocyte nucleus is broken through, and the development of the research field of the hepatocyte nucleus is quickly promoted.

The invention discloses a tissue culture method for reducing the somatic variation rate of bamboo reed by adding chitosan oligosaccharide. According to the method, 50-200 mg / L of chitosan oligosaccharide with the specific molecular weight of (3000-5000 Da) is added into a callus induction culture medium, and 1.5 mg / L of 2, 4-D and 0.2 mg / L of 6-BA are combined, so that the variation rate of somatic cells is reduced from 23.7% to 9.8%. The SSR molecular marker and flow cytometry prove that the genetic stability is remarkably improved, and the biomass of the regenerated plant is increased by 15-20%. The method is suitable for large-scale cloning propagation of energy plants.

The invention discloses a method for creating poplar triploid, and belongs to the technical field of poplar polyploid induction. The creation method comprises the following steps: performing cross pollination on female flower buds of populus japonica by using pollen of a male parent of a poplar to obtain a female inflorescence, treating the female inflorescence at 39-41 DEG C for 2-4 hours, managing the inflorescence, harvesting seeds, sowing and raising seedlings, and identifying seedlings to obtain the populus japonica triploid plant. According to the method, on the basis of systematically analyzing the development process of the female gametes of the populus japonica, a high-temperature induced female gamete chromosome doubling technology is innovatively adopted, a populus japonica female gamete chromosome doubling induction system is successfully constructed, the triploid induction rate is increased by 63.95%, and a solid foundation is laid for populus japonica triploid germplasm creation.

The invention relates to a polyploid forest gene editing type identification method and application thereof, and belongs to the technical field of molecular biology. The invention provides a polyploidy forest gene editing type identification method which comprises the following steps: carrying out a first round of PCR (Polymerase Chain Reaction) amplification by taking genome DNA (Deoxyribonucleic Acid) of a gene editing strain as a template to obtain a first round of amplification product; carrying out second-round PCR amplification by taking the first-round amplification product as a template to obtain a second-round amplification product; and carrying out Hi-TOM sequencing analysis on the second round of amplification product to obtain a Hi-TOM sequencing result. According to the identification method disclosed by the invention, two rounds of PCR amplification are combined with HI-TOM sequencing analysis, so that the identification of the editing type of the polyploidy forest tree gene editing strain can be realized, a homozygous gene editing plant is obtained, the defects of a traditional identification method are overcome, and the method has the characteristics of high efficiency and low cost.

The invention discloses an application of H2A. Z1 protein and related biological materials thereof in regulation and control of plant chromosome doubling. According to the invention, a model species poplar of a woody plant is taken as a research object, and a transgenic poplar with reduced H2A.Z1 gene expression quantity is constructed. Experiments prove that compared with a wild type material, the plant height and the stem node number of the transgenic poplar with reduced H2A.Z1 gene expression quantity are reduced, and the stem node length is increased. In addition, cells in stem and leaf organs of the transgenic poplar with reduced H2A.Z1 gene expression quantity become larger, cell chromosomes are doubled, and diploid is changed into tetraploid. The invention has important significance for researching polyploid breeding work of plants such as vegetables, important flowers, fruit trees and the like.

The application belongs to the technical field of plant breeding, and particularly relates to a method for inducing taro tissue culture seedlings by using herbicides. The method comprises the following steps: S1, treating explants; S2, inducing polyploidy; S3, subculture; and S4, inducing rooting. The method uses an inducer liquid medium to induce taro single buds, and then uses a separation medium to culture, so that polyploid taro single buds which are obviously morphologically changed are separated, then a subculture medium is used for subculture to obtain polyploid multiple buds, and finally a rooting medium is used for rooting culture of the taro single buds on the polyploid multiple buds, so that the taro tissue culture seedlings are finally obtained. The method is simple to operate and easy to realize. In addition, the taro tissue culture seedlings obtained by using the herbicides have high resistance to taro blight, can enrich taro germplasm resources, and promote the development of the taro industry.