298 results about "Hybrid gene" patented technology

The invention discloses a method for identifying seed purity of a muskmelon variety namely green angel hybrid variety based on an EST-SSR (expressed sequence tag-simple sequence repeat) marker. The method comprises the following steps: using the muskmelon variety namely green angel hybrid variety and parental genome DNA (deoxyribonucleic acid) of the muskmelon variety namely green angel hybrid variety as a template; designing 57 pairs of EST-SSR primers by using Primer 3.0 online primer design through 214 muskmelon EST sequences published by a GeneBank database; obtaining a pair of complementary band type primers of which the hybrid variety band type is parental by screening. By virtue of a field verification test, the primer is good in stability, is relatively matched with results of the field test, and can be used for performing purity identification on the muskmelon hybrid variety. A detection method disclosed by the invention can be used for completing seed purity identification within 3 hours, and has the advantages of high speed, low cost, convenience in operation and the like.

The invention discloses primers and a method for purity identification of hybrid seeds of Chinese pumpkin 'Guangmi NO.1'. The primers are VRISSRPMK8-F:5'-TAATTAAGGCGTGAAATTGG-3' and VRISSRPMK8-R:5'-GAAAAGAAACATAAACAGGGC-3'. The identification method is as follows: 1) extracting a genome DNA (deoxyribonucleic acid) of a pumpkin seedling; 2) carrying out PCR (polymerase chain reaction) amplification by taking the genome DNA of the pumpkin as a template and using the SSR primers VRISSRPMK8; 3) carrying out gel electrophoresis on the amplified product; and 4) analyzing the electrophoresis result, wherein only the single plant simultaneously having parent specific bands is a real hybrid, and the plant lacking of any one band is marked as false hybrid. The primers and the method can be used for effectively distinguishing the hybrid seed of the 'Guangmi NO.1' from the female parent and the male parent, and can identify the purity of the hybrid seeds rapidly and accurately. The method has the advantages of being fast and accurate, low in cost and simple in operation, and the like.

An eka-cecropinA-magainin heterozygous gene engineering antibacterial peptide is prepared by adopting No.1-7 amino acid residue on N end of cecropinA CA and No.2-12 amino acid residue on N end of magainin M, design synthesizing heterozygous peptide CA(1-7)-M(2-12) gene by codon, connecting with carrier pPICZ alpha-A, mutating by point mutation technology to obtain mutant pPICZ alpha-CA-Mu-1, pPICZ alpha-CA-Mu-2 and pPICZ alpha-CA-Mu-3, converting SMD1168, and expressing CecA-Mag heterozygous antibacterial peptide to make into use under alcohol hydrogenase starter regulation.

The invention relates to a breeding method and application of a cytoplasmic male sterility restoring line of brassica napus rapeseed and radish. The method comprises the following steps of: choosing rapeseed S1628 with the preservation number of CCTCC NO: P201203, performing continuous inbreeding directional selection for six years to breed an excellent, high-oil and stable inbred line; using the cytoplasmic male sterility restoring line of the brassica napus rapeseed and radish as a female parent and using the bred excellent and stable inbred line as a male parent to select out strains comprising radish cytoplasmic male sterility restoring genes from large amounts of filial generations; using the strains comprising the radish cytoplasmic male sterility restoring genes as the female parent and using Zhongshuang No.2, Huashuang No.3, Huyou 17, Zunyou No.1 and Zheyou 758 of double-low winter rapeseed varieties in China as recurrent male parents to hybridize, performing the continuous inbreeding directional selection for six years to breed the cytoplasmic male sterility restoring line of the double-low and high-oil brassica napus rapeseed and radish. The restoring line has been crossed with hybrid varieties. A gap of producing the hybrid varieties by the cytoplasmic male sterility restoring line of the double-low radish in China is filled, and the application prospect is broad.

The invention provides a primer group for identifying a male parent and a female parent of tachysurus fulvidraco and leiocassis longirostris hybrid species. The primer group comprises a primer pair for amplifying COI genes and a primer pair for amplifying an ITS sequence, wherein the primer pair for amplifying COI genes consists of a primer COIF and a primer COIR; the primer pair for amplifying an ITS sequence consists of a primer ITSF and a primer ITSR. The invention also provides a kit which is for identifying the male parent and the female parent of tachysurus fulvidraco and leiocassis longirostris hybrid species and comprises the primer group. The kit also comprises any one or more of the following reagents: a buffer solution for PCR, dNTP, heat-resistant DNA polymerase, a cloning vector, a DNA ligase, and a buffer solution for ligation. The invention also provides a method for identifying the male parent and the female parent of tachysurus fulvidraco and leiocassis longirostris hybrid species. The primer group, the kit or the method can be used for accurately identifying the male parent and the female parent of the tachysurus fulvidraco and leiocassis longirostris hybrid species, thus effectively managing a breeding plan and monitoring negative influence thereof, and has great significance in sustainable development of aquaculture industry.

The invention discloses a simple sequence repeat-polymerase chain reaction (SSR-PCR)-based hybrid rape seed purity identification method, which comprises the following steps of: performing sprouting culture on hybrid rape sample seeds to be detected, performing alkaline lysis on the cultured seedlings, simultaneously performing ultrasonic disruption treatment, and adding an extracting buffer solution to obtain a genome DNA solution; performing PCR amplification on genome DNA by using an SSR primer sequence; performing voltage stabilizing electrophoretic separation on a PCR amplification product in agarose gel; performing imaging and tape reading on the PCR amplification product subjected to electrophoretic separation in a gel imaging system, comparing band characteristics of the sample seeds with those of parent seeds, counting seeds with the band characteristics of male parent and the band characteristics of female parent in the sample seeds, and obtaining the purity of the hybrid rape seeds to be detected according to a variety purity formula. The identification method has the advantages of quickness, simplicity, convenience, high throughput, low detection cost, high detection efficiency, stable and reliable detection results and the like.

The invention discloses a primer for identifying purity of a broccoli hybrid scarlet pimpernel variety. The primer is a combination of one or more of four pairs of primers: (1), primers Ol12-D05; (2), primers Ra1-F06; (3), primers FITO439; and (4), primers Ol10-A11. The molecular marking method as well as the kit and the primer for identifying purity of broccoli hybrid scarlet pimpernel variety have the beneficial effects of being capable of clearly distinguishing and judging different varieties of broccoli and capable of accurately and quickly distinguishing the variety truth and variety purity, so that reliable technology is provided for variety identification, seed market management, variety protection, seed quality control before seeding, good seed breeding and the like.

The invention discloses a pair of EST-SSR markers ATA157 and ATA154 which are closely interlocked with the disease resistant and susceptible genes of cabbage turnip mosaic virus, and belong to the technical field of biology. The markers are a pair of allelic markers, and the interlocking distance thereof with cabbage TuMV resistance locis is 3.8cM. Wherein the fragment length of the marker ATA157 is 157bp, and the fragment length of the marker ATA154 is 154bp. The invention also discloses a pair of EST-SSR labeled primers HCC259 which are provided with sequences in sequence tables SEQ ID No.3 and SEQ ID No. 4. Through the primers, the genome DNA of a single plant to be tested is adopted as the template and the markers are amplified so as to realize the high-efficiency selection of cabbage TuMV resistance, speed up the breeding of novel TuMV resistant breed strains of cabbage, and establish a novel Chinese cabbage hybrid parent selection system.

The invention discloses a method for identifying hybrid germplasm of actinidia based on genome heterozygosity, relating to the technical fields of plant genetic resources and biology. The method comprises the following steps: (1) performing low-depth genome sequencing on a sample; (2) comparing reference genomes of sequencing data and mining single-base mutation sites; (3) calculating the heterozygosity of genome level of the sample based on the single-base mutation sites; (4) constructing a basic species heterozygosity distribution rule database of a main ancestor of the actinidia; and (5) performing heterozygosity comparison on a to-be-detected sample and the database so as to identify the hybrid germplasm. Compared with the traditional molecular marker technology, the method disclosed by the invention has the advantages that the identification accuracy is greatly improved; and meanwhile, compared with the traditional morphological identification method, the method disclosed by the invention is high-efficiency and rapid, and the identification time and cost can be saved. The method disclosed by the invention has wide applicability on different sequencing platforms, and comprehensive evaluation and application of the germplasm resources of actinidia can be formed.

The invention provides a method for rapidly creating an engineering female sterile line suitable for mechanized seed production by using a genome editing technology. The method comprises the followingthree steps: (1) acquiring a female sterile mutant by virtue of a mutagenesis technology or a gene editing technology; (2) transferring an expression vector carrying three expression cassettes such as female fertile genes, pollen lethal genes and selective marker genes into a receptor material by virtue of a transgenic technology to obtain a transgenic material; and (3) breeding the engineering female sterile line suitable for the mechanized seed production by hybridizing and polymerizing the female sterile mutant and the transgenic material. By adopting the method, an excellent restoration line with same initial planting period of the sterile line can be rapidly converted to the engineering female sterile restoration line suitable for the mechanized seed production. The method is wide inapplication and market prospect in the field of agriculture.

The invention relates to a cultivation method of common wheat capable of stably expressing six HMW-GS (High Molecular Weight-Glutenin Subunits) and belongs to the field of plant chromosome engineering technologies. The method comprises the following steps of: taking common wheat as a female parent and wild emmer wheat as a male parent; pollinating pollen of the wild emmer wheat with four HMW-GS to sextuploid common wheat; and carrying out inter-specific crossing distant hybridization, multi-generation selfing and identification to breed the common wheat capable of stably expressing the six HMW-GS. The novel common wheat with the six HMW-GS, which is cultivated by the method, not only has a strain leave type which is similar with the male parent, but also has a purple stalk property which does not exist in the female parent, and has better grain setting property, output property and excellent quality property when being compared with the female parent; and HMW-GS detection is continuously carried out on grains of F10, F11 and F12 generations of hybrid seeds so as to testify that 1Ay genes can be stably transmitted and expressed generation by generation.

The invention discloses a maize yellow-green leaf gene ygl-1 disclosed as SEQ ID NO.1 and a coded protein thereof disclosed as SEQ ID NO.2. The invention also discloses primers for amplifying the maize yellow-green leaf gene ygl-1, which are disclosed as SEQ ID NO.4 and SEQ ID NO.5. The maize yellow-green leaf gene ygl-1 is detected in maize for the first time, can be used for maize high-quality seed breeding and crossbreeding, and can be used as a molecular marker for germplasm resource identification. After the germplasm material or transformed descendant containing the maize yellow-green leaf gene ygl-1 is selected, the marker can be utilized to accurately select the genotype of the backcross transformed descendant, and can also be used for screening corn germplasm resources to search for the mutant material with more abundant genetic background, thereby having important application value for improving the hybrid seed purity control effect.

The invention provides a breeding method of dedicated silkworm hybrid secreting natural and green sericin with high content, comprising the following steps: taking exarate pupae breeds (with Nd-s genes) as stuff to be hybridized with the breeds with high amount of silk production, and breeding Japanese special original strains with sericin content being more than 99.0% and homozygous genotypes (Nd-s/Nd-s) by adopting the methods of selfing, backcrossing, marker gene selecting, sericin content tracking and detecting and the like; and hybridizing the Japanese special original strains with the Chinese conventional breeds (Gc/Gc) with green cocoon properties to breed the dedicated silkworm hybrid. The method can provide high quality raw materials to such fields as cosmetics, medicines, food, fiber modified materials and the like and improve the product added value.

The invention discloses an exogenous inserting fragment flanking sequence of transgenic corn BBHTL8-1 and an application of the exogenous inserting fragment flanking sequence of the transgenic corn BBHTL8-1. The nucleotide sequences of the flanking sequence are respectively shown as a sequence table SEQID NO:1 and a sequence table SEQID NO:2. The invention further provides a primer pair for detecting a 3' end flanking sequence. The identification of the exogenous inserting fragment flanking sequence of the transgenic pest-resistant herbicide-resistant quality improvement corn BBHTL8-1 disclosed by the invention are suitable for corn BBHTL8-1 including parents, hybrid and descendant relevant materials, and includes detection of plants, tissue, seeds and relevant products.

The invention belongs to the field of breeding techniques, and particularly relate to a method for breeding good corn germplasm high in combining ability. The method comprises the steps of firstly collecting good selfing lines for constructing an initial group, performing genotype identification and heterotic group division, selecting specific hybrids as test hybrids for assessing the combining ability expression of each material in the initial group, and establishing a complete genome selection model; then in the heterotic group, according to the combining ability assessment result, performing breeding step by step to obtain good single plants, and further obtaining homozygosis selfing lines; and finally, performing genotype identification on the homozygosis selfing lines, forecasting thecombining ability according to the complete genome selection model, and reserving the homozygosis selfing lines having high combining ability estimated value namely obtaining the good germplasm highin combining ability. The method disclosed by the invention is high in breeding efficiency and short in breeding cycle, and can accelerate the breeding process of good germplasm.

The invention discloses a microsatellite marker paternity identification primer and method suitable for nile tilapia, blue tilapia and a hybrid thereof, and application thereof, and belongs to the technical field of fish breeding. The method screens 13 pairs of fluorescent marker microsatellite primers for paternity identification. The identification method comprises the following steps of: constructing a complete sibling family of the tilapia, the blue tilapia and the hybrid thereof; extracting genomic DNA of parents and progeny of each family; screening polymorphic microsatellite markers; performing PCR amplification on fluorescent marker microsatellite primers; and performing multiple capillary electrophoresis typing and paternity identification. The invention establishes a microsatellite marker paternity identification method suitable for the tilapia, the blue tilapia and the hybrid thereof for the first time, the identification accuracy rate is 100%, and the identification accuracy rate can reach 99.43% in practical application. The method has high identification accuracy and simple and convenient operation, and can be used for guiding population inheritance and family pedigree management of the three tilapia in the breeding process.

The invention provides a rice fertility regulating gene and its mutant and an application thereof. The invention provides a rice gene GMS1, which has the functions of regulating rice male germ cell development and pollen fertility, a CDS sequence is shown as SEQ ID NO: 2, and an amino acid sequence is shown as SEQ ID NO: 3. The invention provides a radiation mutagenesis mutant and a CRISPR knockout mutant of the GMS1 gene, and provides a molecular marker identification method for the mutant. The rice gene GMS1 provided by the present invention can be used for sterile breeding and production ofrice hybrids, and has great application value and economic value.

The present invention provides an improved and simplified plant chromosome fluorescence in-situ hybridization method, wherein the method does not include a long-time dewatering process of a specimen slide before hybridization and multiple rinsing processes of the specimen slide after hybridization, such that the method is the technology for carrying out rapid in-situ hybridization on the intranuclear chromosome in a biological cell sample. The method mainly comprises: 1) carrying out variable temperature germination, 8-hydroxy quinoline pretreatment, and fixation of the strong cell division tissue in the plant root tip meristem region with a Carnoy fixation liquid to obtain a metakinesis phase specimen with characteristics of clear image and good chromosome dispersion; 2) after mixing fluorescent probes having different labels, carrying out one time denaturation, and carrying out low temperature storage so as to achieve long term repeated use; 3) carrying out denaturation on the sample slide with a NaOH alcohol solution; and 4) during a hybridization reaction process, simplifying the multi-step dewatering, rinsing and other operation steps so as to substantially simplify the whole complex hybridization process. According to the present invention, the method is particularly suitable for the rapid large-scale detection and analysis of the exogenous chromosomes fragment and the chromosome fragment in the plant distant hybridization and close hybridization species, and the morphology and structure authenticity of the specimen chromosome is well maintained.

The invention relates to a breeding and seed producing method for a recessive gene male sterile line of cauliflower. The invention adopts the technical scheme that a recessive gene male sterile mutant strain of the cauliflower is subjected to sister hybrid and progeny selfing separation to breed a sterile strain with the sterile rate of 100 percent and the sterile degree reaching or approximating to 100 percent; the sterile line used as a female parent is hybridized and combined with an excellent selfing line; and a combined hybrid seed is produced by applying a honeybee pollination method. The invention has the advantages that: firstly, a new sterile source for heterosis advantage utilization of the cauliflower is provided and the defects of cytoplasmic sterility are overcome; secondly, the recessive gene male sterile line is maintained and reproduced by using a tissue culture technology and various works for filed isolation and preproduction of the sterile line and a maintenance line are omitted, so that the influence on preproduction of the sterile line caused by weather, season, land and isolating conditions is avoided; and thirdly, a hybrid strain is produced by using the recessive gene male sterile line and honey can be used for pollination, and thus the seed production cost is reduced and the seed production efficiency is improved.

The invention relates to a cultivation method of a high-powdery-mildew-resistance, short-fruit-stem, and thick-flesh cucumber chromosome single segment substitution line. Dwarf powdery mildew susceptible inbred line cucumbers D8 are used as a female parent, a long-bine powdery mildew resistant inbred line JIN5-508 is used as a male parent, and hybridization, backcross and molecular marker-assisted selection technology are adopted, so that a dwarf high-powdery-mildew-resistance cucumber chromosome single segment substitution line D85109267 is obtained through consecutive 12-generation backcross and 4-generation inbred selective breeding. The genetic background of the substitution line is almost identical to that of the powdery mildew susceptible female parent D8, and the only difference lies in that a chromosome single segment with length of only 0.7cM in the powdery mildew resistant male parent JIN5-508 is substituted in the genetic background. The new dwarf powdery mildew resistant cucumber line D85109267 has the advantages of short internode, concentrated flowering period, short fruit stem, thick flesh, concentrated fruit picking, high early yield and income increasing by more than 20%, and can be used for preparing hybrids for the purpose of early-maturing.

The invention discloses a molecular marker closely linked with rice blast Pi9 gene and an application thereof and belongs to the field of molecular marker-assisted breeding. The invention discloses primer pairs for detecting the marker closely linked with rice blast Pi9 gene and a method for detecting whether a rice variety or a plant line contains Pi9 gene or not by virtue of the primer pairs. The method comprises the following steps: (1) extracting genomic DNA of a to-be-detected rice sample; (2) carrying out PCR amplification through the primer pairs; (3) carrying out enzyme digestion on the PCR amplification product obtained in the step (2) with an endonuclease and carrying out electrophoresis; and (4) determining whether the to-be-detected rice sample contains the Pi9 gene or not according to the enzyme digestion results. By the method, whether the to-be-detected rice sample contains the Pi9 gene or not and the contained Pi9 gene is heterozygous or homozygous can be accurately determined and the purity of an F1 hybrid seed of one of the parents containing the Pi9 gene can also be identified and the method has application prospects in Pi9 gene-assisted breeding.

The invention discloses a method for establishing a DNA fingerprint spectrum of eggplant varieties. The method comprises the following steps: extracting eggplant genome DNA; performing SSR primer amplification and screening; performing PCR (Polymerase Chain Reaction) amplification and electrophoresis detection of the product; establishing DNA digital fingerprinting, namely the fingerprint spectrum; and establishing digital two-dimensional codes. The invention further provides a DNA fingerprint spectrum established according to the method and application of the spectrum in purity identificationof seeds of a hybrid eggplant 'Hu Eggplant V'. The method for identifying the seeds by utilizing the DNA fingerprint spectrum of the variety and the digital two-dimensional codes is a molecular marker method by taking polymorphism of DNA as basis, and multiple defects and difficulties in morphological identification of the seed purity as well as isozyme and seed protein electrophoresis identification can be overcome and solved. Specifically, the DNA fingerprint spectrum and digital two-dimensional codes of the facility cultivation variety 'Hu Eggplant V'and parents 'Fu-2' and 'Jiang Eggplant'are established by utilizing the SSR technology, and a scientific basis can be provided for identification of the variety and the purity detection.

The invention discloses a method for converting tomato red fruit material into pink fruit material by gene editing. The invention discloses two sgRNA target sequences edited by CRIPSR/Cas9 gene of SlMYB12 gene. The SlMYB12 gene of tomato was precisely edited by transforming the vector, and the homozygous mutation of SlMYB12 gene of tomato is screened out from the self-bred progeny, and the corresponding hybrid of tomato is bred by transforming the vector into tomato. The experiment proves that the gene editing method of the invention is applicable to different tomato varieties and has high editing efficiency. Compared with the traditional backcross breeding at y locus, this method can greatly shorten the breeding time, realize the fast and accurate transformation of fruit color traits within one year, and is not limited by the factors such as linkage redundancy and transgenic safety, so it has great breeding application prospects and economic value.