48 results about "Heterozygous genotype" patented technology

The invention discloses an SNP (Single Nucleotide Polymorphism) molecular marker for early identification of the cold resistance of an actinidia chinensis tree body in the dormancy period, and the SNP molecular marker is located at the Chr69297479 position on the No.6 chromosome of the actinidia chinensis and is closely associated with the cold-resistant phenotype median lethal temperature (LT50) of the actinidia chinensis. The method can be used for identifying the tree body dormancy period cold resistance of natural population and hybrid filial generation of actinidia chinensis. The homozygous genotype (CC) of the SNP locus corresponds to a high-cold-resistance individual in the dormancy period of an actinidia chinensis tree body, and the heterozygous genotype (TC) of the SNP locus corresponds to a low-cold-resistance individual. The cold resistance prediction accuracy in the F1-generation hybrid population reaches 94.94%, and the prediction accuracy in the natural population reaches 96.15%. The molecular marker detection method disclosed by the invention is convenient and rapid, has good specificity and high accuracy, is not influenced by climate, environment and the like, can rapidly and effectively evaluate the cold resistance of natural resources and hybrid filial generations of the actinidia chinensis, and provides powerful technical support for breeding cold-resistant varieties of the actinidia chinensis.

The invention discloses an SNP (Single Nucleotide Polymorphism) molecular marker related to the tolerance character of white spot syndrome virus (WSSV) of procambarus clarkia, the molecular marker is located in the third intron region of a TRIM gene, is specifically located at 585bp of a DNA (Deoxyribose Nucleic Acid) sequence as shown in SEQ ID NO.1, and is Tgt; the T / C heterozygous genotype is obviously related to the WSSV tolerance character of the procambarus clarkia. The invention provides a favorable marker for breeding of disease-resistant molecules of procambarus clarkii, and can accelerate the breeding process of disease-resistant varieties of procambarus clarkii.

The invention discloses an identification method of agilawood germplasm resources of aquilaria agallocha. The identification method is characterized in that seven pairs of primer sites corresponding to DNA molecular markers of seven genome SNP sites are provided; the method comprises the following steps: carrying out PCR (Polymerase Chain Reaction) amplification and primer effect test, carrying out enzyme digestion on a PCR product, and distinguishing agilawood and aquilaria sinensis according to an enzyme digestion result, and / or distinguishing different varieties of agilawood; if the seven pairs of primer loci are not subjected to enzyme digestion, judging the aquilaria sinensis as aquilaria sinensis. Effective amplification of the agilawood molecular marker is realized by adopting a conventional PCR technology, and the method has universality; genotype identification is completed by integrating an enzyme digestion method with agarose gel, an expensive sequencer is not needed, and the method has the technical advantages of being easy and convenient to operate, low in cost and the like; the polymorphism of genotypes, including homozygous genotypes and heterozygous genotypes, can be accurately identified.

The invention belongs to the technical field of plant molecular biology, and discloses a PCR amplification primer suitable for molecular identification of amorphophallus konjac and amorphophallus albus hybrids, the PCR amplification primer comprises a ZJ20 primer, and the sequence of the ZJ20 primer is F: CAGACCATAATCAGCATG; r is GTCTCCACT GCCGTACCTT, and R is GTCTCCACT The molecular identification method comprises the following steps: step 1, extracting genome DNA of a to-be-detected konjak sample, carrying out amplification by using the PCR primer, then carrying out first-generation DNA sequencing, and if a hybrid genotype A / T is observed at three positions of 170-180bp, 190bp-200bp and 210bp-220bp, determining that a hybrid first filial generation of amorphophallus konjac and amorphophallus albus is obtained, and a reaction system for PCR amplification in the step 2 is as follows: the reaction total volume is 25 microliters, and the reaction time is 30 minutes; a reagent contained in the step 3 is as follows: 12.5 mu L of a 2 * PCR Mix premixed solution, 0.3 mu M of R and F primers and 30 ng of a sample genome DNA solution, finally, sterile ultrapure water is used for complementing the deficient volume, and the amplification process of the PCR reaction in the step 3 is as follows: pre-denaturation at 95 DEG C is performed for 5 minutes, then denaturation at 95 DEG C is performed for 45 seconds, annealing at 56 DEG C is performed for 45 seconds, extension at 72 DEG C is performed for 45 seconds, and 35 amplification cycles are performed in total.

The invention relates to a method for breeding a new variety of Sichuan rural black pigs. Belongs to the technical field of breeding and breeding. The method includes the steps that Duroc pigs are used as male parents, Tibetan pigs are used as female parents, and grading hybridization is carried out until F4 generation hybridization is achieved; crossbreeding the F4-generation boars and the F4-generation sows to establish a zero-generation basic pig herd; carrying out closed breeding on the zero-generation basic swinery, and carrying out breeding on each generation until the fifth generation; wherein the breeding mode of each generation comprises the following steps: breeding the pig individuals of the generation based on the numerical value of the BLUP comprehensive selection index of the individual, carrying out hair color SNP locus genotype detection on the pig individuals of the generation through a Snapshot technology, and rejecting the pig individuals with black heterozygous genotypes in the pig herd; and determining the pig individuals of the fifth generation as the new male parent variety of the Sichuan-rural black pigs. As a male parent, the Sichuan rural black pigs bred by the method have the advantages of pure black hair color, excellent meat quality, high production efficiency and the like.

Mouse models of obesity associated with human MC4R missense mutations are provided. Human mutations introduced with CRISPR / Cas9 into the mouse MC4R gene lead to an severely obese mouse phenotype in mice with a heterozygous genotype.

The invention belongs to the field of molecular genetic breeding, and discloses an InDel molecular marker of a rice salt-tolerant gene STG5, a primer group and application of the InDel molecular marker, and the nucleotide sequence of the InDel molecular marker is as shown in SEQ ID NO.1; aiming at specific difference sites existing in STG5 gene sequences of a salt-tolerant haplotype Hap1 and a salt-sensitive haplotype Hap2, an InDel molecular marker primer STG5UF / STG5UR for distinguishing a salt-tolerant genotype and a sensitive type of a rice gene STG5 is designed, so that an STG5 allele is accurately identified, and a favorable tool is provided for subsequently cultivating a new salt-tolerant rice variety. The invention discloses an intragene molecular marker of a salt-tolerant gene STG5, which is a co-dominant functional marker and can accurately distinguish a salt-tolerant homozygous genotype, a salt-sensitive homozygous genotype and a heterozygous genotype.

The application relates to a co-dominant molecular marker of a potato long-day adaptation gene StCDF1.4 and application thereof. The molecular marker can identify a StCDF1.4 homozygous or heterozygous genotype alone, and makes up the shortcoming that an existing molecular marker can distinguish StCDF1.4 genotype types only by combining multiple pairs of primers.

The invention discloses a molecular marker, detection primer and application of a fast-growing trait of Litopenaeus vannamei. The SNP molecular marker significantly associated with the fast-growing trait is located at the 308bp site of the nucleotide sequence shown in SEQ ID NO.1 (i.e., the P2638 site inside the DAN4-like cell wall protein encoding gene dan4), and the genotype of the site includes G / G, A / A, and G / A types. The prawn individuals with the A / A homozygous genotype of the SNP site have a significantly fast growth rate, the prawn individuals with the G / G homozygous genotype of the SNP site have a relatively slow growth rate, and the prawn individuals with the G / A heterozygous genotype of the SNP site have a medium growth rate. The SNP molecular marker and the molecular marker detection primer of the invention can be used for the evaluation of the characteristics of the growth trait of the population, and can also be used for the selection of prawn individuals with potential growth advantage traits.

The invention relates to a CAPS (cleaved amplified polymorphic sequence) molecular marker for identifying the content of rosa chinensis aroma leaf acetate and application of the CAPS molecular marker, and relates to the technical field of molecular breeding, the marker is located at the 27954680 site of the third chromosome of a rosa chinensis genome, and the nucleotide sequence of the CAPS molecular marker for identifying the content of rosa chinensis aroma leaf acetate is shown as SEQ ID No.1. DNA fragments with the length of 447 bp are obtained through amplification of a specific primer pair, and genotypes are distinguished by combining DdeI enzyme digestion analysis: after enzyme digestion, the generated 447 bp and 274 bp fragments are low-content heterozygous types (T / A), and a 447 bp single band is retained as a high-content homozygous type (T / T). The genetic difference of the leaf acetate content of the Chinese rose can be rapidly identified, the flower fragrance character screening efficiency is remarkably improved, the breeding cost is reduced, an efficient molecular tool is provided for high-fragrance Chinese rose variety breeding, and the method has important application value.

The invention relates to the technical field of plant molecular biology, in particular to an SNP (Single Nucleotide Polymorphism) molecular marker related to a pitted surface of pepper and application. The SNP molecular marker can stably and accurately distinguish pitted surface (G / G), smooth surface (C / C) and heterozygous (G / C) genotypes of capsicum fruits, the marker typing and phenotype consistency is high, the accuracy rate reaches 90% or above, and the SNP molecular marker is remarkably superior to an existing phenotype-dependent observation method. Moreover, the SNP molecular marker can be used for identifying the pitted surface character of the fruit in the early stage of the plant, so that character judgment after fruit ripening is avoided, the breeding period can be effectively shortened, and the breeding efficiency can be improved. The SNP molecular marker can be directly applied to molecular breeding of pepper pitted surface characters, can provide reliable technical support for breeding of high-quality varieties, and has high practical value and popularization and application prospects.

The application discloses a SNP molecular marker related to a white spot syndrome virus (WSSV) tolerance trait of Procambarus clarkii, which is located in a 3rd intron region of a TRIM gene, specifically, a T>C type mutation at 585bp of a DNA sequence shown in SEQ ID NO. 1, wherein a T / C heterozygous genotype has significant correlation with the WSSV tolerance trait of the Procambarus clarkii. The application provides a favorable marker for molecular selection of the Procambarus clarkii, and can accelerate the selection process of the disease-resistant variety of the Procambarus clarkii.

The present application belongs to the technical field of sample pollution evaluation, and discloses a sample cross-contamination evaluation method, device, equipment and medium, comprising: performing quality control on batch sequencing data, and performing mutation site detection and genetic marker typing according to a human reference genome sequence and a site interval file; according to the genetic marker typing result and the site interval file, the number of heterozygous genotype sites is counted and the sample heterozygosity is calculated, according to the sample heterozygosity and a preset threshold, the sample contamination state is determined; according to the copy number estimation of the sample, the allele frequency is diploid standardized correction; based on the allele frequency in the genetic marker typing result and the mutation site detection result, the cross-contamination relationship between each two samples is analyzed by using identification rules to determine the pollution source; the site for calculating the pollution proportion is selected, and the weighted average algorithm is used to calculate the pollution proportion. The sample pollution degree can be accurately estimated and the pollution source sample can be identified.

This invention provides a rice gene Bph54 for resistance to brown planthopper and its application. The rice provided by this invention... Bph54 Genes are of great significance for breeding new rice varieties resistant to brown planthoppers. This invention successfully cloned the rice Bph54 gene using map-based cloning, and performed functional annotation and identification. This gene exhibits complete dominance, and both homozygous and heterozygous genotypes confer equal resistance to brown planthoppers in rice. This characteristic is beneficial for the application of this gene in breeding. Based on this, the Bph54 gene can be introduced into brown planthopper-susceptible plant varieties through traditional breeding techniques such as hybridization and backcrossing, or through genetic engineering, to cultivate new varieties of plants resistant to brown planthoppers with stable genetic traits. This invention provides important germplasm resources for effectively reducing insect damage and achieving increased and stable crop yields.

The invention provides development and application of an anti-pyrithiobac-sodium KASP molecular marker, and belongs to the technical field of genetic typing. The invention provides a KASP typing primer group for detecting the SNP site of the millet anti-pyrithiobac-sodium gene, and verifies that the resistance of millet to pyrithiobac-sodium herbicide is caused by single SNP mutation of an ALS target site. The KASP primers are designed on the basis of SNP loci, the primer pair can be used for accurately distinguishing resistance, susceptibility and heterozygous genotypes, and the method has the advantages of being stable and reliable in result, easy and convenient to operate, rapid and high in automation degree. By adopting the molecular marker to assist breeding, the breeding of the pyrithiobac-sodium-resistant millet matching variety can be accelerated, the breeding period can be shortened, the breeding efficiency can be improved, and the high-throughput identification of the pyrithiobac-sodium-resistant herbicide genotype and the molecular marker to assist the breeding of herbicide-resistant millet are significant.

The invention relates to the technical field of plant genetic engineering, and discloses a rice brown planthopper resistant gene Bph56, a molecular marker closely linked with the same and application of the rice brown planthopper resistant gene Bph56. The rice Bph56 gene provided by the invention is of great significance for cultivating a new variety of brown planthopper resistant rice. The rice Bph56 gene is successfully cloned through a map-based cloning method and is subjected to function annotation and identification, the gene is completely dominant, the homozygous and heterozygous genotypes endow rice with equal brown planthopper resistance, and the characteristic is beneficial to application of the gene in breeding. On the basis, the Bph56 gene can be introduced into a plant variety susceptible to brown planthopper through traditional breeding technologies such as hybridization, backcross and the like or genetic engineering means, and a novel brown planthopper-resistant plant variety with stable hereditary characters is obtained through cultivation. The invention provides important germplasm resources for effectively reducing insect pests and realizing yield increase and stable yield of crops.

This invention discloses a SNP marker, detection primers, and applications for genes related to nitrite resistance in Litopenaeus vannamei. The SNP molecular marker is located at the 193bp site of the sequence shown in SEQ ID NO.2, with mutation types of A / A homozygous, G / G homozygous, and A / G heterozygous. Litopenaeus vannamei with the A / A homozygous SNP molecular marker exhibits significantly higher nitrite resistance than those with the G / G homozygous and A / G heterozygous genotypes. This invention accelerates the breeding process of superior nitrite-resistant Litopenaeus vannamei varieties by identifying SNP markers associated with nitrite resistance and applying these markers to establish a marker-assisted breeding method for nitrite resistance in Litopenaeus vannamei.

This invention belongs to the field of apple molecular genetics and breeding technology, and specifically relates to a gene that increases GABA content in apples, its SNP molecular marker, and its application. This invention, for the first time, utilizes GWAS analysis to identify the GABA-related regulatory gene MdHVA22g in apple fruit. The gDNA sequence of this gene is shown in SEQ ID NO.1, and the CDS sequence is shown in SEQ ID NO.2. A corresponding SNP molecular marker was developed by associating the SNP with position 678 of the gene's gDNA, with polymorphisms of A or C. Apples with a homozygous AA genotype at this site have higher GABA content than apples with homozygous CC and heterozygous AC genotypes. Furthermore, this invention utilizes this SNP to design dCAPS primers, which can effectively and rapidly distinguish the GABA content of apples from different germplasms, facilitating the breeding of high-GABA apple varieties.

A method of genotyping one or more genetic rare variants in a plurality of nucleic acid samples is described. The method comprises running one or more assays of the plurality of nucleic acid samples using at least one microarray comprising a plurality of probesets and generating assay data. Heterozygous genotypes in the plurality of nucleic acid samples are called and evaluated to identify any rare heterozygous genotype calls that are from a probeset having at least one probe, wherein the number of heterozygous genotype calls for the probeset does not exceed a maximum threshold value. Each identified rare heterozygous genotype call is evaluated to determine whether the identified rare heterozygous genotype call comprises a true rare heterozygous genotype call using a support vector machine prediction model or supervised machine learning classification model comprising a plurality of predictor values for identifying a true rare heterozygous genotype call.

The application discloses a method for creating a DH line of eggplant by using anther culture and application thereof, and relates to the technical field of plant breeding. The application establishes a morphological index of a flower bud based on a calyx lobe separation state, replaces a time-consuming microscope examination procedure, and optimizes a combination of heat treatment and low-temperature pretreatment, so that the embryoid induction rate is significantly improved. Finally, through strict ploidy identification and self-pollination purification, a DH line with consistent traits and stable heredity can be obtained, an efficient pure line source is provided for eggplant hybrid breeding, and efficient conversion from a heterozygous genotype plant to a homozygous DH line is realized.

Co-dominant molecular markers for long-day adaptation gene StCDF1.4 in potato and the use thereof, which belongs to the technical field of molecular biology. The co-dominant molecular marker alone may identify homozygous or heterozygous genotypes of StCDF1.4, thus overcoming the limitation of existing molecular markers that require combinations of multiple pairs of primers to distinguish genotypes of StCDF1.4.

The invention belongs to the field of molecular genetic breeding, and discloses a CAPS molecular marker of a rice blast-resistant gene Pi2, a primer group and application, and the nucleotide sequence of the CAPS molecular marker is as shown in SEQ ID NO.1; the SEQ ID NO. 1 contains a Pst I restriction enzyme cutting site. The invention designs a CAPS marker primer Pi2-CF / Pi2-CR and restriction enzyme Pst I for distinguishing a disease-resistant gene Pi2 from other disease-resistant genes and susceptible genes, so that the gene Pi2 is accurately identified, and a favorable tool is provided for subsequently cultivating a broad-spectrum durable rice blast-resistant new rice variety. The invention discloses an intragene molecular marker of a rice blast resistant gene Pi2, which is a co-dominant functional marker, and a disease-resistant homozygous genotype, a susceptible homozygous genotype and a heterozygous genotype can be accurately distinguished by only using one marker.

The invention discloses a molecular marker related to the high lysine character of corn as well as a primer group and application of the molecular marker, and belongs to the technical field of plant molecular breeding. On the basis of whole genome re-sequencing comparison, an InDel molecular marker closely linked with a corn high-lysine key gene is found, and the marker is located in a corn chromosome 7. The invention provides a specific primer group for detecting the marker, PCR (Polymerase Chain Reaction) amplification is carried out by utilizing the primer group, and identification can be carried out through agarose gel electrophoresis according to the length polymorphism of an amplified fragment: an individual amplified with a 835bp band is a high-lysine genotype, an individual amplified with a 610bp band is a common genotype, and an individual amplified with the 610bp band is a high-lysine genotype. And the individual with the 835bp band and the 610bp band simultaneously appearing is a heterozygous genotype. Therefore, the invention provides an efficient and reliable technical tool for molecular marker-assisted selective breeding of high-lysine corn.

The invention relates to the technical field of molecular biology and genetic breeding, and discloses a PARMS primer of an SNP site significantly associated with brassica napus germination period drought tolerance and application of the PARMS primer, the SNP marker provided by the invention is used for identifying rape germplasm resources, and it is found that when the rape germplasm resources are treated for seven days under 22.5% PEG-6000 simulated drought stress conditions, the drought tolerance of the brassica napus can be significantly improved. The average germination rate (59.12%-66.37%) of an AA genotype material is remarkably higher than the average germination rate (29.84%-37.60%) of GG genotype and AG heterozygous genotype materials. The molecular marker can be used for early identification of the drought-enduring character of the rape and molecular marker-assisted selective breeding, the selection efficiency is improved, and the drought-enduring breeding process of the rape is accelerated.

The invention relates to the technical field of genome information analysis, in particular to a local pig optimal heterosis prediction method based on genome information. According to the method, pure parents with clear genetic backgrounds are accurately identified and screened out by utilizing high-density SNP marker information covering a whole genome, a reference population is constructed, large-scale hybridization propagation is simulated, structured virtual population data is formed, and multi-character phenotype data of filial generations are standardized and weighted, so that the genetic backgrounds of the filial generations are optimized, and the genetic backgrounds of the filial generations are optimized. The method comprises the following steps of: dividing a plurality of characteristic groups such as a production performance type and a reproductive performance type, and analyzing the correlation between phenotypic deviation and genotype in each group, so as to separate and quantify dominant effect caused by heterozygous genotype and recessive effect caused by homozygous genotype, construct explicit and recessive weight parameters, and determine the genetic relationship between the dominant effect and the recessive effect; finally, the non-additive effect parameters are superposed to a traditional additive breeding value model, direct quantitative prediction of heterosis is achieved, and therefore the accuracy and efficiency of hybrid combination matching are greatly improved.

The present invention provides a molecular marker identification method related to porcine sperm motility and its application, relating to the field of biotechnology. This application involves applying a substance that detects the SNP molecular marker rs3476761356 to the preparation of a product for predicting porcine sperm motility. The SNP molecular marker rs3476761356 is associated with porcine sperm motility, with the TA genotype being the dominant genotype. Selecting pigs with the TA heterozygous genotype for breeding can effectively improve porcine sperm motility and accelerate the genetic improvement of boar semen quality, providing important insights into pig performance evaluation and genetic breeding.

Provided are a codominant molecular marker of a potato long-day adaptive gene StCDF1.4 and a use of the codominant molecular marker, relating to the technical field of molecular biology. The codominant molecular marker alone is sufficient to identify the homozygous or heterozygous genotype of StCDF1.4, overcoming the shortcoming that existing molecular markers need combination of a plurality of pairs of primers to identify the genotype of StCDF1.4.

The invention belongs to the technical field of apple molecular genetic breeding, and particularly relates to a gene for improving the GABA content of apples and an SNP molecular marker and application thereof. GWAS is used for analyzing and mining the regulating gene MdHVA22g related to the GABA content of the apple fruits for the first time, the gDNA sequence of the gene is shown as SEQ ID NO.1, the CDS sequence of the gene is shown as SEQ ID NO.2, a corresponding SNP molecular marker is developed through associated SNP, the SNP site is the 678th site of the gDNA of the gene, the polymorphism of the SNP site is A or C, and the GABA content of the apple fruits with the homozygous AA genotype is higher than that of the apple fruits with the homozygous CC genotype and the heterozygous AC genotype at the site. Besides, the dCAPS primer is designed by utilizing the SNP, so that the GABA contents of the apple fruits with different germplasm can be effectively and quickly distinguished, and the breeding of the high GABA variety of the apple can be accelerated.

The invention relates to an SNP (Single Nucleotide Polymorphism) site related to the content of tocotrienol in rice or a detection reagent and application thereof, and belongs to the technical field of plant molecular biology. The application of the SNP site related to the content of tocotrienol in rice or the detection reagent of the SNP site comprises the following (I) or (II): (I) application in predicting the content of tocotrienol in rice; (II) application in breeding of a rice line with high tocotrienol content; the SNP site related to the content of the tocotrienol in the rice is located at the 23781761th basic group of the No.7 chromosome of the rice. Based on the mutation that the 23781761 base G of the No.7 chromosome of rice at the SNP site is replaced with T, a three-primer combination with short amplified fragment and high specificity is designed and developed, homozygous T, homozygous G and heterozygous H genotypes can be clearly distinguished, and whether the content of tocotrienol in rice is increased or not is predicted.

The present invention discloses a wheat SNP site and multiple applications thereof, a wheat SNP site, the SNP site corresponds to the 363rd base in the sequence shown in SEQ ID NO.1, and its variation space includes: G / G homozygous genotype, A / A homozygous genotype, G / A heterozygous genotype; the G / G homozygous genotype has a higher wheat spikelet number per ear phenotype than the A / A homozygous genotype; the use is to develop and construct theoretical research and / or wheat breeding applications and / or agricultural industry applications related to the number of spikelets per ear of wheat based on the SNP site. On the basis of the research related to wheat thousand-grain weight, the present invention further develops SNPs related to the trait of the number of spikelets per ear of wheat, which provides important gene resources and technical foundations for the research on the development and regulation mechanism of the number of spikelets per ear of wheat, especially for the directional breeding related to the number of spikelets per ear of wheat.