CN111172315B - A01 chromosome major QTL site with main inflorescence grain weight character of brassica napus, SNP molecular marker and application - Google Patents
A01 chromosome major QTL site with main inflorescence grain weight character of brassica napus, SNP molecular marker and application Download PDFInfo
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Abstract
The invention provides an A01 chromosome major QTL site for the main inflorescence grain weight trait of Brassica napus, which is positioned between 390120 th base and 531852 th base of the A01 chromosome of the Brassica napus. Preferably, the contribution rate to the grain weight character of the main inflorescence of the brassica napus is 13.80%. Closely linked to the first SNP molecular marker, which is located at base 390120 and is either A or G, this mutation results in a polymorphism. Closely linked to the second SNP molecular marker, which is located at base 531852 and is either G or T, the mutation results in a polymorphism. Closely linked to the peak SNP molecular marker, which is at base 399218, and is either C or T, this mutation results in a polymorphism. Also provides related SNP molecular markers and application. The A01 chromosome major QTL locus has high contribution rate to the main inflorescence grain weight character of the brassica napus, plays a key role in regulating and controlling the main inflorescence grain weight of the brassica napus, can be used for map cloning and molecular marker assisted selection, and is suitable for large-scale popularization and application.
Description
Technical Field
The invention relates to the technical field of molecular biology and rape breeding, in particular to the technical field of main inflorescence grain weight traits of brassica napus, and specifically relates to an A01 chromosome major QTL site of the main inflorescence grain weight traits of brassica napus, an SNP molecular marker and application.
Background
The demand of edible oil in China is rigidly increased at the speed of 100 ten thousand tons per year, and the demand of vegetable oil is estimated to be 3.5-4.0 times of the current domestic production capacity by 2025 years, but the arable land resources in China are continuously reduced, so that the improvement of the unit yield of oil crops is a fundamental way for realizing the increase of the total yield of oil crops. Rape is the first large oil crop in China, the annual sowing area exceeds 1 hundred million acres, the total yield is more than 1500 ten thousand tons, and the rape seed oil accounts for more than 55 percent of the total amount of edible vegetable oil in China. At present, the planting area and the yield of rape industry in China continuously decline for years, the industrial safety degree declines year by year, and China is the largest world oil import country. The yield per unit is improved, the production cost is reduced to promote the income of farmers, and the method is a core measure for promoting the development of rape industry in China and ensuring the supply safety of edible vegetable oil.
At present, the rape yield level in China is low, and the planting enthusiasm of farmers is influenced. The regional trial yield level of winter rape variety examined by China in 2001-2016 is 2.26-3.75 tons/hm 2 In between, the test is much lower than that of a new variety in 2013-2015 Canada for 4.06 tons/hm 2 The level of yield of (a); in recent years, the yield per unit of rape field production in China is about 1.92 tons/hm 2 And 3.12 tons/hm in the European Union 2 Canada of 2.24 tons/hm 2 . Because the yield per unit of rape is low and the economic benefit is low, the enthusiasm of farmers for rape planting is influenced, and the normal development of the rape industry is restricted. In the production process, the creation of a new variety of the super-high-yield rape with the unit yield level exceeding that of European Union and Canada is urgently needed.
The variety lack suitable for the whole-process mechanized production causes the production cost to be high. The total rape production cost in China is as high as 640 yuan/mu, wherein the labor input and management is higher than 360 yuan, and the total cost is 60-70%; the total production cost of the rape of European Union and Canada is less than 300 yuan/mu, wherein the labor input and management is only 7.5 yuan/mu, and the total cost is about 2.5 percent. The traditional rape varieties have high lodging, scattered branches and staggered branches, inconsistent maturity and easily cracked siliques, great difficulty is caused to mechanized harvesting, semi-short stalk lodging resistance, compact plant type, good maturity consistency and angle cracking resistance are urgently needed in production, and the rape varieties suitable for mechanized operation are just started in the mechanized breeding of rape in China at present, and are extremely lack of suitable mechanized varieties in production.
One of the rape variety characteristics suitable for mechanized harvesting is suitable for close planting, the close planting can shorten the growth period, realize late sowing and early maturing, can increase the leaf area coefficient and the light interception rate, and fully utilize the light energy. The close planting of rape is realized by increasing the effective pod number of the group. With the increase of density, the yield of a single plant is reduced, the absolute yield of the main inflorescence is also reduced, but the specific gravity of the main inflorescence in the yield is continuously improved from 32% to 60% [ Zhang Uwen, guo Asia Ru. The advantages of the main inflorescence of rape and the utilization thereof are initially analyzed [ J ]. Northwest plant academic newspaper, 1996,16 (6): 126-131]. The major inflorescence grain weight comprises the major inflorescence silique number, the grain number per silique and the thousand grain weight, and is controlled by complex quantitative traits.
Therefore, a main effect QTL site for the main inflorescence grain weight trait of the brassica napus is needed, has high contribution rate to the main inflorescence grain weight trait of the brassica napus, plays a key role in regulating and controlling the main inflorescence grain weight of the brassica napus, and can be used for map cloning and molecular marker assisted selection.
Disclosure of Invention
In order to overcome the defects in the prior art, the invention aims to provide the A01 chromosome major QTL locus for the main inflorescence grain weight trait of the brassica napus, which has high contribution rate to the main inflorescence grain weight trait of the brassica napus, plays a key role in regulating and controlling the main inflorescence grain weight of the brassica napus, can be used for map bit cloning and molecular marker-assisted selection, and is suitable for large-scale popularization and application.
The invention also aims to provide the SNP molecular marker of the A01 chromosome major QTL locus of the main inflorescence grain weight trait of the brassica napus, which can detect the size of the main inflorescence grain weight of the brassica napus, can predict the size of the main inflorescence grain weight of the brassica napus, can effectively select the size of the main inflorescence grain weight of the brassica napus, can be used for molecular marker-assisted breeding of the brassica napus with the large main inflorescence grain weight, accelerates the high-yield breeding process of the brassica napus and is suitable for large-scale popularization and application.
The invention also aims to provide the SNP molecular marker of the major QTL site of the A01 chromosome for the main inflorescence grain weight character of the brassica napus, which has the advantages of ingenious design, simple and quick detection, low cost, no environmental influence and suitability for large-scale popularization and application.
The invention also aims to provide application of the SNP molecular marker of the A01 chromosome major QTL site of the main inflorescence grain weight trait of the brassica napus, which can detect the size of the main inflorescence grain weight of the brassica napus, predict the size of the main inflorescence grain weight of the brassica napus, effectively select the size of the main inflorescence grain weight of the brassica napus, be used for molecular marker-assisted breeding of the brassica napus with the large main inflorescence grain weight, accelerate the high-yield breeding process of the brassica napus and be suitable for large-scale popularization and application.
The invention also aims to provide application of the SNP molecular marker of the A01 chromosome major QTL site of the main inflorescence grain weight character of the brassica napus, which has the advantages of ingenious design, simple and quick detection, low cost, no environmental influence and suitability for large-scale popularization and application.
In order to achieve the above object, in a first aspect of the present invention, there is provided an a01 chromosome major QTL locus for the trait of brassica napus major inflorescence grain weight, characterized in that the a01 chromosome major QTL locus for the trait of brassica napus major inflorescence grain weight is located between 390120 th base and 531852 th base of the a01 chromosome of brassica napus.
Preferably, the contribution rate of the A01 chromosome major QTL site of the main inflorescence grain weight trait of the brassica napus to the main inflorescence grain weight trait of the brassica napus is 13.80%.
Preferably, the A01 chromosome major QTL locus of the main inflorescence grain weight trait of the Brassica napus is closely linked with a first SNP molecular marker, the first SNP molecular marker is located at 390120 th base, the 390120 th base is A or G, and the mutation causes polymorphism.
Preferably, the A01 chromosome major QTL locus of the main inflorescence grain weight trait of the Brassica napus is closely linked with a second SNP molecular marker, the second SNP molecular marker is located at the 531852 th base, the 531852 th base is G or T, and the mutation causes polymorphism.
Preferably, the A01 chromosome major QTL site of the main inflorescence grain weight trait of the cabbage type rape is closely linked with a peak SNP molecular marker, the peak SNP molecular marker is located at 399218 th base of the A01 chromosome of the cabbage type rape, the 399218 th base is C or T, and the mutation causes polymorphism.
In a second aspect of the invention, the SNP molecular marker of the major QTL site of the A01 chromosome of the main inflorescence grain weight trait of the Brassica napus is provided, and is characterized in that the SNP molecular marker is located at 390120 th base of the A01 chromosome of the Brassica napus, the 390120 th base is A or G, and the mutation causes polymorphism.
In a third aspect of the present invention, the application of the SNP molecular marker at the a01 chromosome major QTL locus for the brassica napus major inflorescence grain weight trait described above in detecting the size of the brassica napus major inflorescence grain weight, predicting the size of the brassica napus major inflorescence grain weight, selecting the size of the brassica napus major inflorescence grain weight, or molecular marker-assisted breeding of brassica napus with a large major inflorescence grain weight is provided.
In the fourth aspect of the invention, the SNP molecular marker of the major QTL site of the A01 chromosome of the main inflorescence grain weight trait of the cabbage type rape is provided, and is characterized in that the SNP molecular marker is positioned at the 531852 th base of the A01 chromosome of the cabbage type rape, the 531852 th base is G or T, and the mutation causes polymorphism.
In a fifth aspect of the present invention, the application of the SNP molecular marker at the a01 chromosome major QTL locus for the brassica napus major inflorescence grain weight trait described above in detecting the size of the brassica napus major inflorescence grain weight, predicting the size of the brassica napus major inflorescence grain weight, selecting the size of the brassica napus major inflorescence grain weight, or molecular marker-assisted breeding of brassica napus with a large major inflorescence grain weight is provided.
In a sixth aspect of the invention, the peak SNP molecular marker of the major QTL site of the A01 chromosome of the main inflorescence grain weight trait of the cabbage type rape is provided, and is characterized in that the peak SNP molecular marker is located at the 399218 th base of the A01 chromosome of the cabbage type rape, the 399218 th base is C or T, and the mutation causes polymorphism.
In a seventh aspect of the present invention, an application of the peak SNP molecular marker for the a01 chromosome major QTL locus for the brassica napus major inflorescence grain weight trait described above in detecting the size of the brassica napus major inflorescence grain weight, predicting the size of the brassica napus major inflorescence grain weight, selecting the size of the brassica napus major inflorescence grain weight, or molecular marker-assisted breeding of brassica napus with a major inflorescence grain weight is provided.
The invention has the following beneficial effects:
1. the A01 chromosome major QTL site of the cabbage type rape main inflorescence grain weight trait is positioned between 390120 th base and 531852 th base of the cabbage type rape A01 chromosome, has high contribution rate to the cabbage type rape main inflorescence grain weight trait, plays a key role in regulating and controlling the cabbage type rape main inflorescence grain weight, can be used for map cloning and molecular marker assisted selection, and is suitable for large-scale popularization and application.
2. The SNP molecular markers of the major QTL site of the A01 chromosome of the main inflorescence grain weight character of the cabbage type rape comprise a 390120 th base SNP molecular marker located on the A01 chromosome of the cabbage type rape, a 531852 th base SNP molecular marker located on the A01 chromosome of the cabbage type rape and a peak SNP molecular marker located on the 399218 th base of the A01 chromosome of the cabbage type rape.
3. The SNP molecular marker of the major QTL site of the A01 chromosome of the main flowering grain weight trait of the cabbage type rape comprises an SNP molecular marker of 390120 th base of the A01 chromosome of the cabbage type rape, an SNP molecular marker of 531852 th base of the A01 chromosome of the cabbage type rape and a peak SNP molecular marker of 399218 th base of the A01 chromosome of the cabbage type rape.
4. The application of the SNP molecular marker of the major inflorescence grain weight trait A01 chromosome major QTL locus of the brassica napus comprises the application of the SNP molecular marker of 390120 th base of the A01 chromosome of the brassica napus, the application of the SNP molecular marker of 531852 th base of the A01 chromosome of the brassica napus and the application of the peak SNP molecular marker of 399218 th base of the A01 chromosome of the brassica napus.
5. The application of the SNP molecular marker of the major QTL site of the A01 chromosome of the main inflorescence grain weight character of the cabbage type rape comprises the application of the SNP molecular marker of the 390120 th base of the A01 chromosome of the cabbage type rape, the application of the SNP molecular marker of the 531852 th base of the A01 chromosome of the cabbage type rape and the application of the peak SNP molecular marker of the 399218 th base of the A01 chromosome of the cabbage type rape.
These and other objects, features and advantages of the present invention will become more fully apparent from the following detailed description, the accompanying drawings and the claims, and may be realized by means of the instrumentalities, products and combinations particularly pointed out in the appended claims.
Drawings
FIG. 1 is a schematic diagram of the distribution results of the main inflorescence grain weight trait of Brassica napus of the present invention.
FIG. 2 is a schematic diagram of allelic analysis by using peak SNP molecular markers of the major QTL site of A01 chromosome for the main inflorescence grain weight trait of Brassica napus in the present invention.
Detailed Description
The inventor of the invention discloses an A01 chromosome major QTL locus of a main inflorescence grain weight character of the cabbage type rape and an SNP molecular marker thereof for the first time through deep research, and can effectively and efficiently improve the yield of the cabbage type rape by utilizing the locus major QTL locus.
The A01 chromosome major QTL site of the main inflorescence grain weight character of the brassica napus is positioned between 390120 th base and 531852 th base of the A01 chromosome of the brassica napus.
Preferably, the contribution rate of the A01 chromosome major QTL site for the main inflorescence grain weight trait of the Brassica napus to the main inflorescence grain weight trait of the Brassica napus is 13.80%.
Preferably, the A01 chromosome major QTL locus of the main inflorescence grain weight trait of the Brassica napus is closely linked with a first SNP molecular marker, the first SNP molecular marker is located at 390120 th base, the 390120 th base is A or G, and the mutation causes polymorphism.
Preferably, the A01 chromosome major QTL locus of the main inflorescence grain weight trait of the Brassica napus is closely linked with a second SNP molecular marker, the second SNP molecular marker is located at the 531852 th base, the 531852 th base is G or T, and the mutation causes polymorphism.
Preferably, the A01 chromosome major QTL site of the main inflorescence grain weight trait of the cabbage type rape is closely linked with a peak SNP molecular marker, the peak SNP molecular marker is located at 399218 th base of the A01 chromosome of the cabbage type rape, the 399218 th base is C or T, and the mutation causes polymorphism.
Also provides an SNP molecular marker of the major QTL site of the A01 chromosome of the main inflorescence grain weight trait of the cabbage type rape, which is located at 390120 th base of the A01 chromosome of the cabbage type rape, wherein the 390120 th base is A or G, and the mutation causes polymorphism. Namely the first SNP molecular marker.
The application of the SNP molecular marker of the A01 chromosome major QTL site with the main inflorescence grain weight character of the brassica napus in detecting the size of the main inflorescence grain weight of the brassica napus, predicting the size of the main inflorescence grain weight of the brassica napus, selecting the size of the main inflorescence grain weight of the brassica napus or in molecular marker-assisted breeding of the brassica napus with the large main inflorescence grain weight is also provided.
Also provides an SNP molecular marker of the major QTL site of the A01 chromosome of the main inflorescence grain weight character of the cabbage type rape, which is positioned at the 531852 th base of the A01 chromosome of the cabbage type rape, wherein the 531852 th base is G or T, and the mutation causes polymorphism. Namely the second SNP molecular marker.
The application of the SNP molecular marker of the A01 chromosome major QTL site with the main inflorescence grain weight character of the brassica napus in detecting the size of the main inflorescence grain weight of the brassica napus, predicting the size of the main inflorescence grain weight of the brassica napus, selecting the size of the main inflorescence grain weight of the brassica napus or in molecular marker-assisted breeding of the brassica napus with the large main inflorescence grain weight is also provided.
Also provides a peak SNP molecular marker of the major QTL site of the A01 chromosome of the main inflorescence grain weight character of the cabbage type rape, which is positioned at the 399218 th base of the A01 chromosome of the cabbage type rape, wherein the 399218 th base is C or T, and the mutation causes polymorphism.
Also provides the application of the peak SNP molecular marker of the A01 chromosome major QTL site of the main inflorescence grain weight character of the cabbage type rape in detecting the size of the main inflorescence grain weight of the cabbage type rape, predicting the size of the main inflorescence grain weight of the cabbage type rape, selecting the size of the main inflorescence grain weight of the cabbage type rape or in molecular marker assisted breeding of the cabbage type rape with the large main inflorescence grain weight.
The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The experimental procedures, for which specific conditions are not noted in the following examples, are generally performed according to conventional conditions such as those described in J. SammBrook et al, molecular cloning, A laboratory Manual, third edition, scientific Press, 2002, or according to the manufacturer's recommendations.
Example 1 measurement of phenotype of the Brassica napus Master inflorescence grain weight trait
1. Determination of major inflorescence grain weight phenotype of a related population
(1) Carrying out field seed test analysis on the agronomic and quality traits of 627 parts of core germplasm materials (from a seed bank of the oil-vegetable research institute in Guizhou province), and selecting 300 high-generation brassica napus strains from all over the world to form a natural population, wherein the natural population comprises 98 parts of resources, 110 parts of breeding materials and 92 parts of varieties or parents; the method is divided into regions, wherein 246 parts belong to domestic and 54 parts belong to foreign sources. 3 years of phenotype identification at 2 points is completed in rape bases in Qinghe village, kaiyang county, guiyang city and Weiyuan town, changshun county.
(2) Direct seeding and final singling are adopted, the row spacing is 40cm, the plant spacing is 25cm, the row length is 3.5m, and each cell has 4 rows. And (4) planting protective rows around the test material field.
(3) The grain weight of the main inflorescence: after the plants were normally mature, the kernel weight of all the pods on the main inflorescence of 10 brassica napus was measured per cell in g to the nearest 0.1g. The tabular values for all environments were averaged for 300 parts of material and the results are summarized as follows:
table 1 average value of major inflorescence grain weight phenotype values for all environments of 300 parts material
The weight distribution result of the main inflorescence grains of the associated population shows that the weight characters of the main inflorescence grains are continuously distributed and normally distributed, and the weight characters of the main inflorescence grains are proved to belong to quantitative characters and have main effective gene loci, which are shown in figure 1.
2. Acquisition of related population high quality SNP data set
The method comprises the following steps of (1) extracting total DNA of leaves by adopting a CTAB method, and extracting the total DNA of the leaves of each material of a related population, wherein the specific method comprises the following steps:
rinsing the young and tender leaves in 10% ethanol; then shearing 0.1-0.2g of blades, putting the blades into a bowl mill, quickly milling the blades into powder by using liquid nitrogen, and putting the powder into a 2mL centrifuge tube; adding 700 mu L of preheated DNA extracting solution; mixing, placing in 65 deg.C water bath for 1h, and mixing for 1 time every 10-15 min; adding 700 μ L of the mixture (phenol: chloroform: isoamyl alcohol = 25: 24: 1), and mixing by gentle inversion for 10min; centrifuging at 10000 Xg for 15min at room temperature; sucking the supernatant into a new 2mL centrifuge tube; adding the mixed solution with the same volume (chloroform: isoamylol = 24: 1), reversing and uniformly mixing, standing for 5min,10000 Xg, centrifuging for 15min, and sucking the supernatant into a new centrifugal tube by using a gun; adding 2 times volume of anhydrous ethanol, mixing, standing at-20 deg.C for 1h,10 000 Xg, centrifuging for 10min, and removing supernatant; adding 500 μ L of precooled 75% ethanol, washing the precipitate, and removing the supernatant; washing and precipitating for 2 times continuously, and then airing; adding 100 μ L of 2% RNase A solution, standing at 37 deg.C for 1h, and standing overnight at 4 deg.C; re-extracting DNA solution with equal volume of mixed solution (chloroform: isoamyl alcohol = 24: 1), mixing by inversion, standing for 10min,10 × g, centrifuging for 15 or 20min, removing RNase A, sucking supernatant (about 60 μ L), and centrifuging again for 1min; detecting the concentration, quality and integrity of the DNA by agarose gel electrophoresis (0.8%) and an ultraviolet spectrophotometer; the absorbance 260/280 ratio was determined to be between 1.8 and 2.0 for all DNA samples. The DNA samples were then transported on dry ice to sequencing Inc. (Huada science and technology, inc.), each material having a sequencing depth of about 9X.
After obtaining high quality DNA as described above, the sequencing company (Huada science and technology Co., ltd.) performed 9 Xcoverage depth sequencing and returned data, and first performed sequencing quality evaluation using FastQC software, and then performed adapter and low quality reads filtration on the sequencing sequence. Obtaining cleardata of double-end sequencing of each material, then performing mapping and GATK software for mutation detection by using bw software, performing SNP dataset quality filtration according to the minimum allele frequency of more than or equal to 0.05, the deletion rate of less than or equal to 0.1 and the heterozygosity rate of less than or equal to 0.15 after obtaining a total SNP dataset of a related group, and finally obtaining a high-quality group SNP dataset for subsequent analysis.
3. Whole genome association analysis
The VCF file of the high-quality SNP dataset generated in the last step is subjected to format conversion by using plink software, and thenPerforming whole gene association analysis on the obtained main inflorescence grain weight phenotype and the SNP data set by using EMMAX software to obtain a P value of each site of the main inflorescence grain weight character, wherein when the P value is less than 5 multiplied by 10 -7 The SNP of (single nucleotide polymorphism) is the obvious SNP, the SNP with the minimum P value is the peak SNP, materials are grouped in different allele types of the peak SNP in a group, variance analysis is carried out, and the percentage of the ratio of the variance between the groups to the total variance is the contribution rate of the peak SNP.
Through analysis, the interval of the major QTL site of the main inflorescence grain weight trait of the cabbage type rape is limited between 390120 th base and 531852 th base of the A01 chromosome of the cabbage type rape, the corresponding SNPs are chrA 01-390120 (A/G), chrA 01-531852 (G/T), and the peak SNP is: the contribution rate of the QTL to the grain weight character of the main inflorescence of the brassica napus is 13.80 percent (the materials are grouped according to different allele types of peak SNP, single-factor variance analysis is carried out, and the percentage of the variance between groups divided by the total variance is the contribution rate).
The peak SNP of the main inflorescence grain weight character is as follows: chrA01_399218 (C/T), corresponding to the major inflorescence grain weight phenotype grouping: when the SNP at position chrA01_399218 is CC, the average main inflorescence grain weight of the material is 5.12g; at CT, the average major inflorescence grain weight of the material was 4.21g; at TT, the average major inflorescence grain weight of the material was 3.59g, as shown in FIG. 2.
One of the border SNPs for the major inflorescence grain weight trait is: chrA01_390120 (A/G), corresponding to the main inflorescence grain phenotype grouping: when the SNP at the position chrA 01-390120 is AA, the average main inflorescence grain weight of the material is 3.64g; AG, the average major inflorescence grain weight of the material is 4.30g; GG showed that the average major inflorescence grain weight of the material was 5.06g, and the contribution rate of the border SNP was 11.98%.
Another border SNP for the main inflorescence grain weight trait is: chrA01_531852 (G/T), grouped as corresponding to the major inflorescence grain weight phenotype: when the SNP at the position chrA01_531852 is GG, the average major inflorescence grain weight of the material is 4.99g; GT, the material had an average major inflorescence grain weight of 4.46g; at TT, the average major inflorescence grain weight of the material was 3.23g, and the contribution rate of this border SNP was 8.85%.
The whole genome sequence of Brassica napus has been published, and is shown in http:// www. The sequences (801 bp) of 400bp before and after the chrA 01-390120 (A/G) are shown in SEQ ID NO:1, the sequences (801 bp) of 400bp before and after the chrA 01-531852 (G/T) are shown in SEQ ID NO:2, and the sequences (659 bp) after the sequences (400 bp) before and after the chrA 01-399218 (C/T) are shown in SEQ ID NO: 3. The technicians in the field can adopt a conventional method to design a specific primer for detecting the SNP locus according to the sequence so as to detect the genotype of the SNP locus, thereby being capable of detecting the weight of the main inflorescence grain of the cabbage type rape, predicting the weight of the main inflorescence grain of the cabbage type rape, effectively selecting the weight of the main inflorescence grain of the cabbage type rape, being also capable of being used for molecular marker assisted breeding of the cabbage type rape with the large main inflorescence grain and accelerating the high-yield breeding process of the cabbage type rape.
Therefore, the method detects a main effect QTL site of the main inflorescence grain weight character of the cabbage type rape on the chromosome A01 of the cabbage type rape by phenotype analysis and whole genome re-sequencing of the main inflorescence grain weight character and then whole genome association analysis, and the contribution rate of the main inflorescence grain weight character of the cabbage type rape is 13.80 percent. The main effective QTL locus of the main inflorescence grain weight character of the cabbage type rape is positioned between 390120 th base and 531852 th base of an A01 chromosome of the cabbage type rape, the obvious SNP of the boundary is chrA 01-390120 (A/G), chrA 01-531852 (G/T), and the peak SNP is chrA 01-399218 (C/T), and according to the SNP molecular marker tightly linked with the main effective QTL locus, the main inflorescence grain weight character of the cabbage type rape can be detected, the main inflorescence grain weight of the cabbage type rape can be predicted, the main inflorescence grain weight of the cabbage type rape can be effectively selected, the main inflorescence grain weight of the cabbage type rape can be used for molecular marker assisted breeding, and the high-yield breeding process of the cabbage type rape can be accelerated.
The SNP molecular marker disclosed by the invention is used for carrying out molecular marker-assisted selection, the identification method is simple, the selection efficiency is high, and the main inflorescence grain weight of the brassica napus can be predicted. The selection target is clear and is not influenced by the environment. The individual cabbage type rape with the large main inflorescence grain weight can be identified in the early growth stage of the cabbage type rape, and other individual plants are eliminated.
In conclusion, the A01 chromosome major QTL site of the main inflorescence grain weight trait of the brassica napus has high contribution rate to the main inflorescence grain weight trait of the brassica napus, plays a key role in regulating and controlling the main inflorescence grain weight of the brassica napus, can be used for map cloning and molecular marker assisted selection, and is suitable for large-scale popularization and application.
It will thus be seen that the objects of the invention have been fully and effectively accomplished. The functional and structural principles of the present invention have been shown and described in the embodiments, and the embodiments may be modified without departing from the principles. Therefore, this invention includes all modifications encompassed within the spirit and scope of the claims.
Sequence listing
<110> rape institute of Guizhou province
<120> A01 chromosome major QTL locus of cabbage type rape main inflorescence grain weight character, SNP molecular marker and application
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atgttgtaaa aattaatatt atgtatcttc gtatactttt actaaagtac gtttatatca 180
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Claims (6)
1. An SNP molecular marker of an A01 chromosome major QTL site of a main inflorescence grain weight character of a cabbage type rape is characterized in that a nucleotide sequence of the SNP molecular marker is shown as SEQ ID NO. 1, the 401 th base of the nucleotide sequence is A or G, and polymorphism is caused by mutation.
2. The application of the SNP molecular marker of the major QTL site of A01 chromosome for the main inflorescence grain weight trait of Brassica napus according to claim 1 in detecting the size of the main inflorescence grain weight of Brassica napus, predicting the size of the main inflorescence grain weight of Brassica napus, selecting the size of the main inflorescence grain weight of Brassica napus or molecular marker-assisted breeding of Brassica napus with large main inflorescence grain weight.
3. An SNP molecular marker of an A01 chromosome major QTL site of a main inflorescence grain weight character of a cabbage type rape is characterized in that a nucleotide sequence of the SNP molecular marker is shown as SEQ ID NO. 2, the 401 th base of the nucleotide sequence is G or T, and polymorphism is caused by mutation.
4. The application of the SNP molecular marker of the A01 chromosome major QTL locus for the main inflorescence grain weight trait of Brassica napus according to claim 3 in detecting the size of the main inflorescence grain weight of Brassica napus, predicting the size of the main inflorescence grain weight of Brassica napus, selecting the size of the main inflorescence grain weight of Brassica napus or molecular marker assisted breeding of Brassica napus with large main inflorescence grain weight.
5. A peak SNP molecular marker of an A01 chromosome major QTL site of a main inflorescence grain weight character of a cabbage type rape is characterized in that a nucleotide sequence of the peak SNP molecular marker is shown as SEQ ID NO. 3, the 401 th base from the 5' end of the nucleotide sequence is C or T, and polymorphism is caused by mutation.
6. The application of the peak SNP molecular marker of the A01 chromosome major QTL locus for the main inflorescence grain weight trait of Brassica napus according to claim 5 in detecting the size of the main inflorescence grain weight of Brassica napus, predicting the size of the main inflorescence grain weight of Brassica napus, selecting the size of the main inflorescence grain weight of Brassica napus or molecular marker assisted breeding of Brassica napus with large main inflorescence grain weight.
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