CN111500757B - A02 chromosome major QTL (quantitative trait locus) site of main inflorescence length character of brassica napus, SNP molecular marker and application - Google Patents
A02 chromosome major QTL (quantitative trait locus) site of main inflorescence length character of brassica napus, SNP molecular marker and application Download PDFInfo
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Abstract
The invention provides an A02 chromosome major QTL locus for the main inflorescence length character of Brassica napus, which is positioned between 2060157 base and 2510914 base of the A02 chromosome of Brassica napus. Preferably, the contribution rate to the main inflorescence length character of the brassica napus is 13.03%. Closely linked to the first SNP molecular marker, which is at base 2060157, either G or T, the mutation results in a polymorphism. Closely linked to the second SNP molecular marker, which is at base 2510914, either A or G, this mutation results in a polymorphism. Closely linked to the peak SNP molecular marker, which is at base 2063166, either G or T, this mutation results in a polymorphism. Related SNP molecular markers and applications thereof are also provided. The A02 chromosome major QTL locus has high contribution rate to the characters of the main inflorescence length of the brassica napus, plays a key role in regulating and controlling the main inflorescence length 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 the main inflorescence length character of brassica napus, and specifically relates to an A02 chromosome major QTL site of the main inflorescence length character of brassica napus, an SNP molecular marker and application thereof.
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 the rape industry in China and ensuring the safety of edible vegetable oil supply.
At present, the rape yield level in China is low, and the planting enthusiasm of farmers is influenced. The regional test yield level of the 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 lack of varieties suitable for the whole-process mechanized production causes the production cost to be high. The total cost of rape production in China is up to 640 yuan/mu, wherein the labor input and management is higher than 360 yuan, and the total cost is 60-70%; and the total production cost of the rape in the 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.
The length of the main inflorescence is the main plant character of the rape, the length of the main inflorescence has obvious genetic correlation with the single plant yield, and the length of the main inflorescence has the greatest influence on the single plant yield. The length of the main inflorescence is in extremely obvious positive correlation with the number of the siliques of the single plant and the number of the siliques of the main inflorescence, so that the yield of the single plant can be improved by increasing the length of the main inflorescence [ Dai Xianglai, zhao Jixian, sunny, ren Tingbo, cheng Guoping ], the correlation analysis of the length of the main inflorescence of the cabbage type hybrid rape and the plant characters, chinese agronomy report 2018,34 (5): 42-48].
Therefore, a main effect QTL site for the main inflorescence length character of the brassica napus is needed, has high contribution rate to the main inflorescence length character of the brassica napus, plays a key role in regulating and controlling the main inflorescence length of the brassica napus, and can be used for map location cloning and molecular marker assisted selection.
Disclosure of Invention
In order to overcome the defects in the prior art, the invention aims to provide an A02 chromosome major QTL locus for the length trait of the main inflorescence of the brassica napus, which has high contribution rate to the length trait of the main inflorescence of the brassica napus, plays a key role in regulating and controlling the length of the main inflorescence of the brassica napus, can be used for map 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 A02 chromosome major QTL site of the main inflorescence length character of the brassica napus, which can detect the length of the main inflorescence of the brassica napus, predict the length of the main inflorescence of the brassica napus, effectively select the length of the main inflorescence of the brassica napus, can be used for molecular marker-assisted breeding of the brassica napus with long main inflorescence length, accelerate 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 A02 chromosome major QTL site of the main inflorescence length 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 the application of the SNP molecular marker of the A02 chromosome major QTL site of the main inflorescence length character of the brassica napus, which can detect the length of the main inflorescence of the brassica napus, can predict the length of the main inflorescence of the brassica napus, can effectively select the length of the main inflorescence of the brassica napus, can be used for molecular marker-assisted breeding of the brassica napus with long main inflorescence length, 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 application of the SNP molecular marker of the A02 chromosome major QTL site of the main inflorescence length 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, an a02 chromosome major QTL locus for the brassica napus major inflorescence length trait is provided, which is characterized in that the a02 chromosome major QTL locus for the brassica napus major inflorescence length trait is located between bases 2060157 th and bases 2510914 th of the brassica napus a02 chromosome.
Preferably, the contribution rate of the A02 chromosome major QTL locus of the main inflorescence length trait of the brassica napus to the main inflorescence length trait of the brassica napus is 13.03%.
Preferably, the A02 chromosome major QTL site of the main inflorescence length trait of the Brassica napus is closely linked with a first SNP molecular marker, the first SNP molecular marker is located at 2060157 base, the 2060157 base is G or T, and the mutation causes polymorphism.
Preferably, the A02 chromosome major QTL site of the main inflorescence length trait of the Brassica napus is closely linked with a second SNP molecular marker, the second SNP molecular marker is located at 2510914 base, the 2510914 base is A or G, and the mutation causes polymorphism.
Preferably, the A02 chromosome major QTL site of the main inflorescence length trait of the Brassica napus is closely linked with a peak SNP molecular marker, the peak SNP molecular marker is located at the 2063166 base of the A02 chromosome of the Brassica napus, the 2063166 base is G 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 A02 chromosome of the main inflorescence length character of the Brassica napus is provided, and is characterized in that the SNP molecular marker is located at the 2060157 base of the A02 chromosome of the Brassica napus, the 2060157 base is G or T, and the mutation causes polymorphism.
In a third aspect of the present invention, the application of the SNP molecular marker of the a02 chromosome major QTL locus for the main inflorescence length trait of brassica napus in detecting the length of the main inflorescence of brassica napus, predicting the length of the main inflorescence of brassica napus, selecting the length of the main inflorescence of brassica napus, or in molecular marker-assisted breeding of brassica napus with long main inflorescence length is provided.
In the fourth aspect of the invention, the SNP molecular marker of the major QTL site of the A02 chromosome of the main inflorescence length character of the cabbage type rape is provided, and is characterized in that the SNP molecular marker is located at the 2510914 base of the A02 chromosome of the cabbage type rape, the 2510914 base is A or G, and the mutation causes polymorphism.
In a fifth aspect of the present invention, the application of the SNP molecular marker for the major QTL locus of a02 chromosome for the trait of the length of the main inflorescence of brassica napus is provided for detecting the length of the main inflorescence of brassica napus, predicting the length of the main inflorescence of brassica napus, selecting the length of the main inflorescence of brassica napus, or assisting in breeding with the molecular marker for brassica napus with long main inflorescence length.
In a sixth aspect of the invention, the peak SNP molecular marker of the major QTL site of the A02 chromosome of the main inflorescence length character of the brassica napus is provided, and is characterized in that the peak SNP molecular marker is located at the 2063166 base of the A02 chromosome of the brassica napus, the 2063166 base is G or T, and the mutation causes polymorphism.
In the seventh aspect of the present invention, the application of the peak SNP molecular marker for the a02 chromosome major QTL locus for the main inflorescence length trait of brassica napus in detecting the length of the main inflorescence of brassica napus, predicting the length of the main inflorescence of brassica napus, selecting the length of the main inflorescence of brassica napus, or in molecular marker-assisted breeding of brassica napus with long main inflorescence length is provided.
The invention has the following beneficial effects:
1. the A02 chromosome major QTL site of the main inflorescence length character of the brassica napus is positioned between 2060157 base and 2510914 base of the A02 chromosome of the brassica napus, has high contribution rate to the main inflorescence length character of the brassica napus, plays a key role in regulating and controlling the main inflorescence length of the brassica napus, can be used for map cloning and molecular marker assisted selection, and is suitable for large-scale popularization and application.
2. The SNP molecular marker of the major inflorescence length trait A02 chromosome major QTL site of the brassica napus comprises an SNP molecular marker of 2060157 base positions of the A02 chromosome of the brassica napus, an SNP molecular marker of 2510914 base positions of the A02 chromosome of the brassica napus and a peak SNP molecular marker of 2063166 base positions of the A02 chromosome of the brassica napus.
3. The SNP molecular markers of the major QTL site of the A02 chromosome of the main inflorescence length character of the brassica napus comprise the SNP molecular marker of the 2060157 th base of the A02 chromosome of the brassica napus, the SNP molecular marker of the 2510914 th base of the A02 chromosome of the brassica napus and the peak SNP molecular marker of the 2063166 th base of the A02 chromosome of the brassica napus.
4. The application of the SNP molecular marker of the major QTL site of the A02 chromosome of the main inflorescence length character of the brassica napus comprises the application of the SNP molecular marker of the 2060157 base of the A02 chromosome of the brassica napus, the application of the SNP molecular marker of the 2510914 base of the A02 chromosome of the brassica napus and the application of the peak SNP molecular marker of the 2063166 base of the A02 chromosome of the brassica napus.
5. The application of the SNP molecular marker of the major QTL site of the A02 chromosome of the main inflorescence length character of the cabbage type rape comprises the application of the SNP molecular marker of the 2060157 base of the A02 chromosome of the cabbage type rape, the application of the SNP molecular marker of the 2510914 base of the A02 chromosome of the cabbage type rape and the application of the peak SNP molecular marker of the 2063166 base of the A02 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 length trait of Brassica napus of the present invention.
FIG. 2 is a schematic diagram of allelic analysis by using peak SNP molecular markers of A02 chromosome major QTL sites of the main inflorescence length trait of Brassica napus in the present invention.
Detailed Description
The inventor of the invention discloses an A02 chromosome major QTL site with main inflorescence length 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 site.
The main effect QTL locus of the A02 chromosome of the main inflorescence length character of the brassica napus is positioned between the 2060157 base and the 2510914 base of the A02 chromosome of the brassica napus.
Preferably, the contribution rate of the A02 chromosome major QTL site of the main inflorescence length character of the brassica napus to the main inflorescence length character of the brassica napus is 13.03%.
Preferably, the A02 chromosome major QTL site of the main inflorescence length trait of the brassica napus is closely linked with a first SNP molecular marker, the first SNP molecular marker is located at the 2060157 base, the 2060157 base is G or T, and the mutation causes polymorphism.
Preferably, the A02 chromosome major QTL site of the main inflorescence length trait of the Brassica napus is closely linked with a second SNP molecular marker, the second SNP molecular marker is located at 2510914 base, the 2510914 base is A or G, and the mutation causes polymorphism.
Preferably, the A02 chromosome major QTL site of the main inflorescence length trait of the Brassica napus is closely linked with a peak SNP molecular marker, the peak SNP molecular marker is located at 2063166 base of the A02 chromosome of the Brassica napus, the 2063166 base is G or T, and the mutation causes polymorphism.
Also provides an SNP molecular marker of the major QTL site of the A02 chromosome of the main inflorescence length character of the Brassica napus, which is positioned at the 2060157 base of the A02 chromosome of the Brassica napus, wherein the 2060157 base is G or T, and the mutation causes polymorphism. Namely the first SNP molecular marker.
The application of the SNP molecular marker of the major QTL site of A02 chromosome of the main inflorescence length character of the cabbage type rape in detecting the length of the main inflorescence of the cabbage type rape, predicting the length of the main inflorescence of the cabbage type rape, selecting the length of the main inflorescence of the cabbage type rape or carrying out molecular marker-assisted breeding on the cabbage type rape with long main inflorescence length is also provided.
Also provides an SNP molecular marker of the major QTL site of the A02 chromosome of the main inflorescence length character of the Brassica napus, which is positioned at the 2510914 base of the A02 chromosome of the Brassica napus, wherein the 2510914 base is A or G, and the mutation causes polymorphism. Namely the second SNP molecular marker.
The application of the SNP molecular marker of the major QTL site of A02 chromosome of the main inflorescence length character of the cabbage type rape in detecting the length of the main inflorescence of the cabbage type rape, predicting the length of the main inflorescence of the cabbage type rape, selecting the length of the main inflorescence of the cabbage type rape or carrying out molecular marker-assisted breeding on the cabbage type rape with long main inflorescence length is also provided.
The peak SNP molecular marker of the major QTL site of the A02 chromosome of the main inflorescence length character of the brassica napus is also provided, the base is 2063166 of the A02 chromosome of the brassica napus, the base at 2063166 is G or T, and the mutation causes polymorphism.
Also provides the application of the peak SNP molecular marker of the A02 chromosome major QTL site of the main inflorescence length character of the cabbage type rape in detecting the length of the main inflorescence of the cabbage type rape, predicting the length of the main inflorescence of the cabbage type rape, selecting the length of the main inflorescence of the cabbage type rape or in molecular marker-assisted breeding of the cabbage type rape with long main inflorescence length.
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. SammBruk et al, molecular cloning protocols, third edition, scientific Press, 2002, or according to the manufacturer's recommendations.
Example 1 determination of phenotype of the Brassica napus Master inflorescence Length trait
1. Determination of major inflorescence length phenotype of related populations
(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-year 2-point phenotype identification is completed on rape bases in Weiyuan village and Changxing town of Kaiyang county in Guiyang city.
(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) Length of main inflorescence: after the plants were normally mature, the length of the main inflorescence of 10 brassica napus was measured per cell in centimeters to the nearest 0.1cm. The tabular values for all environments were averaged for 300 parts of material and the results are summarized as follows:
table 1 mean value of dominant inflorescence length phenotype values for all environments of 300 materials
The distribution result of the length of the main inflorescence of the associated population shows that the character expression distribution of the length of the main inflorescence is in continuous distribution and in normal distribution, and the main inflorescence length character is proved to belong to quantitative characters and has main effective gene sites, which is 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 1 hr, 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 mixed solution with the same volume (chloroform: isoamylol = 24: 1), reversing and mixing uniformly, standing for 5min,10000 Xg, centrifuging for 15min, and sucking supernatant liquid 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 clear data of double-end sequencing of each material, then using bwa software to carry out mapping and GATK software to carry out mutation detection, after obtaining a total SNP data set of an associated group, carrying out SNP data set 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, and finally obtaining a high-quality group SNP data set for subsequent analysis.
3. Genome-wide association analysis
Format conversion is carried out on the VCF file of the high-quality SNP data set generated in the last step by using plink software, then EMMAX software is used for carrying out whole-gene association analysis on the obtained main inflorescence length phenotype and the SNP data set, the P value of each site of the main inflorescence length character is obtained, and when the P value is smaller than 5 multiplied by 10 -7 The SNP is the obvious SNP, the SNP with the minimum P value is the peak SNP, the materials are grouped by 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 length character of the cabbage type rape is limited between 2060157 bases to 2510914 bases of an A02 chromosome of the cabbage type rape, corresponding SNPs are chrA02_2060157 (G/T), chrA02_2510914 (A/G), and peak SNPs are as follows: the contribution rate of the QTL to the main inflorescence length character of the brassica napus is 13.03 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 SNPs for the major inflorescence length trait were: chrA02_2063166 (G/T), corresponding to the major inflorescence length phenotype grouping: when the SNP at the position chrA02_2063166 is GG, the average main inflorescence length of the material is 57.93cm; GT, the average major inflorescence length of the material is 62.96cm; at TT, the average major inflorescence length of the material was 71.06cm, as shown in FIG. 2.
One of the border SNPs for the major inflorescence length trait is: chrA02_2060157 (G/T), corresponding to the major inflorescence length phenotype grouping: when the SNP at the position of chrA02_2060157 is GG, the average main inflorescence length of the material is 70.07cm; GT, the average major inflorescence length of the material is 62.66cm; at TT, the average main inflorescence length of the material is 57.88cm, and the contribution rate of the boundary SNP is 12.83%.
Another border SNP for the major inflorescence length trait is: chrA02_2510914 (A/G), grouped corresponding to the main inflorescence length phenotype: when the SNP at the position of chrA02_2510914 is AA, the average main inflorescence length of the material is 58.19cm; AG, the average major inflorescence length of the material is 63.59cm; in GG, the average main inflorescence length of the material is 75.50cm, and the contribution rate of the boundary SNP is 13.00%.
The whole genome sequence of Brassica napus has been published, see http:// www.genoscope.cns.fr/brassicana/. The sequences (801 bp) of 400bp before and after containing chrA02_2060157 (G/T) are shown as SEQ ID NO:1, the sequences (801 bp) of 400bp before and after containing chrA02_2510914 (A/G) are shown as SEQ ID NO:2, and the sequences (801 bp) of 400bp before and after containing chrA02_2063166 (G/T) are shown as 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, so that the length of the main inflorescence of the cabbage type rape can be detected, the length of the main inflorescence of the cabbage type rape can be predicted, the length of the main inflorescence of the cabbage type rape can be effectively selected, and the method can also be used for molecular marker assisted breeding of the cabbage type rape with long main inflorescence length and accelerating the high-yield breeding process of the cabbage type rape.
Therefore, the major inflorescence length trait phenotypic analysis and the whole genome re-sequencing are carried out, then the whole genome association analysis is carried out, a major QTL site of the major inflorescence length trait of the cabbage type rape is detected on the chromosome A02 of the cabbage type rape, and the contribution rate to the major inflorescence length of the cabbage type rape is 13.03%. The main effect QTL locus of the main inflorescence length character of the brassica napus is positioned between 2060157 base pairs to 2510914 base pairs of an A02 chromosome of the brassica napus, the obvious SNP of the boundary is chrA02_2060157 (G/T), chrA02_2510914 (A/G), and the peak SNP is chrA02_2063166 (G/T).
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 length of the main inflorescence 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 long main inflorescence length can be identified in the early growth stage of cabbage type rape, and other individual plants are eliminated.
In conclusion, the A02 chromosome major QTL site of the main inflorescence length character of the brassica napus has high contribution rate to the main inflorescence length character of the brassica napus, plays a key role in regulating and controlling the main inflorescence length 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> research institute for rape in Guizhou province
<120> A02 chromosome major QTL locus of brassica napus main inflorescence length character, SNP molecular marker and application
<160> 3
<210> 1
<211> 801
<212> DNA
<213> Brassica napus (Brassica napus, L.)
<220>
<221> misc_feature
<222> (1)...(801)
<223> genomic sequence comprising 400bp sequences of both before and after chrA02_2060157 (G/T)
<400> 1
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attgctatgt tttagttgag atgtctgaat ccttgtgtgc ttttgctaaa ttaacagttc 120
tatgcttgat ggtgggcgtg gcagagaggg caaagatcac atctctaact caaatggaga 180
tcacagtttt caggtaaaga tgatatatgt tagacatgta ttcttgcata gtatagttca 240
tgtgaaaaaa ctgcgtcttg tacacaggtt aaaggagata ttgaagcgct gattaagact 300
cgtgacctgc ttcatgatga ggtatgaggt cgcttatcat gaatcctttt cacaatttag 360
tataagccaa tggagcaaca gtttccttac ataatcgtgg tagatggaaa aatgattgtc 420
tagtttcgct cttaaggtac tggtgatgga aggcagattg agagcaaaag aagatctgtg 480
cgcagagcta cagcagaagt ttgaggtatg tgtctactat attctacgaa cccaatatgt 540
tttaatttta ctatggtttt gttctcattt gaaatggttt aacacagaca ttggaagatt 600
tattgaacgg tgaaaagaaa ctgacttcac aacgccgaaa agaattggca aaggtaaaca 660
aacataggtt cgggtttcgt ttacaaagct agttttgtct ttgtcgagta agttcgatgt 720
ttggtttatg tttttggtag ctacataaat catattcaag ggtcagagaa tgcagcgata 780
acgtgaaaga ctgcgaacag g 801
<210> 2
<211> 801
<212> DNA
<213> Brassica napus (Brassica napus, L.)
<220>
<221> misc_feature
<222> (1)...(801)
<223> genome sequence comprising 400bp sequences before and after chrA02_2510914 (A/G)
<400> 2
ctgtacttta ttttgtagga aaattacata tatatcaaac tctggttctg gtatatgtat 60
ttcgtatcag cttcttaaat aactttaaga taccaaagaa tatgtttccc taaataaata 120
cgctaacctt agtctgaatc tcaacattaa accatgtaat cataacatta tttattttta 180
ttttttttcc taaaaaaaag ttaaacctga atctattaaa gacatagact aacttccgga 240
tgggaggtgc atcctacgga tagaccttgt ctcagatatt caaatgaacc gaaaaacagt 300
agtccatatc cgtgtaaaat gactaggaaa ataacttagt ttcgcatagt aaatggatca 360
aacctgaaat gtggtgacat cccaaatcat tctccttacc acaaacttcc ggtctatttg 420
ttcgctcttc cttcaattgt ttatatatga ataaaatgat tgttgaaaac cttcttttga 480
gaatcatcct tagaaatgta gatgttaagt aatttccata agtcattgga cagagtaatt 540
acctgcattt tataatatga atgatgtaat atcgttattt catatcttta taaccgagag 600
ggagattttc tttcattata agtggagtgg atgctatatt ggtggcagag gaacaaatag 660
atagatcgat gaagccaaat ttaacattca agaatgcatt tcgtgttgtc aattaaaaag 720
tgataaagat ttgggcgaat ttttcttctt ttggatggaa tttttccatt tcgtgttgtc 780
tatttcaacg tagcaaatct a 801
<210> 3
<211> 801
<212> DNA
<213> Brassica napus (Brassica napus, L.)
<220>
<221> misc_feature
<222> (1)...(801)
<223> genome sequence comprising 400bp sequences before and after chrA02_2063166 (G/T)
<400> 3
ttctctttgc ttctaatact catagtggaa tcctagccgt tgatttggtg gacaagagtg 60
ttagaagaat caactcatgt cctttaggtc cacgtggcac atcctcgtgg agacggtccg 120
gtatgaagct tgtacccgat ccatcaaatc caagtcattt tcggttcatg tttgctgaga 180
tggtaaataa ccgaccggtt ttgtttactt atcattctaa cacaaccacg tggactacta 240
aagaagcaga ggaagtaaat aattgggctt ataagaaaaa caataacgtt ttcttaagct 300
tatcaaaccg acctcacgag agtattgtga tgagcgtgga cgatgggtcg atgatgaatc 360
gtgttccatc gattttgagg ccaaggatga accaggacgc gatcagagga tgcccttcaa 420
gtgttggatt cagcaggaac gatctagaaa gtcagttgtt acatatacat ggagatgaat 480
acaaagtggt tattaggtta gacacaatag agttatcaaa gctgaagaga atgaagagtt 540
tagaggtttg ggagataagc tcaaacggtg agaagtggga actagtgtca agagctccaa 600
gtgaagtgct cagcaacaag ctttgtggtg tgatgatggg gtgcttagag aggagactag 660
gggtgattag tgtggctttg atgacgaacc gtgagggttt atggaatatt atatggttgg 720
attatgataa ggagaaggat aagtgggaat gggtcccttt accacattgt agattcttgc 780
aaggctcaaa tatggccggt a 801
Claims (6)
1. An SNP molecular marker of an A02 chromosome major QTL site with a cabbage type rape main flower sequence length character 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 G or T, and polymorphism is caused by mutation.
2. The application of the SNP molecular marker of the major QTL locus of A02 chromosome for the main inflorescence length of Brassica napus according to claim 1 in detecting the length of the main inflorescence of Brassica napus, predicting the length of the main inflorescence of Brassica napus, selecting the length of the main inflorescence of Brassica napus or molecular marker-assisted breeding of Brassica napus with long main inflorescence.
3. An SNP molecular marker of an A02 chromosome major QTL site of a main inflorescence length 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 A or G, and the mutation causes polymorphism.
4. The application of the SNP molecular marker of the A02 chromosome major QTL locus for the main inflorescence length trait of the Brassica napus according to claim 3 in detecting the length of the main inflorescence of the Brassica napus, predicting the length of the main inflorescence of the Brassica napus, selecting the length of the main inflorescence of the Brassica napus or molecular marker-assisted breeding of the Brassica napus with long main inflorescence length.
5. A peak SNP molecular marker of an A02 chromosome major QTL site of a main flowering length character of a brassica napus is characterized in that a nucleotide sequence of the peak SNP molecular marker is shown as SEQ ID NO. 3, a 401 th base of the nucleotide sequence is G or T, and polymorphism is caused by mutation.
6. The application of the peak SNP molecular marker of the A02 chromosome major QTL site for the main inflorescence length trait of the Brassica napus according to claim 5 in detecting the length of the main inflorescence of the Brassica napus, predicting the length of the main inflorescence of the Brassica napus, selecting the length of the main inflorescence of the Brassica napus or molecular marker assisted breeding of the Brassica napus with long main inflorescence length.
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| CN108239647A (en) * | 2017-11-29 | 2018-07-03 | 华中农业大学 | A kind of gene, molecular labeling and application for controlling rape plant type |
| CN108728575A (en) * | 2018-06-21 | 2018-11-02 | 贵州省油菜研究所 | Main effect QTL site, SNP marker and the application of cabbage type rape Pod length character |
| CN110592251A (en) * | 2019-06-17 | 2019-12-20 | 中国农业科学院油料作物研究所 | Development and application of major QTL (quantitative trait locus) and SNP (Single nucleotide polymorphism) molecular marker for flowering phase characters of brassica napus |
| CN110741929A (en) * | 2019-11-21 | 2020-02-04 | 贵州省油菜研究所(贵州省油菜工程技术研究中心国家油菜改良中心贵州分中心) | Main inflorescence multi-pod cabbage type rape and breeding method thereof |
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| CA3044152A1 (en) * | 2018-05-29 | 2019-11-29 | Agrisoma Biosciences Inc. | Brassica carinata producing seed with reduced glucosinolate content |
| US20190373836A1 (en) * | 2018-06-12 | 2019-12-12 | Agrisoma Biosciences Inc. | Plants and seeds of brassica carinata variety dh-153.113 |
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| CN108239647A (en) * | 2017-11-29 | 2018-07-03 | 华中农业大学 | A kind of gene, molecular labeling and application for controlling rape plant type |
| CN108728575A (en) * | 2018-06-21 | 2018-11-02 | 贵州省油菜研究所 | Main effect QTL site, SNP marker and the application of cabbage type rape Pod length character |
| CN110592251A (en) * | 2019-06-17 | 2019-12-20 | 中国农业科学院油料作物研究所 | Development and application of major QTL (quantitative trait locus) and SNP (Single nucleotide polymorphism) molecular marker for flowering phase characters of brassica napus |
| CN110741929A (en) * | 2019-11-21 | 2020-02-04 | 贵州省油菜研究所(贵州省油菜工程技术研究中心国家油菜改良中心贵州分中心) | Main inflorescence multi-pod cabbage type rape and breeding method thereof |
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