A haplotype bnhapglu for identifying the content of glucosinolates in rapeseed and its application
A haplotype, bn-a09-p3053860 technology, applied in the fields of genetic breeding and molecular biology, to achieve the effect of clear selection target, saving breeding cost and high contribution rate
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Embodiment 1
[0039] Example 1. Determination of Glucosinolate Content in Related Populations
[0040] The population used in this example was 370 Brassica napus materials from major rapeseed producing countries in the world. The determination of glucosinolate content in five environments at two points in three years was obtained by using near-infrared scanning rapeseed. Multi-year multi-point data were processed using optimal linear unbiased estimation (BLUP), and the heritability of glucosinolate was obtained: 97.4%, indicating that the level of glucosinolate content is mainly determined by genetic factors. The results of the distribution of glucosinolate content in the related population showed that the performance distribution of the glucosinolate content trait was a continuous distribution, but the variation distribution was not in a normal distribution, which proved that the glucosinolate content trait was a simple quantitative trait and there was a main effect gene locus (see figure...
Embodiment 2
[0042] Example 2. Extraction of total DNA from associated population leaves
[0043] Use the CTAB method to extract the total DNA of the leaves of the associated population (Doyle J.A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochemical Bulletin, 1987, 19, 11-15), the specific method is:
[0044] a) Rinse the young leaves in 10% ethanol; then cut 0.1-0.2g of leaves and put them in a mortar, quickly grind them into powder with liquid nitrogen, and put them into a 2mL centrifuge tube;
[0045] b) Add 700 μL of preheated DNA extraction solution; after mixing, put it in a 65°C water bath for 1 hour, and mix once every 10-15 minutes;
[0046] c) Add 700 μL of mixed solution (phenol: chloroform: isoamyl alcohol = 25:24:1), gently invert and mix for 10 min; centrifuge at 10 000×g for 15 min at room temperature;
[0047] d) Draw the supernatant into a new 2mL centrifuge tube; add an equal volume of mixed solution (chloroform: isoamyl alcohol = 24:1),...
Embodiment 3
[0052] Example 3. Association group structure and kinship analysis
[0053] Since there are SNPs with obvious population structure and close kinship in the population that may produce false positives in the results of association analysis, it is necessary to conduct population structure and kinship analysis when performing genome-wide association analysis of glucosinolate content. The population structure analysis was carried out using Structure 2.3 software, and the results showed that the associated population was divided into three subgroups ( figure 2 ). Kinship itself is a relative value that defines the genetic similarity between two specific materials and the genetic similarity between arbitrary materials. Tassel5.0 software was used to evaluate the relative kinship, and the matrix (K matrix) of the relative kinship was calculated. The results show that: the total average value of genetic relationship among materials in this group is 0.1077( image 3 ), in which 58....
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