dCAPS molecular marker primer pairs for identifying dehydration rate genotypes of corn kernels and application
A technology of molecular markers and primer pairs is applied in the field of molecular genetic breeding to achieve the effects of speeding up breeding, low cost, and reducing breeding costs
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Embodiment 1
[0030] Example 1 Analysis of corn grain dehydration rate phenotypic traits and candidate gene association analysis
[0031] 1. Test materials
[0032] The test materials used in Example 1 were 132 inbred lines. The inbred lines are natural populations and all come from the Institute of Crop Resources, Jilin Academy of Agricultural Sciences. The list of specific materials is shown in Table 1.
[0033] Table 1 List of 132 corn inbred lines
[0034]
[0035]
[0036] 2. Experimental design and survey method
[0037] The test was carried out in Gongzhuling Experimental Base of Jilin Academy of Agricultural Sciences in 2014, 2015 and 2016. A random block design was adopted, with 5 row blocks, row length 5m, row spacing 0.65m, plant spacing 0.20m, and 3 repetitions. In order to avoid marginal effects, 2 plants at the beginning and end of the two side rows and the middle three rows of each plot were not used for trait determination.
Embodiment 2105
[0070] Example 2105 Genotyping of Maize Inbred Lines
[0071] 105 parts of corn inbred lines were genotyped using the specific primers designed by the present invention, and the specific steps were as follows:
[0072] 1. Genomic DNA extraction
[0073] Take the seedlings or leaves (about 200mg-300mg) of the sample, and quickly grind them into powder under liquid nitrogen. Transfer the powder into a 2.0mL centrifuge tube, add 700μL 65°C preheated CTAB extract, and mix well. Heat in a water bath at 65°C for 45 minutes, and shake gently by inverting constantly. After the water bath, remove the centrifuge tube and cool to room temperature. Add 700 μL of chloroform:isoamyl alcohol (24:1) under the fume hood, and shake it gently for 5 to 10 minutes. Centrifuge at 12000rpm (room temperature) for 10min, transfer the supernatant to a new 2.0mL centrifuge tube with the tip of a decapitated pipette. Add 10 μL RNase solution (10 mg / mL), and incubate at 37° C. for 30 min. Add 700 μL...
Embodiment 3
[0093] Embodiment 3 specific primer pair is applied in breeding
[0094] 1. The corn inbred line Ji A512 with fast dehydration rate (selected department of Jilin Academy of Agricultural Sciences) was used as donor, and the corn inbred line 4287 with slow dehydration rate (female parent of self-fertilized hybrid Jidan 27 of Jilin Academy of Agricultural Sciences) was used as donor ) as acceptor, F 1 , with F 1 As the induced material, "Jiyu 101" is the induced line. After induction, it was doubled with colchicine and screened to obtain 120 haploid (DH) lines after grain identification and field identification.
[0095] 2. Select 24 DH lines with good performance in the field to extract genomic DNA.
[0096] 3. With the genomic DNA of 24 parts of DH lines as a template, PCR amplification is carried out with the primer sequences shown in SEQ ID No.1 and SEQ ID No.2, and the PCR amplification product is detected by electrophoresis, and the amplified product is subjected to TaqI ...
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