Method for Parentage Identification of Cotton Hybrids
A paternity test and hybrid technology, applied in the field of cotton hybrid paternity test, can solve the problems of poor timeliness, confusion in the hybrid market, and being easily affected by environmental and subjective factors.
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Embodiment 1
[0030] Example 1 The acquisition of 36 pairs of SSR core primers distributed in the 26 chromosomes of the cotton genome
[0031] A large number of SSR candidate primers were screened by using 32 main cotton varieties from the three main cotton producing provinces in my country (Xinjiang cotton area, the Yellow River cotton area, and the middle and lower reaches of the Yangtze River) as the core primer screening and evaluation materials. Considering the identification ability of primers, amplification effect and chromosome distribution, 1-2 pairs of primers with high polymorphism and good stability were selected from each chromosome as core primers, a total of 36 pairs (Table 2).
[0032] Table 236 basic information of core primers
[0033] Primer name
Embodiment 2
[0034] Embodiment 2 identifies whether the cotton species to be tested is produced by the F 1 generation seed
[0035]The purpose of the experiment: To identify whether the cotton seed provided by a hybrid seed production base is the F produced by the two parents of 'Zhongmian 48' 1 generation seeds.
[0036] The detection process is as follows:
[0037] 1. DNA extraction
[0038] Randomly pick 5 dry seeds of male parent, female parent and the hybrid to be tested for DNA preparation.
[0039] (1) Shell a single cotton seed, thoroughly pulverize it and transfer it to a 2ml centrifuge tube.
[0040] (2) Add 800μl DNA extraction solution (1% SDS, 0.01mol / L EDTA8.0, 0.7mol / L NaCl, 0.05mol / L Tris-HCl, 0.5% sorbitol, 1% PVP, 1% β-mercaptoethanol ), vortex until fully mixed, then bathe in water at 65°C for 30 minutes, and shake gently at intervals of about 10 minutes.
[0041] (3) After the water bath is over, add an equal volume of 800 μl of a mixture of phenol, chloroform and...
Embodiment 3
[0072] Embodiment 3 identifies whether the cotton species to be tested is produced by the F 1 generation seed
[0073] Purpose of the experiment: To identify whether the cotton seed provided by another hybrid seed production base is the F produced by the two parents of 'Zhongmian 48' 1 generation seeds.
[0074] According to the steps of Example 2, DNA preparation, PCR amplification, electrophoresis detection and silver staining detection were performed.
[0075] Result analysis: Through the above detection, the DNA fingerprints (Table 6) of the two parents and the hypothetical hybrid at 36 sites were obtained, and the results of fingerprint comparison analysis showed that the band pattern performance of the hybrid to be tested at 14 sites As parental exclusion loci, 6 loci showed parental heterozygous loci, and the other 16 loci were consistent with the paternal or maternal band pattern. According to the judgment standard, it shows that the tested hypothetical hybrid has n...
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