Molecular marking method for peanut flowering habit related gene locus and application of molecular marking method
A technology of flowering habits and molecular markers, applied in the fields of crop molecular genetics and molecular breeding
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Embodiment 1
[0046] Example 1: Genetic Mapping of Genetic Loci Related to Flowering Habits in Hybrid Populations of Monogenic Inheritance Models
[0047] (1) Materials and methods:
[0048] (1) Construction of plant materials and genetic populations: The continuous flowering cultivated peanut Pingdu 9616 and the alternate flowering cultivated peanut Florunner preserved in the Peanut Center Laboratory of Qingdao Agricultural University were used as test materials, and the continuous flowering cultivated peanut Pingdu 9616 and alternately flowering cultivated peanut were used as test materials. Flowering cultivated peanut Florunner hybrid, planting hybrid F 1 , selfing to obtain F 2 Separation of populations, continuous self-breeding of multiple generations of strains, and finally the formation of F 7 Recombinant inbred line RIL-PF-F 7 .
[0049] (2) The population genomic DNA was extracted by the improved CTAB method (for details, refer to the "Molecular Cloning Experiment Guide").
[...
Embodiment 2
[0063] Example 2: Development and application of locus-assisted selection markers related to flowering habit in monogenic genetic model populations
[0064] With flatness 9616, Florunner and RIL-PF-F thereof in embodiment 1 7 The genomic DNA of the group individual is used as a template, and a pair of primers or two pairs of primers in the pair of primers labeled PID12-21 and PID12-29 are used to perform PCR amplification on the above template on a PCR machine, and 1-1.5 μL of amplified The amplified product was detected by electrophoresis on an 8% non-denaturing polyacrylamide gel with a voltage of 300V and a current of 280mA for 1-1.5 hours. The length of electrophoresis can be appropriately adjusted according to the size of the target fragment. The gel was fixed, infiltrated, rinsed and developed to develop the gel silver stain.
[0065] The physical position of the marker PID12-21 is located at the end of chromosome 12 of the peanut reference genome Tifrunner, and its cor...
Embodiment 3
[0076] Example 3: Genetic Mapping of Genetic Loci Related to Flowering Habits in Hybrid Populations of Digene Inheritance Model
[0077] (1) Materials and methods:
[0078] (1) The alternate flowering cultivated peanut Xiaohongmao and the continuous flowering cultivated peanut Henan Nanyang preserved in the Peanut Center Laboratory of Qingdao Agricultural University were used as parents, and hybridization was carried out to plant the hybrid F 1 , selfing to obtain F 2 Separation of populations, continuous self-breeding of multiple generations of strains, and finally the formation of F 8 Recombinant inbred line RIL-HN-F 8 .
[0079] (2) The population genomic DNA was extracted by the improved CTAB method (for details, refer to the "Molecular Cloning Experiment Guide").
[0080] (3) Development of polymorphic InDel markers: Based on the resequencing data of Xiaohongmao and Henan Nanyang peanut genomes and the reference genomes of Tifrunner and Shitouqi downloaded from the NC...
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