KASP primer group related to wheat grain weight and application thereof
A technology of wheat kernels and primer sets, used in recombinant DNA technology, biochemical equipment and methods, and the determination/inspection of microorganisms
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Embodiment 1
[0033] Example 1 Screening thousand-grain weight marker interval
[0034] A recombinant inbred line (RIL) population (F 2:8 ), including 282 families. The RIL population and its parents were planted in Liuhe Base of Jiangsu Academy of Agricultural Sciences in two consecutive growing seasons from 2016 to 2017 and 2017 to 2018.
[0035] Augmented design (see literature "Lan C, Zhang Y, Herrera-Foessel S A, Basnet B R, Huerta-Espino J, Lagudah E S, Singh R P. Identification and characterization of pleiotropic and co-located resistance loci to leaf rust and stripe rust inbread wheat cultivar Sujata. Theoretical and Applied Genetics, 2015, 128:549-561"), 2 parents and 30 randomly selected materials were planted for 2 repetitions, and the rest of the materials were planted for 1 repetition, planted in three rows of plots, 60 grains per row , row length 1.6m, row spacing 0.25m, conventional cultivation management. The grains were harvested after they matured, and the thousand-grai...
Embodiment 2
[0038] Example 2 Screening and validation of KASP molecular markers
[0039] In order to better utilize the Qtkw-7A locus screened in Example 1 for breeding, a KASP marker suitable for high-throughput genotyping was developed based on its interval marker sequence. PCR amplification primers were designed according to SNP sites and flanking sequences, and KASP molecular markers were developed.
[0040] KASP primers were designed using Polymarker (http: / / www.polymarker.info / ), and the primers were synthesized by Sangon Bioengineering (Shanghai) Co., Ltd. Two SNP-specific primers (F1 / F2) and one universal primer (R) were designed for each marker, and a specific sequence (SEQ ID NO.10:5'-GAAGGTGACCAAGTTCATGCT-3 '), the front of the F2 primer added a specific sequence capable of combining with HEX fluorescence (SEQ ID NO.11:5'-GAAGGTCGGAGTCAACGGATT-3'). Three SNP markers with lower homology were selected from multiple SNP sites in this marker interval for primer design as shown in...
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