The kasp marker closely linked to the wheat dwarf gene and its application
A technology of wheat and reaction products, applied in the fields of molecular biology and crop breeding, can solve the problems of lack of parental genetic variation and limited yield increase potential in wheat breeding.
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Embodiment 1
[0043] Example 1. Comparison of plant height and stem strength between wild type and dwarf mutant JE0124
[0044] 1. Experimental materials and methods
[0045] The wild-type Jing411 and the dwarf mutant JE0124 were planted in the experimental field of the Crop Science Research Institute of the Chinese Academy of Agricultural Sciences. Fifteen plants were planted in each row, and the row length was 2 meters. The plant height was measured when the wheat was mature and harvested in June. Plant height was measured for three consecutive years from 2016 to 2018, with at least 5 repetitions each time. The comparison of stem strength is determined by the measurement index of stem fracture resistance. The specific measurement method is: cut 20cm from the base of the main stem with stable plant height, and use a digital display push-pull gauge perpendicular to the stem in the central area (10cm position) The fracture resistance of stems was measured, and each material was measured abo...
Embodiment 2
[0048] Example 2. Preliminary mapping of dwarf genes based on 660k SNP chips
[0049] 1. Genetic population construction and experimental methods
[0050] The dwarf mutant JE0124 was reciprocally crossed with the relatively tall backbone parent Nongda 5181 to obtain 2 F 2 groups for subsequent gene mapping analysis. F obtained by crossing Nongda 5181 as female parent and JE0124 as male parent (Nongda 5181 / JE0124) 2 The number of individual plants in the population is 312; the F 2 The number of individual plants in the colony is 282. take two F's 2 DNA was extracted from leaf samples of individual plants in the population for subsequent positioning analysis. After the plant height phenotype was stabilized in the field, the plant height of each individual plant was measured.
[0051] According to the plant height phenotype data, two F 2 20-30 high-stem and short-stalk extreme individuals in the population were mixed with equal amounts of DNA to construct extreme tall-stem...
Embodiment 3
[0054] Example 3. Analysis and verification of genetic linkage mapping of dwarf gene
[0055] 1. Experimental method
[0056] According to the polymorphic sites on the 2D chromosomes of the two parents of Nongda 5181 and JE0124 on the SNP chip, and based on the known flanking sequences, two allele forward primers and one reverse primer with different terminal bases were designed, and tested in the two The 5' ends of the forward primers were respectively connected to sequences with different fluorescent probes. Using KASP Master mix (LGC), the genotype of the sample could be determined according to the fluorescent signal detected after PCR amplification. According to the SNP sites of Nongda 5181 and JE0124 on the 2D chromosome, a total of 48 pairs of KASP marker primers were designed, and 8 pairs of KASP markers with good genotypes were screened and obtained by detecting the genotypes in the parents and some populations, which were used for genetic Linkage map construction.
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