The application of the KASP markers
The fine mapping results showed 6 KASP markers (bropsK49, bropsK18,
bropsK51, bropsK52, bropsK42, bropsK43) were closely associated with the
target gene and, possibly any one of them could be in the target gene
region. These 6 markers were used to screen 92 individuals of the
F2 segregation population derived from the mutant GS and
the wild line PS. The genotyping results of 5 KASP markers (bropsK18,
bropsK51, bropsK52, bropsK42 and bropsK43) were in agreement with the
phenotypic data by more than 98%, while the genotyping result of
bropsK49 was in 100% accordance with the phenotypic data (Figure 3D).
It showed that the six KASP markers based on C9: 59999594 to 60239356
could perform well in genotyping for each plant in the new population
(Figure 3D). Based on the genotypic and phenotypic data of the
F2 individuals, the range of genomic location of
bud-color at low temperature was further narrowed down to 229.649Kb,
located between bropK47 and bropK18, i.e., C9:59999594 ~
60229243, and the gene where bropsK49 locates could probably be the
target gene.
Also, the 6 KASP markers were used to screen forty-two germplasm
materials in Brassica oleracea L. (Figure 3E). Out of the 6 KASP
markers, the genotyping data of two KASP markers (bropsK18 and bropsK49)
matched more than 95% with the phenotyping data and were effective in
predicting whether the buds would turn purple or keep green in cold
temperature in Brassica oleraceaL. Within the 42 germplasm lines,
40 lines showed consistent results for both bropsK18 and bropsK49 with
the genotypic data in conformity with the phenotyping, whereas 2 lines
(1947 and K2145), which would keep green buds at low temperature, lack a
43-bp deletion in bropsK49 where there was a 43-bp InDel, and exhibited
inconsistent results between genotyping and phenotyping.