The invention relates to a development method and application of a KASP marker closely related to the
pungency degree of
hot peppers, and the development method comprises the following steps: constructing two hot pepper
pungency degree difference extreme mixing pools by utilizing an F2 segregation
population obtained by hybridizing HJ01 of high-
pungency hot peppers and HJ02 of low-pungency
hot peppers; according to a BSA-seq
whole genome sequencing result, a candidate region for regulating and controlling the pungency degree of hot peppers is preliminarily screened out,
correlation analysis is carried out on genes in the candidate region in combination with
RNA-seq, it is found that only two significant intervals are located, QTL intervals are located on a
chromosome 2, the starting position range and the ending position range of the QTL intervals are 135670001-136670000 and 136800001-152640000, and the QTL intervals are located on a
chromosome 2. The method is used for the interval range of a key
gene causing the difference between the
capsaicin content in the 'HJ01' high-spicy
wild type material and the
capsaicin content in the 'HJ02' low-spicy
mutant material. KASP molecular markers are developed on the basis of variation sites,
target site genotype detection is carried out on high-spicy materials and low-spicy materials, the high-spicy materials are C: C type materials, the low-spicy materials are G: G type materials, and the markers are co-dominant; kASP is used for carrying out
genotype identification on the F2
population, and the accuracy rate reaches 89.3%.