High oleate peanut is more and more popular in food, oil processing, seed industry and so on be⁃
cause of its high quality of oil components. However, the varieties with high oleate cultivated at recent years have
narrow genetic diversity. It is necessary to develop novel molecular markers and breed high oleate varieties (lines)
with excellent characteristics and wide adaptability, and to broad the genetic background resources of domestic pea⁃
nut. In this study, with respect to the high oleic acid mutant with F458 background, resulting from the MITE inser⁃
tion into peanut AhFAD2B gene, a MITE molecular marker was developed for assisted-selecting of F1 hybrids and
the single plant in the offspring generation.This marker and detection method could distinguish three types of geno⁃
types through 1 round of conventional PCR reaction and common agarose gel electrophoresis. It has the advantages
of easy operation, low cost and good stability, and significantly improved the efficiency of target trait selection. In or⁃
der to breed new germplasm of high oleate peanut, the hybrid combination was made with Yuhua 2 (YH2), a peanut
cultivar with strong salt tolerance and edible oil as the female parent, and Weihua25(WH25), a high oleic acid pea⁃
nut as the male parent. Finally, six (5 with semi-erect type and 1 with erect type) new high oleate lines were ob⁃
tained through screening agronomic traits by pedigree method, and the genotype identification of F1 true hybrids and
erance of the main seeds of these lines were investigated and analyzed.