Development and validation of sex-associatedspecific markers using whole-genome re-sequencing in frog Quasipaa spinosa

Liaoruilin Zhang¹Juan Li²Xiang Li¹Jinrong He¹Jie Zhou¹Jinliang Hou¹Yulu Liu¹Lei Zhang¹Yanfei Huang¹Hong Li¹Xiaolin Liao¹Xinhua Liu¹Yazhou Hu¹Deliang Li¹Jianguo Xiang^1*

• ¹College of Fisheries, Hunan Agricultural University, Changsha, China
• ²Animal Husbandry and Fisheries Affairs Center, Changsha, China

The development of sexsex-specific genetic markers is crucial for addressing monosexual breeding in aquaculture species and for identifying traits that are sexually inherited, especially for elucidating the mechanisms of sex determination in amphibians. In aquaculture, comprehending sex determination is especially vital because the market value of animal products frequently depends on their sex.Quasipaa spinosa (Anura, Dicroglossidea) is a valuable frog species in the aquaculture industry of China and Southeast southeast Asia, yet there exists limited genomic information regarding this organism. Current data indicates that the adoption of all-male breeding techniques in Q. spinosa could substantially benefit the Chinese aquaculture industry, both by augmenting its economic prospects and by ensuring the effectiveness of wildlife reintroduction efforts. The growth rate, adult size, disease resistance, and other traits of male Q. spinosa surpass those of females, making the 带格式的: 行距: 单倍行距 development of all-male breeding a significant focus in the Q. spinosa aquaculture industry. Therefore, it is imperative to establish a marker specific to males. In this research, we used the male Q. spinosa genome as reference and performed whole-genome resequencing on 30 males and 30 females. Subsequently, we exhibited evident sexual differentiation on chromosome 3 and primers were designed for PCR detection of the identified candidate male INDEL loci. Ultimately, two sex-associatedspecific INDELs that could be effectively detected were obtained and validated on the samples collected from the remaining three locations, thereby confirming the robustness of these two INDELs for sex identification in Q. spinosa.