The Asian longhorned tick, Haemaphysalis longicornis, is an ectoparasite of domestic and wild animals, which transmit various pathogens including Severe Fever with Thrombocytopenia Syndrome Virus (SFTSV). They are the predominant hard tick species in Republic of Korea (ROK) and widely distributed throughout ROK. It is known that H. longicornis produce their offspring via two reproductive strategies, bisexual and parthenogenesis. It might affect their population maintenance and vectorial capacity. Parthenogenesis H. longicornis had the insertion of two thymine ‘T’ in mitochondrial 16s rDNA. In this study, we analyzed the proportion of bisexual and parthenogenesis H. longicornis in ROK including thirteen cities: Goseong[GS], Sokcho[SC], Chuncheon[CC], Ganghwa[GH], Samcheok[SCH], Sangju[SJ], Boryeong[BR], Ulsan[US], Gochang[GC], Jinju[JNJ], Jindo[JD], Jeju[JJ], and Seogwipo[SG]. Parthenogenesis individuals predominated from the northeastern are of ROK including Goseong, Sokcho, Chuncheon, Ganghwa, Samcheok, Sangju, Ulsan, and Jinju. Whereas bisexual individuals predominated from the southwestern area in ROK including Boryeong, Gochang, Jindo, Jeju, and Seogwipo. The analysis of haplotype diversity using concatenated sequences of mitochondrial Cytochrome c oxidase subunit I (COI) and Cytochrome B (CytB) revealed that H. longicornis were grouped into two major haplotypes. Two major haplotypes were correlated with bisexual and parthenogenesis, respectively. Likewise, H. longicornis individuals were divided into two clades and each clade were indicated by bisexual and parthenogenesis. The current study provides us an understanding of the genetic characteristics of two reproductive strategies for H. longicornis, which will be led to expand knowledge of the life cycle and population maintaining for H. longicornis.