Indian Journal of Entomology Online published Ref. No. e23633 DoI. No.: 10.55446/IJE.2023.633 mtCO1 ANALYSIS OF THE BUTTERFLY GENUS PAPILIO, FROM INDONESIA Roni KoneRi 1 , SyamSul BachRy 2 * and Beivy Jonathan Kolondam 1 1 Department of Biology, Faculty of Mathematics and Natural Sciences, Sam Ratulangi University. Jl. Kampus Bahu, Manado 95115, North Sulawesi, Indonesia 2 Department of Biology, Faculty of Life Sciences, Pahlawan University, Tuanku Tambusai No.23, Bangkinang, Kec. Bangkinang Kampar, Riau 28412, Indonesia *Email: syamsulbachry89@gmail.com (corresponding author): ORCID ID: ABSTRACT The genus Papilio is a butterfy that has many subspecies with similar phenotypes. So until now, researchers are still studying the genus Papilio based on morphology and genetics. Partial sequences of mitochondrial cytochrome oxidase-I (mtCOI) genes from individuals from 13 species of Papilionidae from North Sulawesi: Indonesia, Papua New Guinea, Malaysia: Negeri Sembilan, India: Assam, South Korea, China, Taiwan: Nantou, and Thailand: Fang, Chiang Mai have been compared in this study. Nucleotide divergences showed that the mean genetic distance between species was 0.023 (2.3%) to 0.723 (72.32%). Phylogenetic analysis revealed that groups of Papilionidae from North Sulawesi: Indonesia formed their own group; Species from Papua New Guinea are closely related to other Papilionidae species from other regions. There are 50 distinct sites or single nucleotide polymorphism (SNP) in the mtCO1 sequence of eleven species. These mtCO1 sequences can reveal the genetic differentiation of closely related species in the genus Papilio. Key words: Butterfy Papilio spp., cytochrome C oxidase-I, North Sulawesi, sequences nucleotide,mtCO1 variations, comparisons, phylogenetic analyses, NcB accessions Butterfies are a group of insects that can be found in tropical and subtropical areas. Butterfy species are insects that have various morphological and colour characteristics that may vary due to environmental habits and geographic location (Simonsen et al., 2012). Due to these butterfies can be used as bioindicators of changes in environmental quality (Basset et al., 2012). The presence of butterfly populations in a habitat ecosystem has begun to decline due to various threats such as uncontrolled exploitation and hunting, as well as land clearing and forest conversion (Basset et al., 2012). Koneri and Maabuat (2016) state that butterfy diversity is very influential in their habitat. Taxonomically, butterflies are included in the order Lepidoptera. Butterfies have an important role in the ecosystem, namely as pollinators of fowers or pollinators (Duara et al., 2014). The classifcation of insect species is very important for basic and applied research. Classifcation based on morphological characteristics creates problems for many insect groups due to their small size, and morphological shapes that change over time (Akhilesh and Sebastian, 2014; Tsao and Yeh, 2008). The Papilionidae family, commonly known as swallowtail butterfies, is large and has a variety of colours. This species can be found in tropical regions and can be found on almost every continent. There are an estimated 550 species in this family. In the 1990s, studies of this type of butterfy began to focus on morphological and molecular systematics (Simonsen et al., 2011). Some morphological evidence in the subfamily Baroniinae, Parnassinae and Papilionidae shows no specific differences. (Vane-Wright, 2003). Based on the cladistic method, Kristensen (1976) found that the Papilionidae family has close ties with several family groups such as Pieridae, Nymphalidae Lycaenidae, and Riodinidae. This genus Papilio contains many cryptic species where phenotypic and evolutionary variability has led to misidentifcation. Apart from that the dimorphism also causes confusion (von-Maria et al., 2011). Taxon identifcation based on DNA sequences has begun to be used to facilitate identifcation and even discover new species (Blaxter, 2005; Hebert et al., 2003a, b). DNA sequences from the mitochondrial cytochrome oxidase 1 (mtCO1) gene can serve to identify and analyze the phylogeny (Hebert et al., 2003a, 2004b; Ward et al., 2005; Huang and Ke, 2015), especially the cryptic or similar species in the tropics (Wilcox et al., 1997; Berkov, 2002; Hebert et al. 2004b; Monaghan et al., 2005; Hajibabaei et al. 2006; Bachry et al. 2019) and insects in different places and at various growth stages (Janzen et al., 2005). In Indonesia, substantial studies