α-solanine is toxic to human health by disturbing digestive and central nervous systems. However, little information has been focused on investigated with respect to α-solanine influence in mammal oocyte maturation and quality. In this study, we investigated the effects of α-solanine on oocyte maturation, quality and possible molecular mechanisms in a pig model. Porcine Cumulus-oocyte complexes (COCs) were treated with increasing concentration (0, 1, 10, 20, 50 μM) of α-solanine subjected to further in vitro maturation culture. The result showed that α-solanine significantly inhibited cumulus cells expansion and increased oocyte death rates when the concentration of α-solanine more than 10 μM. After cell cycle and cytoskeleton analysis, the results showed that α-solanine (10 μM) disturbed meiotic resumption, increased abnormal spindle formation and cortical granules (CGs) distribution rates when compared with the untreated group. α-solanine (10 μM) triggered autophagy by increasing the expression of autophagy-related genes (LC3, ATG7, LAMP2) and accumulation of LC3-specific puncta (an autophagy maker). TUNEL staining assay showed that α-solanine significantly increased apoptosis in porcine oocytes confirmed by up-regulated the levels of BAX and CAPS3 genes. Further study revealed that exposure α-solanine (10 μM) to porcine oocytes induced ROS generation, reduced mitochondrial membrane potential. In addition, our results suggested that α-solanine (10 μM) significantly increased the levels of H3K36me3 and H3K27me3 in porcine oocytes. Taken together, these data indicated that α-solanine toxic impaired oocyte maturation and quality by inhibited cumulus cells expansion, increased abnormal spindle and CGs distribution rates, triggered autophagy/apoptosis occur, accumulated ROS, decreased mitochondrial membrane potential, and changed epigenetic modifications.

In pig, more than half of the recovered cumulus cell-oocyte complexes (COCs) have one or two layers of cumulus cells and are considered morphologically poor. If we could take full advantage of these poor quality COCs, we could potentially improve the efficiency of in vitro embryo production. During in vitro maturation, although some maturation factors are transmitted bidirectionally between the oocyte and cumulus cells of the same COC, transmission also occurs between different COCs. We hypothesized that morphologically poor COCs fail to undergo complete oocyte maturation due to their insufficient secretion of maturation factors. Here, we investigated whether co-culture with morphologically good COCs (having three or more layers of cumulus cells) could improve the maturation and utilization rates of morphologically poor COCs. Our results revealed that the oocyte maturation rate, glutathione level, embryo development capacity, blastocyst quality, and cumulus cell gene expression levels of BCL-2 and PCNA were similar in the co-culture and good quality-groups, and that these levels were all significantly higher than those in the poor quality-group. Our results strongly suggest that the co-culture strategy greatly improved the utilization rate of morphologically poor COCs without reducing their capacity for maturation and subsequent development.

Early pregnancy results in th production of various signal molecules such as steroids, prostaglandins, and many protein factors. The proteins especially produced by the placenta have been used to detect pregnancy for many years in other species. More recently, pregnancy-specific protein B, which is a placental glycoprotein can be measured by RIA or proteomic methods in serum of pregnant cow. And 2D Fluorescence difference gel electrophoresis (DIGE) is an emerging technique for comparative proteomics, which improves the reproducibility and reliability of differential protein expression analysis between samples. For this reason, we are analyzed serum of bovine. The purpose of this study was to apply DIGE technique for identification of bovine pregnancy-specific proteins using bovine pregnant and non-pregnant serum samples. Serums of 2 pregnant Holstein dairy cattle at day 21 after AI and those of 2 non-pregnant were used in this study. The molecular weight and charge matched cyanine dyes enable pre-electrophoretic labeling of non-pregnancy and pregnant serum proteins which are then mixed and labeled with Cy2 were used as an internal standard. Two pools of proteins are labeled with Cy3 and Cy5 fluorescent dyes, respectively. Labeled proteins with Cy2, Cy3 and Cy5 mixed together and separated in same gel and then were detected by fluorescence image analyzer. The 2D DIGE analysis using fluorescence CyDye flour showed higher sensitivity and better reproducible results than conventional 2D gel electrophoresis. Approximately 1,500 protein spots were detected by 2D DIGE. The differentially expressed proteins were identified by MALDI-TOF Mass spectrometer. Total 16 protein spots differentially expressed in the pregnant serum were detected, among which 7 spots were up-regulated proteins identified as conglutinin precursor, modified bovine fibrinogen, IgG1 etc, and 6 spots were down-regulated proteins identified as hemoglobin, complement component 3, bovine fibrinogen, IgG2a etc. These results indicated that DIGE system could be advantageous for the analysis of serum proteomics diversified by physiological conditions.

The number of reported mitochondrial genomes (mitogenomes) from the monotypic Lasiocampoidea has been limited until recently. In this study, we sequenced the complete mitogenome of the lappet moth, Kunugia undans (Lepidoptera: Lasiocampidae), and compared it to those of other lasiocampid species and macroheteroceran superfamilies (59 species in six superfamilies). The 15,570-bp long K. undans genome had the typical set of genes found in animal mitogenomes, with the exception of one additional trnR that are located between trnA and trnN loci. Considering that the two trnR copies are located in tandem with proper secondary structures and identical anticodons, a gene duplication event might be responsible for the presence of the two tRNAs. In summary, the general mitogenome characteristics of Lasiocampoidea did not differ greatly from the remaining macroheteroceran superfamilies, but it did exhibit some unique features.

Currently, only limited number of mitochondrial genomes (mitogenome) is available from Odonata. In order to extend current mitogenome data for comparative biology and phylogeny we sequenced complete mitogenomes of two endangered dragonfly species, Libellula angelina and Nannophya pygmaea (Ododana: Libellulidae). The whole genomes were 15,233 bp in L. angelina and 15,112 bp in N. pygmaea and included a typical set of genes (13 protein-coding genes, two rRNA genes, and 22 tRNA genes) and one major non-coding A+T-rich region. The arrangement of the genomes was identical to typical one found in insects. Phylogenetic reconstruction using concatenated sequences of 13 PCGs and two rRNAs of Odonata (17 species in eight families in three suborders) using both Bayesian Inference (BI) and Maximum Likelihood (ML) methods have shown a strong support for monophyletic Zygoptera (BI, BPP = 1 and ML, 100%). Currently, further scrutinized analysis is under progress.

The spotted-wing drosophila (SWD), Drosophila suzukii (Diptera: Drosophilidae), is an economically damaging pest that feeds on most thin-skinned fruits. In this study, we sequenced portions of the mitochondrial (mt) COI and ND4 genes from a total of 195 individuals collected mainly from Korea. A total of 139 haplotypes were obtained from the concatenated COI and ND4 sequences. A dataset combining GenBank sequences with our own data identified a total of 94 worldwide COI haplotypes with a maximum sequence divergence of 5.433% (32 bp). A rough estimate of genetic diversity in each country showed higher diversity in ancestral distributional ranges, but the invasion over Asian countries seems to have been substantial because haplotype diversity was only 2.35-3.97-fold lower in the USA, Canada, and Italy than that in the populations ancestral ranges.

In order to understand evolutionary characteristics of gene rearrangement in Lepidoptera, we collected all available complete mitogenome (mitogenome) sequences registered in GenBank (274 mitogenomes from 44 families in 23 superfamilies as of August 6, 2015). It turned up six rearrangements that differ from the arrangement of ancestral insects, including that of the gelechioid Mesophleps albilinella that we sequenced in this study. The M. albilinella mitogenome has a unique gene arrangement among the Gelechioidea: ARNESF (the underline signifies an inverted gene) at the ND3 and ND5 junction, as opposed to the ARNSEF that is found in ancestral insects. Most of the rearrangements can be explained by the tandem duplication-random loss model, but inversion, which requires recombination, is also found in two cases, including M. albilinella. Excluding the MIQ rearrangement at the A+T-rich region and ND2 junction, which is found in nearly all Ditrysia, most of the remaining rearrangements found in Lepidoptera appear to be independently derived in that they are automorphic at several taxonomic scales. Current mitogenomic data are limited, particularly for congeneric data. Thus, future research focused on congenerics could clarify evolutionary independency at the generic level also.

The Gelechioidea is the second most species-rich group of Lepidoptera, but only limited number of mitochondrial genome (mitogenome) sequences is available. Thus, we sequenced the complete mitogenome of a gelechioid Hieromantis kurokoi (Lepidoptera: Stathmopodidae) to use the data for future study for the higher phylogeny of Ditrysia in Lepidoptera. The arrangement of the genome was identical to typical one found in Ditrysia (trnM-trnI-trnQ) (underline for inverted gene). The COI began with CGA, which has been designated as the start codon for majority of lepidopteran species, whereas other protein-coding genes (PCGs) began with the typical ATN codon. The 360-bp long A+T-rich region harbored the conserved sequence blocks Phylogenetic analysis using the 13 PCGs both by Bayesian inference (BI) and Maximum-likelihood (ML) methods indicated that H. kurokoi belonging to the family Stathmopodidae grouped together with within-familial species Atrijuglans hetaohei with the highest nodal support (BI, 1.0; ML, 100%).

The lycanid butterfly, Shijimiaeoides divina (Lepidoptera: Lycaenidae) is listed as the second-degree endangered wild animal in Korea from 2012. The 15,259-bp long complete mitochondrial genome of the species consisted of a typical set of genes (13 protein-coding genes, two rRNA genes and 22 tRNA genes) and one major non-coding A+T-rich region, with the typical arrangement found in majority of Lepidoptera. The 15,259-bp long S. divina mitogenome is well within the range found in Lycaenidae and has typical sets of 37 genes and a major non-coding A+T-rich region as 379 bp. As other lycanid butterflies S. divina COI also started with CGA. The gene arrangement of S. divina is identical to that of the Ditrysia in Lepidoptera that has the order trnM-trnI-trnQ (underline for inverted gene) between the A+T-rich region and ND2. Comparison of the skewness between the PCGs encoded in major and minor strand indicates a substantial difference between them in GC skewness (0.261 ~ 0.340 in minor strand vs. -0.081 ~ -0.115 in major strand). The 151-bp intergenic spacer sequence of the S. divina mitogenome is spread over 16 regions ranging in size from 1-53 bp. The longest one (53 bp) located between trnQ and ND2 shows substantially high sequence homology to neighboring ND2 may indicating the origination of the region by a partial duplication of the ND2 gene. One of the unusual features of the S. divina mitogenome is the presence of a trnK-like sequence that is encoded at the major strand of the genome in the A+T-rich region.

Immunoprecipitation-based high performance liquid chromatography (IP-HPLC) is a type of modified enzyme-linked immunosorbent assay (ELISA) that uses protein A/G (or antibody)-conjugated beads instead of the antibody-conjugated wells used in ELISA. In order to determine the fidelity of IP-HPLC, the author used 83 antisera to identify protein expression changes caused by cisplatin treatment in KB human oral cancer cells. KB cells were cultured for 12 or 24 hours with 10 ug/mL cisplatin. The results obtained by IP-HPLC were comparable with published cisplatin data, although ELISA was not conducted in the present study. Cisplatin dominantly reduced the levels of proteins associated with cell proliferation, transcription factors, growth factors, cytoskeletal proteins, and cellular differentiating factors, but on the other hand, apoptosis-related factors, oncogenes, and protective proteins were usually up-regulated, presumably to address cisplatin-induced DNA damage. In particular, cisplatin directly inactivated genomic DNA by down-regulating histone H1 and demethylase and by up-regulating deacetylase. Cisplatin also rapidly induced p53 overexpression and mitochondria-mediated endogenous apoptosis occurred after 12 hours of cisplatin treatment, although this was almost completely replaced by FASL/FAS-mediated exogenous apoptosis after 24 hours. This preliminary study was conducted to investigate the anticancer effect of cisplatin on the KB human oral cancer cells and to determine the fidelity of IP-HPLC data. It was concluded that IP-HPLC is useful for identifying profile changes of genome wide essential proteins and signaling changes of major molecular pathways.

Poly(vinyl alcohol) (PVA)에 대하여 가교제로써 glutaraldehyde (GA), maleic anhydride (MA)를 이용하여 제조한 코팅용액을 알칼리로 가수분해 시킨 poly acrylonitrile (PAN) 중공사 막표면에 코팅하여 막을 제조하였다. 제조된 막의 투과 특성평가를 위해서 물/에탄올 혼합액에 대한 투과증발 실험을 수행하였다. 60°C의 90 wt%의 물/에탄올 혼합액에 대하여 반응온도 및 가교제의 농도변화에 따른 투과도 및 선택도를 측정하였다. 일반적으로 반응온도, 가교제 농도가 증가할 경우 투과 도는 낮아지고, 선택도는 증가하는 경향을 보여주었다. 가교제로 GA의 대표적 결과는 반응온도 120°C, GA 11 wt%로 투과 도는 165 g/m 2 hr 선택도는 81이고, MA는 반응온도 120°C, MA 11 wt%로 투과도는 174 g/m 2 hr 선택도는 73의 결과를 얻을 수 있었다.

We used the three kinds of mosquito traps (Black-hole with UV light, CO2-baited Mos-hole with the newly developed attracting-solvent, CO2-baited DMS; Digital Mosquito Monitoring System) to know their collecting efficiency for the female mosquitoes in Korean rural areas. The Black-hole mosquito trap caught many kinds of insects including only few female mosquitoes. The Black-hole trap has the UV-light and the light seemed to attract other terrestrial and aquatic insects, such as the common flies, May flies, and the stone flies. Even though the trap was developed to collect mosquitoes, the trap caught only few of female mosquitoes less than 1% of all insects caught. Their selective efficiency to collect the female mosquitoes was relatively lower than other two kinds of traps. The Mos-hole and CO2-baited DMS traps had the collecting efficiency of over 80% to collect the female mosquitoes. The two traps caught the relatively lower number (less than 3% of total insects) of other insects, such as few Coleoptera and Diptera, and their collecting efficiency for the female mosquitoes was very higher. Generally speaking, mosquitoes disliked the UV light but they relatively preferred CO2 gas including the attracting-solvent. They had also been attracted the acidic solvent with CO2 gas. If we could use the efficient and selective mosquito traps with the fully understanding about the mosquito habits, we could assume that we can keep the biodiversity high around the mosquito habitats as well as to save money for the insect pest control.

This snag-dwelling arthropod community study was conducted for the ecological evaluation of dead woods at Korean fir stand in Mt. Woonak in Pocheon-si, Gyeonggi-do, from April 2010 to August 2011. We put a windows trap and an emergence trap on the trunk of each snag, and we selected the six snags during the study periods. We collected 3930 individuals (5 class, 21order, 52 families) but we didn’t include the number of unidentified larva. We separated those individuals into the three functional groups and we found out the proportion and number of each functional group from the total individuals: herbivores (27.6%, 1083) predators (10.9%, 430), detritivores (61.4%, 2413), etc(0.1%, 4). We found out the proportion and number of each taxon group for herbivores, Armadillidae (0.15%, 6), Acarina (2.57%, 101), Psocoptera (0.25%, 10), Hemiptera (0.46%, 18), Mecoptera (0.05%, 2), Hymenoptera (not ant) (5.14%, 202), Aphididae (3.82%, 150), Cicadellidae (0.4%, 16), Curculionidae (3.61%, 142), Chrysomelidae (0.23%, 9), Elateridae (3.36%, 132), Erotylidae (3.16%, 124), Nitidulidae (2.6%, 102), Pyrochoroidae (0.08%, 3), Scarabaeidae (0.31%, 12), and Cetoniidae (0.13%, 5). Predators were consisted of the following taxa groups: Araneae (2.9%,114), Chilopoda (0.31%, 12), Formicidae (4.25%, 167), Carabidae (0.08%, 3), Staphylinidae (1.09%, 43), Cleridae (0.05%, 2), Pselaphidae (0.1%, 4), Colydiidae (0.38%, 15), Harpalidae (0.1%, 4), Histeridae (0.36%, 14), and Dermestidae (0.8%, 31). Detritivores were consisted of the following taxa groups: Millipedes (0.92%, 36), Archaeognatha(1.6%, 63), Diptera (7.81%, 307), Collembola (35.47%, 1394), Protura (0.03%, 1), Dermaptera (0.1%, 4), Tettigoniidae (0.08%, 4), Raphiidophoridae (0.03%, 1), Ipidae (14.12%, 555), Silphidae (0.15%, 6), Cuculidae (0.15%, 6), Cerambycidae (0.38%, 15), Oedeeridae (0.03%, 1), Lucamnidae (0.03%, 1), Stenotrachelidae (0.05%, 2), Buprestidae (0.13%, 5), Tenebrionidae (0.23%, 9), and Mordellidae (0.1%, 4), etc. Conclusively, the snag plays an important roll as the diverse arthropods’ habitats in the Korean fir forest ecosystem.

The pear psylla, Cacopsylla pyricola Foerster (Homoptera: Psyllidae), is a serious insect pest of commercial pear crops. The species, which resides on pear trees throughout its life cycle, is rapidly spreading in some regions of the world. Given the life cycle, it is unclear how such a rapid spread has been facilitated. Presently, the population genetic structure of the species including genetic diversity and gene flow was studied to understand the nature of dispersal and field ecology of the species. Pear psylla was collected from several pear orchards in Korea. The 658-bp region of mitochondrial COI gene and the 716-bp long complete internal transcribed spacer 2 (ITS2) of the nuclear ribosomal DNA were sequenced. Unlikely other previously studied insect pests, the COI-based genetic diversity of the pear psylla was extremely low (maximum sequence divergence of 0.15%). This finding allowed us to conclude that the species may have been introduced in Korea relatively recently, possibly with the phenomenon of genetic bottlenecks. ITS2 sequence-based analyses of phylogeny, population differentiation, gene flow, and hierarchical population structure all concordantly suggested that the pear psylla populations in Korea are neither genetically isolated nor hampered for gene flow. These genetic data are concordant with the dispersal of an overwintering winterform morph outside the non-pear habitat in the fall and the possibility of subsequently longer distant dispersal.

Diamondback moth (Plutella xylostella L.) belonging to genus Lepidoptera is a notorious pest of cruciferous crops worldwide. We evaluated the bioinsecticidal activity of the liquid cultures (LB and NB) of a bacterial strain, Serratia sp. EML-SE1, isolated from a diseased diamondback moth. The pathogenicity of a bacterial strain to diamondback moth was confirmed by the following procedures: treatment of liquid culture on cabbage leaves, ingestion of inoculated cabbage and mortality response. For the test, twenty 3rd instar larvae of diamondback moth were placed on the Chinese cabbage leaf in a round plastic cage (Ø 10 × 6 cm) and sprayed with the liquid cultures. After 72 hours, insecticidal activity of LB and NB cultures of Serratia sp. against P. xylostella larvae showed 91.7% and 88.3%, respectively. In addition, the bioinsecticidal activity on potted cabbage with 14 leaves in a growth cage (165 × 83 × 124 cm) also was similar to that of plastic cage experiment. Summarized, the Serratia sp. EML-SE1 may be a potent candidate as a bioinsecticidal agent to control diamondbac kmoth.

We have determined the complete mitochondrial genome of the yellow-spotted long horned beetle, Psacothea hilaris (Coleoptera: Cerambycidae), an endangered insect species in Korea. The 15,856-bp long P. hilaris mitogenome harbors gene content typical of the animal mitogenome and a gene arrangement identical to the most common type found in insect mitogenomes. As with all other sequenced coleopteran species, the 5-bp long TAGTA motif was also detected in the intergenic space sequence located between tRNASer (UCN) and ND1 of P. hilaris. The 1,190-bp long non-coding A+T-rich region harbors an unusual series of seven identical repeat sequences of 57-bp in length and several stretches of sequences with the potential to form stem-and-loop structures. Furthermore, it contains one tRNAArg-like sequence and one tRNALys-likes equence. Phylogenetic analysis among available coleopteran mitogenomes using the concatenated amino acid sequences of PCGs appear to support the sister group relationship of the suborder Polyphaga to all remaining suborders, including Adephaga, Myxophaga, and Archostemata. Among the two available infraorders in Polyphaga, a monophyletic Cucujiformia was confirmed, with the placement of Cleroidea as the basal lineage for Cucujiformia. On the other hand, the infraorder Elateriformia was not identified as monophyletic, thereby indicating that Scirtoidea and Buprestoidea are the basal lineages for Cucujiformia and the remaining Elateriformia.

The complete nucleotide sequences of the mitochondrial genome (mitogenome) from the white-spotted flower chafer, Protaetia brevitarsis (Coleoptera: Cetoniidae) was determined. The 20,319-bp long circular genome is the longest among the completely sequenced arthropods. This extraordinary length of the genome stemmed from 5,654-bp long A+T-rich region composed of twenty-eight 117-bp tandem repeats, seven 82-bp tandem repeats, and each two 19-bp and 38-bp tandem repeats. The P. brevitarsis contains a typical gene complement, order, and arrangement identical to most common type found in insects. The P. brevitarsis COI gene does not have typical ATN codon. Thus, we also designated it as AAC (asparagine), which is found in the start context of all sequenced Polyphaga within Coleoptera. All tRNAs showed stable canonical clover-leaf structure of other mt tRNAs, except for tRNASer (AGN), DHU arm of which could not form stable stem-loop structure. The 5bp-long motif sequence (TAGTA) that has been suggested to be the possible binding site for the transcription termination peptide for the major-strand also was found betweent RNASer (UCN) and ND1, as have been detected in all sequenced coleopteran insects.

The 15,389-bp long complete mitogenome of the endangered red-spotted apollo butterfly, Parnassius bremeri (Lepidoptera: Papilionidae) was determined. This genome has a gene arrangement identical to those of all other sequenced lepidopteran insects, which have the gene order of tRNAMet, tRNAIle, and tRNAGln at the beginning. Due to the uncertainty the start codon for COI gene in insect has been discussed extensively. We propose the CGA sequence as the start codon for COI gene in lepidopteran insects, based on complete mitogenome sequences of lepidopteran insects including our P. bremerii and additional sequences of the COI start region from a diverse taxonomic range of lepidopteran species (a total of 51 species belonging to 15 families). As has been suggested in other sequenced lepidopteran insects the 18 bp-long poly-T stretch and the downstream conserved motif ATAGA that were previously suggested to serve as a structural signal for minor-strand mtDNA replication also was found at the 3’-end region of the P. bremerii A+T-rich region. In an extensive search to find out tRNA-like structure in the A+T-rich region, each one tRNATrp-like sequence and tRNALeu (UUR)-like sequence were found in the P. bremeri A+T-rich region, and most of other sequenced lepidopteran insects were shown to have tRNA-like structure within the A+T-rich region, thereby indicating that such feature is frequent in the lepidopteran A+T-rich region. Phylogenetic analysis using the concatenated 13 amino acid sequences and nucleotide sequences of PCGs of the four macrolepidopteran suferfamilies together with Tortricoidea and Pyraloidea well recovered a monophyly of Papilionoidea and a monophyly of Bombycoidea. However, Geometroidea and Noctuoidea were unexpectedly clustered as one group and placed this group to the sister group to Bombycoidea, instead of Papilionoidea in most analyses.

Invertebrate mitochondrial genome contains 13 protein-coding genes and major start codons for them are ATA (Met) and ATG (Met). However, alternative start codons such as ATT (Ile), ATC (Ile), TTG (Leu), and GTG (Val) also have been suggested from a diverse organism. Approximately 120 complete mitochondrial genome reported showed that the start codon for COI gene evidences an array of diverse designation of COI start codon such as typical ATN, tetranucleotide TTAG and ATAA, newly proposed AAT and AAC and so on. In the case of Lepidoptera, many completely sequenced species showed no typical start codon at the start context of COI and even within the neighboring tRNATyr. In order to clarify, we newly sequenced the beginning context of COI gene, encompassing the neighboring tRNATyr and start region of COI gene from 39 species belonging to eight lepidopteran families. We found the newly sequenced 39 species and 14 available complete lepidopteran mitochondrial genomes all possessed CGA (arginine), which is the first non-overlapping in-frame codon in COI gene. Furthermore, this CGA is highly well aligned in terms of both nucleotide and amin o acid sequences with neighboring region. Thus, the CGA (arginine) may be synapomorphic character for Lepidoptera, functionally constrained. We, therefore, propose the CGA sequence as the start codon for COI gene in lepidopteran insects.

We analyzed a portion of mitochondrial COI gene sequences (658 bp) to investigate the genetic diversity and geographic variation of the swallowtail butterfly, Papilioxuthus L., and the cabbage butterfly, Pieris rapae (Lepidoptera: Papilionidae). P. xuthus showed a moderate level of sequence divergence (0.91% at maximum) in 15 haplotypes, whereas P. rapae showed a moderate to high level of sequence divergence (1.67% at maximum) in 30 haplotypes, compared with other relevant studies. Analyses of population genetic structure showed that most populations are not genetically differentiated in both species. The distribution pattern of both species appears to be consistent with category IV of the phylogeographic pattern sensu Avise (Avise et al. 1987): a phylogenetic continuity, an absence of regional isolation of mtDNA clones, and extensive distribution of close clones. The observed pattern of genetic diversity and geographic variation of the two butterfly species seems to reflect the abundant habitats, abundant host plants, and flying abilities in connection with the lack of historical biogeographic barriers.