Monochamus alternatus (M. alternatus) and Monochamus saltuarius (M. saltuarius) are major vectors for Bursaphelenchus xylophilus in South Korea. When an adult, they are easily distinguishable by several morphological classification. However, it is difficult to identification between M. alternatus and M. saltuarius when they are larvae as they have very similar morphological characters. Thus, they are not easily distinguishable without expertise about Cerambycidae taxonomy. Furthermore, during epidemiological investigation, sometimes, adults or larvae would not be founded in death pine trees. For these reasons, in this experiment, we are able to identified between M. alternatus and M. saltuarius by mitochondrial 12S rRNA gene primers that are specific to 12S rRNA gene fragment of M. alternatus using larvae tissue and frass. Moreover, we had examined whether vectors that were already escaped from dead pine tree have Bursaphelenchus xylophilus or not by multiplex PCR using larva frass that was remained in dead pine tree.

Monochamus alternatus (M. alternatus) and Monochamus saltuarius (M. saltuarius) are major vectors for Bursaphelenchus xylophilus in South Korea. When an adult, they are easily distinguishable by several morphological classification. However, it is difficult to identification between M. alternatus and M. saltuarius when they are larvae as they have very similar morphological characters. Thus, they are not easily distinguishable without expertise about Cerambycidae taxonomy. Furthermore, during epidemiological investigation, sometimes, adults or larvae would not be founded in death pine trees. For these reasons, in this experiment, we are able to identified between M. alternatus and M. saltuarius by mitochondrial 12S rRNA gene primers that are specific to 12S rRNA gene fragment of M. alternatus using larvae tissue and frass. Moreover, we had examined whether vectors that were already escaped from dead pine tree have Bursaphelenchus xylophilus or not by multiplex PCR using larva frass that was remained in dead pine tree.

The Japanese oak silkmoth, Antheraea yamamai Guérin-Méneville 1861 (Lepidoptera: Saturniidae), is one of the important natural resources possessing industrial value for silk fiber production. In this study, ten microsatellite markers and two mitochondrial DNA (mtDNA) gene sequences (COI and ND4) were used to investigate the genetic variation and geographic structure of A. yamamai populations in South Korea. Two mtDNA gene sequences revealed very low total genetic variation and resultant low geographic variation, validating to use further variable molecular markers. Population-based FIS, FST, RST, and global Mantel test consistently support that A. yamamai populations are overall well interconnected with a relatively high gene flow. Nevertheless, STRUCTURE analysis using microsatellite data and mtDNA sequences coincidently indicate the presence of two genetic pools in many populations.

The tiny dragonfly, Nannophya pygmaea (Odonata: Libellulidae) is one the smallest dragonflies in the world and listed as a second-degree endangered wild animal in Korea. We developed microsatellite markers and applied selected markers to South Korean populations to understand population genetic characteristics, along with two mitochondrial DNA (mtDNA) gene sequences (COI and ND5). Two mtDNA-based population genetic analysis indicates substantially reduced genetic diversity in an island population (Muuido) compared to others. On the other hand, population-based FST and RST consistently support that N. pygmaea populations are overall well interconnected with a relatively high gene flow. These results may collectively indicate that N. pygmaea populations in South Korea may have rather larger population size than we previously acknowledged based on a single-locus mtDNA sequence and field observation.

The red-spotted apollo butterfly, Parnassius bremeri Bremer, 1864 (Lepidoptera: Papilionidae), has been listed as an endangered species in Korea. We developed microsatellite markers by nest-generation sequencing (NGS), selected 12 markers, and applied the markers to available South Korean populations to understand population genetic characteristics. The genotyping of 40 P. bremeri individuals from three localities in South Korea showed that at each locus, the observed number of alleles ranged from 17 to 43, the observed and expected heterozygosities were 0.84722-0.90556 and 0.76045-0.79208, respectively, and FIS was –0.155 to –0.121. STRUCTURE analysis supported the presence of two genetic pools in all three populations, although an immediate reason for this subdivision is not known. The population based FST, RST, and front wing length collectively suggest that at least Samchuk population in Gangwon Province has a significant distance. Further scrutinized analysis is undergoing.

We developed single nucleotide polymorphism (SNP) markers and are establishing diagnostic systems to distinguish disease resistance- and susceptible-strains of honey bees using the SNPs. For development of SNP markers, whole genome was sequenced each from 20 individuals of “disease resistance-strain” and “susceptible-strain” of Apis mellifera ligustica using the Illumina HiSeq 2000 sequencer. Approximately, 344 and 294 million sequence reads were mapped to the honeybee reference assembly (Amel_4.5) for each strain, respectively. Among the total 2,246,428 SNPs yielded, 33 were found to be fixed between the two strains with all homozygosity. Sixteen of them were casually amplified and sequenced from randomly selected each 10 individual of honey bees from each strain and presented strain specific SNPs. These ten SNPs were used to diagnose the two strains either by original size difference, caused by indel-accompanying SNP, typical PCR-RFLP, or AS PCR.

We collected four species of Osmia spp. consisting of O. cornifrons, O. pedicornis, O. taurus and O. satoi in Yeongwol, Bonghwa and Yechon locations. O. pedicornis was the dominant species, which accounts for 59.6% to 76.4% in the three locations. The sex ratio of Osmia spp. were from 1:1 to 1: 2.8 in O. cornifrons and from 1:2.8 to 1: 5.5 in O. pedicornis. The sex ratio of O. cornifrons and O. pedicornis was different in 3 locations. The female and male cocoons of O. cornifrons were the long elliptical-shaped and grey-brown color. The female and male cocoons of O. pedicornis were the long elliptical-shaped and polished dark-brown color. In both species of O. cornifrons and O. pedicornis, the cocoon size of the females was significantly larger than that of the male. Between those two species, the latter had a relatively bigger size than the former.

In the age of sexual maturity of queen, mating occurred at 6.7% immediately after eclosion and it was the highest as 85.0% at 10 days after emergence. However, a remarkable decrease was occurred at 20 days after the emergence. With regard to oviposition rate, the highest rate was observed as 81.3-81.8% at 6 to 10 days of eclosion. The rate of colony foundation and progeny-queen production were the highest as 43.8% and 37.5%, respectively, at 8 days of eclosion. In the age of sexual maturity of male, mating rate was as high as 38.3% immediately after the eclosion, the highest as 80.0% at 25 days of eclosion, and 76.6% at 8 days of eclosion. The oviposition rate was the highest as 76.9% at 6 days of eclosion, and decreased to 75.0% at 8 days and 72.7% at 10 days of eclosion. Summarized, our results indicate that sexual maturity for mating of B. terrestris is most favorable 6-8 days after eclosion for queen and 6-10 days after for male.

Due to their environmental and economical consequences, invasive species have become a major concern worldwide. Among them, the yellow-legged hornet, Vespa velutina, is a keen hunter of domestic honeybees. They use olfactory cues from the prey to assess food information. In this study, we investigated the attraction behavior of V. velutina to honeybee pheromone under outdoor conditions. Nine honeybee pheromones were tested and compared in order to find the best attractant. When testing specific compounds, the honeybee queen pheromone, homovanillyl alcohol, proved highly attractive.

Ethyl formate (EF) and phosphine (PH3) is an alternative fumigant to methyl bromide (MB). For applying import sweet pumpkins in Korea, the efficacy of EF and PH3 was evaluated on Tetranychus urticae adult and egg stage which is pest in sweet pumpkins. The eggs of T. urticae were more tolerance than adults in both EF and PH3. When T. urticae eggs were treated with EF for 4 hrs, LCT99=107.63 mg/L and LCT99=45.37 mg/L at 5°C and 20°C, respectively. PH3 treatment at 5°C and 20°C for 24 hrs, eggs were controlled LCT99=49.44 mg/L and LCT99=17.23 mg/L, respectively. The mixed treatment of EF and PH3 showed no significant synergistic effect on T. urticae. However, EF (80 mg/L) and PH3 (4 mg/L) treatment did not cause any external phytotoxicity damages in sweet pumpkin even when treated with the maximum amount of fumigant at 5℃ and 20℃ for 24hrs.

Pseudococcus orchidicola and Pseudococcus longispinus, a notorious cosmopolitan pest species of mealybugs, known to be distributed indoors in Korea. The effect of temperature on the development of P. orchidicola and P. longispinus was studied. The duration of nymphal period of the two mealybugs at 24 and 28℃ have been recorded. Developmental duration of P. longispinus was longer than P. orchidicola in both females and males. The adult longevity was estimated to 74.9 and 91.1 days at 24℃ for P. orchidicola and P. longispinus, respectively. The average fecundity per female was greater at 28℃. The survival of P. orchidicola and P. longispinus during winter at outdoor was failed.

Ethyl formate (EF) and phosphine (PH3) is an alternative fumigant to methyl bromide (MB). The egg, nymph, and adult stages of Frankliniella occidentalis in asparagus were examined for the fumigation activity of EF and PH3. The eggs of F. occidentalis were more tolerance than other stages (adults and nymphs) in both EF and PH3. When the EF was treated for 4 hrs at 5℃ and 20℃, the eggs of F. occidentalis were LCT99=98.70 mg/L and LCT99=61.13 mg/L, and adults were LCT99=11.50 mg/L and LCT99=3.18 mg/L, respectively. However, at 5℃ and 20℃, the eggs were LCT99=83.76 mg/L and LCT99=53.6 mg/L, and adults were LCT99=4.58 mg/L and LCT99=3.44 mg/L, respectively for 4 hrs treatment of PH3. The PH3 was not any external phytotoxic damages at the maximum dose (4 mg/L), but the EF caused significant phytotoxicity in asparagus.

Entomopathogenic fungi, widely distributed in nature, have been developed as biopesticides of agricultural pests. However, little is known about isolation method for obtaining a variety of highly virulent strains from the soil. In this work, the previously established method using mealworm (larvae of Tenebrio molitor) as a baiting system was used to efficiently isolate fungal strains from soil, and the isolated fungal library in Korea was established. To isolate entomopathogenic fungi, soil samples from 10 regions of Korea such as Iksan, Imsil, Gunsan, Yeosu, Suncheon, Gwangyang, Jinju, Hamyang, Boseong and Jangseong were collected. Of the collected soils, 21 and 34 isolates of entomopathogenic fungi were isolated from Iksan and Gunsan, respectively. The isolates were classified as genera of Metarhizium, Beauveria and Isaria. This work can suggest an effective insect pathogenic fungal isolation method

Entomopathogenic fungi have been known as promising candidates in biological control. However, a large number of conidia with higher stability should be produced for industrialization. In this work investigated the possibility of several grains for the production of thermotolerant Isaria javanica and I. fumosoroseus conidia. The fungal conidia were produced on ten grains for 14 days in Petri dish conditions. Of the ten granular substrates, millet, perilla seed and barley showed the high conidial production, 15.7 ± 8.4, 17.3 ± 5.2 and 13.5 ± 1.9 × 109 conida/g in I. javanica and 15.9 ± 0.6, 15.0 ± 0.5 and 15.0 ± 0.7 ×109 conida/g in I. fumosoroseus, respectively. However, the two strains showed relatively low thermal stability regardless of substrates, when exposed to 45℃ for 1, 2 and 3 hours. This work suggests a possible substrate for conidial production of entomopathogenic fungi with low cost.

Oriental fruit fly, Bactrocera dorsalis, is one of the serious pests of fruit throughout the world. Two regional populations (Bangladeshi and Malaysia) were collected using methyl eugenol lure and one laboratory population was obtained from Thailand. Three populations were compared in cytochrome b (CYB) cytochrome oxidase I (COI) sequences. COI sequences were highly homologous. However, CYB sequences were polymorphic and used to discriminate these regional populations by a phylogenetic analysis. In CYB phylogeny analysis, Malaysia and Thailand populations were clustered and Bangladeshi population was separated

The objectives of this study are to examine the genetic variation in oriental fruit flies (Diptera: Tephritidae) and to use it as a barcode for the origin elucidation, in preparation for the their incursions into Korea. About 1,600 specimens of B. dorsalis sp. complex and B. correcta were collected from 10 countries, the Indochina peninsula, the Philippines, Taiwan and South China. A total of 182 cytochrome c oxidase (COI) sequences were obtained and aligned from these regional specimens. Three sequences from the Genebank reference were also included. Six hundreds base pair fragments were aligned and trimmed and used for a barcode. The phylogenetic tree was generated using the neighbor-joining method with 1,000 bootstrap replicates. There were two distinct groups in the phylogenetic tree, Bactrocera dorsalis sp. complex and B. correcta. Three specimen, intercepted in the hand-carried mango at the airport inspection and collected in the confiscated mango, smuggled from Vietnam were included for a test. The DNA from the airport were 100% identical to that of one of various Vietnam specimens, and that from the confiscated mango was mostly similar to those of Vietnam, suggesting the usefulness of the barcode for the origin elucidation tool of oriental fruit fly.

The oriental fruit fly, Bactrocera dorsalis (Hendel), has potential of serious damage to various vegetables or fruits, especially genus of Citrus, such as mandarin orange and tangerine in Korea, where its larvae do damages. Animal and Plant Quarantin Agency of Korea and Animsl Systematics Laboratory of Kunans National University have collected the samples of oriental fruit fly in East Asian countries, Vietnam, Philippines, Myanmar, China, Cambodia, India and Nepal. To confirm genetic differences and structure of B. dorsalis population samples collected from those countries, we analyzed 192 individuals from 28 locations over 7 countries using 15 microsatellite loci. In total samples, number of different alleles, number of effective alleles and Shannon's Information Index were 6.421±0.364 (standard error, SE), 3.664±0.137 (SE) and 0.902±0.030 (SE), respectively. Grand means (±SE) of observed and expected heterozygosity over all loci and populations were 0.534 (±0.018) and 0.597 (±0.017), respectively. Among all populations, Fst values ranged from 0.016 to 0.705 with averaging 0.194 (±0.026).