Two new Palaearctic species of Xynobius Foerster (Hymenoptera, Braconidae, Opiinae).

Two new and very similar species of the genus Xynobius Foerster, 1863 are described and illustrated, X.subparallelus Han & van Achterberg, sp. nov. from Japan (Honshu) and X.setosiscutum van Achterberg, sp. nov. from Norway. Three species are newly reported from Norway: Xynobiusaciculatus (Thomson, 1895), X.comatus (Wesmael, 1835), and X.polyzonius (Wesmael, 1835). X.polyzonius (Wesmael, 1835) and X.sapporanus (Fischer, 1963) are new combinations. Identification keys to the Xynobius species known from Norway and Japan are added.

Population Genetics of the Blueberry Gall Midge, Dasineura oxycoccana (Diptera: Cecidomyiidae), on Blueberry and Cranberry and Testing Invasion Scenarios.

We compared the population genetic structure between populations of the blueberry gall midge-Dasineura oxycoccana (Johnson) (Diptera: Cecidomyiidae)-from blueberry and cranberry and determined the genetic relationships among geographical subgroups by genotyping 632 individuals from 31 different populations from their native USA regions (New Jersey, Michigan, and Georgia) and from invaded Korean regions using 12 microsatellite loci. Our population genetic analyses showed a clear separation between the two host-associated D. oxycoccana populations from blueberry and cranberry. Using data from only the blueberry-associated D. oxycoccana populations, we identified five genetically isolated subgroups. An analysis of the approximate Bayesian computation suggests that the invasive D. oxycoccana population from Korea appears to have been introduced from an unsampled source population rather than directly from its native range. Our findings will allow for an easier identification of the source of D. oxycoccana into newly invaded regions, as well as to determine their association with blueberry and cranberry, which based on our results can be considered as two distinct species.

Brief Pollination Assessment of a Critically Endangered Food-Deceptive Orchid (Cypripedium guttatum) Using a Network Approach.

The translocation of orchids (Orchidaceae) cannot be successful if one is unaware of their effective pollinators and plant-pollinator interactions. Cypripedium guttatum is a generalized food-deceptive orchid, which is highly threatened in the Republic of Korea, thus, requiring immediate translocation actions. Although effective pollinators of the orchid are well known in China, little is known about the pollinators in the Republic of Korea and the ecological context in which the orchid can be successfully pollinated. To briefly assess the pollination of C. guttatum prior to translocation, we conducted a one-month survey of general pollination and the community-wide plant-pollinator network properties. Over 21 h of observation, we found that an effective pollinator of the orchid was the sweat bee Lasioglossum virideglaucum. The network was significantly specialized and modular, but not significantly nested. L. virideglaucum (pollinator) and Arabis gemmifera (plant) were determined to be keystone species, based on network metrics. A total of six network modules were identified and the flower colors of the plant species belonging to the C. guttatum module were purple, white, and yellow. After comparing the daily network patterns, we found that pollination of the orchid was accomplished when various flowering plant species bloom, and the nestedness value was high. This study revealed that high plant and pollinator richness could increase the chance that the deceptive orchid would be pollinated. Our study suggests that the network properties of this food-deceptive orchid community could provide useful insight into understanding the ecologically suitable habitat for the translocation of the highly threatened orchid species C. guttatum.

Population Genetics for Inferring Introduction Sources of the Oriental Fruit Fly, Bactrocera dorsalis: A Test for Quarantine Use in Korea.

To infer the introduction sources of the oriental fruit fly, Bactrocera dorsalis, we used a mitochondrial marker to reconstruct the haplotype network and 15 microsatellite loci to reveal genetic structure and relationships between the geographically or temporally different collections from Asia. We performed Approximate Bayesian computations to infer a global origin and a source of the quarantine collections found in Korea. As a result, the 40 populations were divided into three groups, of which genetic similarity is not related to the geographic vicinity. Korean samples had a similar genetic structure to Taiwan and Thailand ones. Our results suggest that the place of origin of the B. dorsalis specimens found in Korea's border quarantine is likely to be Taiwan or Thailand. As the global origin of B. dorsalis, we estimated that Taiwan and Thailand were most likely the global origins of Southeast Asian populations by testing hypothetical scenarios by the approximate Bayesian computation analyses. Our results will allow easier identification of the source region of the forthcoming invasion of quarantined B. dorsalis specimens.

Tracing the Origin of Korean Invasive Populations of the Spotted Lanternfly, Lycorma delicatula (Hemiptera: Fulgoridae).

Lycorma delicatula (White) suddenly arrived in Korea where it rapidly spread out in the central region of Korea and caused serious damage to grape vineyards. To trace the source region of its invasiveness, population genetic structures were compared between the native region, China, and the introduced regions, Korea and Japan. We examined 762 individuals from 38 different population collections using 15 microsatellite loci. Both principal coordinate and structure analyses displayed that the Chinese populations were separated into three subgroups which were located significantly far apart from each other. Among them, the Shanghai population was located closest to most Korean populations. Based on the genetic relationships and structures, it was revealed that the multiple introductions into Korea occurred at least three times. In addition, the Shanghai population was strongly estimated to be a source of initial invasive populations of Korea. In addition, analysis of the approximate Bayesian computation suggested simultaneous spread from two distant locations early in the invasion by artificial transportation of the host plants bearing egg masses. Our population genetics study can provide a precedent case with regards to identifying spreads by anthropogenic outcomes in other invasive regions.

Minimization of energy transduction confers resistance to phosphine in the rice weevil, Sitophilus oryzae.

Infestation of phosphine (PH3) resistant insects threatens global grain reserves. PH3 fumigation controls rice weevil (Sitophilus oryzae) but not highly resistant insect pests. Here, we investigated naturally occurring strains of S. oryzae that were moderately resistant (MR), strongly resistant (SR), or susceptible (wild-type; WT) to PH3 using global proteome analysis and mitochondrial DNA sequencing. Both PH3 resistant (PH3-R) strains exhibited higher susceptibility to ethyl formate-mediated inhibition of cytochrome c oxidase than the WT strain, whereas the disinfectant PH3 concentration time of the SR strain was much longer than that of the MR strain. Unlike the MR strain, which showed altered expression levels of genes encoding metabolic enzymes involved in catabolic pathways that minimize metabolic burden, the SR strain showed changes in the mitochondrial respiratory chain. Our results suggest that the acquisition of strong PH3 resistance necessitates the avoidance of oxidative phosphorylation through the accumulation of a few non-synonymous mutations in mitochondrial genes encoding complex I subunits as well as nuclear genes encoding dihydrolipoamide dehydrogenase, concomitant with metabolic reprogramming, a recognized hallmark of cancer metabolism. Taken together, our data suggest that reprogrammed metabolism represents a survival strategy of SR insect pests for the compensation of minimized energy transduction under anoxic conditions. Therefore, understanding the resistance mechanism of PH3-R strains will support the development of new strategies to control insect pests.

Developing molecular diagnostics for detection of red imported fire ants using two genes, Sinv11108 and Sinv11977.

Aggressive red imported fire ants (RIFAs) are expanding their habitat due to active international trade and global warming. To prevent infestation and settlement, RIFAs must be removed during the quarantine process. Because RIFAs are social insects and have different morphological characteristics depending on their castes, non-ant taxonomists have difficulty confirming RIFAs based on their morphological characteristics alone. The disadvantages of previously reported RIFA molecular diagnostics are that they require additional steps, such as restriction enzyme digestion followed by agarose gel electrophoresis separation or DNA sequence verification for polymerase chain reaction (PCR)-amplified products. To overcome these drawbacks, two RIFA-specific genes were selected and used to develop diverse PCR-based RIFA molecular diagnostic techniques. We found that RIFAs could be confirmed by conventional PCR targeting of two RIFA-specific genes followed by agarose electrophoresis separation. In addition, TaqMan probe real-time PCR methods had the advantage of confirming RIFAs immediately after the reactions were completed by observing fluorescence indexes. Finally, multiplex PCRs enhanced RIFA specificity and sensitivity. The new molecular diagnostic methods developed in this study had the advantages of reducing false positive and negative results together with high specificity and sensitivity for RIFAs.

DNA barcoding in quarantine inspection: a case study on quarantine insect monitoring for Lepidoptera obtained through quarantine inspection on foreign vessels.

We conducted quarantine insect species diversity monitoring using DNA barcoding with 517 lepidopteran samples that were obtained from quarantine inspections of foreign vessels entering Korea. For species delimitation and species identification of the analyzed samples, we applied a 2% cutoff rule. Consequently, 145 (368 samples) were considered taxonomically identified. Therefore the number of samples that were identified to the species level was relatively low, at approximately 71%. Thirty of 145 species were not known in Korea, three, i.e., Noctua pronuba (Noctuidae), Orthosia hibisci (Noctuidae), and Pieris brassicae (Pieridae), were checked as 'Regulated pests' in Korea.

Genetic structure and demographic history of Lymantria dispar (Linnaeus, 1758) (Lepidoptera: Erebidae) in its area of origin and adjacent areas.

We analyzed the population genetic structure and demographic history of 20 Lymantria dispar populations from Far East Asia using microsatellite loci and mitochondrial genes. In the microsatellite analysis, the genetic distances based on pairwise FST values ranged from 0.0087 to 0.1171. A NeighborNet network based on pairwise FST genetic distances showed that the 20 regional populations were divided into five groups. Bayesian clustering analysis (K = 3) demonstrated the same groupings. The populations in the Korean Peninsula and adjacent regions, in particular, showed a mixed genetic pattern. In the mitochondrial genetic analysis based on 98 haplotypes, the median-joining network exhibited a star shape that was focused on three high-frequency haplotypes (Haplotype 1: central Korea and adjacent regions, Group 1; Haplotype 37: southern Korea, Group 2; and Haplotype 90: Hokkaido area, Group 3) connected by low-frequency haplotypes. The mismatch distribution dividing the three groups was unimodal. In the neutral test, Tajima's D and Fu's FS tests were negative. We can thus infer that the Far East Asian populations of L. dispar underwent a sudden population expansion. Based on the age expansion parameter, the expansion time was inferred to be approximately 53,652 years before present (ybp) for Group 1, approximately 65,043 ybp for Group 2, and approximately 76,086 ybp for Group 3. We propose that the mixed genetic pattern of the inland populations of Far East Asia is due to these expansions and that the inland populations of the region should be treated as valid subspecies that are distinguishable from other subspecies by genetic traits.

Bees of the Lasioglossum series (Hymenoptera: Halictidae) in South Korea, with an illustrated keys to species.

The South Korean fauna of the bee genus Lasioglossum Curtis (Halictidae: Halictini) belonging to the Lasioglossum series (i.e., those with the second submarginal crossvein strong) are reviewed. A total of 12 species are recognized for the country. Lasioglossum circularum Fan & Ebmer is recorded for the first time from the Korean Peninsula and the following species are newly recorded from South Korea: L. denticolle (Morawitz), L. formosae (Strand), L. kansuense (Blüthgen), L. occidens (Smith), L. sutshanicum Pesenko, and L. upinense (Morawitz). Bionomical data as well as illustrated keys to females and males of South Korean species are provided. DNA sequences including a part of barcode region are given for L. kansuense and L. occidens.

DNA Barcoding of the Korean Lymantria Hübner, 1819 (Lepidoptera: Erebidae: Lymantriinae) for Quarantine Inspection.

DNA barcoding and morphological analyses of Korean Lymantria (Erebidae, Lepidoptera) were conducted for quarantine inspection. In DNA barcoding, Lymantria dispar identified through quarantine inspection was distinguished as three species, L. dispar asiatica, L. albescens, and L. xylina. Lymantria monacha, which is known as a single species in Korea, is revealed as containing three species, L. monacha, L. minomonis, and L. sugii. At the subspecies level, L. dispar dispar formed a single cluster, whereas L. d. asiatica and L. d. japonica formed a cluster containing both subspecies. In morphological re-examination on DNA barcoding results, L. dispar was distinguished from L. albescens by wing pattern, and from L. xylina by papillae anale. L. monacha and the related species were hard to be distinct from each other by using wing pattern, but it was easily distinct through comparison of genitalia. Therefore, DNA barcoding led to accurate identification in species level, but in subspecies level, only a taxon showing geographically far distance was discriminated from the others. These results may provide a taxonomic outline of the Korean Lymantria fauna and may be used as an identification reference for Lymantria species during quarantine inspection.