Genetic differentiation and species diversification of the Adoxophyes orana complex (Lepidoptera: Tortricidae) in East Asia.

The Adoxophyes tea tortrix (Lepidoptera: Tortricidae) is a group of leaf rollers that cause enormous economic losses on tea and apple crops. In East Asia, taxonomic ambiguity of the Adoxophyes orana complex (AOC), which consists of A. orana, A. dubia, A. honmai, and A. paraorana, has persisted for decades because of vague diagnostic characters. In this study, differences in the AOC were examined to improve species identification, determine genetic variations, and develop control strategies. Analyses revealed that A. orana comprised 2 lineages, a widely distributed Palearctic lineage and an East Asian lineage that was nested with other Asian species. Genetic divergence of >3% is proposed to confirm the AOC species that would benefit subsequent taxonomic revision. The monophyletic Taiwanese A. sp. with 2.8-4% from other AOC species appeared to suggest it as an independent taxon, and low interspecific divergence between A. honmai and A. dubia of 0.3% indicated possibility of recent divergence or intraspecific variations. Our result further suggested that the Z9-14:Ac ratio of semiochemicals could be a reference for the reblending of pheromone attractants in Taiwanese tea plantations. Moreover, the AOC species appeared to have a tendency of specific geographic distributions, with A. dubia and A. honmai in Japan and China, A. paraorana in Korea, and A. sp. in Taiwan. Maintaining the unique genetic composition of Adoxophyes species in each geographic region and preventing the possible invasions into those AOC-free countries through the transportation of host plants are essential in managing the AOC in East Asia.

Genetic Profile of the Parasitic Varroan Mite Varroa destructor (Arachnida: Mesostigmata: Varroidae) in Taiwan: a New Taiwanese Haplotype Intermediate Between the Highly Virulent Russian and Less Virulent Japanese Types Identified in the Honey Bee Host Apis cerana.

The Modern beekeeping industry is being challenged by the varroan mite and its transmitted pathogens. Various types of Varroa destructor exhibit different levels of virulence toward honey bees, but only the Japanese (J) and Russian (R) types were found to infect Apis mellifera. Type R was more highly virulent against A. mellifera in comparison with type J. Examining the genetic profile of Varroa species is therefore of crucial importance in apiary management. In this study, maternally inherited mitochondrial cytochrome oxidase I (COI) and bisexual nuclear internal transcribed spacer (ITS) sequences of V. destructor individuals from Taiwan were determined. All 168 COI sequences observed in populations obtained from A. mellifera were identical and belonged to type J, with one base difference to that of populations collected from A. cerana; the new type is named 'T type' (Taiwan type). ITS sequences of V. destructor and its sister species V. jacobsoni were identical. A network analysis based on 611 COI sequences compiled from references indicated the presence of 27 haplotypes in V. destructor. Epidemic history and relationship analyses of V. destructor showed that the basal haplotypes were those from A. cerana and many R-extending haplotypes infesting A. mellifera involving amino acid substitutions. Calibration dating based on COI analysis revealed that V. destructor differentiated from its sibling lineage (occurring in Sri Lanka) prior to 1.3 million years ago (Mya). The ancestral haplotype retention and drift in V. destructor that occurred locally during 0.10-0.64 Mya might be relevant to its host A. cerana, which had been isolated geologically. The highly virulent type R was spreading quickly and could gradually outcompete the common and less virulent type J. Type T, being intermediate between types R and J, ought to be studied to better understand the pathogenic mechanism of V. destructor in A. mellifera. Moreover, for areas where type R does not occur, such as Taiwan, quarantine requirements are crucial for reducing invasion risks.

Composition and Dynamics of Hexapod Communities on Yushan Bamboo (Yushania niitakayamensis) in the Subtropical Montane Areas of Taiwan.

Plant communities that colonize high-elevation zones generally have short growing seasons which lead to specialized adaptations in such zones. In montane areas of Taiwan, Yushan bamboo (YB, Yushania niitakayamensis) is dominant at elevations ranging from 2500 to 3300 m and grows in a grassland-like open habitat. In this study, hexapods were collected from YB bimonthly between 2009 and 2012 by using a sweeping net. The composition of and several bioindices for the hexapods were determined, and multivariate analyses were conducted to explore the dynamics and seasonal distribution of the hexapods. A total of 32,000 individuals belonging to 11 orders and 113 families were collected, with adult individuals being collected more frequently in warmer seasons (from June to October). Of the sampled individuals, 90% belonged to the orders Collembola (42%), Hemiptera (35%), and Hymenoptera (13%). The number of individuals belonging to Hemiptera were stable in all seasons, and the number of hymenopteran wasps was influenced by temperature and exhibited a stable dynamic pattern. The number of individuals belonging to Collembola fluctuated dramatically. The multivariate analyses revealed that the collected hexapods could be divided into two major family groups according to survey season (i.e., summer and winter groups). Several families were collected only in summer, but a few were collected only in winter. Eigenvalues obtained from a principal component analysis revealed that the families Chironomidae, Delphacidae, Entomobryidae, Hypogastruridae, Sminthuridae, and Thripidae (all dominant) were the major contributors to the winter group. These families were abundant all year, although some were more abundant during winter. The three dominant orders Collembola, Hemiptera, and Hymenoptera, each of which has a distinct community structure and dynamic pattern, may have their own adaptive mechanisms in the subtropical regions of Taiwan. Hemiptera individuals, which feed on YB, were most abundant in the adult stage in summer and in the nymphal stage in winter. The abundance of parasitic hymenopteran wasps, which had stable dynamic patterns, was associated with that of their host insects and temperature. The drastic fluctuations in the abundance of Collembola may have been caused by abiotic factors, such as precipitation and microhabitat factors. The early onset of spring and the late onset of winter might also affect the dynamics of the studied hexapods.

Global population genetic structure and lineage differentiation of the stable fly, Stomoxys calcitrans.

The bloodsucking fly, Stomoxys calcitrans (Diptera: Muscidae), is a cosmopolitan pest that transmits potential pathogens mechanically. We conduct phylogeographic analyses of S. calcitrans to resolve its global population genetic structure for establishing baseline of molecular studies. Results from mitochondrial gene suggested that the major divergence of S. calcitrans predominantly occurred 0.32-0.47 million years ago (Mya) and the subsequent diversifications took place during 0.13-0.27 Mya. The Ethiopian region was deduced as the most likely origin of S. calcitrans and the Nearctic lineages were considered to have originated from Oriental or Palaearctic regions. Our results further revealed that each biogeographic region of S. calcitrans likely maintains its genetic specialty, and yet, those non-monophyletic relationships were possibly caused by ancestral retention, dispersal with mammals, long-distance migration, and the international livestock industries. Moreover, the three highly diverged Ethiopian lineages may be putative cryptic species that require clarification of their veterinary importance. Unravelling the genetic structure of stable fly and preventing gene flow among biogeographic regions through anthropogenic activities are thus pivotal in livestock industry administration, particularly genetic exchange among differentiated lineages that might lead to the consequence of ecological trait alterations.

Molecular epidemiology of Akabane virus in Taiwan.

BACKGROUND: Akabane virus (AKAV) is a teratogenic and neuropathogenic arbovirus that infects livestock and wild animals. AKAVs are endemic arboviruses from dairy farms in Taiwan in 1989, and the first sequence was detected in cattle with nonsuppurative encephalitis in 1992. OBJECTIVES: This study aims to understand the epidemiological relationships of the akabane viruses between Taiwan and nearby places. METHODS: In this study, 17 specimens were identified or isolated from vector insects, and ruminant fetuses collected from 1992 to 2015 were sequenced and analysed. RESULTS: Sequence analyses revealed all Taiwanese AKAVs belonged to genogroup Ia but diverged into two clusters in the phylogenetic trees, implying that at least two invasive events of AKAV may have occurred in Taiwan. CONCLUSIONS: The two clusters of AKAVs could still be identified in Taiwan in 2015, and a reassortment event was observed, indicating that the two clusters of AKAVs are already endemic in Taiwan.

Differentiation patterns of emperor moths (Lepidoptera: Saturniidae: Saturniinae) of a continental island: divergent evolutionary history driven by Pleistocene glaciations.

Background: On the basis of molecular dating, Pleistocene glaciations have been proposed as the major driving force of biota speciation in the Palearctic and the pre-Quaternary origin of Amazonian taxa. However, the major driving factors in East Asia remain unclear. All 16 saturniine species inhabiting Taiwan with congeners of populations, subspecies, or species in East Asia constitute research objects for addressing the mode of speciation because of the repeated formation and disappearance of a landbridge from the Asian mainland to Taiwan during glacial cycles. Methods: The genetic divergences of mitochondrial cytochrome c oxidase subunit I (COI) and 16S rDNA and the nuclear 28S rDNA of the saturniine species from Taiwan and the Asian mainland were assessed to determine the monophyly of each genus and species of Saturniinae. Moreover, 519 saturniine COI sequences of 114 taxa from adjacent East and Southeast Asian populations and closely related species were retrieved from GenBank and analyzed. The differentiation timing and possible origination of the insular saturniines were elucidated based on phylogenetic relationships, haplotype networks, and lineage calibrations. Results: Approximately 90% of intraspecific COI divergence was <2%; all divergences exceeding 2% originated from comparisons between allopatric populations or subspecies. Relationship analyses revealed that multiple introductions likely occurred in insular saturniines and that some East Asian saturniines were paraphyletic as deduced by analyzing endemic insular species. Calibration dating revealed that Taiwanese endemic saturniines split from sibling Asian species 0.2-2.7 million years ago (Mya), whereas subspecific-level and population-level splitting events occurred 0.1-1.7 Mya and 0.2-1.2 Mya, respectively. Moreover, phylogenetic patterns combined with geographical distributions revealed that hill-distributed Taiwanese saturniines are closely related to those from southern China and Southeast Asia, whereas saturniines inhabiting altitudes higher than 1,500 m in Taiwan have siblings distributed in temperate Northeast Asia. Discussion: The Global DNA Barcoding Initiative was successfully applied to study the population genetic structure in species. Most Formosan saturniines are distinct and monophyletic, reflecting the vicariant barrier of the Taiwan Strait; Pleistocene glacial cycles provided opportunities for insular saturniines to experience repeated isolation from and secondary contact with the continental mainland. Each insular saturniine may have evolved with a unique differentiation timing pattern that possibly emerged in the Early, Middle, or Late Pleistocene with these patterns differing from the consistent pattern that occurred in the temperate Palearctic and tropical Amazonian regions. Moreover, multiple migrations or artificial genetic admixtures may have also occurred, as suggested by the coexistence of two divergent lineages in a few Taiwanese saturniines.

Drugstore Beetle Damage Pattern of Glue Paste-Lined Oil Paintings.

A glue paste lining is a traditional conservation treatment used to reinforce the aged canvas of oil paintings. Several insect infestation cases concerning glue paste-lined oil paintings have been reported around the world, particularly in European countries. In 2008, Chimei Museum, a private museum located in Tainan, Taiwan, was affected by a severe beetle infestation of oil paintings. To confirm the infesting insects and to quantify the damage pattern for further development of control and monitoring methods, three severely damaged paintings were examined along with the restoration procedure. A total of four insect species were collected. The drugstore beetle, Stegobium paniceum (L.) (Coleopter: Ptinidae), was the primary pest found in all three paintings investigated and was identified based on morphological and genetic data. Thrips and booklice were considered secondary pests, and a Pteromalid wasp was speculated to have a parasitic relationship with the drugstore beetle. Drugstore beetle larvae mainly bored into the glue paste layer and original canvas and required only 5.94 ± 1.38 mm3 of feed to grow from egg to pupa. Their bores were not evenly distributed, and most of them were found in the shaded area covered by the stretcher and outer frame. The body length of drugstore beetles varied, ranging from 1.67 to 2.75 mm, which may explain the various sizes of exit holes on gummed tape surrounding the frame. Detailed and quantified information on drugstore beetle's pattern of damage provided in this study will be beneficial for further developing conservation practices and inspection methods.

Molecular Characteristics of Fat Body Protein 1 in the Oriental Fruit Fly, Bactrocera dorsalis.

Bactrocera dorsails fat body protein 1 (Bdfbp1) cDNA was cloned (GenBank accession no. MT514270), and the complete 3,749-bp cDNA encoded a 1,152-amino acid protein. The phylogenetic relationship of dipteran fbp1s was analyzed. The sequence XP_028900815 from the insect genome project for Zeugodacus cucurbitae (LOC105219342) was proposed that two fbp1 genes were present in the sequence. The developmental transcriptional expression profiles were determined. In the larval stages, Bdfbp1 mRNA had significantly higher expression in the late third instar larvae compared with first, second, and early third instar larvae. In the pupal stages, the highest expression of Bdfbp1 mRNA was found in the newly pupated pupae and then decreased with age. In the fat body of female adults, Bdfbp1 was highly expressed in newly emerged samples and decreased rapidly over the following three days. In the fat body of male adults, Bdfbp1 was highly expressed in newly eclosed samples. RNAi treatment decreased the expression level of Bdfbp1 without statistical difference. However, RNAi treatment significantly decreased the rate of eclosion. These results suggest that Bdfbp1 may function as a storage protein and be associated with adult eclosion.

Ancestral Haplotype Retention and Population Expansion Determine the Complicated Population Genetic Structure of the Hilly Lineage of Neolucanus swinhoei Complex (Coleoptera, Lucanidae) on the Subtropical Taiwan Island.

The present study demonstrates that the complicated genetic structure of the hilly lineage of the Neolucanus swinhoei complex was driven by its biological features and habitat requirements as well as hindrance by the CMR during periodical Pleistocene glaciations. The results revealed a tendency of geographical differentiation and major and sub- lineage divergences before and after the Riss glaciation, followed by stable population growth during Würm glaciation. At least four refugia were inferred for N. swinhoei during the Riss-Würm glaciations. The ancestral haplotype retention in the cytochrome oxidase subunit I (COI) gene and compensated substitution in 16S rRNA gene is a possible evolutionary scenario resulting in the inconsistent evolution pattern between COI and 16S rRNA gene coupled with the long-distance dispersal of N. swinhoei. Although the CMR did hinder the dispersal of N. swinhoei, its ancestors may have dispersed to eastern Taiwan through the northern and southern low mountains of the CMR before the Riss glaciation. Our finding suggests that the population growth in the Würm glaciation led a dispersal back to western Taiwan, which is contrast to the more common dispersal scenario from western Taiwan to eastern populations proposed in other studies.

Influence of gap distance between bone and plate on structural stiffness and parallel interfragmental movement in far-cortical locking technique - a biomechanical study.

Sufficient interfragmental movement is the key to successful fracture healing in the theory of secondary bone healing. The far-cortical locking technique enables both stiffness reduction and parallel motion for ideal callus formation and fracture healing, but the influence of plate-bone gap on the performance of far-cortical locking technique remains unclear. The current study conducted a series of finite element analyses with mechanical validation to clarify this issue. Plate-bone gaps were assigned by 1, 2, 3, and 4 mm in a simulated mid-shaft fracture model fixed with locking plate and six semi-rigid locking screws. Axial compressive load to 500 N was applied to the fixation structure to evaluate the structural stiffness, pattern of interfragmental movement (parallel motion), and stresses on the screws. Results revealed the increased plate-bone gaps reduced the structural in order (315.3, 288.8, 264.9, and 243.4 N/mm). Tilting angles for determining parallel interfragmental movement (1.58°-1.85°) and stresses on semi-rigid screws for evaluating implant safety were not severely altered. Greater stresses were found on the screws adjacent to the fracture site in all simulated models. The current study suggested that 1 mm gap between the locking plate and the bone shall be ideal in view of parallel motion achieved balanced callus formation in far-cortical locking technique. Issue of reducing structural stiffness with limited plate-bone gap distance should be further investigated.

Rapid identification of the invasive fall armyworm Spodoptera frugiperda (Lepidoptera, Noctuidae) using species-specific primers in multiplex PCR.

The fall armyworm (FAW), Spodoptera frugiperda (Smith), is a major pest native to the Americas. A recent invasion of FAWs from Africa eastward to South Asia, the Indochina Peninsula, and mainland China has received much attention due to the considerable economic losses in agriculture. FAWs can rapidly colonise a new area, likely due to the wide range of host plants, good flying capability, and high egg production. Therefore, a convenient, quick, and accurate tool for FAW identification is urgently required to establish a FAW invasion management strategy. In this study, FAW-specific primers were designed to recognise FAWs on the basis of internal transcribed spacer 1 (ITS1). The results revealed the accurate FAW recognition of the three congeneric species and eight common corn lepidopteran pests, especially at their larval stage. Furthermore, species-specific primers have confirmed their efficacy by using 69 FAW specimens from Taiwan, Thailand, and the United States, with a 96% success rate, excluding 3 decayed specimens. By using the simple, reliable, and convenient FAW-specific primers, a pest management programme can be developed not only to reduce sequencing costs and experimental time from 2 days to 4 h, but eradicate the FAW as soon as it enters a new area.

Invasive and Quarantine Risks of Cacopsylla chinensis (Hemiptera: Psyllidae) in East Asia: Hybridization or Gene Flow Between Differentiated Lineages.

Pear psyllids are major pests and the causal agents of pear decline disease in orchards. In the past two decades, their outbreaks have raised issues pertaining to invasions and taxonomic identification of the dimorphic Cacopsylla chinensis (Yang and Li) in East Asia. The present study elucidated, as an aid to quarantine management, the invasive origins, differentiation history, and putative gene flow and hybridization between C. chinensis and its sibling species Cacopsylla jukyungi (Kwon). Analyses revealed that the ancestors of C. jukyungi might have diverged from C. chinensis approximately 3.5 million yr ago (Mya) and that differentiation between C. chinensis lineages I and II probably occurred 1.5 Mya. The known overlapping distribution of C. chinensis and C. jukyungi in northeastern China and the two C. chinensis lineages in the Bohai Rim region and Taiwan could be attributed to recent population expansion after the Last Glacial Maximum and/or anthropogenic activities. Analyses of the nuclear gene demonstrated that frequent gene flow between the two C. chinensis lineages and the paraphyletic relationship between C. chinensis and C. jukyungi might be caused by incomplete lineage sorting or hybridization events. On the basis of the current distribution, it is evident that C. jukyungi is not present in middle-southern China, whereas C. chinensis is not distributed in Japan and Korea. Preventing new invasions of Cacopsylla psyllids among geographic regions through the transportation of pear scions is thus pivotal in East Asia, particularly for the possible genetic exchanges among differentiated lineages after secondary invasion events.

Cephalomalthinus simplicicornis (Wittmer, 1993) rev. stat. et comb. n.: a resurrected soldier beetle (Coleoptera, Cantharidae) from Taiwan based on morphological and molecular data.

The genus Cephalomalthinus Pic, 1921 is a diverse group of soldier beetles known from East Asia, with modifications on the male antennae, which are traditionally used for species delimitation. However, antennae are uniform in some Cephalomalthinus groups, thus making species identification difficult. In this study, both morphological and molecular data are applied to solve the generic classification and species status of Micropodabrus simplicicornis Wittmer, 1993, which was recently synonymized with Cephalomalthinus formosanus (Pic, 1910). We analyzed holotypes, multiple specimens and DNA barcodes to re-examine the validity of this synonymy. The results suggest that M. simplicicornis should be resurrected from the synonymy with C. formosanus, and a new combination, Cephalomalthinus simplicicornis (Wittmer, 1993) rev. stat. et comb. n., is proposed accordingly. The results highlight the value of multisource taxonomy and the discordance between molecular and morphological data.

Identification of Subterranean Termite Genus, Reticulitermes (Blattodea: Rhinotermitidae) in Taiwan.

Species identification in the genus Reticulitermes is often difficult because of ambiguous morphological characters. Challenges in Reticulitermes spp. recognition have also been encountered in East Asia, including Taiwan. Because of unknown Reticulitermes taxa in Taiwan and the possible origin of alien Reticulitermes kanmonensis Takematsu in Japan and Korea, reexamining Reticulitermes fauna in Taiwan is imperative. To clarify the Reticulitermes fauna in Taiwan, this study applied two mitochondrial genes (cytochrome oxidase subunit II [COII] and 16S rDNA) and morphological characters for species delimitation. Reticulitermes specimens collected from 63 localities across the main and adjacent islands of Taiwan were analyzed. Phylogenetic analyses, morphological comparisons, and ecological traits suggested the existence of three species in Taiwan: Reticulitermes flaviceps (Oshima), R. kanmonensis, and Reticulitermes leptomandibularis Hsia and Fan. Altitudinal distributions among the three Reticulitermes termites tended to differ: R. flaviceps adapted to low hills, but R. kanmonensis and R. leptomandibularis occurred in medium mountainous areas. The combined data, including haplotype diversities and distribution range, suggest that 1) R. flaviceps is an endemic species and only found in Taiwan; 2) R. kanmonensis and R. leptomandibularis are both native species in Taiwan and China; 3) Japanese R. kanmonensis populations originated from southern China and/or Taiwan and that Korean populations were possibly introduced from Japan.

Nicotinic acetylcholine receptor subunit α6 associated with spinosad resistance in Rhyzopertha dominica (Coleoptera: Bostrichidae).

The lesser grain borer, Rhyzopertha dominica, which is a primary pest of stored products, breaks up whole grains and makes them susceptible to secondary infestation by other pests. Insecticide application is the main control measure against this borer. A resistant strain of R. dominica against the insecticide, spinosad, was selected in the laboratory. The full-length cDNA of the target site of spinosad, nicotinic acetylcholine receptor subunit α6, from R. dominica (Rdα6) was cloned and analyzed using reverse transcription PCR and rapid amplification of cDNA ends. The complete 2133-bp cDNA contains the open reading frame of 1497 bp encoding a 498-amino-acid protein. There are four predicted transmembrane (TM) regions, and six extracellular ligand-binding sites at the N-terminus, upstream from the first TM in Rdα6. Three mutations have been found in the resistant strain compared with the susceptible one: (1) a 181-bp fragment truncated at the N-terminus, resulting in the appearance of a premature stop codon, (2) one missing bp at the position 997, causing a frame-shift mutation, and (3) an 87-bp fragment truncated in the TM2 region. In addition, real-time quantitative PCR was applied to detect the transcriptional expression of Rdα6 in both the susceptible and resistant strains. The results indicated that the expression of Rdα6 was significantly lower in then resistant strain than in susceptible one. In conclusion, mutation of Rdα6 may cause R. dominica resistant to spinosad due to target site insensitivity.

Subspecific Differentiation Events of Montane Stag Beetles (Coleoptera, Lucanidae) Endemic to Formosa Island.

Taxonomic debates have been carrying on for decades over Formosan stag beetles, which consist of a high proportion of endemic species and subspecies featuring morphological variations associated with local adaptation. With the influence of periodical Pleistocene glaciations and the presence of several mountain ranges, the genetic differentiation and taxonomic recognition, within this medium-size island, of two endemic subspecies for each of four montane stag beetles, i.e. Lucanus ogakii, L. kanoi, Prismognathus davidis, and Neolucanus doro, has been an appealing issue. Based on monophyletic lineages and population structure, possible divergent scenarios have been proposed to clarify the subspecific status for each of the above mentioned stag beetles. Phylogenetic inferences based on COI+16S rDNA+28S rDNA of 240 Formosan lucanids have confirmed most species are monophyletic groups; and the intraspecific (<2%) and interspecific (>2%) genetic distances of the two mitochondrial genes could be applied concordantly for taxonomic identification. On account of Bayesian-based species delimitation, geographic distribution, population structure, and sequence divergences, the subspecific status for L. ogakii, L. kanoi, and Pri. davidis are congruent with their geographic distribution in this island; and the calibration time based on the mitochondrial genes shows the subspecific split events occurred 0.7-1 million years ago. In addition, a more complicated scenario, i.e. genetic differentiation including introgression/hybridization events, might have occurred among L. ogakii, L. kanoi, and L. maculifemoratus. The geological effects of mountain hindrance accompanied by periodical glaciations could have been vital in leading to the geographical subspecific differentiation of these montane stag beetles.

Revision of Mandarella Duvivier from Taiwan, with a new species, new synonymies and identities of highly variable species (Insecta, Chrysomelidae, Galerucinae, Alticini).

Taiwanese species of Mandarella Duvivier are compared on the basis of morphological and molecular evidence. Only three of eleven morphospecies are considered to be valid. Mandarella uenoi (Kimoto, 1969) is transferred from the genus Luperus Geoffroy. Stenoluperus taiwanus Kimoto, 1991 and Stenoluperus kimotoi Döberl, 2001 are synonymized with Mandarella uenoi. Taiwanese records of Stenoluperus tibialis Chen, 1942, Stenoluperus nipponensis Laboissière, 1913, and Stenoluperus potanini (Weise, 1889) were based on misidentifications and represent Mandarella uenoi. The Taiwanese population previously erroneously identified as Stenoluperus pallipes Gressitt and Kimoto, 1963 is here described as a new species, Mandarella tsoui sp. n., Stenoluperus esakii Kimoto, 1969, Stenoluperus matsumurai Takizawa, 1978, and Mandarella taiwanensis Medvedev, 2012 are synonymized with Mandarella flaviventris (Chen, 1942).

A NONOVARY-SPECIFIC VITELLOGENIN RECEPTOR FROM THE ORIENTAL FRUIT FLY, Bactrocera dorsalis (HENDEL).

The yolk protein precursor, vitellogenin (Vg), is absorbed into growing oocytes via receptor-mediated endocytosis for embryonic development. In this study, a Vg receptor (VgR) cDNA of the oriental fruit fly (Bactrocera dorsalis Hendel) was cloned via RT-PCR and RACE (GenBank accession no. KR535603) and its expression analyzed. The BdVgR cDNA has a length of 6,585 bp encoding 1,923 amino acids. It has a conserved motif arrangement with other insect VgRs, and showed high identity to the B. cucurbitae VgR (91.4%). The expression of BdVgR mRNA and proteins was shown in both ovary and fat body. This is the first report on a nonovary-specific VgR from a nonsocial insect. In ovary, the expression of BdVgR mRNA and proteins was inconsistent, with the transcription, but not protein, level high on D0. In fat body, the expression levels of BdVgR mRNA and proteins were high on days 5 and 6. The function of BdVgR in the fat body is not clear. However, it may be involved in reuptake of yolk proteins from the hemolymph as an amino acid reservoir or as autocrine regulation of yolk protein expression.

Taxonomy and biology of a new ambrosia gall midge Daphnephila urnicola sp. nov. (Diptera: Cecidomyiidae) inducing urn-shaped leaf galls on two species of Machilus (Lauraceae) in Taiwan.

Recent field surveys show that galls induced by Daphnephila spp. (Cecidomyiidae) on Machilus spp. (Lauraceae) are common in Taiwan, yet only five species, four leaf-gall inducers and one stem-gall inducer on M. thunbergii, have been named in the past. Here we describe a new species, Daphnephila urnicola sp. nov. Chiang, Yang & Tokuda, inducing urn-shaped galls on leaves of both M. zuihoensis and M. mushaensis. Comparisons of D. urnicola populations on M. zuihoensis and on M. mushaensis, indicate that they belong to one species, a result supported by gall midge morphology, life-history traits, gall shape and structure, the developmental process of gall tissues, fungal associations, and DNA-sequencing data. Size and structure of the gall operculum was found to differ between M. zuihoensis and M. mushahaensis.

cDNA CLONING AND TRANSCRIPTIONAL REGULATION OF THE CECROPIN AND ATTACIN FROM THE ORIENTAL FRUIT FLY, Bactrocera dorsalis (DIPTERA: TEPHRITIDAE).

We described the cDNA cloning of two antimicrobial peptides (AMPs), cecropin (BdCec), and attacin C (BdAttC), from the oriental fruit fly, Bactrocera dorsalis (Hendel), a serious insect pest of fruit trees. Using rapid amplification of cDNA ends, fragments encompassing the entire open reading frames of BdCec and BdAttC were cloned and sequenced. The complete 425 bp cDNA of BdCec encodes a protein of 64 amino acids with a predicted molecular weight of 6.84 kDa. The 931 bp cDNA of BdAttC encodes a protein of 239 residues with a predicted molecular weight of 24.97 kDa. Real-time quantitative RT-PCR demonstrated that the developmental transcription profiles of BdCec and BdAttC were similar in each larvae, pupae, and adults. The constitutive expression levels of both AMPs were high in the first-instar and late third-instar larvae, suggesting that their antimicrobial activity is active in the newly hatched larvae and just before pupation. The basal expression levels were not significant different in adult fat bodies. The expression of BdCec and BdAttC was upregulated after bacterial challenge in adult fat bodies. The ratio of inducible expression to constitutive expression was lower in males compared to females.