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Bombyx mori is a key insect in the sericulture industry and one of the very important economic animals that are responsible for not only the livelihood of many farmers internationally but also expended biomedical use. The National Institute of Agricultural Sciences of the Rural Development Administration of Korea (NIAS, RDA, Korea) has been collecting silkworm resources with various phenotypic traits from the 1960s and established breeding lines for using them as genetic resources. And these breeding line strains have been used to develop suitable F1 hybrid strains for specific use. In this study, we report the whole-genome sequences of 37 breeding line B. mori strains established over the past 60 years, along with the description of their phenotypic characteristics with photos of developmental stages. In addition, we report the example phenotypic characteristics of the F1-hybrid strain using these breeding line strains. We hope this data will be used as valuable resources to the related research community for studying B. mori and similar other insects. Measurement(s) Larval period • Cocoon yield • Single cocoon weight • Egg color • Cocoon shape • Number of cocoons per liter • Voltinism • Moltinism • Cocoon color• Larva pattern Technology Type(s) Clock Device • scale • Eye • amount Factor Type(s) Artificial feed • Target Season Sample Characteristic - Organism Bombyx mori Sample Characteristic - Environment house Sample Characteristic - Location South Korea [ABSTRACT FROM AUTHOR]

The black-veined white, Aporia crataegi (Lepidoptera: Pieridae), is distributed from northwest Africa throughout most of Europe and temperate Asia to far eastern Russia and Japan and is presumed to be extinct in South Korea, with only a few dried specimens remaining. A common conservation practice for such near-extinct species is the launch of an introduction program, but prior genetic analysis of donor and donee populations may be essential for the success of such a program. In the present study, we investigated the genetic diversity and the relationships of A. crataegi populations from South Korea and nearby Asian countries, using 11 newly developed A. crataegi-specific microsatellite markers and mitochondrial DNA (mtDNA) sequences from two genes (a total of 1317 bp from COI and CytB). mtDNA-based analyses, using a total of 115 individuals collected from China, Russia, Mongolia, and Japan and museum specimens from South Korea consistently showed long-held, close genetic relationships between A. crataegi populations from Mongolia, China, and Russia and those from South Korea, with the highest diversity detected in the Mongolian population. Principal coordinate analyses and STRUCTURE analyses, based on microsatellite markers, also suggested that A. crataegi from South Korea had a similar gene pool to populations from China, Mongolia, and Russia, with a similar degree of genetic diversity among these populations. Taken together, the mtDNA and microsatellite data suggest that the northern populations, particularly those from Mongolia, can be considered the most suitable population as an immediate donor for the near-extinct South Korean population of A. crataegi. [ABSTRACT FROM AUTHOR]

Sacbrood virus (SBV) caused significant colony collapse in Korean Apis cerana. Therefore, breeding of resistant bees to counter this viral disease is urgently needed. Considering that hygienic behavior in honey bees confers colony-level resistance against brood diseases, we utilized this trait for selecting A. cerana colonies. In addition, the brood survival rate was evaluated after colonies were SBV-inoculated. Over four selective generations, dead brood removal and brood survivorship in selected colonies were higher than those in the controls, which were not selectively bred for those traits (P < 0.01, 99.3 vs. 89.9% for removal of pin-killed pupae; P < 0.01, 99.0 vs. 63.9% for removal of SBV-killed larvae; and P < 0.01, 70.0 vs. 9.2% for brood survivorship). Following SBV inoculation, selected colonies showed an increase in the number of surviving pupae and adults, whereas control colonies collapsed mostly. Our results confirm the feasibility of selecting SBV-resistant A. cerana. [ABSTRACT FROM AUTHOR]

The tiny dragonfly, Nannophya pygmaea (Odonata: Libellulidae), has been listed as an endangered insect in South Korea. We sequenced the complete 15,112-bp-long mitochondrial genome (mitogenome) of the species. The genome included a typical set of genes (13 protein-coding genes [PCGs], two rRNA genes, and 22 tRNA genes) and one non-coding region with an arrangement identical to that found in most insects. Among the 13 PCGs, only ND1 started with the atypical TTG. The 441-bp-long A+T-rich region possessed the highest A/T content (84.6%) in the genome. N. pygmaea was placed as the sister to Orthetrum species belonging to Libellulidae. Unlike conventional phylogenetic results, the suborders Anisozygoptera and Zygoptera formed a strong sister group in both Bayesian inference (BI) and maximum likelihood (ML) methods (BI, BPP = 1 and ML, 88-94%), justifying the use of different types of molecular markers for phylogenetic analysis. [ABSTRACT FROM AUTHOR]

The tiny dragonfly, Nannophya pygmaea (Odonata: Libellulidae), is listed as a second-degree endangered wild animal in South Korea. The application of molecular markers to assess genetic diversity and population relationships can provide information necessary to establish an effective conservation strategy. In this study, we developed 12 microsatellite markers specific to N. pygmaea using the NextSeq 500 platform. Forty individuals of N. pygmaea collected from three currently known localities in South Korea were genotyped to validate these markers and to preliminarily assess population genetic characteristics. The observed number of alleles, observed heterozygosity, and expected heterozygosity at a locus ranged from 2 to 9, 0.421-1.0, and 0.508-0.766 in a population with the largest sample size (20 individuals), respectively, thereby validating the suitability of the markers for population analysis. Five of 12 loci showed significant deviation from the Hardy-Weinberg equilibrium in the population. Our preliminary data indicate an absence of inbreeding in all populations and an absence of obvious genetic difference. The microsatellite markers developed in this study will be useful for studying the population genetics of N. pygmaea collected from other regions worldwide, including additional sites in South Korea. [ABSTRACT FROM AUTHOR]

We sequenced the entire mitochondrial genome (mitogenome) of two gelechioids, Mesophleps albilinella and Dichomeris ustalella, and compared their genome organization and sequence composition to those of available gelechioid mitogenomes for an enhanced understanding of Gelechioidea genomic characteristics. We compared all available lepidopteran mitogenome arrangements, including that of M. albilinella, which is unique in Gelechioidea, to comprehend the extensiveness and mechanisms of gene rearrangement in Lepidoptera. The genomes of M. albilinella and D. ustalella are 15,274 and 15,410 bp in size, respectively, with the typical sets of mitochondrial (mt) genes. The COI gene begins with CGA (arginine) in all sequenced gelechioids, including M. albilinella and D. ustalella, reinforcing the feature as a synapomorphic trait, at least in the Gelechioidea. Each 353- and 321-bp long A + T-rich region of M. albilinella and D. ustalella contains one ( D. ustalella) or two ( M. albilinella) tRNA-like structures. The M. albilinella mitogenome has a unique gene arrangement among the Gelechioidea: ARN ESF (the underline signifies an inverted gene) at the ND3 and ND5 junction, as opposed to the ARNSE F that is found in ancestral insects. An extensive search of available lepidopteran mitogenomes, including that of M. albilinella, turned up six rearrangements that differ from those of 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 MI Q 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, although current mitogenomic data are limited, particularly for congeneric data. [ABSTRACT FROM AUTHOR]

The geographic relationships and biogeography of Apis cerana have been studied extensively, but Korean populations have not been investigated thoroughly. We sequenced the non-coding region between the tRNA and COII mitochondrial (mt) genes (herein named NC2) of Korean samples, along with the samples from seven Asian localities (China, Vietnam, and Thailand). Four undiscovered haplotypes were found in Korea and China, respectively. A phylogenetic analysis confirmed that Korean A. cerana belonged to the Mainland Asian group. Dominance of Japan1 haplotype in Mainland Asia including Korea suggests extensive gene flow and a common genetic origin. A newly developed non-coding region between the tRNA and tRNA mt genes (named NC1) provided nine haplotypes with twice the number of variable positions compared to those in NC2. A NC1-based phylogenetic analysis revealed the presence of two phylogenetic groups in Apis cerana from Korea suggesting two different sources. [ABSTRACT FROM AUTHOR]

We newly sequenced mitochondrial genomes of Spodoptera litura and Cnaphalocrocis medinalis belonging to Lepidoptera to obtain further insight into mitochondrial genome evolution in this group and investigated the influence of optimal strategies on phylogenetic reconstruction of Lepidoptera. Estimation of p-distances of each mitochondrial gene for available taxonomic levels has shown the highest value in ND6, whereas the lowest values in COI and COII at the nucleotide level, suggesting different utility of each gene for different hierarchical group when individual genes are utilized for phylogenetic analysis. Phylogenetic analyses mainly yielded the relationships (((((Bombycoidea + Geometroidea) + Noctuoidea) + Pyraloidea) + Papilionoidea) + Tortricoidea), evidencing the polyphyly of Macrolepidoptera. The Noctuoidea concordantly recovered the familial relationships (((Arctiidae + Lymantriidae) + Noctuidae) + Notodontidae). The tests of optimality strategies, such as exclusion of third codon positions, inclusion of rRNA and tRNA genes, data partitioning, RY recoding approach, and recoding nucleotides into amino acids suggested that the majority of the strategies did not substantially alter phylogenetic topologies or nodal supports, except for the sister relationship between Lycaenidae and Pieridae only in the amino acid dataset, which was in contrast to the sister relationship between Lycaenidae and Nymphalidae in Papilionoidea in the remaining datasets. [ABSTRACT FROM AUTHOR]

In an effort to gain greater insight into the nature of the population genetic structure of the pest insect Spodoptera litura (Lepidoptera: Noctuidae), tobacco cutworms were collected from six Korean and five Chinese localities, and their mitochondrial A+T-rich region and nuclear internal transcribed spacer 2 (ITS2) were cloned and sequenced. The A+T-rich region and nuclear ITS2 provided a maximum sequence divergence of 2.88 and 1.82%, respectively. Overall, a low level of genetic fixation ( F = 0-0.02965 in the A+T-rich region and 0-0.34491 in ITS2) and no discernible isolated population were noted among most S. litura populations. Along with these results, the absence of genetic variance between Korea and China indicates a profound interrelation of the S. litura populations in the two countries, consistent with the current notion that S. litura has sufficient flight capacity for dispersal. [ABSTRACT FROM AUTHOR]

The silkworm ( Bombyx mori) is of great economic value from an industrial perspective. Casual discrimination and accumulation of genetic information from a diverse variety of silkworms are essential for practical utilization and longterm conservation. In this study, a total of 54 silkworm strains preserved in Korea were typed for nine polymorphic microsatellite loci. We determined per-locus numbers of alleles ranging from 3 to 17 with an average value of 7.5, per-locus observed heterozygosity ranging from 0.04 to 0.98, and per-locus polymorphic information content (PIC) ranging from 0.06 to 0.86, thereby indicating that some of these loci are profoundly variable. Phylogenetic analysis using the nine concatenated microsatellite loci showed no clustering on the basis of known strain characteristics and origin. A total of 17 strain-specific apomorphic alleles, which discriminate 14 among 54 silkworm strains, were obtained from eight loci. These strain-specific alleles can, therefore, be casually utilized to discriminate between applicable strains, without any further typing of other loci. Furthermore, a substantial number of homozygote strains, represented by 24 among 67 alleles in nine loci, were also detected. These results collectively implicate silkworm microsatellite DNA as useful and potentially important molecular markers for the eventual discrimination of silkworm strains, hundreds of which are currently preserved in Korea. [ABSTRACT FROM AUTHOR]

The molecular phylogenetic relationships among true butterfly families (superfamily Papilionoidea) have been a matter of substantial controversy; this debate has led to several competing hypotheses. Two of the most compelling of those hypotheses involve the relationships of (Nymphalidae + Lycaenidae) + (Pieridae + Papilionidae) and (((Nymphalidae + Lycaenidae) + Pieridae) + Papilionidae). In this study, approximately 3,500 nucleotide sequences from cytochrome oxidase subunit I (COI), 16S ribosomal RNA (16S rRNA), and elongation factor-1 alpha (EF-1α) were sequenced from 83 species belonging to four true butterfly families, along with those of three outgroup species belonging to three lepidopteran superfamilies. These sequences were subjected to phylogenetic reconstruction via Bayesian Inference (BI), Maximum Likelihood (ML), and Maximum Parsimony (MP) algorithms. The monophyletic Pieridae and monophyletic Papilionidae evidenced good recovery in all analyses, but in some analyses, the monophylies of the Lycaenidae and Nymphalidae were hampered by the inclusion of single species of the lycaenid subfamily Miletinae and the nymphalid subfamily Danainae. Excluding those singletons, all phylogenetic analyses among the four true butterfly families clearly identified the Nymphalidae as the sister to the Lycaenidae and identified this group as a sister to the Pieridae, with the Papilionidae identified as the most basal linage to the true butterfly, thus supporting the hypothesis: (Papilionidae + (Pieridae + (Nymphalidae + Lycaenidae))). [ABSTRACT FROM AUTHOR]

To identify novel bioinsecticidal agents, a bacterial strain, Serratia sp. EML-SE1, was isolated from a dead larva of the lepidopteran diamondback moth ( Plutella xylostella) collected from a cabbage field in Korea. In this study, the insecticidal activity of liquid cultures in Luria-Bertani broth (LBB) and nutrient broth (NB) of a bacterial strain, Serratia sp. EML-SE1 against thirty 3rd and 4th instar larvae of the diamondback moth was investigated on a Chinese cabbage leaf housed in a round plastic cage (Ø 10×6 cm). 72 h after spraying the cabbage leaf with LBB and NB cultures containing the bacterial strain, the mortalities of the larvae were determined to be 91.7% and 88.3%, respectively. In addition, the insecticidal activity on potted cabbage containing 14 leaves in a growth cage (165×83×124 cm) was found to be similar to that of the plastic cage experiment. The results of this study provided valuable information on the insecticidal activity of the liquid culture of a Serratia species against the diamondback moth. [ABSTRACT FROM AUTHOR]

Cecropin is a well-studied antimicrobial peptide that is synthesized in fat body cells and hemocytes of insects in response to hypodermic injury or bacterial infection. A 503 bp cDNA encoding for a cecropin-like peptide was isolated by employing annealing control primer (ACP)-based differential display PCR and 5′-RACE with immunized Papilio xuthus larvae. The open reading frame of the isolated cDNA encoded for a 62-amino acid prepropeptide with a putative 22-residue signal peptide, a 2-residue propeptide, and a 38-residue mature peptide with a theoretical mass of 4060.89 Da. The deduced amino acid sequence of the peptide evidenced a significant degree of identity with other lepidopteran cecropins. This peptide was named papiliocin. RTPCR results revealed that the papiliocin transcript was detected at significant levels after injection with bacterial lipopolysaccharide (LPS). On the basis of the deduced amino acid sequence of papiliocin, a 38-mer mature peptide was chemically synthesized via the Fmoc method, and its antimicrobial activity was analyzed. The synthetic papiliocin peptide evidenced a broad spectrum of activity against fungi, Gram-positive and Gram-negative bacteria, and also evidenced no hemolytic activity against human red blood cells. [ABSTRACT FROM AUTHOR]

Hemolin has been known to play a key role in insect innate immunity. In an attempt to examine expression pattern of the Hemolin gene in the diamondback moth, Plutellea xylostella, the full-length cDNA of Hemolin was cloned using 5′-RACE PCR technique. The cDNA contained a 5′ untranslated region of 48 nucleotides and a 3′ untranslated region of 198 nucleotides, including a stop codon (TAA) and a poly (A) tail. It consists of 1,401 bp with an open reading frame of 1,245 bp, encoding 414 amino acids. The deduced amino acid sequence of PxHemolin has relatively low identities (35−42%) to various insect Hemolins. However, it has high three-dimensional structural similarity to Hemolin. Interestingly, analysis of spatial expression pattern of PxHemolin shows that it was highly expressed in the Malpighian tubule and the silk gland although it was also detected in fat body and gut. Furthermore, PxHemolin mRNA was highly induced 3 hr after immune-challenging with lipopolysaccharide and was gradually up-regulated after laminarin treatment. These data suggest that PxHemolin may play a role in innate immune responses although it remains to further elucidate the precise biological functions in P. xylostella. [ABSTRACT FROM AUTHOR]

The 15,389-bp long complete mitogenome of the endangered red-spotted apollo butterfly, Parnassius bremeri (Lepidoptera: Papilionidae) was determined in this study. The start codon for the COI gene in insects has been extensively discussed, and has long remained a matter of some controversy. Herein, we propose that the CGA (arginine) sequence functions as the start codon for the COI gene in lepidopteran insects, on the basis of complete mitogenome sequences of lepidopteran insects, including P. bremeri, as well as additional sequences of the COI start region from a diverse taxonomic range of lepidopteran species (a total of 53 species from 15 families). In our extensive search for a tRNA-like structure in the A+T-rich region, one tRNA-like sequence and one tRNA (UUR)-like sequence were detected in the P. bremeri A+T-rich region, and one or more tRNA-like structures were detected in the A+T-rich region of the majority of other sequenced lepidopteran insects, thereby indicating that such features occur frequently in the lepidopteran mitogenomes. Phylogenetic analysis using the concatenated 13 amino acid sequences and nucleotide sequences of PCGs of the four macrolepidopteran superfamilies together with the Tortricoidea and Pyraloidea resulted in the successful recovery of a monophyly of Papilionoidea and a monophyly of Bombycoidea. However, the Geometroidea were unexpectedly identified as a sister group of the Bombycoidea, rather than the Papilionoidea. [ABSTRACT FROM AUTHOR]

The bumblebee species, Bombus, is an invaluable natural resource for greenhouse pollination. Low levels of genetic variation of Bombus ardens have been reported in a previous mitochondrial (mt) gene study. In this study, we sequenced the complete internal transcribed spacer 2 (ITS2) of the nuclear rDNA obtained from 100 B. ardens individuals collected from several Korean localities, in an effort to assess its usefulness in characterizing the genetic diversity and relationships among populations of B. ardens. The ITS2 sequences of B. ardens were shown to be longest among known insects, ranging in size from 1,971-1,984 bp. The sequences harbor four duplicated repeats-≈27 bp repeats, ≈20 bp repeats, ≈33 bp repeats, and ≈34 bp repeats-which have never before been reported in other insect ITS2 rDNA. The maximum sequence divergence of 1.01% among 96 sequence types confirmed the applicability of this molecule to the study of intraspecific variation, revealing higher sequence variation as compared to the previously studied mt COI gene. Overall, a very high per generation migration ratio ( Nm = 5.83 ≈ infinite) and a very low level of genetic fixation ( FST =0 -0.08) were noted to exist among populations of B. ardens. The high estimation of gene flow among most populations-in particular, between the remote island Ulleungdo and several inland populations-suggest that historical events may be more responsible for the contemporary population structure of B. ardens. The finding of the lowest genetic diversity (π) in the population on Ulleungdo Island (π = 0.007434) may be reflective of a relatively small population size and the geographical isolation of the population as compared with other inland populations. [ABSTRACT FROM AUTHOR]

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 tRNA(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 tRNA-like sequence and one tRNA-like sequence. 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. [ABSTRACT FROM AUTHOR]