In order to understand the nature of population genetic structure of the notorious pest insect, Spodoptera litura (Lepidoptera: Noctuidae), a total of 159 individuals of the tobacco cutworm collected from six Korean and five Chinese localities were cloned and sequenced their mitochondrial A+T-rich region. A total of 90 haplotypes ranged 324 bp ~ 372 bp in length and 0.30% ~ 4.85% in sequence divergence. Compared to the preliminary mitochondrial COI gene sequence data that provided nearly identical sequences in all samples, the A+T-rich region provided substantially high sequence variation, validating the applicability of this molecule to the study of intraspecific variation. Overall, a very high per generation migration ratio (Nm = 5.91 ~ infinite), a very low level of genetic fixation (FST = 0 ~ 0.077796), and no discernable isolated population were noted to exist among populations of S. litura, although some significant genetic differentiations were found between several pairs of populations. These results collectively suggest that the S. litura populations in both Korea and China are very well interrelated to each other. The structural analysis revealed that the S. litura A+T-rich region harbored two stretches of the [TA(A)]n sequence, the poly-A stretch, and a stem-and-loop structure that are well found in other lepidopteran A+T-rich region. Furthermore, the region harbors the 13 ~ 19-bp long T stretch and 5-bp long motif that have been suggested to function as a possible recognition site for the initiation of replication of the minor strand of mtDNA also were found.

The bumblebee, Bombus ignitus (Hymenoptera: Apidae), is a valuable natural resource that is one of the most notably utilized for greenhouse pollination in Korea. In order to understand the nature of genetic relationships, gene flow, and population structure of the species we sequenced a partial COI gene of mitochondrial DNA (mtDNA) corresponding to “animal barcode” region and the complete internal transcribed spacer 2 (ITS2) of the nuclear ribosomal DNA (nrDNA) with the individuals collected from Korean localities. Although the 658-bp long mtDNA sequence provided only six haplotypes with the maximum sequence divergence of 0.456% (3 bp, from 91 individuals), but the ITS sequences provided 33 sequence types with the maximum sequence divergence of 0.78% (16 bp, from 35 individuals), confirming better applicability of the ITS sequences to the study of intraspecific variation. The ITS2 sequences of B. ignitus were shown to be the longest among known insects, ranging in size from 2,034 bp ~ 2,045 bp and harbored two duplicated repeats, indicating unusual structure of B. ignitus ITS2 sequences compared with other insect ITS sequences. Overall, a very high per generation migration ratio, a very low level of genetic fixation, and no discernable hierarchical population/population group were noted to exist among populations of B. ignitus on the basis of both molecules, thus suggesting that the B. ignitus populations on the Korean peninsula are panmictic, which is consistent with our understanding of the dispersal capability.

Up to now only 17 coleopteran mitogenomes have been fully or near fully sequenced, but this chart does not include any complete mitogenome sequence of the coleopteran infraorder Scarabaeiformia, in which the monotypic Scarabaeoidea is included. In this study, we firstly present three scarabaeoid mitogenomes, two of which are completed and one is near completed. The complete nucleotide sequences of the white-spotted flower chafer, Protaetia brevitarsis (Scarabaeiformia: Scarabaeidae), was determined to be 20,319 bp, the two-spotted stag beetle, Metopodontus blanchardi (Scarabaeiformia: Lucanidae), was 21,678 bp, and the incomplete garden chafer, Polyphylla laticollis (Scarabaeiformia: Scarabaeidae), which lacks sequence information from whole A+T-rich region and a partial srRNA gene was 14,473 bp. Thus, the two complete mitogenome sequences are longest among the completely sequenced insect mitogenomes, followed by the 19,517 bp-long dipteran Drosophila melanogaster. Phylogenetic analysis using the concatenated 13 amino acid sequences of PCGs of the 13 coleopteran insects including three newly sequenced scarabaeoid successfully recovered a monophyly of suborder Polyphaga, a monophyly of infraorder Cucujiformia, a monophyly of infraorder Elateriformia, and also a monophyly of infraorder Scarabaeiformia within Polyphga. However, the Scarabaeiformia represented by three newly sequenced species was unexpectedly placed as a sister group to the Cucujiformia, instead of the placement to the sister to the Cucujiformia and Elateriformia group.

Eumenis autonoe, a member of the lepidopteran family Nymphalidae (superfamily Papilionoidea) is an endangered species, and is found only on one isolated remote island, Jeju in South Korea, on Halla Mt., at altitudes higher than 1,400 meters. In this study, we report the complete mitochondrial genome (mitogenome) of E. autonoe. The 15,489-bp long E. autonoe genome evidenced the typical gene content found in animal mitogenomes, and harbors the gene arrangement identical to all other sequenced lepidopteran insects, which differs from the most common type found in insects, due to the movement of tRNAMet to a position 5’-upstream of tRNAIle. As has been observed in many other lepidopteran insects, no typical ATN codon for the COI gene is available. Thus, we also designated the CGA (arginine) found at the beginning of the COI gene as a lepidopteran COI starter, in accordance with previous suggestions. The 678-bp long A+T-rich region, which is second longest in sequenced lepidopteran insects, harbors 10 identical 27-bp long tandem repeats plus one 13-bp long incomplete final repeat. Such a repeat sequence has been, thus far, only rarely detected in lepidopteran mitogenomes. The E. autonoe A+T-rich region harbors a poly-T stretch of 19 bp and a conserved ATAGA motif located at the end of the region, which have been suggested to function as structural signals for minor-strand mtDNA replication.

The experimental work was conducted to investigate the effects of the application level of composted cattle manure (CCM) on forage productivity using whole crop barely (WCB)-whole crop rice (WCR) double cropping system for 3 years. Main plot was consisted of application level of CCM such as 150%, 200% and control plot. The total DMY of forages under WCB and WCR in treatment of CF significantly decreased as compared with that of treatment of CCM 150%. However, NDF, ADF and CP content of WCB and WCR were hardly influenced by CCM application.

Gochujang samples with essential oils were evaluated for sensory including taste and, smell during various storage periods with the aim of developing improved high value-added products. The sensory testing was performed on Gochujang with 0.005% Zanthoxylum piperitum and Chrysanthemum indicum essential oils, extracted through steam distillation. During storage at 4 and 20℃ for 8 weeks, sensory testing was performed at 0, 1, 2, 4 and 8 weeks by trained panelists. Overall quality decreased in Gochujang samples containing Z.piperitum essential oil and increased in samples containing Cindicum essential oil, at 4 and 20℃. Samples stored at 4℃ were quality more then those stored at 20℃. Sensory evaluation was highest for Gochujang with Cindicum essential oil stored at 4℃. It is concluded that functional Gochujang can be developed by adding small quantities of essential oils. Increasing the amount of essential oil added is likely to affect the innate taste and flavor of Gochujang.

This study examined the effects of adding Pleurotus ostreatus, Agaricus bisporus and Flammulina velutipes on the quality characteristics of cookies and multiple purposes ofwheat powders. The water content, crude protein content, mineral content， color value and sensory evaluation of mushroom cookies and multiple purposes ofwheat powders were measured. In the case of cookies, the water content was 1.10-1.24% and crude protein content was 8.20-9.80%. General preferences， such as appearance, color, taste and texture， increased when mushroom was added to the cookies, especially flavor, which showed a much higher preference. In the studies where mushroom was added to wheat powders, the water content was 0.57-0.92% and crude protein content was 11.90-12.80%. The addition of mushroom to both of the cookies and multiple purposes of wheat powders resulted in a remarkable increase in the mineral content, especially Fe, K. In addition, an increase in the amount of added mushrooms resulted in an increase in the hunter L and a values; however, this also resulted in a decrease in the b value.

We assessed the environmental risk of herbicide resistant transgenic rice (Protox) on non-target herbivore, grasshoppers (Oxya japonica japonica Thunberg). We conducted life-history experiments of grasshoppers with measuring their body weight, body length, eating amount, and feces amount between non-transgenic rice (nTR; Dongjin rice) and transgenic rice (TR; Protox rice) under laboratory conditions (Temp. 25Ð, R.H. 50-70%, Photoperiod L16:D8) in 2007. The growth of grasshoppers appeared to increase at each measuring date. We also compared the growth rate of grasshoppers between nTR and TR to examine the transgenic impact on the herbivore and we found there was no statistically signifi cant difference between the two plant types (P>0.05). We found that body weight and body length for grasshoppers were highly correlated at each of the two types of plants, nTR (0.962) and TR (0.960). The correlation of eating amount and feces amount of grasshoppers were higher nTR (0.830) than TR (0.782). The energy effi ciency of the grasshopper was not a signifi cant between nTR and TR (P> 0.05). But the molt timing of the grasshoppers for TR difference was faster than for nTR. Conclusively life-history of the grasshoppers but molt timing was not a signifi cant difference between nTR and TR. Therefore, we could conclude there was not any environment risk on herbivore from our result.

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.

Eumenis autonoe belonging to a lepidopteran family Nymphalidae (superfamily Papilionoidea) is an endangered species in Korea. Historically, the species was distributed in Europe and Asian region including a wide region in Korean peninsula. However, in Korean peninsula, the species is found only in two isolate dregions: South in a remote island Jeju, where altitude is higher than1, 400 meter on Halla Mt. and North in far northern Korean peninsula around Mt. Bekdu. In this study, we report the complete mitochondrial genome of the endangered E. autonoe collected from Mt. Halla. The 15,489-bp long E. autonoe genome has a typical gene content found in animal mitochondrial genomes and contains the gene arrangement identical to all other sequenced lepidopteran insects, which differs from the most common type found in insects, as the result of the movement of tRNAMet to a position 5’-upstream of tRNAIle. As seen in many other lepidopteran insects, no typical ATN codon for COI gene is available. Thus, we tentatively designated the CGA (arginine) found at the beginning of the COI gene, as has been suggested for lepidopteran COI starter. The intergenic spacer sequence located between tRNASer (UCN) and ND1 of E. autonoe mitogenome also contains the ATACTAA motif which is conserved in all sequenced lepidopteran species. The 678-bp long A+T-rich region, which is longest in sequenced lepidopteran insects contains ten identical tandem repeats composed of 27 bp plus one 13-bp long identical incomplete final repeat. Such repeat sequence is rare in the lepidopteran mitogenomes known so far. The E. autonoe A+T-rich region also contains a poly-T stretch located at the end of the region as 19 bp and also contains the downstream conserved motif ATAGA that were previously suggested to serve as a structural signal for minor-strand mtDNA replication. Phylogenetic analysis using the concatenated 13 amino acid sequences of PCGs among available six lepidopteran superfamilies (Tortricoidea, Pyraloidea, Papilionoidea, Bombycoidea, Geometroidea, and Noctuoidea) rooted with three dipteran species with BI and ML analyses supported the following topology: ((((Bombycoidea + Geometroidea +Noctuoidea) + Papilionoidea) + Pyraloidea) + Tortricoidea). Within Papilionoidea, a closer relationship between Lycaenidae and Pieridae, excluding Nymphalidae was observed. Further fruitful information will be available after more analysis is done.

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 bumblebees, Bombus species are valuable natural resources being utilized for greenhouse pollination. Low level of genetic variation of Bombus species has been reported previously. In this study, we sequenced complete internal transcribed spacer 2 (ITS2) of the nuclear rDNA from 100 individuals of B. ardens collected from seven localities in Korean peninsula. The ITS2 sequence of B. ardens is longest, ranging in size from 1,940 bp-1,954 bp among known in insects, which ranges approximately from 241 bp-1,728 bp. The ITS2 sequences have ~51% of C+C content and contain each two 27 bp repeats, 20 bp repeats, 33 bp repeats, and 34 bp repeats at the beginning. Such repeats were not found in other insects. Uncorrected pairwise distance among 96 sequences that were obtained from 100 individuals revealed a maximum sequence divergence of 1.03%. Genetic diversity (π) of each population ranged from 0.007801 to 0.009627, and the lowest diversity was obtained from islet population of Ulleungdo, indicating possibly small, isolation of the population. Significant level of genetic distance was only found when Ulleungdo population was compared to two other mainland populations. Except for this, overall, a very high rate of per generation migration ratio (Nm=7.1-infinite) and a very low level of genetic fixation (FST=0-0.06546) were detected between pairs of localities. Analysis of hierarchical relationships among localities consistently revealed no statistically significant structure among populations. Taken these together, the B. ardens populations on the Korean peninsula are panmictic this is consistent with our understanding of the dispersal capability.

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.

The bumblebee, Bombus ardens, is a valuable natural resource, and is most notably utilized for greenhouse pollination. In order to gain a greater understanding of the population genetic structure and the genetic diversity of this species, we sequenced a portion of the mitochondrial COI gene corresponding to the “DNA Barcode” region (658 bp) from 160 individuals collected over 15 Korean localities. Uncorrected pairwise distances among the eight haplotypes suggested low intraspecific genetic diversity, with a maximum sequence divergence of 0.3%. Such a low level of intraspecific genetic diversity was further reflected in local populations, particularly to islet populations, such as those of Youngheungdo, Jakyakdo, and Ulleugdo, which had zero genetic diversity. Geographically, one haplotype (BARBA01) was widespread and dominant, with a frequency of 90.6% (145 among 160 individuals). Other haplotypes were restricted to one to three localities and had low frequency. Overall, a very high rate of per generation female migration ratio (Nm= 4.6 to infinite) and a very low level of genetic fixation (FST= 0 to 0.099) were detected between pairs of localities, suggesting that the B. ardens population sonthe Korean peninsula are panmictic, which is consistent with our understanding of their dispersal capability.