The role of Cl channels in regulatory volume decrease (RVD) in human salivary gland acinar cells was examined using a whole-cell patch clamp technique. Human tissues were obtained from healthy volunteers or from patients with oromaxillofacial tumors. During the measurements, K+-free solutions were employed to eliminate contamination of whole-cell conductance by K+ currents. When the cells were exposed to a 70% hypotonic solution, outward-rectifying currents, which were not observed in the resting state, were found to have significantly increased both in human labial and parotid gland acinar cells. The amplitudes of the currents were reduced in a low CI bath solution. Furthermore, the addition of 100μM 5-Nitro-2- (3-phenyl propylamino) benzoic acid (NPPB) or 100μM 4,4'-diisothio cyanatostilbene-2,2'-disulphonic acid (DIDS), known to partially block Cl channels, significantly inhibited these currents. Its outward-rectifying current profile, shift in reversal potential in a low Cl bath solution and pharmacological properties suggest that this is a Cα2+ independent, volume activated Cl current. We conclude therefore that volume activated Cl channels play a putative role in RVD in human salivary gland acinar cells.

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.

The experimental work was conducted to investigate the effects of the application level of composted cattle manure (CCM) on forage productivity using Italian ryegrass-WCR double cropping system for 3 years (2006~2008). Main plot was consisted of application level of CCM such as 150%, 200% and control plot. The total day matter yield (DMY) of Italian ryegrass and WCR in treatment of CCM 150% and CCM 200% significantly decreased as compared with that of treatment of chemical fertilizer (CF). However, NDF, AND and CP content of Italian rye grass and WCR were hardly influenced by CCM application.

Sweetpotato whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae), is a serious pest of many economically important crops. The insect has developed resistance to chemical insecticides. Therefore, the development of microbial agent is necessary. Among the several entomopathogenic fungi, Lecanicillium lecanii Btab01 which has high insecticidal activity was carried out this experiments. To develop mass culture, we subcultured L. lecanii Btab01 on PDA, TSA, SDA+Y, RA and GSA media at 25℃ incubator to select the optimal solid culture medium. Hyphal growth was measured every 3 or 4 days. L. lecanii Btab01 grew fastest in RA, followed GSA, SDA+Y, PDA and TSA. L. lecanii Btab01 was cultured on PDB, TSB, SDB+Y, RB, GSB media at 25℃, 180rpm shaking incubator to select the optimal liquid medium. Spore germination was measured by spread plate method every 12 or 24 hours. Spore germination appeared 7.8×108 CFU/ml after 4 days in RB, followed GSB (5.5×108 CFU/ml), SDB+Y (2.7×108 CFU/ml), TSB (1.7×108 CFU/ml) and PDB (0.6×108 CFU/ml).

Reactive oxygen species (ROS) is toxic to living organisms, because its high reactivity causes oxidative damage to proteins, nucleic acids, and lipids. Superoxide dismutase (SOD) is an enzyme facilitating the removal of superoxide anions from living organisms. This study focused on the cloning of MnSOD cDNA from Hyphantria cunea and its induction upon bacterial infection and various stresses. The open reading frame of MnSOD is composed of 645 bp, encoding 215 amino acid residues. The theoretical molecular mass and pI of putative MnSOD was evaluated to be 24276 Da and 9.14, respectively. The MnSOD from H. cunea is highly similar to human MnSOD (59.5%) as well as Bombyx mori MnSOD (76.2%). MnSOD showed no big induction upon bacterial infection and stresses, compared to that of Cu/ZnSOD.

Apolipophorin-Ⅲ (apoLp-Ⅲ) is a hemolymph protein whose function is to facilitate lipid transport in an aqueous medium in insect. Recently, apolipophorin-Ⅲ in Galleria mellonella and Hyphantria cunea was shown to play an unexpected role in insect immune activation. We show here a novel possible function/role of apoLp-Ⅲ in insects. To investigate the genes which have a relationship with apoLp-Ⅲ in fall webworm larvae, we reduction of endogenous Hc apoLp-Ⅲ mRNA levels in larvae via RNA interference (RNAi). The RNAi-mediated Hc apoLp-Ⅲ reduction resulted in the reduction of antioxidants, like MnSOD, catalase, and glutathione S transferase as well as immune proteins. In particular, expression of MnSOD commonly decreased in fat body, midgut, and hemocytes following the knockdown of Hc apoLp-Ⅲ, which induced an elevated level of superoxide anion in H. cunea larvae. The observed effect of Hc apoLp-Ⅲ RNAi suggests that Hc apoLp-Ⅲ is related to the action/expression of antioxidants.

Innate immunity responses are triggered by the immune challenge and therefore involve signaling processes. The cellular response is initiated by hemocytes and mainly involves phagocytosis and encapsulation of intruders by these cells. To address whether Hc-STAT is activated upon bacterial challenge, we examined the subcellular location of STAT protein in hemocyte by immunostaining. A new insect member of the STAT family of transcription factors (Hc-STAT) has been cloned from the lepidopteran, Hyphantria cunea. The domain involved in DNA interaction and the SH2 domain are well conserved. The gene is transcribed at a low level during all stages of development, and the protein is present in hemocytes, fat body, midgut, epidermis, and Malphigian tuble (Mt). Especially, hemocytes and Mt showed transcriptional activation of Hc-STAT upon Gram (-) bacteria and fungal challenge. Gram (-) bacteria and fungal challenge specifically results in nuclear translocation of Hc-STAT protein and induction of DNA-binding activity that recognizes a STAT target site in H. cunea hemocyte. In vitro treatment with pervanadate translocates Hc-STAT to the nucleus in hemocyte cells. Here we report the first evidence for the involvement hemocyte JAK/STAT pathway upon microbial infection in lepidopteran insect.

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 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.