The glycoprotein hormone family represents a class of heterodimers, that includes the placental hormone equine chorionic gonadotropin (eCG) and the anterior pituitary hormones‐ follitropin (FSH), lutropin (LH), and thyrotropin (TSH). The 4 hormones are heterodimers, with a common α‐subunit and unique β‐subunits. eCG is the most heavily glycosylated of the known pituitary and placental glycoprotein hormones. Recent observations using single chain glycoprotein hormone analogs in which, the β‐and α‐subunits are linked, implied that heterodimeric‐like quaternary configuration is not a prerequisite for receptor/signal transduction. To study the function and signal transduction of tethered rec‐eCG, a single chain eCG molecule was constructed and rec‐eCG protein was produced. Molecular mass of the single chain is about 45 kDa. All mice were ovulated by tethered rec‐eCG treatment. The dual activity of tethered rec‐eCG was determined in receptor cell lines of nonequid species; in fact, this dual activity was proven in species other than horse. Tethered rec‐eCG in equids does not bind to FSH receptors, suggesting that eCG is primarily an LH‐like hormone in the horse. Taken together, these data suggest that tethered rec‐eCG has dual activity in nonequid species in vitro. However, it has only LH‐like activity in equid species in vitro.
The Classical Swine Fever Virus (CSFV) is a member of the Pestivirus genus of the Flaviviridae. The genome of CSFV is a positive single-stranded RNA molecule 12.3 kb and contains a single large open reading frame (ORF). The polyprotein composed of eight nonstructural and four structural proteins (nucleocapsid protein C and three envelope glycoprotein E0, E1 and E2). E2, the most immunogenic of the CSFV glycoproteins, induces a protective immune response in swine. To determine the characteristics of the CSFV, LOM strain, we investigated the nucleotide sequence of the glycoprotein E0, E1 and E2. Comparison of the LOM with the other strains revealed nucleotide sequence identity ranging from 97 to 98%. Expression of the glycoprotein E2 was identified by SDS-PAGE and Western blot analysis using anti-CSFV E2 monoclonal antibodies in Sf21 cells. The expression levels of glycoprotein E2 were observed from day 3 and 5 days maximum. In addition, its expression efficiency by media and cell line was investigated. The result showed that High-Five cells and Grace’s insect media for Sf21 were the best conditions for the expression of the glycoprotein E2.
Entomopathogenic fungi were isolated directly from a cadaver of adult M. saltuarius (infected with white fungi) supporting fungal sporulation, to develop biological control of pine wilt disease vector, M. saltuarius which was the most abundant in the middle to northern part of Korea and caused enormous damage to native pine tree in Korea, Japan and other regions of Asia. Pathogenicity of each fungus was tested using oak longicorn beetle, Moechotypa diphysis, as substitutive insect. As the result, only one of them showed pathogenic to adults of M. diphysis, with up to 100% mortality within 13 days of inoculation. Selected fungus was named as MsW1 and identified by Beauveria bassiana using microscopic examination, B. bassiana-specific PCR primers and genetic sequencing of the ITS region analysis. Pathogenicity test were conducted with various concentration of conidial suspensions of this isolate on M. saltuarius (3rd instar larvae and adults). Mortality rates varied from 57.1% to 100.0% and from 16.7% to 100.0% of M. saltuarius (3rd instar larvae and adults), respectively at 30 days. This is the first report of natural infection of M. saltuarius by B. bassiana.
In agricultural fields, the entomopathogenic fungal species have been investigated for their potential as the biological control agents due to their role of natural enemies for insects. Until recent times, most of the studies for these fungi have been based on isolation from insect cadaver or soil. However, these methods, especially isolation from soil, might cause a problem involving differential isolation of the each entomopathogenic fungi. The purpose of this study is to determine the optimal isolation medium for entomopathogenic fungi using dodine, chitin, and skim milk. The growth rates of entomopathogenic fungi and non-entomopathogenic fungi were compared on dodine agar medium. The medium for this experiment was modified Veen semiselective medium which consisted of based on SDA (Sabouraund Dextrose Agar), 100 mg/ml for chloramphenicol, 50 mg/ml for streptomycin and adjusted dodine to 40, 50, 70 and 100 mg/ml. As a result, optimal concentration of dodine for isolation of entomopathogenic fungi was 50 mg/ml and 168 positive entomopathogenic fungi were isolated in 470 soil samples and 11 cadavers of insect. In addition, the isolates had significantly greater chitinase and protease activity than non-entomopathogenic fungi. The isolation method described represents a valuable tool for rapid and simple isolation of entomopathogenic fungi. These positive entomopathogenic fungi may have potential against variety pests in agriculture.
The potato tuber moth (PTM, Phthorimaea operculella (Zeller)) is a serious pest of potato (Solanum tuberosum L.) plants in the field in regions with warm temperate to tropical climates, and of tubers stored under ambient temperatures post-harvest. Although PTM can be a minor to serious pest depending on the area and year, this pest is getting more and more serious to potato fields in Korea according to accidental surveys. Son (1979) represented 12.5℃ of average annual temperature as northern limit of distribution where corresponding to Youngdeog, Dalseong, Boseong and Jindo regions. Soon after, Choi and Park (1980) reported that the northern limit of PTM distribution coincided with -8℃ isotherm for average annual minimum temperature of January. In recent years, concerns on PTM occurrence and damage to potato are increasing due to climate change resulted in global warming. We carried out an extensive survey of PTM distribution using sex pheromone traps at 24 sites across the nation in 2009. As a result, we confirmed that the northern limit of PTM distribution climbed to southern Gyeonggi and middle Gangwon regions from Gyeongbuk and Chungbuk regions indicated by Choi and Park (1980). Hereafter, we are trying to investigate why drastic going northward of PTM happened in Korea. Here is one presumption that a sudden rise in average temperature has occurred during 30 years, from 6.34℃ in 1973 to 7.62℃ in 2007, especially around Daegwallyeong area where is a typical highland with 800 m high.
Major fruit moths in commercial apple orchards are known as Carposina sasakii and Grapholita molesta. In addition, recently G. molesta is dominant species compared with C. sasakii in apple orcahrds.
In case of Japan, the fruit damage occurred a lot unexpectedly in apple orchards of the northern Nagano during the harvesting season in 2004. As the result of identifying after emerging the larvae collected from the damaged fruit, Grapholita dimorpha was found out. The morphology of Grapholita dimorpha are almost similar to that of Grapholita molesta, and the shape of the damaged fruits was too similar to distinguish them.
We installed sex pheromone traps of Grapholita dimorpha in apple orchards to check if there were Grapholita dimorpha in Korean apple orchards and to survey seasonal occurrence.
As a result of survey by installing sex pheromone traps of G. dimorpha at one apple orchard in Uiseong-Gun and Gunwi-Gun area respectively, we could find high population density of G. dimorpha in sex pheromone traps. The peak of the first generation of G. dimorpha was dated in early May. The first generation occurrence of G. molesta in sex pheromone traps was lower than that of Grapholita molesta. However, it had higher occurrence of sex pheromone traps than G. molesta since it increased after mid and late July.
Also, as a result of identifying in G. molesta traps, there were 673 G. molesta and 8 G. dimorpha. As a result of survey of 1,102 G. dimorpha in its sex pheromone traps, there were 1,099 G. dimorphaand 3 G. molesta.
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 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.