Pleurotus eryngii is one of the most commercially important mushrooms cultivated in Korea. However, the shelf-life of the fruiting body is short, limiting its export. A new hybrid strain H17 of P. eryngii was developed to extend the shelf-life by mono-mono crossing between monokaryotic strains derived from DanBi and KNR2774. Although the cultivation period of H17 was slightly longer than that of the reference cultivar Kenneutari No.2, the quality did not change and remained normal after a period of 65.0 days at 4°C. This result was significantly different from that of the reference cultivar Kenneutari No.2. Analysis of the genetic characteristics of the new hybrid strain H17 revealed a different profile from that of the parental and reference cultivars when random amplification of polymorphic DNA (RAPD) primers was used. These results demonstrate that H17 is a new cultivar with improved storability after harvesting.

In abalones, interspecific hybridization has been suggested as a possible means to increase production and desired traits for the industry. In Korea, Haliotis gigantea is considered a species with a larger size and higher temperature tolerance than H. discus hannai. However, H. discus hannai is considered the most valuable and popular fishery resource due to its better acceptance and higher market prices. Thus, viable interspecific hybrids have been produced by artificial inseminating H. gigantea eggs with H. discus hannai sperm. However, the reciprocal hybrid cross was not successful. In this study, the hybridity and the growth and thermal tolerance performance of the interspecific hybrids were examined. A combination of various assays revealed maximum growth occurrence at 21℃ and the higher growth rate in the hybrids than that of H. discus hannai parent. In addition, the growth and survival at high-temperature (28℃) of the hybrids was equivalent to that of the highly tolerant H. gigantea parent, suggesting new possibilities to overcome the mass mortality in H. discus hannai during high temperature periods of summer season in Korea. Furthermore, the induced interspecific hybrid status was confirmed by the presence of species-specific bands for each parental species of the random amplified polymorphic DNA (RAPD) profiles using universal rice primer (URP), which could be used as speciesspecific markers to distinguish the hybrids and their parental species.

The entomopathogenic fungus Metarhizium anisopliae B is a powerful biological control agent against Monochamusalternatus, a crucial mediator of the pinewood nematode Bursaphelenchus xylophilus. In this study, production of destruxins(dtxs), insecticidal cyclic hexadepsipeptides, was monitored in the submerged culture of M. anisopliae B. Three typesof dtxs, i.e., destruxin A, B, and E, were produced during the culture. Among the three dtxs, the production yield ofdestruxin A was best, followed by destruxin B and E. Destruxin A production was increased when pH was controlledat 6.0, whereas production of destruxin E was not affected by the pH control. The highest yield of dtxs A, B, and Ewere 16.4, 7.3, and 6.1 mg L-1, respectively. Considering that process for dtxs production has not been optimized, M.anisopliae B has more powerful implication as a biocontrol agent.

We report on the dispersion state of partially reduced graphene oxide (PRGO) in organic solvents, namely methyl ethyl ketone, ethyl acetate, methylene chloride, toluene, and xylene, by controlling the carbon to oxygen (C/O) atomic ratio of the PRGOs. A two-phase solvent exchange method is also proposed to transfer PRGO from water to an aprotic solvent, such as methyl ethyl ketone. We achieve relatively good dispersion in aprotic and non-polar solvents by controlling the C/O atomic ratio of the PRGOs and applying the two-phase solvent exchange method. There is an increase in the glass transition temperatures with the dispersion of PRGOs into amorphous polymers, in particular a 4.4°C increase for poly(methyl methacrylate) and 3.0°C increase for polycarbonate. Good dispersion of PRGO in a nonpolar polymer, such as linear low density polyethylene, is also obtained.

Oocyte is the central factor in the bi-directional communication axis in the ovarian follicles. It controls the cumulus or granulosa cells to perform functions which are beneficial for its own development via secreting paracrine growth factors, including GDF9 and BMP15. The aim of this study was to investigate whether the recombinant GDF9 and BMP15 are able to promote meiotic resumption and cumulus expansion of canine COCs during IVM, as well as to demonstrate the actions of GDF9 and BMP15 in regulating the expression of connexin transcripts in the ovarian granulosa cells. As results, GDF9 and BMP15 significantly improved the meiotic resumption rate and cumulus expansion by activating ERK1/2 signaling. Treatments with GDF9 significantly improved the expression of CyclinB1 but inhibited the expression of Cx43 transcripts. In addition, cumulus expansion genes (MAPK1, Ptgs2, Tnfaip6 and Ptx3) were differentially improved by GDF9 and BMP15. In the ovarian granulosa cells, GDF9 suppressed the expression of Cx43 transcripts by binding ALK4/5/7 receptors and activation Smad2/3 signaling, whereas, BMP15 stimulated the expression of Cx43 transcripts by binding ALK2/3/6 receptors and activating Smad1/5/8 signaling. In conclusion, by regulating functions of granulosa/cumulus cells, oocyte has the potential to enhance the growth and maturation of itself.

Oocyte is the central factor in the bi-directional communication axis in the ovarian follicles. It controls the cumulus or granulosa cells to perform functions which are beneficial for its own development via secreting paracrine growth factors, including GDF9 and BMP15. The aim of this study was to investigate whether the recombinant GDF9 and BMP15 are able to promote meiotic resumption and cumulus expansion of canine COCs during IVM, as well as to demonstrate the actions of GDF9 and BMP15 in regulating the expression of connexin transcripts in the ovarian granulosa cells. As results, GDF9 and BMP15 significantly improved the meiotic resumption rate and cumulus expansion by activating ERK1/2 signaling. Treatments with GDF9 significantly improved the expression of CyclinB1 but inhibited the expression of Cx43 transcripts. In addition, cumulus expansion genes (MAPK1, Ptgs2, Tnfaip6 and Ptx3) were differentially improved by GDF9 and BMP15. In the ovarian granulosa cells, GDF9 suppressed the expression of Cx43 transcripts by binding ALK4/5/7 receptors and activation Smad2/3 signaling, whereas, BMP15 stimulated the expression of Cx43 transcripts by binding ALK2/3/6 receptors and activating Smad1/5/8 signaling. In conclusion, by regulating functions of granulosa/cumulus cells, oocyte has the potential to enhance the growth and maturation of itself.

Somatic cell nuclear transfer (SCNT) has been considered for preserving genetically valuable or endangered animals. Sapsaree is a Natianal Monument in Korea to maintian a pure pedigree. The aim of this study was to produce azoospermia Sapsaree using SCNT and identify normal reproductive ability of cloned azoospermia Sapsaree. Ear skin biopsy was performed on a thirteen-year-old azoospermia Sapsaree and ear skin fibroblasts were isolated for SCNT as donor cells. The fibroblasts were injected into enucleated in vivo matured oocytes, the couplets were electrical fused by two pulse of direct current (55 V for 15 μs) using titanium and platinum fusion needle and activated by calcium ionophore. Cloned embryos were surgically transferred into oviducts of natuarally estrus cycle synchronized recipient dogs. The fusion rate of platinum needle was 70%, which was higher than those of titanium needle (64.1%). Developmental rate to the 8 cells and 10 cell stages was higher in platinum needle group (24% and 16%, respectively) than those of platinum needle group (14.8% and 3.1%, respectively). Total 35 SCNT embryos were transferred into oviducts of 3 recipient dogs and one recipient finally delivered a puppy by caesarean section. As results, this study demonstrated that platinum fusion needle could be successfully make the reconstructed embryos and improve the efficiency of canine SCNT. Cloning azoospermia Sapsaree may contribute to conserve genetically valuable and unique pedigree. And further study should be confirm whether cloned live dog is azoospermia.

Prolonged communication between oocytes and the surrounding somatic cells is one of the unique reproductive physiology in canine. Paracrine Kit ligand (KITL) signaling is a well-known communication between granulosa cells and the oocyte. KITL is a cytokine growth factor secreted by granulosa cells that signals via the c-kit receptor expressed by oocytes. Paracrine factors, including growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15), exert their effects by binding with the kinase receptors expressed on the granulosa cells. However, the regulations of GDF9 and BMP15 in the canine KITL expression are currently poorly understood. Therefore, we investigated the effects of GDF9 and BMP15 on the expression of KITL in canine ovarian granulosa cells in vitro. In Annexin V assay recombinant GDF9 and BMP15 did not induce apoptosis in the cultured ovarian granulosa cells. When treated, FSH significantly increased KITL expression, and hCG suppressed its expression. When both FSH and hCG were treated, the expression of KITL was affected by GDF9 and BMP15 in dose and time dependent manner in the luteal granulosa cells. GDF9 (10 ng/mL) significantly decreased KITL expression after12 h. BMP15 (10 ng/mL) significantly also decreased KITL expression after 24 h. Western blot and immunochemistry results indicate that GDF9 activated Smad2/3. After blocking ALK 4/5/7 receptors by SB, GDF9 failed to activate Smad2/3, also BMP15 did not activate Smad1/5/8 after blocking ALK 2/3/6 receptors by DM. So GDF9 exerts its effects via using ALK 4/5/7 receptors to activate SMAD2/3 signaling, and BMP15 binds ALK 2/3/6 receptors to activate SMAD1/5/8 signaling. The expression of KITL was not changed by SB or DM treatment. However, the effect of GDF9 and BMP15, which decreased the expression of KITL, was suppressed by SB or DM treatment. In conclusion, this study provides the first evidence that recombinant GDF9 and BMP15 decrease KITL expression in canine ovarian granulosa cells.

The cancer and Parkinson's disease associated protein DJ-1 is multifunctional protein that involves in diverse cellular process. DJ-1 protein has a cellular protective role and promoted cell survival under an oxidative stress. However, the cellular protective mechanism of DJ-1 is not fully understand, and we needs to be further study their functions in novel organisms. In the present study, we investigated the protective role of DJ-1 against induced oxidative stress in canine cell line. On the basis of these experiments, canine DJ-1 overexpressing and null cell lines were established. The stable overexpression and down regulation of DJ-1 efficiency confirmed by the western blot analysis. Subsequently, the DJ-1 gene transfected cell lines and control cells were subjected to induced the oxidative stress, and then cell viability, cell proliferation assay, cellular apoptosis detection analysis (Annexin V and TUNEL assay), intracellular ROS and mitochondrial activity were measured appropriately. The results showed that DJ-1 overexpressed cells were up-regulated cell viability under oxidative stress conditions induced by the rotenone and hydrogen peroxide (H2O2), whereas loss of DJ-1 cells were down-regulated the cell survival activity. Additionally, overexpression of DJ-1 cells increased cell resistance to oxidative stress and inhibited the elevation of cell death and cellular ROS induced apoptosis. Moreover, DJ-1 overexpressed cells was increased mitochondrial functions by using confocal microscopy with MitoTracker staining. On the contrary to this, DJ-1 null cells show defective cellular protection and mitochondria activity against oxidative stress conditions. Our data indicate that canine DJ-1 protein attenuates cellular apoptosis and ROS generation, enhances the cellular survival activity and promote mitochondrial function under the oxidative stress, likewise other mammalian cells. Importantly, DJ-1 overexpression may be an important part of a protective strategy as a sensor for oxidative stress.

There are over 3,000 different species of mosquitoes throughout the world; currently 56 species are reported in Korea. Mosquitoes can act as vectors for many disease-causing viruses and parasites during blood sucking. Various species of mosquitoes are estimated to transmit various types of disease to more than 700 million people annually in Africa, South America, Central America, Mexico, Russia, and much of Asia, with millions of resultant deaths. At least two million people annually die of these diseases, and the morbidity rates are many times higher still. To prevent the spread of diseases, KNIH (Korea National Institute of Health) used three categorized methods; identifying or taxonomic analysis of mosquitoes, detecting virus caring mosquitoes, and detecting malaria from Anopheline mosquitoes. We have proved that taxonomic analysis using DNA barcording method (COI gene) is useful to complement identification of mosquito species. In detecting virus, we have reported Cx. orientalis as a new potential Japanese encephalitis virus (JEV) vector. We also reported that the Anopheline mosquito species composition and Plasmodium vivax infection rates in malaria hot spot in Korea.

This study was conducted from October 2014 to May 2015 to explore forage production and feed values of Italian ryegrass, Rye and whole crop barley as winter forage crops in the Southern region of Korea. The experimental location was over 10 points for each species and each sampling point area was 1 m² (Width: 1 m × Length: 1 m). Air mean temperature and rainfall in the Southern region of Korea during the experimental period was 6.95 ± 5.75℃ and 70.45 ± 54.68 mm, respectively. Fresh forage yield of Italian ryegrass, the most cultivated forage in the Southern region of Korea, was 44.4 ± 7.0 ton/ha. The percentage of dry matter for whole crop barley was 28.9 ± 7.0%. Crude protein (CP) was higher in Italian ryegrass (10.7 ± 5.3%) while total digestible nutrient (TDN) had the highest value in whole crop barley. Crude protein was not significantly different by location. However, the neutral detergent fiber (NDF), acid detergent fiber (ADF) and total digestible nutrient value of forage from Jeonbuk province were higher than in forage from Gyeongnam province.

2007년부터 2011년 사이 우리나라로 수입된 식물류에서 검출된 113종의 깍지벌레 목록을 정리하였으며, 이들 종들의 검출횟수, 기원, 기주식물 및 분포에 대한 자료도 추가하였다. 한편, 동 자료의 분석을 통해 국내 유입시 식물에 피해를 줄 것으로 예상되는 깍지벌레 종에 대한 정보를 제공하고자 한다.

For more than two decades, the intracytoplasmic sperm injection (ICSI) technique has been used as a valuable tool to provide opportunities for studying fertilization, treating human infertility, and producing transgenic animals. Not only in facilitating fertilization but also in propagating mammalian species, ICSI has enhanced the potential of assisted reproductive technologies in human. Polyspermic fertilization has been one of major problems in pig reproduction, but the ICSI helped to solve the problem, and used widely to generate transgenic piglets. Although the ICSI technique is considered to be a very useful tool in assisted reproductive technologies, including generation of transgenic animals, there are some disadvantages using the technique. In this review, we describe the ICSI technique and its application in animal production and human infertility, and discuss advantage and disadvantage of the technique in mammals.