The objective of this study was to investigate to influence of glutathione (GSH) on development and antioxidant enzyme activity in tetraploid porcine embryos. Tetraploid embryos were produced using parthenogenetic 2-cell embryo by electrofusion method. Tetraploid embryo development was observed every 24 hours and intracellular antioxidant enzyme activity was measured at 120 hours after electrofusion. The 4-cell to 16-cell stage tetraploid embryos was increased in 100 and 500 μM GSH-treated groups compared control group at 48 hours (P < 0.05) but cleavage rates were not significantly different among the GSH treatment groups at 48, 72, 96, and 120 hours. Blastocyst formation was significantly increased by 300 and 500 μM GSH at 120 hours in tetraploid embryos (P < 0.05). But blastocyst cell number were not significantly different among the GSH treatment groups (16.4 ± 0.8, 16.8 ± 2.6, 18.5 ± 2.8 and 17.5 ± 1.8). The intracellular antioxidant enzyme level was increased in 500 μM GSH compared to 0 and 100 μM GSH (P < 0.05). We suggest that GSH may be improve development of tetraploid embryo in pigs.

As a one of unsaturated fatty acid, polyunsaturated fatty acids (PUFAs) have multiple actions: as precursor of prostaglandins (PGs), steroid hormone synthesis and energy production in animal reproduction. PUFAs, which include omega-3 (n-3) and omega-6 (n-6), are derived from the diet and changed by diet, species, breed and season. The plasma membrane of spermatozoa in mammals contain various PUFAs. These composition of PUFAs regulate the membrane fluidity and cause lipid peroxidation via generation of reactive oxygen species (ROS). Induced lipid peroxidation by ROS decreased viability and motility of spermatozoa, and it is reduced by addition of antioxidant and low concentration of PUFAs. Because oocytes of animal have a high lipid components, process of oocyte maturation and embryo development are influenced by PUFAs. In in vitro study, oocyte maturation, embryo development, intracellular cAMP and MAPK activity were increased by treatment of n-3 α-linolenic acid (ALA) during maturation, whereas n-6 linoleic acid (LA) negatively influenced. Also, inhibition of fatty acid metabolism in oocyte influenced blastocyst formation of cattle. PGs are synthesized from PUFAs and various PUFAs influence PGs via regulation of PG-endoperoxide synthase (PTGS). Steroid hormone synthesis from cholesterol is regulated by expression of steroid acute regulator (StAR) protein and mRNA. Exogenous n-3 and n-6 PUFAs altered sex hormone in animal through stimulate or inhibit StAR activity. Because PUFAs altered PG and steroid hormone synthesis, follicular development was influenced by PUFAs. This effect of unsaturated fatty acid could provide information for improvement of reproductive ability in animals.

The oocyte undergoes various events during maturation and requires many substances for the maturation process. Various intracellular organelles are also involved in maturation of the oocyte. During the process glucose is essential for nuclear and cytoplasmic maturation, and adenosine triphosphate is needed for reorganization of the organelles and cytoskeleton. If mitochondrial function is lost, several developmental defects in meiotic chromosome segregation and maturation cause fertilization failure. The endoplasmic reticulum, a store for Ca2+, releases Ca2+ into the cytoplasm in response to various cellular signaling molecules. This event stimulates secretion of hormones, growth factors and antioxidants in oocyte during maturation. Also, oocyte nuclear maturation is stimulated by growth factors such as epidermal growth factor. This review summarizes roles of organelles with focus on the Golgi apparatus during maturation in oocyte.

Objective of this study was to investigate the effect of nicotinic acid (NA) on the characteristics in fresh semen of miniature pig. We evaluated viability, acrosome reaction and mitochondrial integrity of sperm on 0, 3, 7 and 10 days during storage period with nicotinic acid. As results, the survival rate of sperm in 15 mM NA (day 3, 87.8 ± 1.2%; day 5, 84.0 ± 2.7%; day 7, 82.2 ± 0.9%) and 30 mM NA (day 3, 87.7 ± 0.3%; day 5, 84.4 ± 2.5%; day 7, 82.3 ± 0.7%) groups were higher than control and 5 mM NA groups in 3, 7 and 10 days of semen storage. The NA-treated sperm on 10 day was used day for observing acrosome integrity. The survival sperm with acrosome reaction was higher in 30 mM NA group (day 3, 2.7 ± 0.2%; day 5, 3.3 ± 0.6%; day 7, 11.4 ± 0.3%) than in the control, significantly (P<0.05). Moreover, the live sperm with mitochondrial integrity was higher in whole treatment groups of NA than control group, significantly (P<0.05). Specially, most mitochondrial integrity on 10 day of semen storage was significantly higher in 30 mM NA group (90.2 ± 1.6%) than other treatment groups (control, 81.8 ± 3.1%; 5 mM NA, 83.4 ± 3.0%; 15 mM NA, 89.1 ± 0.7%, P<0.05). In conclusion, supplement of NA in liquid semen of miniature pig can improve and maintain semen quality, such as viability, acrosome reaction, and mitochondria integrity.

L-Carnitine is an antioxidant for the transport of fatty acids in mitochondria and breakdown of lipids for metabolic energy. Some studies have suggested that carnitine improves sperm motility in mammals. The objective of this study was to investigate the effect of L-carnitine on the characteristics in fresh semen of miniature pigs. The collected fresh semen was stored in modena B medium with L-carnitine (0, 1.0, 2.0, and 4.0 mg/ml) for 10 days at 18℃. The semen quality of viability, acrosome reaction and mitochondria integrity was analyzed on 0, 3, 7, and 10 day of semen storage. The percentages of live and dying sperm were not different among treatment groups with different concentrations of L-carnitine during the storage period. In acrosome reaction analysis, when the sperm stored for 7 day, the percentages of live sperm with acrosome reaction were significantly (p<0.05) lower in 1 (9.0±0.9%), 2 (7.6±0.2%) or 4mg/ml (7.9±0.8%) L-carnitine-treated groups than the control group (0 mg/ml L-carnitine) (11.12±0.2%). However, there were no difference in percentages of live sperm with acrosome reaction for 3 and 10 days of storage with each concentrations of L-carnitine. When sperm was stored for 3 and 10 days, the percentages of live sperm with mitochondria integrity were significantly higher in 2 mg/ml of L-carnitine-treated group than control group (p<0.05). In conclusion, the L-carnitine has a positive effect on acrosome reaction and mitochondria integrity in liquid state of fresh semen in miniature pigs.

We tested the identification ability of DNA barcodes comparing with morphological data using the Korean butterflies. The 921 samples (4.6 samples per species) for 202 resident Korean species except migratory species were used. The obtained samples were morphologically identified based on wing patterns. In a result, genetic divergence to the nearest-neighbouring taxon varied from 0 to 28.2%, with an average of 13.4 per cent. The neighbour joining (NJ) tree profile showed that sequence data for 185 of the 202 species formed distinct barcode clusters. Thus, our results indicated that 91.6 percent of the species were possible to allow the reliable identification using DNA barcoding. The rest 17 species (8.4%) consist of following four cases: clustering separated from each species by less than 1% branch length (two species pairs), paraphyletic clustering (two species pairs and one triple species pair), polyphyletic clustering with sharing barcodes (three species pairs), and clustering separated from existing species by the deep branch divergence (four clusters). However, it was not easy to interpret these ambiguous cases only using our current taxonomic evidences. Therefore, we are performing integrative taxonomy on these cases using other additional evidences such as examination on male genitalia and analysis of other gene regions.

In traditional taxonomy on the family Cantharidae, color pattern of the body and the structure of the male genitalia have been often used as diagnostic characters in identification of the specific level. However, these characters caused the difficulty in identifying the female in case a species was described only by male specimens or has the several color types among individuals. In this study, we attempted to evaluate the species reality of Asiopodabrus fragiliformis which was often difficult to be identified due to individual variation in color pattern and lack of information of female, through searching for new morphological diagnostic characters as well as DNA barcoding analysis, including their closely relative species from Russia and Japan. The results showed that A. fragiliformis was represented as three clusters strongly supported by high value of boots trap (>99%) and over 3% branch length. The pairwise distances between species of Asiopodabrus were detected larger, ranged from 3.4–9.5%, than the intragroup distance ranged from 0–2.9% indicating presence of a barcoding gap. And then, the three clusters were respectively determined as A. fragiliformis, A. kurvatovi and a new species through the analysis of morphology and COI gene. Therefore, we suggest that the species delineation on polymorphic species and the female specimens of closely resembling species would be more exactly and effectively determined if DNA barcoding and the traditional taxonomy are used as complementary methods for identification.

The correct development of male gametophytes (pollen grains) in flowering plants is essential for proliferate in gamete production. Here we have taken a map-based cloning approach using Arabidopsis male gametophytic mutant, named gemini pollen3 (gem3) to identify and characterize key gene that is expressed gametophytically for the completion of microgametogenesis focusing on genes which control cell division and cell fate determination. Previously reported gem1 and gem2 mutants with similar characteristics to gem3 that are disturbed at asymmetric division and cytokinesis at pollen mitosis I (PMI) in Arabidopsis. However, gem3 was mapped to a different genetic locus, and pollen developmental analysis revealed that gem3 exert an effect at meiosis and mitosis causing complete sterility. We also discovered that gem3 homozygous lines produce aberrant pollen grains, arising from incomplete cytokinesis during male meiosis with sporophytic phenotypes of twisted-shape leaves, large flowers. This mutation shows reduced genetic transmission of gem3 allele through male gametophyte. In previous results, the gem3 locus was confirmed by mapping to the region located on chromosome 5. To further confirm strong candidate gene, we performed sequencing and genetic complementation analysis. Currently, we are performing functional studies of the gem3 gene for the better understanding of molecular mechanisms that control asymmetric division at meiosis and mitosis during pollen development.

The purpose of this study was to determine the free amino acid contents, total phenolic contents and antioxidant activity in raw Sesamum indicum seeds (cv. Kopum and cv. Mihuk) and their sprouts germinated for 7 days. Total free amino acid contents of sprouts (29.34±3.3 mg/g DW) were significantly higher than that of raw seeds (6.85±0.39 mg/g DW). All individual free amino acid, including asparagine, alanin, arginine, and leicine were significantly increased in the sesame sprout. And also germinated sprout significantly increased the total phenolic contents (2.2±0.3 mg GAE/g) that resulted in the increased DPPH (2,2-diphenyl-1-picrylhydrazyl) free radical scavenging capacity. Subsequently, two varieties of sesame and its sprouts were analyzed for their phenolic constituents using high-performance liquid chromatography (HPLC). Catechin, sinapic acid and salicylic acid were identified as the major phenolic acid presented in sesame sprout. However, the major biological constituents sesamin and sesamolin content were significantly decreased during germination.

The perilla [Perilla frutescens (L.)], which belongs to the family Lamiaceae, have been used as a not only important traditional source of oil but also used traditional herbal medicine for treating various disease including depression, anxiety, tumor, cough, bacterial and fungal infections, allergy, detoxication and some intestinal disorders in east asian countries. In this context, luteolin isolated from the P. frutescens inhibited the linoleic acid peroxidation catalyzed by soybean lipoxygenase-1 (EC 1.13.11.12, Type 1) with an IC50 of 5.0 μM. To investigate the inhibitory effect of luteolin on dioxygenase enzyme, we assayed soybean lipoxygenase-1 activity with the inhibitor. Soybean lipoxygenase-1 showed time-dependent inhibition in the presence of luteolin. Increasing luteolin concentrations led to the decrease in both the initial velocity (vi) and the steady-state rate (vs) in the progress curve. Thus luteolin showed a simple reversible noncompetitive slow-binding inhibition against soybean lipoxygenase-1 with kinetic parameter (k3 = 0.056 μM-1min-1, k4,= 0.006 μM-1min-1, Ki app = 0.106 μM).

The influenza neuraminidase (E.C. 3.2.1.18) is an antiviral target of high pharmaceutical interest because of its essential role in cleaving sialic acid residues from surface glycoproteins and facilitating release of virions from infected cell. In this context, polyphenolic compounds including luteolin, rosmarinic acid, and apigenin from Perilla frutescens were evaluated for their inhibitory effect on recombinant virus H1N1 neuraminidase. Among the test compounds, luteolin and rosmarinic acid inhibited the rvH1N1 neuraminidase with an IC50 of 8.4 and 46.7 μM, respectively. The inhibition kinetics analyzed by Dixon plots indicates that luteolin and rosmarinic acid are noncompetitive inhibitor and inhibition constant, Ki, were established as 14.3 and 43.9 μM, respectively. Subsequently, we also analyzed the rosmarinic acid and luteolin contents of 383 accessions of perilla seed germplasms by high performance liquid chromatography (HPLC). The rosmarinic acid and luteolin contents of perilla seeds were ranged from 15.7 μg/g to 2,894.9 μg/g and from 1.6 μg/g to 949.1 μg/g, respectively.

This study was carried out to investigate the variation of triterpene soyasapogenol A and B in soybean cultivars. Soyasapogenol A and B were isolated from acid hydrolysis of 80% aqueous ethanol extract of soybean and the structures of these soyasapogenols were characterized by 1H and 13C NMR analysis. And the distribution of soyasapogenol A and B were determined by high-performance liquid chromatography (HPLC) with variable wavelength detector (VWD). The soyasapogenol A and B contents of soybean cultivars ranged form 49.9 ㎍/g (Geomjeongsaeol) to 701.5㎍/g (Sorog) and from 91.4㎍/g (Danmi) to 2,315.9 ㎍/g (Daewonkong), respectively. Sorog cultivar showed the highest total soyasapogenol contents (2,773.8 ㎍/g), whereas Geomjeongkong 3 cultivar displayed the lowest (83.5㎍/g). The average content ratio of soyasapogenol B (1,061.4㎍/g) was significantly higher than that of soyasapogenol A (292.5 ㎍/g).

Anthocyanins of black soybean may play an important role in physiological functions related to human health such as antioxidant, anti-inflammatory, and anticancer. The objective of this study was to investigate the profiles of anthocyanin in fourteen black soybean varieties using high-performance liquid chromatography (HPLC) with photodiode array detector (PAD). In all of the cultivars analyzed totally nine anthocyanins including, CatCy3glc, Dp3gal, Dp3glc, Cy3gal, Cy3glc, Pt3glc, Pg3glc, Pn3glc, and Cy were found. Cy3glc was the major anthocyanin content, represention 69.5% of anthocyanin, followed by Dp3glc (23.0%), Pt3glc (4.9%), Pn3glc (1.2%), and Pg3glc(1.1%), respectively. In contrast, the other five anthocyanins were in very low amounts, below 0.3 % in all varieties. Comparing cultivars and anthocyanin compositions, Geomjeong 2, Cheongja 2, and Cheongja 3 were found to content 9 anthocyanins. Geomjeong 1 and Seonheuk contented CatCy3glc, Cy3gal, Cy3glc, and Pn3glc. The variation of total anthocyanin concentration were significant for soybean cultivars. Geomjeong 2 showed highest average total anthocyanin content (17,937.8 ㎍/g seed coat), and Tawonkong and Heugcheong had the lowest total anthocyanin values (2,835.7 and 2,853.1 ㎍/g seed coat, respectively).

Nitrogen (N) fertilization is essential for alleviating nutrient deficiencies of the world’s population by increasing rice production, one of the most important food crops of our time. Here we established an in vivo hydroponics rice seedling culture system to investigate the physio-biochemical and molecular responses of various rice genotypes to low nitrogen application. Yoshida’s nutrient solution (YS) was used to grow rice seedlings, and at three-week-old the seedlings manifested highly stable and reproducible symptoms, such as reduced shoot growth and length. Out of 12 genetically selected or tested genotypes, almost all (11 genotypes) showed varied degrees of growth reduction response to applied nitrogen (4 and 40 ppm N for treatment and control, respectively), but SR19663-B-B-34-3-3-3-1 showed similar growth as the control though its leaf width was smaller than the control. The leaves of a 11 representative low nitrogen-responsive genotype as BG90-2 were sampled for revealing the protein profiles between low and normal (control) nitrogen application by using two-dimensional gel electrophoresis (2-DGE) followed by staining of separated proteins with silver. Fifty differentially expressed silver stained protein spots were excised from 2-D gels and 41 proteins identified using high-throughput mass spectrometry (MS) using matrix-assisted laser desorption/ionization-time of flight-MS and nano electrospray ionization liquid chromatography tandem MS. These proteins could be assigned as major (energy metabolism, photosynthesis and oxidative stress) and minor functional categories, revealing many novel low N-responsive proteins, including those having energy/photosynthesis, and defense/stress, and iron homeostasis-related functions.