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 analyzed a portion of mitochondrial COI gene sequences (658 bp) to investigate the genetic diversity and geographic variation of the swallowtail butterfly, Papilioxuthus L., and the cabbage butterfly, Pieris rapae (Lepidoptera: Papilionidae). P. xuthus showed a moderate level of sequence divergence (0.91% at maximum) in 15 haplotypes, whereas P. rapae showed a moderate to high level of sequence divergence (1.67% at maximum) in 30 haplotypes, compared with other relevant studies. Analyses of population genetic structure showed that most populations are not genetically differentiated in both species. The distribution pattern of both species appears to be consistent with category IV of the phylogeographic pattern sensu Avise (Avise et al. 1987): a phylogenetic continuity, an absence of regional isolation of mtDNA clones, and extensive distribution of close clones. The observed pattern of genetic diversity and geographic variation of the two butterfly species seems to reflect the abundant habitats, abundant host plants, and flying abilities in connection with the lack of historical biogeographic barriers.

We conducted a year-long survey in 2000 to examine seasonal fluctuations in the abundance of the demersal copepod Pseudodiaptomus sp., the dominant copepod in the Seomjin River estuary, where the spring tide strongly affects changes in salinity gradients.

Angelica tenuissima, also known as Ligusticum tenuissimum, is classified as a food-related plant and has been used as traditional medicines treating headache and anemia in Asia. However, its anti-melanogenic effect has not been reported in detail. When the extract of Angelica tenuissima (ATE) was prepared by the extraction with 70% EtOH at 80°C (final yield = 22%), the contents of decursin and Z-ligustilide in ATE were determined 0.06% and 8.43%, respectively. Total flavonoid and phenolic content in mg ATE were 5.52±0.07 ㎍ quercetin equivalents and 237.27±13.24 ㎍ gallic acid equivalents, respectively. Antioxidant capacity of ATE determined by DPPH and ABTS assay was increased with a dose dependent manner up to 1000 ㎍/㎖. The amount of melanin synthesis followed by α-melanocyte stimulating hormone on B16F10 cells were significantly reduced in the presence of ATE (250 to 1000 ㎍/㎖, p<0.05). ATE (125 to 1000 ㎍/㎖, p<0.05) suppressed the tyrosinase activity but did not show any significant effect on α-glucosidase activity at the same condition. Taken together, ATE possesses tyrosinase inhibitory potential with significant antioxidant capacities. These effects of ATE might be involved in suppression of melanin synthesis, at least, in B16F10 cells. The anti-melanogenic potential of ATE will provide an insight into developing a new skin whitening product.

EP was obtained through 20% ethanol extraction of Pueraria lobata root, and the fermented form of EP, FEP, was prepared from the EP after incubating with Lactobacillus rhamnosus vitaP1. There was no significant toxicity by EP and FEP up to 1000 ㎍/㎖ in NIH-3T3, HaCaT, and B16F10 cells. In addition to antioxidant potentials of EP and FEP determined by DPPH and ABST assays, we confirmed increase of procollagen type I and elastin synthesis by supplementation of the EP and FEP at the concentration of 50 ㎍/㎖ using ELISA kits. The protein expression levels of matrix metalloprotease (MMP)-1, -3, and -9, those are involved in the degradation of collagen or other skin matrix proteins, were remarkably suppressed while their inhibitory protein metallopeptidase inhibitor 1 (TIMP-1) was greatly up-regulated by supplementation of the EP and FEP at a concentration of 50 ㎍/㎖. Taken together, both EP and FEP supplementation could be involved in the suppression of the skin wrinkle formation through inhibiting degradation of collagen and stimulating the synthesis of collagen and elastin. The results showed that the anti-wrinkle potential of the EP and FEP will be a promising candidate for developing cosmeceutical compounds or products.

Background : Agrimonia pilosa (A. pilosa) Ledebour has been registered in The Korean Herbal Pharmacopeia (KHP). In the recent study, A. Coreana showed antioxidant and anti-inflammatory effect. However, Studies of components in Agrimonia coreana (A. Coreana) Nakai was not much. So, we compared A. pilosa and A. coreana by High performance liquid chromatography (HPLC). we perfomed Thin layer chlromatography (TLC) including the anatomical characteristics by using microscope. Methods and Results : The anatomical characteristics of A. pilosa were similar to them of A. coreana. But, fascicular fivers of A. Coreana was broader than it of A. pilosa. TLC were performed to identify the Rutin and Apigenin-7-glucuronide compound. In the extract of Agrimoniae Herba, they were identified on the spot of Rf 0.2, 0.4 in Ethyl acetate - Formic acid – Water (8 : 1 : 1). The Rutin and Apigenin-7-glucuronide were analysed by HPLC/UV with Thermo Column (5 μm, 4.6 × 250 mm, C18), the column temperature at 40 ℃ and a diode-array detector (DAD) seted at 255 nm and 338 nm. The mobile phase was composed of water and acetonitrile containing 0.1% formic acid with the flow rate 1 mL/min. All compounds showed good linearity (R2>0.999) within test ranges. Agrimoniae herba was extrated by four kinds of extraction methods: MeOH, 50% MeOH, EtOH and water. The highest extraction rate occurred, when it was treated with 50% Methanol for refluxing extraction (60min). Content of Rutin was found to be 0.07±0.00 mg/g in A. pilosa and 0.02±0.00 mg/g in A. coreana. Content of Apigenin-7-glucuronide was found to be 0.12±0.00 mg/g in A. pilosa and 0.11±0.00 mg/g in A. coreana. Conclusion : The anatomical characteristics of A. pilosa were similar to them of A. coreana. Contents of Rutin and Apigenin-7-glucuronide in A. pilosa was higher than them in A. coreana slightly. But there were observed the similar patterns of Agrimonia pilosa Ledebour and Agrimonia coreana Nakai on the finger print anelysis.

Exposure to ionizing radiation is regarded as a kind of abiotic stresses that can change the expression of genes in living organisms. This study aimed on investigating the variations in gene expressions induced by two different types of irradiations with different doses, which were low linear energy transfer (LET) gamma rays (100, 200, and 400 Gy) and high LET ion-beams (20, 40, and 80 Gy) on rice. RNA sequencing was carried out using the Illumina HiSeq-2500 platform. The average amount of reads were 4.8 Gb per individual, and 5 to 8% of the reads were removed after quality control. More than 90% of the RNA-seq reads were mapped to the rice reference genome sequence (IRGSP-1.0). A total of 247 differentially expressed genes (DEGs) were identified by comparison of the gene expression levels between the wildtype and the irradiated individuals. The 247 DEGs were divided into five modules and 27 intra-modular hub genes were found using the weighted correlation network analysis (WGCNA) method. The MEturquiose module had the most number of genes with 75 related to carbohydrate and small molecule metabolic processes. The co-expression network reconstructed using ARACNE (algorithm for reconstruction of accurate cellular networks) showed specific up- or down-regulation of the genes in each module according to the types and doses of radiation. This study will contribute to understanding the gene expression responses to ionizing irradiation.

Mungbean (Vigna radiata) is a fast-growing, warm-season legume crop that is primarily cultivated in developing countries of Asia. We constructed a draft genome sequence of mungbean to facilitate genome research into the subgenus Ceratotropis and to enable a better understanding of the evolution of leguminous species. The draft genome sequence covers 80% of the estimated genome, of which 50.1% consists of repetitive sequences. In total, 22,427 high confidence protein-coding genes were predicted. Based on the de novo assembly of additional wild mungbean species, the divergence of what was eventually domesticated and the sampled wild mungbean species appears to have predated domestication. Moreover, the de novo assembly of a tetraploid Vigna species (Vigna reflexo-pilosa var. glabra) provided genomic evidence of a recent allopolyploid event. To further study speciation, we compared de novo RNA-seq assemblies of 22 accessions of 18 Vigna species and protein sets of Glycine max and Cajanus cajan. The species tree was constructed by a Bayesian Markov chain Monte Carlo method using highly confident orthologs shared by all 24 accessions. The present assembly of V. radiata var. radiata will facilitate genome research and accelerate molecular breeding of the subgenus Ceratotropis.

After vasectomy (VAX), the flux and composition of the epididymal fluid are modified, leading to sequelae to the epididymis. In an effort to understand molecular pathophysiology of the epididymis following VAX, we investigated the changes of gene expression and sperm functions in the epididymis of vasectomized mice. After VAX, the epithelial cell height was significantly decreased in cauda epididymis, resembling the those of vas deference. This suggests that VAX evokes alteration of segmental characteristics of epididymal epithelium. Of note, these was an increase in luminal diameter in corpus and cauda epididymis, indicating the alteration of fluid homeostasis in epididymal lumen and protein synthesis and secretion. Also, the formation of sperm granuloma and infiltration of inflammatory cells were noted in lumen of epididymis at 8 weeks postvasectomy, indicating the activation of immune response in epididymis. The serum TNF-α levels and epididymal TNF-α and IL-1β mRNA levels were significantly increased in VAX mice. Microarray analysis demonstrated that claudin (CLDN) 10 and cystic fibrosis transmembrane conductance regulator (CFTR) were downregulated after vasectomy. In contrast, angiotensin II receptor type 2 (Agtr2) was up-regulated after vasectomy. Taking into account the functional importance of angiotensin system in epididymal epithelia and muscle tissue, VAX may alter the secretory function of epithelia and sperm transport via alteration in angiotensin system. Importantly, the IgG type of anti-sperm antibody (ASA) were markedly increased in the blood of vasectomized mice. Together this indicates that VAX provokes local inflammation in epididymis as well as systemic inflammation, which in turn change the blood epididymal barrier, an important element for epididymal immunological privilege. In addition, the number of sperm in cauda epididymis was increased but the sperm motility was decreased after vasectomy. The spontaneous acrosome reaction was increased by vasectomy after capacitation. This suggests that VAX affects sperm functions as well as sperm maturation. In conclusion, bilateral vasectomy lead to local immune response of epididymis, causing immunologic infertility in men.

The Egr family of zinc finger transcription factors is rapidly induced by various mitogens and regulates cell growth, differentiation, and apoptosis. While it is clear that loss of Egr1 leads to anovulatory infertility due to LHβ deficiency in female mice, molecular function of Egr1 in male reproduction has not been clearly investigated. Here, we demonstrate that Egr1 acts as an intrinsic transcription factor in Leydig cells to regulate their proliferation and steroidogenesis in the testis as well as an extrinsic factor for male reproduction via LHβ transcription in the pituitary. Egr1 is predominantly expressed in spermatogonia and Leydig cells in immature testes and later detected in some of these cell types in mature testes. The fertility potential of Egr1(-/-) male mice is relatively deteriorated even at 2 month-old age and aggravated with aging. The incidence of abnormalities of seminiferous tubules such as Sertoli cell only was dramatically increased with aging. The number and mean size of Leydig cells were significantly reduced in Egr1(-/-) testes. The impairment of Leydig cells is consistent with significant reduction in levels of testosterone and expression of factors critical for steroidogenesis such as StAR in Egr1(-/-) testes. Exogenous administration of hCG rapidly and transiently induced Egr1 expression in Leydig cells culture in vitro. hCG could reinstate reduced mean size of Leydig cells but not reduced number of Leydig cells and aberrantly low StAR expression, suggesting that Egr1 has critical functions for Leydig cell proliferation and their steroidgenesis. In addition, daily sperm production and in vitro fertilization (IVF) competence were significantly reduced, and apoptosis was facilitated in these mice. Furthermore, hCG administration to compensate for relatively low LH levels in Egr1(-/-) males could not restore the compromised reproductive phenotypes such as IVF competence and apoptosis in these mice. Interestingly, expression of Egr2, a member of Egr family, is significantly elevated in Egr1(-/-) Leydig cells suggesting that genetic compensation of Egr2 may alleviate phenotypic aberration of Egr1(-/-) male testes. Collectively, these results suggest that Egr1 act as an intrinsic transcription factor required for proliferation and steroidogenesis of Leydig cells to govern spermatogenesis in the testis.

VitE (tocotrienols and tocopherols) are micronutrients with antioxidant properties synthesized by photosynthetic bacteria and plants that play important roles in animal and human nutrition. A new mutant line, T1001-1, was isolated from in vitro mutagenized population by ionizing radiation and shown to have increased VitE contents. The total VitE content was 26% increased in the T1001-1 mutant seeds compare with cv. Dongan (wild-type). In addition, we showed that the mutant confers retarded seedling growth during the early seedling growth stage in rice. To study the molecular mechanism of VitE biosynthesis, we used the rice microarray to identify genes that are upor down-regulated in T1001-1 mutant. In addition, we identified differentially regulated pathway using MapMan analysis, which provides deep insight into changes in transcript and metabolites. Our results enhanced the transcription of genes involved in starch and lipid metabolism in T1001-1 mutant. To identify the molecular mechanisms of the events involving transcription factors in tocopherol accumulation, we compared the expression patterns of transcription factors. The AP2-EREBP, WRKY, C2H2 transcription factor were up-regulated, whereas the MYB family was down-regulated in T1001-1 mutant. Our results demonstrate change of important transcript in high level of VitE accumulating rice mutant.

Molecular genetic markers have been widely used as powerful tools for analyzing the genome. In particular, SSR markers have practical applications in breeding systems because they can be used in high-throughput analyses for genetic mapping, heritable diversity testing, purity analysis, and marker-assisted selection. Currently, due to technical advances in DNA sequencing, large sequence databases are available for large-scale SSR mining and marker development. Here, we describe an automated method, the SSR Finder program, for SSR discovery in the onion sequence database, and primer design for amplifying the detected SSRs. A total of 1,049 SSR primers were obtained for genetic purity testing, and 100 SSR sets were analyzed in 14 bulb onion breeding lines using clustering analysis. A total of 20 selected markers from screening of all 1,049 SSR primers, were finally applied for genetic purity testing in three breeding lines, NW1, NW9, and NW10. The initial tests showed that 15%, 71%, and 97% of individuals within NW1, NW9, and NW10, respectively, were genetically homogeneous. These markers produced using the SSR Finder will be useful for investigating the genetic purity of onion breeding lines.

Aquaporin 5 (AQP5) implicated in the generation of saliva, tears, and pulmonary secretions functions as a water-specific channel. Epididymal epithelial cells actively reabsorb water, ions and proteins. Large quantity of testicular fluid movement across the epididymal tubules generates high osmotic milieu which is important for sperm maturation. In an effort to understand the fluid homeostasis and its regulation by sex steroids in male reproductive tract, the expression of AQP5 was examined in different regions of mouse epididymis during postnatal development. The effect of androgen on the expression of epididymal AQP5 was examined in ORX model. AQP5 mRNA levels were the highest in corpus region in which drastic increase was noted during sexual maturation. Epididymal AQP5 immunoreactivity was largely found in apical as well as basal region of luminal epithelia. Moderate immunoreactivity for AQP5 was found in in smooth muscle cells in both immature and mature mice. Epididymal lumen of ORX mice showed shrinkage together with decrease in AQP5 expression. Alteration of AQP5 expression in ORX epididymis was partially recovered by androgen injection. AQP5 mRNA was induced at 10uM 5α-DHT in organ cultured epididymis. Chromatin immunoprecipitatin (ChIP) showed that 5α-DHT induced recruitment of androgen receptor (AR) to the －4635 to －4453 bp region of the AQP5 gene promoter in adult epididymis. Taken together, axial regulatory mechanism may control transcription of AQP5 along the length of epididymal tubule. Water transport through AQP5 is important for late sperm maturation and storage in epididymis. Androgen may directly induce AQP5 gene transcription via activation of AR in epididymis.

Ionizing radiation is known to cause chromosomal alterations such as inversions and deletions and affects gene expression within the plant genome. To monitor the genome-wide transcriptome changes by ionizing radiation, we used rice Affimetrix GeneChip microarray to identify genes that are up- or down regulated by gamma-ray (200 Gy, 60Co source), cosmic-ray and ion beam (40 Gy, 220 MeV carbon ion). The overall expression patterns between gamma-ray and ion beam were similar but cosmic-ray was regulated differently. Combined results from all 3 radiations identified 27 up-regulated genes and 188 down regulated genes. These results mean the induction of similar mechanism changes in treatments of gamma ray and ion beam. However the different expression in treatment of cosmic-ray might be due to the other environmental conditions. Among the commonly up- or down- regulated genes, we chose highly up- or down- regulated several genes and confirmed its regulation in response to ionizing radiation exposure by RT-PCR analysis. Moreover, we showed that specific co-expression networks of candidate radio marker genes by ARACNE algorithm. Our results present profiles of gene expression related to different ionizing radiation and marker gene to predict sensitivity to ionizing radiation, such as GS (glutelin subunit) and FBX322.

Foxi1, a forkhead family of transcription factor, in narrow and clear cells in epididymis is required for male fertility through regulating transcription of vacuolar H+-ATPase. To understand the regulation of Foxi1 gene activation in epididymis, the effects of steroids and their receptor antagonists and testicular factors on the expression of Foxi1 in epididymal segments were examined in mouse. Epididymis were sampled from adult mice following injections of ICI 182,780 (5mg/head, 2 times for 15 days), dexamethasone (DEX, 0.1,1,10ug/kg/day for 5 days) or oral administration of flutamide (FLM, 100mg/kg/day for 10 days). Otherwise, adult mice were orchidectomized (ORX), rested for 2 weeks, and received testosterone propionate(TP, 3mg/kg/day) for 7 days. In addition, adult male mice were subjected to efferent duct ligation (EDL) and epididymis was collected after 15 days. To study estrogen regulation of Foxi1 gene activation via estrogen receptor α (ESR1), Foxi1 expression was examined in ESR1 knock-out mice epididymis. Expression and subcellular localization of Foxi1 was analyzed by realtime RT-PCR and immunohistochemistry. To search transcription factor binding in the mouse Foxi1 gene promoter, in silico analysis was performed using TESS, TFSEARCH, and Gene-Regulation. ICI 182,780 significantly decreased Foxi1 mRNA levels in caput and corpus but increased in cauda epididymis. Foxi1 mRNA levels in caput epididymis of ESR1 KO mice were significantly lower than those of WT mice, but no significantly changed in corpus and cauda epididymis. Taken together, estrogen differentially regulates Foxi1 gene expression in epididymis. In ORX mice, Foxi1 mRNA levels were significantly increased in epididymis, and which was abrogated by TP. Though FLM did not significantly alter the Foxi1 mRNA levels, androgen may affect Foxi1 gene expression in epididymis. DEX significantly decreased Foxi1 mRNA levels in caput and corpus epididymis at 0.1ug/kg/day and in cauda epididymis at 1ug/kg/day, suggesting that glucocorticoid may negatively regulate Foxi1 gene expression. No significant change in Foxi1 mRNA levels was found after EDL. Foxi1 immunoreactivity was found in the nuclei of narrow cells of caput epididymis including initial segment and clear cells of corpus and cauda epididymis. Of note, in ORX mice, Foxi1-positive narrow cells and clear cells were increased, and which was abrogated by TP. In silico analysis revealed the presence of putative binding sequences for ESR1, AR, and GR in the 5’ upstream region from the Foxi1 promoter. In conclusion, the expression of Foxi1 in narrow cells in caput epididymis might be positively regulated by estrogen via ESR1, which was different from estrogen–ESR signaling in clear cells in corpus and cauda epdididymis. Androgen and glucocorticoid may negatively regulate expression of Foxi1 in all epdididymial segments.