Tricholoma matsutake is an ectomycorrhizal fungus, related with the host of Pinus densiflora. Most of studies on T. matsutake have focused on mycelial growth, genes and genomics, phylogenetics, symbiosis, and immune activity of this strain. T. matsutake is known for its unique fragrance in Eastern Asia. The most major component of its scent is (R)-(-)-1-octen-3-ol and is biosynthesized from the substrate linoleic acid by the sequential reaction of lipoxygenase and peroxide lyase. Here, we report for the first time the biosynthesis of (R)-(-)-1-octen-3-ol of T. matsutake using the yeast Saccharomyces cerevisiae as a host. In this study, cDNA genes correlated with these reactions were cloned from T. matsutake, and expression studies of theses genes were carried out in the yeast Saccharomyces cerevisiae. The product of these genes expression study was carried out with Western blotting. The biosynthesis of (R)-(-)-1-octen-3-ol of T. matsutake in recombinant Saccharomyces cerevisiae was subsequently identified with GC-MS chromatography analysis. The biosynthesis of (R)-(-)-1-octen-3-ol with S. cerevisiae represents a significant step forward.
This study was performed to determine the effects of dietary fat sources, i.e., beef tallow, soybean oil, olive oil and coconut oil (each 3% in feed), on the growth performance, meat quality and gene expression in growing-finishing pigs. A total of 72 crossbred pigs (Landrace${\times}$Large White${\times}$Duroc) were used at $71{\pm}1$ kg body weight (about 130 d of age) in 24 pens ($320{\times}150$ cm) in a confined pig house (three pigs per pen) with six replicate pens per treatment. The growing diet was given for periods of $14{\pm}3$ d and the finishing diet was given for periods of $28{\pm}3$ d. The fat type had no significant effect either on growth performance or on chemical composition or on meat quality in growing-finishing pigs. Dietary fat type affected fatty acid composition, with higher levels of unsaturated fatty acids (UFAs) and monounsaturated fatty acids (MUFAs) in the olive oil group. Microarray analysis in the Longissimus dorsi identified 6 genes, related to insulin signaling pathway, that were differentially expressed among the different feed groups. Real time-PCR was conducted on the six genes in the longissimus dorsi muscle (LM). In particular, the genes encoding the protein kinase, cAMP-dependent, regulatory, type II, alpha (PRKAR2A) and the catalytic subunit of protein phosphatase 1, beta isoform (PPP1CB) showed the highest expression level in the olive oil group (respectively, p<0.05, p<0.001). The results of this study indicate that the type of dietary fat affects fatty acid composition and insulin signaling-related gene expression in the LM of pigs.
Background: Panax ginseng is an important crop in Asian countries given its pharmaceutical uses. It is usually harvested after 4-6 years of cultivation. However, various abiotic stresses have led to its quality reduction. One of the stress causes is high content of heavy metal in ginseng cultivation area. Plant growth-promoting rhizobacteria (PGPR) can play a role in healthy growth of plants. It has been considered as a new trend for supporting the growth of many crops in heavy metal occupied areas, such as Aluminum (Al). Methods: In vitro screening of the plant growth promoting activities of five tested strains were detected. Surface-disinfected 2-year-old ginseng seedlings were dipping in Rhizobium panacihumi DCY116T suspensions for 15 min and cultured in pots for investigating Al resistance of P. ginseng. The harvesting was carried out 10 days after Al treatment. We then examined H2O2, proline, total soluble sugar, and total phenolic contents. We also checked the expressions of related genes (PgCAT, PgAPX, and PgP5CS) of reactive oxygen species scavenging response and pyrroline-5-carboxylate synthetase by reverse transcription polymerase chain reaction (RT-PCR) method. Results: Among five tested strains isolated from ginseng-cultivated soil, R. panacihumi DCY116T was chosen as the potential PGPR candidate for further study. Ginseng seedlings treated with R. panacihumi DCY116T produced higher biomass, proline, total phenolic, total soluble sugar contents, and related gene expressions but decreased H2O2 level than nonbacterized Al-stressed seedlings. Conclusion: R. panacihumi DCY116T can be used as potential PGPR and "plant strengthener" for future cultivation of ginseng or other crops/plants that are grown in regions with heavy metal exposure.
Objective: This study was conducted to investigate the effects of age and diet forms on growth-development patterns, serum metabolism indicators, and parameters of body fat deposition in Cherry Valley ducks. Methods: According to the hatching age and initial weight, a total of 150 1-day-old male SM3 Cherry Valley ducks were randomly assigned to two diet forms (pellet vs powder form). Each treatment had with 5 replicates per treatment and 15 meat ducks per replicate. The study lasted 42 d, which was divided into two periods (1 to 21 vs 22 to 42 d). Results: Our results showed that compared with powder group, ducks in pellet group had greater growth performance during different period (p<0.05). The inflection point was 24 d and was not numerically affected by diet forms. Increasing age (42 vs 21 d) significantly increased the weight of body fat and hepatic fat metabolism related enzyme activities in ducks (p<0.05), meanwhile, increasing age (42 vs 21 d) improved serum metabolism indicators and decreased mRNA expression levels of fat metabolism-related genes in liver (p<0.05). Ducks fed different diets (pellet vs powder form) increased growth performance as well as the weight of body fat and improved serum metabolism indicators (p<0.05). In addition, interactions were found between age and diet forms on the levels of serum metabolism indicators in ducks (p<0.05). Conclusion: In conclusion, powder feed reduced growth performance of ducks, and the day of inflection point was 24 days old. Ducks with higher age or fed with pellet diet showed higher fat deposition. The effect of age and feed forms on body fat deposition might result from changes in the contents of serum metabolism indicators, key enzyme activity of lipid production, and hepatic gene expressions.
Kim, Dong Hwi;Yong, Hyo Jeong;Mander, Sunam;Nguyen, Huong Thi;Nguyen, Lan Phuong;Park, Hee-Kyung;Cha, Hyo Kyeong;Kim, Won-Ki;Hwang, Jong-Ik
Biomolecules & Therapeutics
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제29권3호
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pp.331-341
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2021
Liver cancer is a common tumor and currently the second leading cause of cancer-related mortality globally. Liver cancer is highly related to inflammation as more than 90% of liver cancer arises in the context of hepatic inflammation, such as hepatitis B virus and hepatitis C virus infection. Despite significant improvements in the therapeutic modalities for liver cancer, patient prognosis is not satisfactory due to the limited efficacy of current drug therapies in anti-metastatic activity. Therefore, developing new effective anti-cancer agents with anti-metastatic activity is important for the treatment of liver cancer. In this study, SP-8356, a verbenone derivative with anti-inflammatory activity, was investigated for its effect on the growth and migration of liver cancer cells. Our findings demonstrated that SP-8356 inhibits the proliferation of liver cancer cells by inducing apoptosis and suppressing the mobility and invasion ability of liver cancer cells. Functional studies revealed that SP-8356 inhibits the mitogen-activated protein kinase and nuclear factor-kappa B signaling pathways, which are related to cell proliferation and metastasis, resulting in the downregulation of metastasis-related genes. Moreover, using an orthotopic liver cancer model, tumor growth was significantly decreased following treatment with SP-8356. Thus, this study suggests that SP-8356 may be a potential agent for the treatment of liver cancer with multimodal regulation.
To investigate the effect of GyeongshinhaeGihwan 1(GGT1) frequently used as an anti-obesity herbal medicine in oriental medicine on the expression of obesity-related genes, we measured the changes in mRNA levels of these genes by GGT1 in human growth hormone transgenic (hGHTg) obese female rats, and these effects by GGT1 were compared with those of reductil (RD), an anti-obesity drug approved by FDA. Rats received once daily oral administrations of autoclaved water, RD, or GGT1 for 8 weeks. At the end of study, rats were sacrificed and tissues were harvested. Total RNA from adipose tissue, liver and kidney was prepared and the mRNA levels for LPL (lipoprotein lipase), $PPAR{\gamma}$ (peroxisome proliferator activated receptor-gamma), $PPAR{\delta}$ (peroxisome proliferator activated receptor-delta), leptin, $TNF{\alpha}$ (tumor necrosis factor-alpha), and internal standard G3PDH (glyceraldehyde-3-phosphate dehydrogenase) were analyzed by RT-PCR. Compared with control group, $PPAR{\gamma}$ mRNA levels of liver and kidney were decreased in both RD and GGT1 groups, and the effects were more prominent in GGT1 group than in RD group, suggesting that GGT1 is effective in the inhibition of lipid storage by decreasing the $PPAR{\gamma}$ expression. $PPAR{\delta}$ mRNA levels of adipose tissue were increased by RD and GGT1 compared with DW, and the magnitude of increase were higher in GGT1 group than in RD group, indicating that GGT1 stimulates fatty acid oxidation and energy metabolism by activating $PPAR{\delta}$ expression. GGT1 group had higher concentrations of serum leptin, a well-known inhibitor of appetite, than control and RD groups. However, The mRNA levels of leptin, LPL, and $TNF{\alpha}$ were not changed by GGT1. These results indicate that GGT1 can prevent obesity in hGHTg obese female rats by down-regulating and up-regulating the mRNA expression of $PPAR{\gamma}$ and $PPAR{\delta}$, respectively, and that this anti-obesity effects were more pronounced in GGT1 group compared with RD group. In addition, GGT1 seems to inhibit obesity by increasing the circulating leptin levels.
Background: Korean ginseng (Panax ginseng Meyer) is a perennial herb prone to various root diseases, with Phytophthora cactorum being considered one of the most dreaded pathogens. P. cactorum causes foliar blight and root rot. Although chemical pesticides are available for disease control, attention has been shifted to viable, eco-friendly, and cost-effective biological means such as plant growth-promoting rhizobacteria (PGPR) for control of diseases. Methods: Native Bacillus amyloliquefaciens strain HK34 was isolated from wild ginseng and assessed as a biological control agent for ginseng. Leaves from plants treated with HK34 were analyzed for induced systemic resistance (ISR) against P. cactorum in square plate assay. Treated plants were verified for differential expression of defense-related marker genes using quantitative reverse transcription polymerase chain reaction. Results: A total of 78 native rhizosphere bacilli from wild P. ginseng were isolated. One of the root-associated bacteria identified as B. amyloliquefaciens strain HK34 effectively induced resistance against P. cactorum when applied as soil drench once (99.1% disease control) and as a priming treatment two times in the early stages (83.9% disease control). A similar result was observed in the leaf samples of plants under field conditions, where the percentage of disease control was 85.6%. Significant upregulation of the genes PgPR10, PgPR5, and PgCAT in the leaves of plants treated with HK34 was observed against P. cactorum compared with untreated controls and only pathogen-treated plants. Conclusion: The results of this study indicate HK34 as a potential biocontrol agent eliciting ISR in ginseng against P. cactorum.
Kim, Joonki;Lee, Hye-Jung;Nogoy, Franz Marielle;Yu, Dal-A;Kim, Me-Sun;Kang, Kwon-Kyoo;Nou, Illsup;Cho, Yong-Gu
Journal of Plant Biotechnology
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제43권3호
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pp.332-340
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2016
Seed size traits are controlled by multiple genes in crops and determine grain yield, quality and appearance. However, the molecular mechanisms controlling the size of plant seeds remain unclear. We performed functional analysis of BrPATL4 encoding Sec14-like protein to determine the genetic architecture of seed size, shape and their association analyses. We used 60 $T_3$ transgenic rice lines to evaluate seed length, seed width and seed height as seed size traits, and the ratios of these values as seed shape traits. Pleiotropic effects on general architecture included small seed size, erect panicles, decreased grain weight, reduced plant height and increased sterility, which are common to other mutants deficient in gibberellic acid (GA) biosynthesis. To test whether BrPATL4 overexpression is deleterious for GA signal transduction, we compared the relative expression of GA related gene and the growth rate of second leaf sheath supplied with exogenous $GA_3$. Overexpression of BrPATL4 did not affect GA biosynthesis or signaling pathway, with the same response shown under GA treatment compared to the wild type. However, the causal genes for the small seed phenotype (D1, SRS1, and SRS5) and the erection of panicles showed significantly decreased levels in mRNA accumulation compared to the wild type. These results suggest that the overexpression of BrPATL4 can control seed size through the suppression of those genes related to seed size regulation. Although the molecular function of BrPATL4 is not clear for small seed and erect panicles of BrPALT4 overexpression line, this study provides some clues about the genetic engineering of rice seed architecture.
Kim, Hyungkuen;Jeon, Eek Hyung;Park, Byung-Chul;Kim, Sung-Jo
Asian-Australasian Journal of Animal Sciences
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제32권11호
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pp.1789-1800
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2019
Objective: Although alveolar macrophages play a key role in the respiratory immunity of livestock, studies on the mechanism of differentiation and survival of alveolar macrophages are lacking. Therefore, we undertook to investigate changes in the lipid metabolism and survival rate, using 3D4/31 macrophages and Dudleya brittonii which has been used as a traditional asthma treatment. Methods: 3D4/31 macrophages were used as the in vitro porcine alveolar macrophages model. The cells were activated by exposure to phorbol 12-myristate 13-acetate (PMA). Dudleya brittonii extraction was performed with distilled water. For evaluating the cell survival rate, we performed the water-soluble tetrazolium salt cell viability assay and growth curve analysis. To confirm cell death, cell cycle and intracellular reactive oxygen species (ROS) levels were measured using flow cytometric analysis by applying fluorescence dye dichlorofluorescein diacetate and propidium iodide. Furthermore, we also evaluated cellular lipid accumulation with oil red O staining, and fatty acid synthesis related genes expression levels using quantitative polymerase chain reaction (qPCR) with SYBR green dye. Glycolysis, fatty acid oxidation, and tricarboxylic acid (TCA) cycle related gene expression levels were measured using qPCR after exposure to Dudleya brittonii extract (DB) for 12 h. Results: The ROS production and cell death were induced by PMA treatment, and exposure to DB reduced the PMA induced downregulation of cell survival. The PMA and DB treatments upregulated the lipid accumulation, with corresponding increase in the acetyl-CoA carboxylase alpha, fatty acid synthase mRNA expressions. DB-PMA co-treatment reduced the glycolysis genes expression, but increased the expressions of fatty acid oxidation and TCA cycle genes. Conclusion: This study provides new insights and directions for further research relating to the immunity of porcine respiratory system, by employing a model based on alveolar macrophages and natural materials.
This study identified hepatic differentially expressed genes (DEGs) affecting the marbling of muscle. Most dietary nutrients bypass the liver and produce plasma lipoproteins. These plasma lipoproteins transport free fatty acids to the target tissue, adipose tissue and muscle. We examined hepatic genes differentially expressed in a differential-display reverse transcription-polymerase chain reaction (ddRT-PCR) analysis comparing high- and low-marbled Hanwoo steers. Using 60 arbitrary primers, we found 13 candidate genes that were upregulated and five candidate genes that were downregulated in the livers of high-marbled Hanwoo steers compared to low-marbled individuals. A BLAST search for the 18 DEGs revealed that 14 were well characterized, while four were not annotated. We examined four DEGs: ATP synthase F0, complement component CD, insulin-like growth factor binding protein-3 (IGFBP3) and phosphatidylethanolamine binding protein (PEBP). Of these, only two genes (complement component CD and IGFBP3) were differentially expressed at p<0.05 between the livers of high- and low-marbled individuals. The mean mRNA levels of the PEBP and ATP synthase F0 genes did not differ significantly between the livers of high- and low-marbled individuals. Moreover, these DEGs showed very high inter-individual variation in expression. These informative DEGs were assigned to the bovine chromosome in a BLAST search of MS marker subsets and the bovine genome sequence. Genes related to energy metabolism (ATP synthase F0, ketohexokinase, electron-transfer flavoprotein-ubiquinone oxidoreductase and NADH hydrogenase) were assigned to BTA 1, 11, 17, and 22, respectively. Syntaxin, IGFBP3, decorin, the bax inhibitor gene and the PEBP gene were assigned to BTA 3, 4, 5, 5, and 17, respectively. In this study, the in silico physical maps provided information on the specific location of candidate genes associated with economic traits in cattle.
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