• The Plant Pathology Journal
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• v.25 no.2
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• pp.193-198
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• 2009

A full-length cDNA of OgGRP gene encoding a glycinerich cell wall protein was isolated from wild rice (Oryza grandiglumis). Deduced amino acid sequences of OgGRP are composed of 148 amino acids (16.3 kDa), and show 85.9% homology with Osgrp-2 (Oryza sativa). RT-PCR analysis showed that RNA expression of OgGRP was regulated by defense-related signaling chemicals, such as cantharidin, endothall, jasmonic acid, wounding, or yeast extract treatment. In relation to pathogen stress, the function of OgGRP was analyzed in OgGRP over-expressing Arabidopsis thaliana. Overexpression of OgGRP in Arabidopsis contributed to moderate resistance against fungal pathogen, Botrytis cinerea, by lowering disease rate and necrosis size. In the analysis of the transgenic Arabidopsis lines to check the change of gene expression profile, induction of PR1, PR5 and PDF1.2 was confirmed. The induction seemed to be caused by the interaction of ectopic expression of OgGRP with SA-and JA-dependent signaling pathways.

• Mycobiology
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• v.45 no.4
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• pp.362-369
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• 2017

We assessed the regulation of cryparin, a class II hydrophobin, using three representative mitogen-activated protein kinase (MAPK) pathways in Cryphonectria parasitica. Mutation of the CpSlt2 gene, an ortholog of yeast SLT2 in the cell wall integrity (CWI) pathway, resulted in a dramatic decrease in cryparin production. Similarly, a mutant of the CpBck1 gene, a MAP kinase kinase kinase gene in the CWI pathway, showed decreased cryparin production. Additionally, mutation of the cpmk1 gene, an ortholog of yeast HOG1, showed decreased cryparin production. However, mutation of the cpmk2 gene, an ortholog of yeast Kss1/Fus3, showed increased cryparin production. The easy-wet phenotype and accumulation of the cryparin transcript in corresponding mutants were consistent with the cryparin production results. In silico analysis of the promoter region of the cryparin gene revealed the presence of binding motifs related to downstream transcription factors of CWI, HOG1, and pheromone responsive pathways including MADS-box- and Ste12-binding domains. Real-time reverse transcriptase PCR analyses indicated that both CpRlm1, an ortholog of yeast RLM1 in the CWI pathway, and cpst12, an ortholog of yeast STE12 in the mating pathway, showed significantly reduced transcription levels in the mutant strains showing lower cryparin production in C. prasitica. However, the transcription of CpMcm1, an ortholog of yeast MCM1, did not correlate with that of the mutant strains showing downregulation of cryparin. These results indicate that three representative MAPK pathways played a role in regulating cryparin production. However, regulation varied depending on the MAPK pathways: the CWI and HOG1 pathways were stimulatory, whereas the pheromone-responsive MAPK was repressive.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.8
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• pp.1178-1187
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• 2000

Many immunostimulating substances have been developed to improve immunity of domestic animals, although their exact mode of action and effects are not clearly defined, and they are now widely used in feed industry. Bacterial lipopolysaccharides, called endotoxin, in particular may have a profound effect not only on the immune system but also on macrophages of the reticuloendothelial system. Glucans from a variety of yeast cell wall have been shown to stimulate both specific and non-specific immune responses and to increase growth performance in pigs. Recently, there has been great interest in the role of complex carbohydrates in disease prevention and treatment. Mannanoligosaccharide is a glucomannoprotein complex derived from the cell wall of yeast. Generally, it was also known that the deficiencies of some major vitamins (vitamin A, E and C) and minerals (chromium and selenium) lead to impaired immune system and, as a result, immune function is depressed and recovery delayed. On the other hand, many researchers suggested that one possible reason for the superior performance observed in pigs fed plasma protein may be because of the presence of biologically active plasma proteins (e.g., immunoglobulins) which are known to contribute to the health of the starter pig. And, immunoglobulins present in plasma protein have been implicated as contributing to the overall immunocompetence of the newborn pig. Other immunostimulants, lactoferrin and lysozyme, mainly found in milk and egg white, have been known as having bacteriocidal and bacteriolytic effect. When considering practical use of immunostimulants, the concept of using immunostimulants is new to many people and, in most cases, it is poorly understood how and why such compounds act, and how they should be used in practice. Therefore, in order to clarify the reason for discrepancies in results, special attention should be paid to the dose/response relationship of immunostimulants and the duration of the effect.

• Asian-Australasian Journal of Animal Sciences
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• v.33 no.4
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• pp.579-587
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• 2020

Objective: This study evaluated the effect of yeast products on growth performance, visceral organ weights, endogenous enzyme activities, ileal nutrient digestibility and meat yield of broiler chickens fed diets containing autolyzed whole yeast (WY) and yeast cell walls (YCW) at varying levels of inclusion. Methods: Nine dietary treatments consisting of WY or YCW included at 0.5, 1.0, 1.5, or 2.0 g/kg diet and a control diet without yeast supplementation was used in the experiment. Each of the nine treatments was replicated six times with nine birds per replicate. Birds were housed in cages, in climate-controlled rooms and fed starter, grower and finisher diets. Results: There was an improvement (p<0.05) in body weight gain and feed conversion ratio on d 10, 24, and 35 for birds fed 1.0 to 2.0 g/kg WY or YCW diet. Small intestine weight was heavier on d 10 and 24 for birds on higher levels of WY and YCW compared to the control group. On d 10 and 24, there was a significant increase (p<0.05) in tissue protein content and pancreatic enzyme activities (trypsin and chymotrypsin) of birds on 1.5 to 2.0 g/kg WY and YCW diets compared to the control group. Compared to the control group, birds on WY (2.0 g/kg diet) and YCW (at 1.5 and 2.0 g/kg diet) had better (p<0.05) protein digestibility on d 24. On d 35, there was significant improvement (p<0.05) in percentage of carcass, absolute and relative breast weight for broiler chickens fed WY and YCW mostly at 2 g/kg diet compared to birds on the control diet. Conclusion: Supplementation of diets with autolyzed WY and YCW products especially at 1.5 to 2.0 g/kg diet improved broiler chicken performance and meat yield through their positive effects on ileal protein digestibility and pancreatic enzyme activities.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.5
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• pp.674-680
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• 2016

This study investigated the effects of feeds naturally contaminated with mycotoxins on growth performance, serum biochemical parameters, carcass traits, and splenic heat shock protein 70 (Hsp70) mRNA expression levels in broiler chickens. The efficacy of yeast cell wall (YCW) adsorbent in preventing mycotoxicosis was also evaluated. Three hundred 1-d-old Arbor Acres broiler chicks were randomly allotted to 3 treatments in completely randomized design for 42 d. Each treatment group had 5 replicate pens with 20 birds. The treatments were as follows: i) basal diet (control), ii) naturally contaminated diet (NCD), and iii) NCD+0.2% YCW adsorbent (NCDD). The NCD decreased average daily gain (ADG) (p<0.01) of 0 to 21 d, 22 to 42 d, and 0 to 42 d, and increased feed conversion ratio (p<0.01) of 22 to 42 d and 0 to 42 d. Both the breast meat percentage and thigh meat percentage of the NCD group were significantly higher (p<0.01) than that of the control group on d 21. The NCD group showed significantly increased levels of triglycerides (p<0.05) and cholesterol (p<0.05) on both d 21 and d 42 compared to the control group. However, the NCD significantly reduced (p<0.01) the high-density lipoprotein (HDL) on d 42 compared to controls. Compared with the NCD, supplementation with YCW significantly improved (p<0.01) the ADG of 0 to 21 d and 0 to 42 d, and increased (p<0.01) concentrations of HDL on d 42, and on d 21, and triglycerides (p<0.05) on d 21 and d 42. Supplementation with YCW reduced (p<0.01) the breast meat percentage, the thigh meat percentage, the concentrations of cholesterol (p<0.01) and the low-density lipoprotein (p<0.05) on d 21, and improved (p<0.01) the splenic Hsp70 mRNA expression levels compared with the NCD group. The results of this study indicated that feeding NCD for 42 d had adverse effects on broiler chickens, and that YCW might be beneficial in counteracting the effects of mycotoxins.

• Journal of Microbiology and Biotechnology
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• v.25 no.11
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• pp.1856-1862
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• 2015

In order to improve the stability of endoglucanase under thermal and acidic conditions, the endoglucanase gene was fused to the N-terminus of the Saccharomyces cerevisiae pir gene, encoding the cell wall protein PIR. The fusion gene was transformed into Pichia pastoris GS115 for expression. A resulting strain with high expression and high activity was identified by examining resistance to Geneticin 418, Congo red staining, and quantitative analysis of enzyme activity. SDS-PAGE analysis revealed that the endoglucanase was successfully displayed on the yeast cell surface. The displayed endoglucanase (DEG) showed maximum activity towards sodium carboxyl methyl cellulose at approximately 275 IU/g cell dry weight. DEG exhibited greater than 60% residual activity in the pH range 2.5-8.5, higher than free endoglucanase (FEG), which had 40% residual activity at the same pH range. The highest tolerated temperature for DEG was 70℃, much higher than that of FEG, which was approximately 50℃. Moreover, DEG showed 91.1% activity at 65℃ for 120 min, while FEG only kept 77.8% residual activity over the same period. The half-life of DEG was 270 min at 65℃, compared with only 150 min for FEG. DEG could be used repeatedly at least three times. These results suggest that the DEG has broad applications as a yeast whole-cell biocatalyst, due to its novel properties of high catalytic efficiency, acid-thermal stabilities, and reusability.

• The Plant Pathology Journal
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• v.29 no.4
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• pp.454-459
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• 2013

The Potexvirus Alternanthera mosaic virus (AltMV) has multifunctional triple gene block (TGB) proteins, among which our studies have focused on the properties of the TGB1 protein. The TGB1 of AltMV has functions including RNA binding, RNA silencing suppression, and cell-to-cell movement, and is known to form homologous interactions. The helicase domains of AltMV TGB1 were separately mutated to identify which regions are involved in homologous TGB1 interactions. The yeast two hybrid system and Bimolecular Fluorescence Complementation (BiFC) in planta were utilized to examine homologous interactions of the mutants. Helicase motif I of AltMV TGB1 was found to be critical to maintain homologous interactions. Mutations in the remaining helicase motifs did not inhibit TGB1 homologous interactions. In the absence of homologous interaction of TGB1, subcellular localization of helicase domain I mutants showed distinctively different patterns from that of WT TGB1. These results provide important information to study viral movement and replication of AltMV.

• Proceedings of the Microbiological Society of Korea Conference
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• 2000.10a
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• pp.39-45
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• 2000

It hab been proposed that CHS3-mediated chitin synthesis during the vegitative cell cycle is regulated by CHS4. To investigate direct protein-protein interaction between their coding products, we used yeast two hybrid system and found that a domain of Chs3p was responsible for interaction with Chs4p. This domain, termed MIRC3-4 (maximum interacting region of chs3p with chs4p), spans from 647 to 700 residues. It is well conserved among CHS3 homologs of various fungi such as Candida albicans, Emericella nidulans, Neurospora crassa, Magnaporthe grisea, Ustilago maydis, Glomus versiforme, Exophiala dermatitidis, Rhizopus microsporus. A series of mutaion in the MIRC3-4 resulted in no appearance of chitin ring at the early G 1 phase but did not affect chitin synthesis in the cell wall after cytokinesis. Absence of chitin ring could be caused either by delocalization of Chs3p to the septum or by improper interaction with Chs4p. To discriminate those two, not mutually exclusive, alternatives, mutants cells were immunostained with Chs3p-specific antibody. Some exhibited localization of chs3p to the septum, while others failed. These results indicate that simultaneous localization and activation Chs3p by Chs4p is required for chitin ring synthesis.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.6
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• pp.812-818
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• 2009

This study evaluated the effects of live yeast and yeast cell-wall mannan-oligosaccharide supplementation onperformance and nutrient digestibility during early lactation in cows fed a diet based on a mixture of corn silage and alfalfa hay as forage sources. Eight multiparous Holstein dairy cows (average days in milk, 27${\pm}$6) were used in a replicated 4${\times}$4 Latin square design. Diets contained 45% forage and 55% concentrate on a dry matter (DM) basis and treatments were: i) basal diet without additive (Control), ii) basal diet with 32 g/d of mannan-oligosaccharides (MOS), iii) basal diet with $1.2{\times}10^{10}$ colony forming units per day (cfu/d) of live yeast (Saccharomyces cerevisiae CNCM 1-1077; SC), and iv) basal diet with a mixture of MOS (32 g/d) and SC ($1.2{\times}10^{10}$ cfu/d; MOS+SC). Treatments had no effect (p>0.05) on DM intake and yields of milk, 3.5% fat-(FCM) and energy-corrected milk (ECM), and on milk fat percentage, body condition score and blood metabolites. Compared with the Control, only supplementation of SC resulted in numerically higher yields of FCM (41.9 vs. 40.1 kg/d) and ECM (41.8 vs. 40.3 kg/d), and milk fat percentage (3.64 vs. 3.43%). While the MOS diet had no effects on performance compared to the Control, the combination treatment MOS+SC increased milk protein percentage (p<0.05). Also, the MOS supplementation, both alone or in combination with SC, numerically increased milk fat percentage. The SC supplementation increased apparent digestibility of DM and crude protein while the MOS supplementation did not affect digestibility. Concentrations of total volatile fatty acids (VFA) and ruminal pH were similar across treatments. Overall results indicated that supplementation of MOS produced variable and inconsistent effects on rumen metabolism and performance, whereas SC supplementation improved nutrient digestibility and numerically increased FCM and ECM yields, which could not be enhanced by the combined supplementation of MOS+SC. According to our experimental condition, there was no effect of MOS alone or in combination with SC on dairy cow performance.

• Journal of Microbiology and Biotechnology
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• v.30 no.1
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• pp.62-70
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• 2020

Zygosaccharomyces rouxii is an important yeast that is required in the food fermentation process due to its high salt tolerance. In this study, the responses and resistance strategies of Z. rouxii against salt stress were investigated by performing physiological analysis at membrane level. The results showed that under salt stress, cell integrity was destroyed, and the cell wall was ruptured, which was accompanied by intracellular substance spillover. With an increase of salt concentrations, intracellular Na⁺ content increased slightly, whereas intracellular K⁺ content decreased significantly, which caused the increase of the intracellular Na⁺/K⁺ ratio. In addition, in response to salt stress, the activity of Na⁺/K⁺-ATPase increased from 0.54 to 2.14 μmol/mg protein, and the ergosterol content increased to 2.42-fold to maintain membrane stability. Analysis of cell membrane fluidity and fatty acid composition showed that cell membrane fluidity decreased and unsaturated fatty acid proportions increased, leading to a 101.21% rise in the unsaturated/saturated fatty acid ratio. The results presented in this study offer guidance in understanding the salt tolerance mechanism of Z. rouxii, and in developing new strategies to increase the industrial utilization of this species under salt stress.