• Title/Summary/Keyword: Osmotic stress

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Proteomic and Phenotypic Analyses of a Putative Glycerol-3-Phosphate Dehydrogenase Required for Virulence in Acidovorax citrulli

  • Kim, Minyoung;Lee, Jongchan;Heo, Lynn;Lee, Sang Jun;Han, Sang-Wook
    • The Plant Pathology Journal
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    • v.37 no.1
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    • pp.36-46
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    • 2021
  • Acidovorax citrulli (Ac) is the causal agent of bacterial fruit blotch (BFB) in watermelon, a disease that poses a serious threat to watermelon production. Because of the lack of resistant cultivars against BFB, virulence factors or mechanisms need to be elucidated to control the disease. Glycerol-3-phosphate dehydrogenase is the enzyme involved in glycerol production from glucose during glycolysis. In this study, we report the functions of a putative glycerol-3-phosphate dehydrogenase in Ac (GlpdAc) using comparative proteomic analysis and phenotypic observation. A glpdAc knockout mutant, AcΔglpdAc(EV), lost virulence against watermelon in two pathogenicity tests. The putative 3D structure and amino acid sequence of GlpdAc showed high similarity with glycerol-3-phosphate dehydrogenases from other bacteria. Comparative proteomic analysis revealed that many proteins related to various metabolic pathways, including carbohydrate metabolism, were affected by GlpdAc. Although AcΔglpdAc(EV) could not use glucose as a sole carbon source, it showed growth in the presence of glycerol, indicating that GlpdAc is involved in glycolysis. AcΔglpdAc(EV) also displayed higher cell-to-cell aggregation than the wild-type bacteria, and tolerance to osmotic stress and ciprofloxacin was reduced and enhanced in the mutant, respectively. These results indicate that GlpdAc is involved in glycerol metabolism and other mechanisms, including virulence, demonstrating that the protein has pleiotropic effects. Our study expands the understanding of the functions of proteins associated with virulence in Ac.

The CsSTE50 Adaptor Protein in Mitogen-Activated Protein Kinase Cascades Is Essential for Pepper Anthracnose Disease of Colletotrichum scovillei

  • Jong-Hwan, Shin;Byung-Seong, Park;Kyoung Su, Kim
    • The Plant Pathology Journal
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    • v.38 no.6
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    • pp.593-602
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    • 2022
  • Anthracnose, caused by the ascomycete fungus Colletotrichum scovillei, is a destructive disease in pepper. The fungus germinates and develops an infection structure called an appressorium on the plant surface. Several signaling cascades, including cAMP-mediated signaling and mitogen-activated protein kinase (MAPK) cascades, are involved in fungal development and pathogenicity in plant pathogenic fungi, but this has not been well studied in the fruit-infecting fungus C. scovillei. Ste50 is an adaptor protein interacting with multiple upstream components to activate the MAPK cascades. Here, we characterized the CsSTE50 gene of C. scovillei, a homolog of Magnaporthe oryzae MST50 that functions in MAPK cascades, by gene knockout. The knockout mutant ΔCsste50 had pleiotropic phenotypes in development and pathogenicity. Compared with the wild-type, the mutants grew faster and produced more conidia on regular agar but were more sensitive to osmotic stress. On artificial and plant surfaces, the conidia of the mutant showed significantly reduced germination and failed to form appressoria. The mutant was completely non-pathogenic on pepper fruits with or without wounds, indicating that pre-penetration and invasive growth were both defective in the mutant. Our results show that the adaptor protein CsSTE50 plays a role in vegetative growth, conidiation, germination, appressorium formation, and pathogenicity in C. scovillei.

Profiling Metabolites Expressed Corn Root Under Waterlogging

  • Jae-Han Son;Young-Sam Go;Hwan-Hee Bae;Kyeong-Min Kang;Beom-Young Son;Seonghyu Shin;Tae-Wook Jung
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.289-289
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    • 2022
  • Waterlogging tolerance of corn is one of the important factor for cultivate in paddy soil condition to increase cultivation area and self-sufficiency of corn in Korea. In order to develop elite waterlogging tolerance corn, the new corn lines bred by crossing wild corn, Teosinte, and cultivated corn inbred lines. Five accessions among the 2 species, Zea mays sub spp. mexicana and Zea mays spp. parviglumis, of 81 Teosinte were selected through the waterlogging treatment. The waterlogging treatments were implemented for 7 days at the seedling(V3) stage. The inbred lines were developed by crossing 5 teosinte accessions and cultivated corn lines and they were estimated waterlogging tolerance. It was screened and analyzed the metabolites extracted from roots of 19KT-32(KS141 × teosinte) that was treated waterlogging. We selected 8 of 180 metabolites like as γ-aminobutyric acid(GABA), putrescine, citrulline, Gly, and Ala that expression was remarkably changed over 2.5-times, 7 metabolites increased and 1 metabolite decreased in waterlogging, respectively. Glutamate decarboxylase(GAD) catalyzing GABA accumulation gene have 10 haplotypes, and exon1 was highly conserved, but identified to 135 SNPs after the first intron. Among the 135 SNPs, the number of transversion mutations (52) surpassed the number of transition mutations (38). Most of metabolites were related to abiotic stress in plant that it regulated to pH, osmotic pressure K+/Ca++ and ATPase activity. We are analyzing the association using these results for increase breeding efficiency.

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Importance and Application of Amphibian Sperm Cryopreservation (양서류 정자 동결보존 기술 개발의 중요성 및 적용)

  • Ji-Eun Lee;Jun-Kyu Park;Yuno Do
    • Journal of Wetlands Research
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    • v.25 no.4
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    • pp.257-266
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    • 2023
  • Amphibian populations are declining globally, pushing many species to the brink of extinction. To promote biodiversity and sustainable management, countries are actively researching amphibian reproductive ecology. Sperm cryopreservation is a crucial assisted reproductive technology that aids in preserving the genetic diversity of amphibians. However, because amphibian sperm cells are sensitive to osmotic stress, the optimal cryopreservation method therefore differs from species to species. This literature review offers an overview of the significance of developing cryopreservation techniques for amphibian conservation and highlights the need to create optimal cryopreservation methods and the introduction of long-term monitoring (e.g., fertilization success and offspring reproduction) to advance cryopreservation technology development. This review can be used as basic research data for amphibian conservation methods.

Effect of Soil Salinity on Nitrate Accumulation of Lettuce (토양 염류집적이 상추의 Nitrate 축적에 미치는 영향)

  • Jin, Sun-Jae;Cho, Hyun-Jong;Chung, Jong-Bae
    • Korean Journal of Soil Science and Fertilizer
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    • v.37 no.2
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    • pp.91-96
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    • 2004
  • Accumulation of nitrate in edible crops is undesirable due to potential risks to human health. Since nitrate has a role in the osmotic regulation of plants, salt accumulation in soil is expected to stimulate nitrate accumulation in plants. Lettuce (Lactuca sativa L.) was grown in soils of different salinities, 9.69 and $4.49dS\;m^{-1}$, in a greenhouse, and the effect of soil salinity on nitrate accumulation in lettuce was investigated. Content of nitrate in the lettuce increased significantly as soil salinity increased under low light intensity and ample supply of nitrate in root media. Soluble sugar and oxalate contents in lettuce were also significantly higher in the soil of higher salinity. Phosphate, Cl, and $SO_4$ contents in lettuce were not significantly different in soils of different salinities. Among the cations, K content in lettuce was significantly higher in the soil of higher salinity, but Na, Ca, and Mg comtents were not much influenced. Comparing to the lettuce grown in low salinity soil, although the growth of lettuce was decreased by 9% in the soil of higher salinity, nitrate accumulation in the lettuce was increased by 18.6%. These results indicate that higher nitrate content in lettuce of higher salinity soil is a positive accumulation to adapt to the water stress condition. The nitrate accumulation of vegetables grown in plastic film houses is known to be due to the heavy fertilization and low light intensity, but salt accumulation in the soil, which can lower soil water potential, is expected to stimulate the nitrate accumulation further.

Change of Quality of Life in Children with ADHD after 12 Weeks OROS-Methylphenidate Treatment (12주 OROS-Methylphenidate 약물 치료 후 ADHD 아동의 삶의 질 변화)

  • Kim, Hyung-In;Kim, Bung-Nyun;Cho, Soo-Churl;Shin, Min-Sup;Yoo, Hee-Jeong;Kim, Jae-Won;Kim, Ji-Hoon;Son, Jung-Woo;Shin, Yun-Mi;Chung, Un-Sun;Han, Doug-Hyun
    • Journal of the Korean Academy of Child and Adolescent Psychiatry
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    • v.23 no.3
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    • pp.103-108
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    • 2012
  • Objectives : The aim of this study is to investigate the effectiveness of treatment with osmotic-release oral system methylphenidate (OROS-MPH) on quality of life (QOL) in children with attention-deficit hyperactivity disorder (ADHD). Another aim is to assess the relationship between change in QOL and other factors including children's symptoms and academic performance or parents' depression and parenting stress. Methods : A total of 111 medication-naive children with ADHD in a multicenter, open-label, 12-week trial of OROS-MPH completed an evaluation using diverse rating scales at two time points; at baseline and after 12 weeks of treatment. Scales for investigation of children included the Parent Report Form-Children's Health and Illness Profile-Children's Edition (PRF-CHIP-CE) on QOL, the ADHD Rating Scale-IV on symptoms, and the Academic Performance Rating Scale (APRS). The Beck Depression Inventory and Parenting Stress Index were used for assessment of their parents. Results : Total scores for mean PRF-CHIP-CE increased from $207.9{\pm}26.7$ at baseline to $226.3{\pm}25.9$ after 12 weeks of treatment (p<.001). The change of APRS showed the strongest correlation with the increment of PRF-CHIP-CE scores (Pearson coefficient=0.561, p<.001), even after controlling for other factors (partial correlation coefficient=0.420, p<.001). Conclusion : Treatment with MPH-OROS results in improvement of QOL in children with ADHD in Korea. The advance in academic performance plays a key role in this change of QOL.

Effect of Water Temperature, Salinity and Anaesthetic of Olive Flounder (Paralichthys olivaceus) to Live Transportation (넙치(Paralichthys olivaceus)의 활어 수송 시 수온, 염분 및 마취제 영향)

  • Yong Hyun, Do;Jae-Hye, Song;Si-Woo, Lee;Jung Yeol, Park;Jun Wook, Hur
    • Journal of Marine Life Science
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    • v.7 no.2
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    • pp.205-212
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    • 2022
  • Transport is essential in the farming process of farmed fish and is one of the physical stress factors such as sorting. The effect of water temperature and anesthesia during low salinity transport was confirmed. In the experimental group, the water temperature was set to 20℃ (natural water temperature, NWT), 15℃ (cooling water temperature, CWT) respectively, in water with a salinity concentration of 35‰, 15‰ and an anesthetic (anesthesia, Anes., Sigma USA) was diluted and mixed to 50 ppm. A styrofoam box (66×42×20 cm) was used as a transport container, and 8 flounder were accommodated and transported in a plastic bag injected with 3 ℓ of seawater and liquid oxygen. As a result of the study, the concentration of cortisol before transport increased significantly from 2.4±0.1 ng ml-1 in the experimental groups except for the CWT+35‰ group (16.7±12.8 ng ml-1). The K+ concentration slightly increased from 3.1±0.0 mEq l-1 before transport to 4.5±1.1 mEq l-1 in the NWT+15‰ group, showing no difference, and significantly increased in all other experimental groups. There was no effect on changes in blood characteristics, and water temperature and anesthetic had a negative effect on osmotic control due to stress. AST and ALT were not affected.

Hypertonicity Down-regulates the $1{\alpha},25(OH)_2$ Vitamin $D_3$-induced Osteoclastogenesis Via the Modulation of RANKL Expression in Osteoblast

  • Jeong, Hyun-Joo;Yushun, Tian;Kim, Bo-Hye;Nam, Mi-Young;Lee, Hyun-A;Yoo, Yun-Jung;Seo, Jeong-Taeg;Shin, Dong-Min;Ohk, Seung-Ho;Lee, Syng-Ill
    • International Journal of Oral Biology
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    • v.30 no.1
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    • pp.23-30
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    • 2005
  • Bone remodeling is a process controlled by the action of two major bone cells; the bone forming osteoblast and the bone resorbing osteoclast. In the process of osteoclastogenesis, stromal cells and osteoblast produce RANKL, OPG, and M-CSF, which in turn regulate the osteoclastogenesis. During the bone resorption by activated osteoclasts, extracellular $Ca^{2+}/{PO_4}^{2-}$ concentration and degraded organic materials goes up, providing the hypertonic microenvironment. In this study, we tested the effects of hypertonicity due to the degraded organic materials on osteoclastogenesis in co-culture system. It was examined the cellular response of osteoblastic cell in terms of osteoclastogenesis by applying the sucrose, and mannitol, as a substitute of degraded organic materials to co-culture system. Apart from the sucrose, mannitol, and NaCl was tested to be compared to the effect of organic osmotic particles. The addition of sucrose and mannitol (25, 50, 100, 150, or 200 mM) to co-culture medium inhibited the number of tartrate-resistant acid phosphatase (TRAP) positive multinucleated cells induced by 10 nM $1{\alpha},25(OH)_2vitaminD_3$ ($1{\alpha},25(OH)_2D_3$). However, NaCl did exert harmful effect upon the cells in this co-culture system, which is attributed to DNA damage in high concentration of NaCl. To further investigate the mechanism by which hypertonicity inhibits $1{\alpha},25(OH)_2D_3$-induced osteoclastogenesis, the mRNA expressions of receptor activator of nuclear factor (NF)-kB ligand (RANKL) and osteoprotegerin (OPG) were monitored by RT-PCR. In the presence of sucrose (50 mM), RANKL mRNA expression was decreased in a dose-dependent manner, while the change in OPG and M-CSF mRNA were not occurred in significantly. The RANKL mRNA expression was inhibited for 48 hours in the presence of sucrose (50 mM), but such a decrement recovered after 72 hours. However, there were no considerable changes in the expression of OPG and M-CSF mRNA. Conclusively, these findings strongly suggest that hypertonic stress down-regulates $1{\alpha},25(OH)_2D_3$-induced osteoclastogenesis via RANKL signal pathway in osteoblastic cell, and may playa pivotal role as a regulator that modulates osteoclastogenesis.

Influence of denitrified biofloc water on the survival rate and physiological characteristics of Pacific white shrimp juveniles, Litopenaeus vannamei (바이오플락 탈질수가 어린 흰다리새우, Litopenaeus vannamei의 생존율 및 생리특성에 미치는 영향)

  • Kim, Su-Kyoung;Jang, Jin Woo;Jo, Yong Rok;Kim, Jun-Hwan;Kim, Su Kyoung
    • Korean Journal of Environmental Biology
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    • v.37 no.2
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    • pp.136-143
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    • 2019
  • This study investigates the effect of denitrified biofloc water on changes in the water quality parameters and the physiological characteristics of shrimps. Biofloc rearing water contains a large number of microorganisms and can rapidly stabilize the water quality and energy saving if reusable due to high water temperatures. Rearing water contain floating bacteria with both aerobic and anaerobic bacteria. Therefore, when the carbon source is added in limited air supply, the anaerobic state is activated and the denitrification process is possible. In this study, the denitrification water had the following properties: ammonia (6.9 mg L-1), nitrite (0.3 mg L-1), nitrate concentration (9.2 mg L-1), high pH (8.42) and alkalinity (590 mg L-1). The experimental group consisted of seawater (SW, control), a mixture of Seawater and denitrified biofloc water (DNW) in the ratio of 3:1, 1:1 and DNW only. All experiments were done in triplicate. As a result, the survival rate never changed even when 100% of the denitrification water was utilized. However, a body fluid analysis showed that creatine and BUN were increased due to index of stress and the tissue damage resulting from the high denitrified water content. Body fluid ions (Na+, K+, and Cl-) significantly decreased as the denitrified water content increased. It was recommended that the denitrification water be mixed with a certain ratio (less than 50%) in the future as it may affect the osmotic pressure control in shrimps.

Seminal Plasma Heparin Binding Proteins Improve Semen Quality by Reducing Oxidative Stress during Cryopreservation of Cattle Bull Semen

  • Patel, Maulikkumar;Gandotra, Vinod K.;Cheema, Ranjna S.;Bansal, Amrit K.;Kumar, Ajeet
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.9
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    • pp.1247-1255
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    • 2016
  • Heparin binding proteins (HBPs) are produced by accessory glands. These are secreted into the seminal fluid, bind to the spermatozoa at the time of ejaculation, favour capacitation, acrosome reaction, and alter the immune system response toward the sperm. The present study was conducted with an objective to assess the effect of purified seminal plasma-HBPs (SP-HBPs) on cross bred cattle bull sperm attributes during two phases of cryopreservation: Pre freezing and freezing-thawing. SP-HBPs were purified from pooled seminal plasma by heparin affinity chromatography. Three doses of SP-HBPs i.e. 10, 20, $40{\mu}g/mLs$ semen were standardized to find out the optimum dose and $20{\mu}g/mLs$ was found to be an optimum dose. Semen as such and treated with SP-HBPs was diluted with sodium citrate-egg yolk diluter and cryopreserved as per the standard protocol. Sperm parameters i.e. motility, viability, Hypo-osmotic swelling test (HOST), acrosome damage, in vitro capacitation and lipid peroxidation were evaluated in SP-HBP treated and untreated (control) semen at both phases of cryopreservation. A considerable variation in percent sperm motility, viability, membrane integrity (HOST), acrosome damage, acrosome reaction and lipid peroxidation was observed at both phases among the bulls irrespective of the treatment. Incubation of neat semen with $20{\mu}g/mL$ SP-HBP before processing for cryopreservation enhanced the average motility, viability, membrane integrity by 7.2%, 1.5%, 7.9%, and 5.6%, 6.6%, 7.4% in pre-frozen and frozen-thawed semen in comparison to control. There was also an average increase of 4.1%/3.9% in in vitro capacitation and acrosome reaction in SP-HBPs-treated frozen-thawed semen as compared to control. However, binding of SP-HBPs to the sperm declined acrosome damage and lipid peroxidation by 1.3%/4.1% and 22.1/$32.7{\mu}M$/$10^9$ spermatozoa in SP-HBP treated pre-frozen/frozen-thawed semen as compared to control, respectively. Significant (p<0.05) effects were observed only in motility, HOST and in vitro acrosome reaction. It can be concluded that treatment of neat semen with SP-HBPs before cryopreservation minimized the cryoinjury by decreasing the generation of reactive oxygen species.