• Title/Summary/Keyword: calmodulin

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Phylogenetic analysis and biological characterization of Stemphylium species isolated from Allium crops

  • Chang-Gi Back;You-Kyung Han;Walftor Bin Dumin;Jong-Han Park;Ji-Won Han;Yeoung-Seok Bae
    • Korean Journal of Agricultural Science
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    • v.49 no.1
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    • pp.113-120
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    • 2022
  • Leaf blight disease caused by Stemphylium species is an important disease in Allium crops, specifically onion, garlic and welsh onion. In 2018, leaf blight symptoms were severe and damaged onion and garlic in Jeonnam province in Korea. In addition, small purple spots on garlic burbs were observed in a post-harvest storage warehouse. Several Stemphylium isolates were isolated from diseased leaves from the field and from garlic bulb samples and were analyzed in terms of homology and the phylogenetic relationship based on the internal transcribed spacer region and calmodulin gene sequence. The results showed that among three Stemphylium species identified, S. vesicarium is most prevalent on onion and garlic. S. eturmiunum was for the first time identified as pathogenic to onion and garlic, whereas S. solani was found in welsh onion crops. Although these isolates grew well at the optimum temperature at 20 - 25℃, they could also grow at low temperatures of 10 - 15℃. A pathogenicity test was conducted using S. vesicarium and S. eturmiunum on onion and garlic respectively. These results showed that two Stemphylium species were highly virulent with cross pathogenicity in onion and garlic. The results of this study can support the biological characterization of Stemphylium species in Korea. Moreover, further research will need to develop fungicide application strategies for onion and garlic crops.

First Report of Charcoal Rot Caused by Macrophomina phaseolina on Peanut Plants in Korea (땅콩에서 Macrophomina phaseolina에 의한 균핵마름병 발생 보고)

  • Soo Yeon Choi;You Kyoung Lee;Chang Ok Geum;Shinhwa Kim;Hyunjung Chung;Sang-Min Kim;Yong Hoon Lee
    • The Korean Journal of Mycology
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    • v.51 no.4
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    • pp.383-387
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    • 2023
  • Peanut plants showing mild wilt were found in fields of Iksan, Korea, in August 2021. The diseased peanut plants were collected, and the causal pathogens were isolated using potato dextrose agar (PDA) medium. The isolated IS-1 strain formed white mycelia on PDA, which turned black with age. Sclerotia were produced on the PDA and barley leaves laid on water agar 7 d after incubation at 30℃. The sequences of both the internal transcribed spacer (ITS) region and calmodulin gene of IS-1 showed a 100% similarity with that of Macrophomina phaseolina. A phylogenetic tree constructed using the ITS regions of fungal pathogens causing disease in peanut plants indicated that the IS-1 stain belongs to M. phaseolina. The inoculation of IS-1 sclerotia into peanut seedlings resulted in yellowing and wilt symptoms in aboveground plants and brown to dark rots in roots 35-40 d after inoculation. Overall, the morphological characteristics, molecular identification, and pathogenicity of IS-1 indicate that the causal pathogen is M. phaseolina. This is the first report of charcoal rot caused by M. phaseolina on peanut plants in Korea. Further study is needed to develop the control measures for charcoal rot in peanut plants.

Panaxcerol D from Panax ginseng ameliorates the memory impairment induced by cholinergic blockade or Aβ25-35 peptide in mice

  • Keontae Park;Ranhee Kim;Kyungnam Cho;Chang Hyeon Kong;Mijin Jeon;Woo Chang Kang;Seo Yun Jung;Dae Sik Jang ;Jong Hoon Ryu
    • Journal of Ginseng Research
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    • v.48 no.1
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    • pp.59-67
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    • 2024
  • Background: Alzheimer's disease (AD) has memory impairment associated with aggregation of amyloid plaques and neurofibrillary tangles in the brain. Although anti-amyloid β (Aβ) protein antibody and chemical drugs can be prescribed in the clinic, they show adverse effects or low effectiveness. Therefore, the development of a new drug is necessarily needed. We focused on the cognitive function of Panax ginseng and tried to find active ingredient(s). We isolated panaxcerol D, a kind of glycosyl glyceride, from the non-saponin fraction of P. ginseng extract. Methods: We explored effects of acute or sub-chronic administration of panaxcerol D on cognitive function in scopolamine- or Aβ25-35 peptide-treated mice measured by several behavioral tests. After behavioral tests, we tried to unveil the underlying mechanism of panaxcerol D on its cognitive function by Western blotting. Results: We found that pananxcerol D reversed short-term, long-term and object recognition memory impairments. The decreased extracellular signal-regulated kinases (ERK) or Ca2+/calmodulin-dependent protein kinase II (CaMKII) in scopolamine-treated mice was normalized by acute administration of panaxcerol D. Glial fibrillary acidic protein (GFAP), caspase 3, NF-kB p65, synaptophysin and brainderived neurotrophic factor (BDNF) expression levels in Aβ25-35 peptide-treated mice were modulated by sub-chronic administration of panaxcerol D. Conclusion: Pananxcerol D could improve memory impairments caused by cholinergic blockade or Aβ accumulation through increased phosphorylation level of ERK or its anti-inflammatory effect. Thus, panaxcerol D as one of non-saponin compounds could be used as an active ingredient of P. ginseng for improving cognitive function.

Effects of Tree-spray of Calcium Agent, Coating Agent, GA4+7 + BA and Paper Bagging on Russet Prevention and Quality of 'Gamhong' Apple Fruits (칼슘제, 피막제, GA4+7 + BA의 수체살포 및 봉지씌우기에 의한 '감홍' 사과의 동녹 방지와 과실품질)

  • Moon, Young-Ji;Nam, Ki-Woong;Kang, In-Kyu;Moon, Byung-Woo
    • Horticultural Science & Technology
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    • v.34 no.4
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    • pp.528-536
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    • 2016
  • This study was conducted to examine the effect of 0.4% of $CaCl_2$, $2H_2O$, $1mg{\cdot}L^{-1}$ of Calmodulin (CaM)-SH, 250-folds of coating agent (WE-36), 1,000-folds of $GA_{4+7}+BA$ and 3 types of paper bagging treatments on russet incidence and fruit quality attributes of 'Gamhong' apple. The pattern of russet occurrence was slightly different for 4 years (from 2012 to 2015) in 'Gamhong' apple. The russet occurrence was lowest in $GA_{4+7}+BA$ treatment at 20 days after full bloom (DAFB), compared with other treatments. The $GA_{4+7}+BA$ treatment increased fruit weight at 20 DAFB, while the other fruit quality attributes were not influenced. The russet occurrence was lower not only in a single bag application than in untreated ones but also in yellow bagging and discolored bagging applications than in a white bagging application. The russet occurrence in a bagging application was lower at 20 DAFB than at 30 and 40 DAFB, while fruit quality attributes were not affected by bagging applications. The russet incidence was lower in $GA_{4+7}+BA$ twice treatments at 20 and 30 DAFB, and calcium coated bag at 30 DAFB after $GA_{4+7}+BA$ treatment at 20 DAFB than in untreated fruit. The rate of russet incidence was lowest at equator region in $GA_{4+7}+BA$ treatment, compared with the other fruit regions. Overall, the results suggest that one and/or two applications of $GA_{4+7}+BA$ (1,000-folds) treatment at 20 DAFB should reduce the risk of russet incidence in 'Gamhong' fruit.

THE EXPRESSION OF NITRIC OXIDE SYNTHETASE IN THE EXPERIMENTAL TOOTH MOVEMENT IN RATS (백서의 실험적 치아이동시 Nitric Oxide Synthetase의 발현 양상)

  • Park, Dong-Kwon;Kim, Sang-Cheol
    • The korean journal of orthodontics
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    • v.31 no.1 s.84
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    • pp.107-120
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    • 2001
  • Nitric oxide(NO) has been reported to be one of the mediators relating to bone remodelling. Nitric oxide is synthesized from L-arguinine by nitric oxide synthetase(NOS), which is largely divided Into two groups. One group which is composed of $NOS_1\;and\;NOS_3$, is dependent of calcium or calmodulin. The other consisted of $NOS_2$, which is independent of calcium or calmodulin. NOS is thought to be a possible intermediate affecting in the course of tooth movement. This study was designed to evaluate the expression of nitrous oxide synthetase(NOS) in periodontal tissue during the experimental movement of rat incisors, by LSAB(labelled streptavidine biotin) immunohistochemical staining for $NOS_2\;and\;NOS_3$. Twenty seven Sprague-Dawley rats were divided into a control group(3 rats), and 6 experimental groups(24 rats), to which 75g of force was applied, with helical springs across the maxillary incisors. Rats of experimental groups were sacrificed at 12 hours, 1, 4, 7, 14 and 28 days after force application, respectively. After that, the tissues of the control group and experimental groups were studied immunohistochemically. The results were as follows: 1. In control group, the expression of $NOS_3$ was rare in gingiva, dentin, periodontal ligament and alveolar bone, and was mild in the capillaries of pulp and intermaxillary suture. And the expression of $NOS_2$ showed similar pattern to that of $NOS_3$. 2. There were no differences in the expression of $NOS_2\;or\;NOS_3$ in dentin, gingiva, cementum, cementoblast and odontoblast, between control and experimental groups, regardless of the duration of the force application. 3. The expression of $NOS_3$ began to increase at 4 days and showed to the highest degree at 7 days after force application, in the apical region of pressure side of periodontal ligament in experimental groups. 4. The expression of $NOS_3$ in alveolar bone was rare until 7 days, after which it increased to mild degree at 14 days through 28 days in experimental group. But there was no difference between pressure and tension side of periodontal ligament. 5. The expression of $NOS_2$ in periodontal ligament was mild from 7 days after force application, regardless of the side of periodontium, which was generally more evident than that of $NOS_3$. 6. The expression of $NOS_2$ in alveolar bone increased to mild degree at 14 days after force application, and it was evident in osteoblasts, osteoclasts and osteocytes. And the expression of $NOS_2$ was little more stronger in the tension side than that of pressure side of alveolar bone.

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Effect of Neurogranin Phosphorylation on Oxidative Stress by Hydrogen Peroxide in Early Onset of Batten Disease (과산화수소에 의한 산화스트레스가 영아형 바텐병에서 neurogranin의 인산화에 미치는 영향)

  • Yoon, Dong-Ho;Kim, Han-Bok;Park, Joo-Hoon;Kim, Sung-Jo
    • Journal of Life Science
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    • v.19 no.4
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    • pp.520-525
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    • 2009
  • Early onset of Batten disease (EBD), one of the most lethal neurodegenerative storage disorders of childhood, is caused by inactivating mutations in the Ceroid Lipofuscinosis, Neuronal (CLN1) gene. Neurogranin, a calmodulin-binding protein, is expressed in the brain and participates in the protein kinase C (PKC) signaling pathway. While oxidative stress is the suggested cause of neurodegeneration in EBD, its molecular mechanism(s) remains obscure. In this research, we examined the levels of neurogranin in the brain mRNA of wild-type (WT) mice and EBD knockout (KO) mice, as well as the proteins. We also performed neuronal cultures to measure the expression levels of neurgranin and phosphorylated-neurogranin with or without oxidative stress inducers and anti-oxidants. Results showed that neurogranin in both EBD KO mice brain mRNA and protein extracts decreased in an age dependent manner. However, high amounts of phosphorylated-neurogranin were detected in the 6-month brain. This pattern was also confirmed by cultured neurospheres samples. Moreover, neurospheres treated with $H_2O_2$, an oxidative stress inducer, showed increased phosphorylated-neurogranin patterns. Interestingly, this pattern returned to normal status when treated with N-acetyl-L-cystein, an anti-oxidant, after $H_2O_2$ treatment was performed. Our results suggest that the phosphorylation of neurogranin is affected by oxidative stress status in EBD, and appropriate anti-oxidant treatment will relieve hyper-phosphorylation of neurogranin.

Cell death phenotype of vacuole Ca2+-ATPase11 (ACA11) transgenic plant in Arabidopsis (애기장대에서 액포막 존재 Ca2+-ATPase11 (ACA11) 형질전환제의 세포사멸 표현형 분석)

  • Lee, Sang-Min;Hoang, My-HanhThi;Kim, Kyung-Eun;Chung, Woo-Sik
    • Journal of Plant Biotechnology
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    • v.36 no.1
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    • pp.59-63
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    • 2009
  • Calcium ion ($Ca^{2+}$) is thought to play the important role as a second messenger for signal transduction that results in various physiological responses to cope with developmental programs and environmental changes in plant. In plant cells, the central vacuole functions as a major calcium store, which is important for both signal transduction and preventing cytotoxicity. Although there is evidence for the biochemical characterizations of a calmodulin-regulated $Ca^{2+}$-ATPase (ACA11) localized to vacuole membrane, the biological function to ACA11 in plant has not been verified. In this study, we show that the cell death as the hypersensitive response (HR) in mature leaves is induced in transgenic plant of a vacuole ACA-type $Ca^{2+}$-ATPase, ACA11. Evidence that cell death phenotype is the result of ACA11 gene silencing is provided by Western blot assay using membrane fraction proteins extracted from transgenic plant. The 3, 3'-diaminobenzidine (DAB) staining study provides that the cell death is caused by the increase of reactive oxygen species (ROS) in mature leaves of transgenic plants.

Molecular Cloning of Plasmodium vivax Calcium-Dependent Protein Kinase 4

  • Choi, Kyung-Mi;Kim, Jung-Yeon;Moon, Sung-Ung;Lee, Hyeong-Woo;Sattabongkot, Jetsumon;Na, Byoung-Kuk;Kim, Dae-Won;Suh, Eun-Jung;Kim, Yeon-Joo;Cho, Shin-Hyeong;Lee, Ho-Sa;Rhie, Ho-Gun;Kim, Tong-Soo
    • Parasites, Hosts and Diseases
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    • v.48 no.4
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    • pp.319-324
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    • 2010
  • A family of calcium-dependent protein kinases (CDPKs) is a unique enzyme which plays crucial roles in intracellular calcium signaling in plants, algae, and protozoa. CDPKs of malaria parasites are known to be key regulators for stage-specific cellular responses to calcium, a widespread secondary messenger that controls the progression of the parasite. In our study, we identified a gene encoding Plasmodium vivax CDPK4 (PvCDPK4) and characterized its molecular property and cellular localization. PvCDPK4 was a typical CDPK which had well-conserved N-terminal kinase domain and C-terminal calmodulin-like structure with 4-EF hand motifs for calcium-binding. The recombinant protein of EF hand domain of PvCDPK4 was expressed in Echerichia coli and a 34 kDa product was obtained. Immunofluorescence assay by confocal laser microscopy revealed that the protein was expressed at the mature schizont of P. vivax. The expression of PvCDPK4-EF in schizont suggests that it may participate in the proliferation or egress process in the life cycle of this parasite.

Regulatory Mechanisms of Angiotensin II on the $Na^+/H^+$ Antiport System in Rabbit Renal Proximal Tubule Cells. II. Inhibitory Effects of ANG II on $Na^+$ Uptake

  • Han, Ho-Jae;Park, Soo-Hyun;Koh, Hyun-Ju
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.4
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    • pp.425-434
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    • 1997
  • Many reports represent that angiotensin II (ANG II) caused a dose dependent biphasic effects on fluid transport in the proximal tubule. However, respective roles of different signaling pathways in mediating these effects remain unsettled. The aim of the present study was to examine signaling pathways at high doses of ANG II on the $Na^+$ uptake of primary cultured rabbit renal proximal tubule cells(PTCs) in hormonally defined serum-free medium. High concentrations of ANG II $(>10^{-9}\;M)$ inhibited $Na^+$ uptake and increased $[Ca^{2+}]_i\;level$ in the PTCs. However, low concentrations of $(<10^{-11}\;ANG\;II)$ stimulated $Na^+$ uptake and did not affect $[Ca^{2+}]_i\;level$. 8-(N, N-diethylamino)-octyl-3,3,5- trimethoxybenzoate (TMB-8), ethylene glycol-bis$({/beta}-amino\;ethyl ether)-N,N,N'$, N'-tetra acetic acid (EGTA), and nifedifine partially blocked the inhibitory effects of ANG II on $Na^+$ uptake. When ANG II and bradykinin (BK) were treated together, $Na^+$ uptake was further reduced $(88.47{\pm}1.98%\;of\;that\;of\;ANG\;II,\;81.85{\pm}1.84%\;of\;that\;of\;BK)$. In addition, W-7 and KN-62 blocked the ANG II-induced inhibition of $Na^+$ uptake. Arachidonic acid reduced $Na^+$ uptake in a dose-dependent manner. When ANG II and arachidonic acid were treated together, inhibitory effects on $Na^+$ uptake significantly exhibited greater reduction than that of each group, respectively. When PTCs were treated by mepacrine $(10^{-6}\;M)$ and AACOCF3 $(10^{-5}\;M)$ for 1 hr before the addition of $(<10^{-9}\;ANG\;II)$, the inhibitory effect of ANG II was reversed. In addition, econazole $(>10^{-6}\;M)$ blocked ANG II-induced inhibition of $Na^+$ uptake. In conclusion, the $[Ca^{2+}]_i$ (calcium-calmodulin-dependent kinase) and phospholipase $A_2\;(PLA_2)$ metabolites are involved in the inhibitory effects of ANG II on $Na^+$ uptake in the PTCs.

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Integrative Study on PPARGC1A: Hypothalamic Expression of Ppargc1a in ob/ob Mice and Association between PPARGC1A and Obesity in Korean Population

  • Hong, Mee-Suk;Kim, Hye-Kyung;Shin, Dong-Hoon;Song, Dae-Kyu;Ban, Ju Yeon;Kim, Bum Shik;Chung, Joo-Ho
    • Molecular & Cellular Toxicology
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    • v.4 no.4
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    • pp.318-322
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    • 2008
  • Obesity is an increasing worldwide health problem that is strongly related to the imbalance of food intake and energy metabolism. It was well-known that several substances in the hypothalamus regulate food intake and energy metabolism. We planned an integrative study to elucidate the mechanism of the development of obesity. Firstly, to find candidate genes with the marvelous effect, the different expression in the hypothalamus between ob/ob and 48-h fasting mice was investigated by using DNA microarray technology. As a result, we found 3 genes [peroxisome proliferator activated receptor, gamma, coactivator 1 alpha (Ppargc1a), calmodulin 1 (Calm1), and complexin 2 (Cplx2)] showing the different hypothalamic expression between ob/ob and 48-h fasting mice. Secondly, a genetic approach on PPARGC1A gene was performed, because PPARGC1A acts as a transcriptional coactivator and a metabolic regulator. Two hundred forty three obese female patients with body mass index (BMI)${\geq}$25 and 285 control female subjects with BMI 18 to<23 were recruited according to the Classification of Korean Society for the Study of Obesity. Among the coding single nucleotide polymorphisms (cSNPs) of PPARGC1A, 2 missense SNPs (rs8192678, Gly482Ser; rs3736265, Thr612Met) and 1 synonymous SNP (rs3755863, Thr528Thr) were selected, and analyzed by PCR-RFLP and pyrosequencing. For the analysis of genetic data, chi-square ($X^2$) test and EH program were used. The rs8192678 was significantly associated with obese women (P<0.0006; odds ratio, 1.5327; 95% confidence interval, 1.2006-1.9568). Haplotypes also showed significant association with obese women ($X^2$=33.28, P<0.0008). These results suggest that PPARGC1A might be related to the development of obesity.