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Dietary Administration of Probiotics, Bacillus sp. IS-2, Enhance the Innate Immune Response and Disease Resistance of Paralichthys olivaceus against Streptococcus iniae (Probiotics를 양식넙치에 투여시 Streptococcus iniae에 대한 면역반응 및 병저항성)

  • Jang, Ik-Soo;Kim, Dong-Hwi;Heo, Moon-Soo
    • Korean Journal of Microbiology
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    • v.49 no.2
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    • pp.172-178
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    • 2013
  • The strains were added to the feed in the concentration of $10^3$, $10^5$, and $10^7$ CFU/kg and 2% of fishes were given the feed twice a day (8 AM and 5 PM) for 12 weeks. In result of the nonspecific immune response study to examine Respiratory burst activity, Lysozyme activity and Phagocytosis activity every two weeks until the end of the study, all test samples showed greater activities than control samples and improved immune activity with Bacillus sp. IS-2. The mortality test performed by artificial infection using Streptococcus iniae, a pathogenic bacterium, after the completion of this study also showed over 55% greater survival rate in all test samples. In result of performing PCR using the universal primer to verify that the probiotic stays in the intestines of the fishes, all test samples showed PCR product of 1,465 bp. Based on the above findings, it was concluded that Bacillus sp. IS-2 in the feed improved farmed flatfish's immune system and resistance against diseases as the probiotics. Also, the physiological indicators discovered by this study would be useful for identifying the mechanisms of probiotics.

Identification, Mycological Characteristics and Response to fungicides of Anthracnose Pathogen Isolated from Pepper and Boxthorn in Cheongyang (청양 지역 고추와 구기자에서 분리한 탄저병균의 동정, 균학적 특징 및 살균제 저항성)

  • Kim, Gahye;Kim, Joohyeng;Kim, Heung Tae
    • The Korean Journal of Pesticide Science
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    • v.19 no.3
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    • pp.335-344
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    • 2015
  • It was conducted to identify all 47 isolates obtained from infected fruits of pepper and boxthorn, and to investigate the mycological characteristics and the response to fungicides. All of 11 isolates from pepper were identified as Colletotrichum acutatum included into A2 group. Among 36 isolates from boxthorn, 14 isolates were identified as C. gloeosporioides, and the others were done as C. acutatum, which were composed as A1 group with 15 isolates and A3 with 7 isolates. After incubating the isolates on PDA at $25^{\circ}C$ for 10 days, the colony color of C. acutatum was greyish white, while that of C. gloeosporioides was orange at center of colony and was gradually turned into an greyish white to the periphery. The rate of conidia showing ellongated ellipsoidal shape with round ends was over 95% in C. acutatum isolated from pepper. However, C. acutatum isolated from boxthorn produced ellongated ellipsoidal conidia with the rate of 75%, and the others were pointed at one or both ends. Regardless of species of Colletotrichum, all isolated used in this study was showed an optimal temperature at $25^{\circ}C$. $EC_{50}$ values of all isolates of Colletotrichum spp. to 2 fungicides as carbendazim and the mixture of carbendazim and diethofencarb was investigated by an agar dilution method. With C. acutatum isolates from pepper belonged to A2 group, the mean of $EC_{50}$ value to carbendazim and the mixture of carbendazim and diethofencarb was 0.68 and $3.16{\mu}g/ml$, respectively. In the case of C. acutatum isolates from boxthorn, which were divided into 2 groups as A1 and A3 group, that to carbendazim was 0.21 at A1 and $0.24{\mu}g/ml$ at A3, while that to the mixture was 1.52 and $3.35{\mu}g/ml$. Isolates of C. gloeosporioides showed the mean of $EC_{50}$ value was $0.12{\mu}g/ml$ to carbendazim and $0.92{\mu}g/ml$ to the mixture. The value of resistant factor was higher in the isolates of C. acutatum obtained in boxthorn than those from pepper.

Resistance of Chili Pepper Cultivars to Isolates of Phytophthora capsici (Phytophthora capsici 균주들에 대한 고추 품종들의 저항성)

  • Jo, Su-Jung;Shim, Sun-Ah;Jang, Kyoung Soo;Choi, Yong Ho;Kim, Jin-Cheol;Choi, Gyung Ja
    • Horticultural Science & Technology
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    • v.32 no.1
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    • pp.66-76
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    • 2014
  • Resistance of one hundred commercialized cultivars of chili pepper to four isolates of Phytophthora capsici was evaluated under controlled environmental conditions. The cultivars are commercialized as resistant (59%) and susceptible (41%) to Phytophthora blight in Korea. Mean disease severities of the cultivars on P. capsici MY-1, KPC-1, JHAI1-7, and KPC-7 isolates were 37, 55, 60, and 74%, respectively. In addition, 38 for MY-1, 48 for KPC-1, 56 for JHAI1-7, and 76 cultivars for KPC-7 showed susceptibility. To P. capsici MY-1, the weakest pathogenicity isolate among them, 59 cultivars represented high resistance. By contrast, only six cultivars showed high resistance to P. capsici KPC-7, the strongest isolate. Furthermore, resistance of most cultivars except for three cultivars was negatively correlated with the virulence of P. capsici isolates. And isolate-specific resistance of the chili pepper cultivars could not be found. Among them, six cultivars showing resistance to all the tested isolates were selected for further study. The development of Phytophthora blight on the six cultivars according to inoculum density ($5{\times}10^4$ to $1.5{\times}10^6$ sporangia/pot) and incubation temperature (25 to $30^{\circ}C$) after inoculation of P. capsici was tested. Resistance of the cultivars to P. capsici KPC-1 and JHAI1-7, moderately pathogenic isolates, was hardly affected. But to KPC-7 isolate, the highly resistant cultivars showed susceptiblility or moderate resistance when the seedlings were inoculated with inoculum density of $1.5{\times}10^6$ sporangia/pot and incubated at 28 to $30^{\circ}C$. From these results, it is likely that resistance of chili pepper cultivars to Phytophthora blight is affected by the virulence of P. capsici isolate.

QTL Analysis Related to the Palatability Score According to Rice-polishing (도정정도에 따른 식미치 관련 QTL 분석)

  • Park, Young-hie;Kim, Kyung-Min
    • Journal of Life Science
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    • v.28 no.3
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    • pp.314-319
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    • 2018
  • We analyzed QTLs for alkali-related digestion by using 120 population crossed Cheongcheong and Nagdong derived from anther culture (CNDH). The DNA markers located in the QTLs gene were selected and applied to existing cultivars. As a result of the investigation of the alkali decay degree, brown rice of Cheongcheong and Nagdong was 1.9 and 1.6, respectively, and the CNDH was $3.79{\pm}2.01$, and the distribution of variance was distributed to 7.0-1.0. The milled rice of Cheongcheong and Nagdong was 5.6 and 4.1, respectively. The mean of the CNDH was $4.86{\pm}1.55$, and the distribution of variance was distributed to 7.0-2.0. Variation distribution curves showed continuous variation that was close to non-normal distribution. In the QTLs analysis, qBRA2, qBRA6, and qBRA11 were mapped in 1-2 replications of brown rice. QHRA2-1, qHRA2-2, qHRA2-3, qHRA3, and qHRA8 were mapped in the first replication. QHRA2-1, qHRA2-2, qHRA2-3 and qHRA3 were mapped in the second replicates. And mapped to qHRA5 in 4 replicates. These were found on chromosome 2, 3, 6, 8 and 11, respectively. The phenotypic variations of qBRA2, qBRA6, and qBRA11 on the chromosomes of brown and milled rice were 1-9%. The polymorphism was analyzed for 12 types of the japonica type and six types of the indica type, based on the nine markers found in the QTLs analysis of alkali digestion. Chromosome 11, RM27258, was selected to determine the segregation ratio, which shows the difference in size by the band pattern. The results of this study will be used as basic data for the development of high-quality rice cultivars.

QTL Analysis of Concerned on Ideal Plant Form in Rice (벼의 이상적인 초형에 관여하는 QTL 분석)

  • Chung, Il Kyung;Kim, Kyung-Min
    • Korean Journal of Plant Resources
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    • v.30 no.2
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    • pp.213-218
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    • 2017
  • Rice is the staple food of at least half of the world's population. Due to global warming, the weather is difficult to forecast nowadays. Therefore, it is necessary to breed various breeding to respond to such changes in the environment. This study was conducted to analyze the QTL about plant form, culm length, ear number and ear length by using 120 lines by anther culture, a cross between the Indica variety Cheongcheong and Japonica variety Nagdong. DNA marker was selected on the QTLs gene, and the following results were obtained. CNDH (Cheongcheong Nagdong Doubled Haploid) lines frequency distribution table curves about culm length, ear number and ear length exhibited showed a continuous variation close to a normal distribution. QTL analysis result, on culm length qPlL1-1 and qPlL1-2 were detected on the chromosome 1 and qPlL5 was detected on the chromosome 5. However, on ear length qPL2, qPL3 and qPL10, were detected on the chromosome 2, 3 and 10, while on ear number qPN1-1 and qPN1-2 were detected on the chromosome 1, qPN9 was detected on the chromosome 9. The QTLs related to culm length was found to chromosomes 5 and LOD scores were 3.81. The QTLs related to ear length was found to chromosomes 2 and 3 LOD scores were 7.13 and 3.20. The QTLs related to ear number was found to chromosome 9 and LOD scores were 4.27. Twenty two (22) Japonica cultivars and 12 Indica cultivars were analyzed polymorphisms, using selected 9 markers from the result about plant form analysis. RM5311, RM555 and RM8111 about the culm length, the ear length and number of ear were selected on the standard of Cheongcheong and Nagdong. Each rate of concordances about the culm length, the ear length and number of ear are 44.11%, 41.17% and 44.11%.

Isolation of Agarivorans sp. KC-1 and Characterization of Its Thermotolerant β-Agarase (한천분해세균 Agarivorans sp. KC-1의 분리 및 내열성 β-아가라제의 특성 규명)

  • Min, Kyung-Cheol;Lee, Chang-Eun;Lee, Dong-Geun;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.28 no.9
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    • pp.1056-1061
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    • 2018
  • This article reports an agar-degrading marine bacterium and characterizes its agarase. The agar-degrading marine bacterium, KC-1, was isolated from seawater on the shores of Sacheon, in Gyeongnam province, Korea, using Marine Broth 2216 agar medium. To identify the agar-degrading bacterium as Agarivorans sp. KC-1, phylogenetic analysis based on the 16S rRNA gene sequence was used. An extracellular agarase was prepared from a culture medium of Agarivorans sp. KC-1, and used for the characterization of enzyme. The relative activities at 20, 30, 40, 50, 60, and $70^{\circ}C$ were 65, 91, 96, 100, 77, and 35%, respectively. The relative activities at pH 5, 6, 7, and 8 were 93, 100, 87, and 82%, respectively. The extracellular agarase showed maximum activity (254 units/l) at pH 6.0 and $50^{\circ}C$ in 20 mM of Tris-HCl buffer. The agarase activity was maintained at 90% or more until 2 hr exposure at $20^{\circ}C$, $30^{\circ}C$ and $40^{\circ}C$, but it was found that the activity decreased sharply from $60^{\circ}C$. A zymogram analysis showed that Agarivorans sp. KC-1 produced 3 agar-degrading enzymes that had molecular weights of 130, 80, and 69 kDa. A thin layer chromatography analysis suggested that Agarivorans sp. KC-1 produced extracellular ${\beta}$-agarases as it hydrolyzed agarose to produce neoagarooligosaccharides, including neoagarohexaose (21.6%), neoagarotetraose (32.2%), and neoagarobiose (46.2%). These results suggest that Agarivorans sp. KC-1 and its thermotolerant ${\beta}$-agarase would be useful for the production of neoagarooligosaccharides that inhibit bacterial growth and delay starch degradation.

Influence of Hypoxia on Catecholamine Secretion Evoked by DMPP, McN-A-343, Excess $K^+$ and ACh from The Perfused Rat Adrenal Gland (저산소증이 흰쥐 관류부신에서 DMPP, McN-A-343, Excess $K^+$ 및 Ach의 카테콜아민 분비작용에 미치는 영향)

  • Lim Dong-Yoon;Heo Jae-Bong;Park Yoo Han
    • The Korean Journal of Pharmacology
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    • v.31 no.1 s.57
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    • pp.63-74
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    • 1995
  • It has been known that, during hypoxia, the adrenal medulla is activated to release catecholamines (CA) while hypoxia also inhibits high $K^+$ -induced CA secretion in the cultured bovine adrenal chromaffin cells. The present study was attempted to examine the effect of hypoxia on CA secretion evoked by chlinergic stimulation and membrane-depolarization from the isolated perfused rat adrenal glands and also to clarify its mechanism of action. For this purpose, using the isolated rat adrenal glands, the effects of hypoxia on CA release evoked by nicotinic ($N_1$) and muscarinic ($M_1$) receptor agonists, membrane-depolarizing agent, $Ca^{++}$-channel activator, intracellular $Ca^{++}$-releaser and ACh were determined. Experiments were carried out, perfusing Krebs solution pre-equilibrated with a gas mixture of 95% N_2$ and 5% $CO_2$. Hypoxia was maintained for $3{\sim}4$ hours through the experiments. Hypoxia gradually caused a time-dependent seduction in CA secretion evoked by DMPP ($100{\mu}M$), McN-A-343 ($100{\mu}M$), ACh (5.32 mM), Bay-K-8644 ($10{\mu}M$) and high $K^+$ (56 mM) respectively. How-ever, it did not affect CA secretion evoked by cyclopiazonic acid ($10{\mu}M$). Hypoxia itself also did fail to produce any influence on spontaneous secretory response of CA. These experimental results suggest that hypoxia depresses CA release evoked by both cholinergic stimulation and membrane-depolarization from the isolated rat adrenal medulla, and that this inhibitory activity may be due to the result of the direct inhibition of $Ca^{++}$ influx into the chromaffin cells without any effect on the calcium mobilization from the intracellular store.

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Improvement of Productivity by Forest Tree Breeding Work in Korea (우리나라에서의 임목육종(林木育種)에 의(依)한 생산성(生産性) 증가(增加))

  • Ryu, Jang Bal;Shim, Sang Yung
    • Journal of Korean Society of Forest Science
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    • v.77 no.4
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    • pp.382-388
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    • 1988
  • Improvement of productivity by forest tree breeding work in Korea was estimated for a few important tree species. Progenies of 17 plus trees of red pine (Pines densiflora) outgrew by 57 percentage compared with progenies of unselected trees at age 15. If best three families are selected among the 17, more than double in volume grow-th is expected. The hybrid Pinus rigida ${\times}$ P. taeda showed more than double volume growth compare to P. rigida at a southern plantation at age 15. However, the superiority of the hybrid decreased at northern plantations, mainly because of low coldhardiness of the hybrid. At a northern plantation, the hybrid grew less than the P. rigida on upper hill, while the hybrid grew much better than the P. rigida on flat area. Another hybrid Populus alba ${\times}$ P. glandulosa grew faster than both parents by two to two and half times according to planting sites at age 10. Introduction of Pinus rigida also showed increased volume growth. Volume increase by selection of best five provenances among 45 at age 12 was estimated as 53 percent compare to progenies of plus trees in Korea, Additional 19 percent of volume increase was expected by selection of the best families within the best provenances. Annual production of chestnuts reached about 70,000 M/T by planting resistant clones to chestnut gall wasp (Dryocosmus kuriphilus), which killed almost all susceptible trees. Although polyploid trees and mutants have been produced by colchicine treatments in over 10 tree species, none of them is economically important Remarkable improvement of productivity is expected by biotechnology in future through selection, hybridization, introduction of foreign genes at cell, cell organelle and gene level, and gene transformation. At present, mass propagation of superior planting materials by tissue culture will increase the productivity.

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Biodiesel production using lipase producing bacteria isolated from button mushroom bed (양송이 배지에서 유래한 Lipase 생산균을 이용한 바이오디젤 생산)

  • Kim, Heon-Hee;Kim, Chan-Kyum;Han, Chang-Hoon;Lee, Chan-Jung;Kong, Won-Sik;Yoon, Min-Ho
    • Journal of Mushroom
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    • v.13 no.1
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    • pp.56-62
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    • 2015
  • A lipase producing bacterium was isolated from button mushroom bed, which showing high clear zone on agar media containing Tributyrin as the substrate. The strain was identified as Burkholderia cepacia by analysis of 16S rDNA gene sequence. Crude lipase (CL) was partially purified from 70% ammonium sulfate precipitation using the culture filtrate of B. cepacia. Immobilized lipases were prepared by cross-linking method with CL from B. cepacia and Novozyme lipase (NL) onto silanized Silica-gel as support. Residual activitiy of the immobilized CL (ICL) and immobilized NL (INL) was maintained upto 61% and 72%, respectively. Biodiesel (Fatty acid methyl ester, FAME) was recovered by transesterification and methanolysis of Canola oil using NaOH, CL and ICL as the catalysts to compare the composition of fatty acids and the yield of FAME. Total FAME content was NaOH $781mg\;L^{-1}$, CL $681mg\;L^{-1}$ and ICL $596mg\;L^{-1}$, in which the highest levels of FAME was observed to 50% oleic acid (C18:1) and 22% stearic acid (C18:0). In addition, the unsaturated FAME (C18:1, C18:2) decreased, while saturated FAME (C16:0, C18:0) increased according to increasing the reaction times with both CL and ICL, supporting CL possess both transesterification and interesterification activity. When reusability of ICL and INL was estimated by using the continuous reaction of 4 cycles, the activity of ICL and INL was respectively maintained 66% and 79% until the fourth reaction.

Survival and Cross-contamination of Escherichia coli O157:H7 on Various Agricultural Product-Contact Surfaces (농산물 접촉 표면 재질에 따른 Escherichia coli O157:H7의 생존 및 상추로의 교차오염도 조사)

  • Kim, Se-Ri;Choi, Song-Yi;Seo, Min-Kyoung;Kim, Won-Il;Chung, Duck-Hwa;Ryu, Kyoung Yul;Yun, Jong-Chul;Kim, Byung-Seok
    • Journal of Food Hygiene and Safety
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    • v.28 no.3
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    • pp.272-278
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    • 2013
  • To evaluate the effect of surface contaminated with Escherichia coli O157:H7 (E. coli O157:H7) on the microbiological safety of lettuce, this study was conducted to investigate the attachment, biofilm producing, survival, and cross-contamination of E. coli O157:H7 on stainless steel and polyvinyl chloride (PVC). The attachment rate of E. coli O157:H7 on PVC was 10 times higher than that on stainless steel after exposure 1 h in cell suspension. However, there was not a difference between two types of surface after exposure for 6 h and 24h. The biofilm producing of E. coli O157:H7 was TSB > 10% lettuce extracts > 1% lettuce extracts > phosphate buffer. When two kinds of materials were stored at various conditions ($20^{\circ}C$ and $30^{\circ}C$, relative humidity (RH) 43%, 69%, and 100%), the numbers of E. coli O157:H7 at $30^{\circ}C$, RH 43% or RH 69% were reduced by 5.0 log CFU/coupon within 12 h regardless of material type. Conversely, the survival of E. coli O157:H7 at RH 100% was lasted more than 5 days. In addition, the reduction rate of E. coli O157:H7 was decreased in the presence of organic matter. The transfer efficiency of E. coli O157:H7 from the contaminated surface to lettuce was dependent upon the water amount of the surface of lettuce. Especially, the transfer rate of E. coli O157:H7 was increased by 10 times in the presence of water on the lettuce surface. From this study, the retention of E. coli O157:H7 on produce contact surfaces increase the risk cross-contamination of this pathogen to produce. Thus, it is important that the surface in post harvest facility is properly washed and sanitized after working for prevention of cross-contamination from surface.