• Title/Summary/Keyword: strains inoculation

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Purtification of Parasporal Protein Crystals of Bacillus thuringiensis (Bacillus thuringiensis의 내독소 단백질의 분리1)

  • Kim, Yeong-Hun;Kim, Sang-Hyeon;Gang, Seok-Gwon
    • Journal of Sericultural and Entomological Science
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    • v.33 no.1
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    • pp.32-36
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    • 1991
  • The study has been carried out to acquire some basic informations about Bacillus thuringiensis for developing the microbial pesticide. Three strains of Bacillus thuringiensis var. Kurstaki, dendrolimus and aizawai, were used in the experiments as follows. Growth characteristics of each strain were examined and parasporal protein crystals were isolated from the mixtures of spores and protein crystals by the new method of centrifugation in two-layer cushion of Renograffin using a fixed angle rotor. The results are as follows. 1. No difference was shown in growth characterestics among three strains of B. thuringiensis. In growth curves, all strains reached to exponential phase by 2 hr and stationary phase by 7-8 hr after inoculation. 2. The pH of the culture media during exponential growth stage decreased about 1.4 of a pH unit at the beginning of sporulation, but recovered during the early stage of sporulation and then remained nearly constant during the later stage. 3. As 10$m\ell$ sample was applied to two-layer cushion of Renograffin and then centrifuged for 1hr at 27,000g a fixed angle rotor, the purity and recovery ratio was 99.9% and 5.8%, respectively. It has been shown that the new method for the isolation of parasporal protein crystals was more efficient than any from the estabilished methods.

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Basic Physiological Activities of Bifidobacterium infantis Maeil-K9 and Lactobacillus plantarum KCTC3099 Selected by Anticarcinogenic Activities. (항암 활성능이 우수한 Bifidobacterium infantis Mneil-K9과 Lactobacillus plantarum KCTC3099의 기초 생리활성)

  • 김응률;정병문;김지연;김서영;정후길;이형주;전호남
    • Microbiology and Biotechnology Letters
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    • v.31 no.4
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    • pp.348-354
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    • 2003
  • This study was conducted to confirm the availability of lactic acid bacteria as probiotics haying inhibitory effects to cancer cell line. Five lactic acid bacteria showing anti-cancer activity were compared by acid tolerance, bile tolerance, antibiotics resistance, milk fermentation, stability, and cell adherence activity to colon epithelial cell. The results obtained are as follows : In acid tolerance, all strains did not have a resistance below pH 3.0 and 3.5 except Lactobacillus plantarum KCTC3099. In antibiotics resistance, Lactococcus lactis and L. plantarum KCTC3099 were resistant to cotrimoxazol (128 mg/1), and Bifidobacterium adolescentis Maeil-K8 and B. infantis Maeil-K9 were resistant to doxycylin and gentamycin (4 mg/1). In case of cell adherence ability to Caco-2 cell, B. infantis Maeil-K9 was found to be superior to others as 3.1%, while the others were less than 0.5%. When the strains were cultured to milk base, viable counts of the strains tested increased more 1 log cycle than inoculation, but acid production was very low except L. plantarum KCTC3099. Also, L. plantarum KCTC3099, B. adolescentis Maeil-K8, and B. infantis Maeil-K9 were stable in fermented milk base during storage. In conclusion, L. plantarum KCTC3099 and B. infantis Maeil-K9 were confirmed to be superior for the availability as probiotics.

Reduced Sensitivities of the Pear Scab Fungus (Venturia nashicola) Collected in Ulsan and Naju to Five Ergosterol-biosynthesis-inhibiting Fungicides (울산과 나주에서 채집한 배나무 검은별무늬병균의 ergosterol 생합성 저해제 5종에 대한 감수성 저하)

  • Kwon, Soo-Mean;Yeo, Moo-Ill;Choi, Se-Hoon;Kim, Gun-Woong;Jun, Kyung-Jin;Uhm, Jae-Youl
    • Research in Plant Disease
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    • v.16 no.1
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    • pp.48-58
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    • 2010
  • In Korea, EBI fungicides which are highly effective for control of pear scab and rust but of high risk for development of resistance have been frequently sprayed by majority of pear growers. To detect any possible resistance or reduced sensitivity in the field strains of scab fungus to five EBIs, difenoconazole, fluquinconazole, flusilazole, fenarimole and hexaconazole, sensitivity tests were conducted with fungal specimens collected in Ulsan and Naju where scab usually occurs and EBIs have been intensively sprayed for many years. As the strains for which $EC_{50}$ values of the EBIs were largely shifted from those of base-line were occasionally found, the resistant or less sensitive strains were supposed to be distributed. In the activity test for the EBIs by artificial inoculation, in which EBI-treated pear leaves on the potted seedlings were inoculated with fungal spores collected in the two regions, development of resistances to EBIs were confirmed. Since the fungal spores collected at 4 and 2 orchards in Naju and Ulsan, respectively, produced much higher disease incidence on the leaves treated with hexaconazole than those on the untreated control, those fungal specimens were determined as resistant to hexaconazole. Similar results were also obtained with two specimens from Ulsan for flusilazole.

Growth Characteristics and Optimal Culture Conditions of Bacterial Strains Degrading Ethylene Glycol and Terephthalic Acid in Polyester Weight Loss Wastewater (Polyester 감량폐수 중에 존재하는 Ethylene Glycol과 Terephthalic Acid를 분해하는 Bacteria 균주들의 성장특성과 최적 배양조건)

  • 김정목;김재훈조무환
    • KSBB Journal
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    • v.8 no.2
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    • pp.156-163
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    • 1993
  • Strains degrading ethylene glycol(EG) and terephthalic acid(TPA) were isolated from water systems, and identified as Pseudomonas sp. They were named as Pseudomonas sp. EAW for EG and as Pseudomonas sp. TS2 for TPA. The optimal culture conditions of temperature, pH and nitrogen source were found to be $35^{\circ}C$, 7.5 and ammonium sulfate, respectively. The growth of strains and removal efficiency was slightly promoted by trace elements such as niacin and biotin in case of EG, and by trace elements such as $Na_2MoO_4{\cdot}2H_2O$ and thiamin i case of TPA. With increasing inoculation sloe for batch culture, the removal efficiency of EG by the strain EAW was conspicuously increased, while the removal efficiency of TPA by the strain TS2 was not changed as much as that of EG. The growth rate of the strain EAW was much more decreased than that of the strain TS2 in the enrichment medium, as the frequency of repeated-batch culture in the rich-medium increased. in case of real wastewater, growth rate and removal efficiencies of EG and TPA were lower than those in the enrichment medium. $COD_{Mn}\;and\;COD_{Cr}$ removal efficiencies after 48 hrs batch culture in real wastewater were 89% and 93%, respectively. The specific growth rate was inhibited when the initial concentration of EG or TPA was more than 25g/L.

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Microbial Inhibition of Lactic Strains isolated from Kimchi (김치에서 분리한 젖산균의 미생물 생육 저해)

  • Park, Yun-Hee;Kwon, Jung-Joo;Jo, Do-Hyun;Kim, Su-Il
    • Applied Biological Chemistry
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    • v.26 no.1
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    • pp.35-40
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    • 1983
  • The inhibitory activity of 20 Lactic strains from Kimchi was tested against Escherichia coli and other microorganisms. Of the lactic strains investigated, A7 (Pediococcus cerevisiae) and C4(Leuconostoc spp.) were the most effective in restricting the growth of test organisms. The mixed culture inoculation of each selected lactic strain and Escherichia coli resulted in a drastic reduction in the plate count of Escherichia coli after 24 hours. Similar results were obtained when Staphylococcus aureus and Bacillus cereus were used as test organisms. For all test organisms, the presence of A7 caused a higher death rate constant than that of C4. Addition of catalase in the mixed culture did not prevent inhibition, suggesting that hydrogen peroxide did not cause the inhibition. The filtrate of A7 culture added to Escherichia coli showed identical inhibitory action, however heat treatment of filtrate at $80^{\circ}C$ 30min. destroyed the inhibitory activity. A7 filtrate treated with trypsin substantially lost the inhibitory effect, but not by pepsin. The results imply that the protein-like compound(s) is the principal inhibitor produced by this lactic strain.

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Sequence analysis and expression of groE gene encoding heat shock proteins of Brucella abortus isolates (Brucella abortus 국내 분리주의 Heat Shock Protein 암호 groE 유전자의 염기서열 분석과 발현)

  • Kim, Tae-Yong;Kim, Ji-Young;Chang, Kyung-Soo;Kim, Myung-Cheol;Park, Chang-Sik;Han, Hong-Ryul;Jun, Moo-Hyung
    • Korean Journal of Veterinary Research
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    • v.45 no.1
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    • pp.45-53
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    • 2005
  • GroE that is a heat shock protein composed of GroEL and GroES is known as an immunodominant target of both the humoral and cellular immune responses in bovine brucellosis. This study was carried out to characterize groE gene encoding heat shock proteins of B. abortus isolated in Korea and to evaluate the immunogenicity of the GroE protein expressed in E. coli system. In PCR the specific signals with the size of 2,077 bp were detected in five strains isolated from the mammary lymphnodes of the dairy cattle that were serologically positive and the reference strains. In comparison of the sequences of nucleotides and amino acids among the strains, GroES showed 100% identity in both sequences. GroEL was evaluated 99.0~99.9% in nucleotides and 98.0~100% homology in amino acids. The groE gene including groES and groEL was inserted into pET29a vector and constructed pET29a-GroE recombinant plasmids. The inserted groE was confirmed by digestion with Nco1 and EcoR1 endonucleases and nucleotide sequencing. E. coli BL (DE3) was transformed with pET29a-GroE, named as E. coli BL (DE3)/pET29a-GroE. In SDS-PAGE, it was evident that the recombinant plasmid effectively expressed the polypeptides for GroES (10 kDa) and GroEL (60 kDa) in 0.5, 1 and 2 hours after IPTG induction. The immuno-reactivity of the expressed proteins were proved in mouse inoculation and Western blot analysis.

Phosphate Solubilizing Activity of Pseudomonas sp. CL-1 and Kluyvera sp. CL-2 (Pseudomonas sp. CL-1 및 Kluyvera sp. CL-2 균주의 인산가용화 특성)

  • Kwon, Jang-Sik;Suh, Jang-Sun;Weon, Hang-Yeon;Kim, Wan-Gyu;Noh, Hyung-Jun
    • Korean Journal of Soil Science and Fertilizer
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    • v.40 no.6
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    • pp.442-446
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    • 2007
  • From the rhizoplane and rhizosphere of pepper, tomato, lettuce, pasture, and grass, unsoluble inorganic phosphate solubilizing bacterial strains were isolated using plate base assay on Pikovskaya's medium. Two strains, CL-1 and CL-2, which produced largest halo on plates (indicative of phosphate solubilization)were selected for further studies. Based on these biochemical and 16S rRNA analysis strains CL-1, CL-2 were found to be as species of Pseudomonas sp. and Kluyvera sp., respectively. In broth assay Pseudomonas sp. CL-1 and Kluyvera sp. CL-2 solubilized insoluble phosphate by 193.4 mg and $493.6P\;mg\;L^{-1}$, respectively after $3^{rd}$ day inoculation. These effecient phosphate solubilizing bacteria have a potential to be developed as microbial based fertilizer in future.

Effect on the Inoculation of Bacillus on the Growth of Chinese Cabbage and Sesame and on Microbial Flora in Soils (Bacillus subtilis 접종이 배추 및 참깨의 생장(生長)과 토양(土壤) 미생물상(微生物相)에 미치는 영향(影響))

  • Kim, Kwang-Sik;Lee, Jae-Pyeong;Kim, Yong-Woong;Rhee, Young-Hwan;Kim, Yeong-Yil
    • Korean Journal of Soil Science and Fertilizer
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    • v.26 no.4
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    • pp.271-277
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    • 1993
  • An antagonistic bacteria was isolated from rhiaosphere of pepper and corn and identified as Bacillus (B.) subtilis. These B. subtilis B-5 was transformed and marked with the plasmid pCPP4 which possess neomycine resistan. gene. The marked stranins showed growth inhibition to Rhizoctonia (R.) solani, Fusarium (F.) solani, and F. oxysporum in vitro, and were used in studying growth promoting effects on sesame and cabbage. All the identified strains utilize glucose, sucrose, fructose, lactose, mannitol and sorbitol as carbon source, but not rhamnose, and the marked strains also showed characteristics similar to wild-type strains. Germination rate of chinese cabbage and sesame seeds was increased by about 10% or more in the plot to which these strains were inoculated and the effect was higher in soil than in petri dish. The early growth promoting effects of these strains appeared higher, as compared with control plot, in the plots to which B. subtilis B-5 and pathogenic fungi was inoculated together. When the marked strains, B. subtilis B-5NEOr, were inoculated in the rhizosphere of chinese cabbage and sesame with $1.1{\times}10^8CFU/g$ dry soil, the number of inoculated strain was decreased slowly to the level of $10^5{\sim}10^6CFU/g$ dry soil after 4 weeks and the number of Pseudomonas spp. maintanied the level of $10^5CFU/g$ dry soil throught total period, but the number of fungi was decreased rapidly from the early level of $10^8CFU/g$ dry soil to $10^3CFU/g$ dry soil after 4 weeks.

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Dual Inoculation Response of Soybean with Rhizobium And Mycorrhiza (콩에 대한 근류균과 균근균의 혼합 접종효과)

  • Kang, Ui-Gum;Park, Hyang-Mee;Lee, Jae-Saeng;Ko, Jee-Yeon;Lee, Yong-Hwan;Jeon, Weon-Tae;Kim, Min-Tae;Joa, Jae-Ho
    • Korean Journal of Soil Science and Fertilizer
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    • v.45 no.3
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    • pp.325-331
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    • 2012
  • The dual inoculation response of soybean with rhizobium and mycorrhiza was examined in pot vermiculite and field soils. In order to select a symbiotically compatible mycorrhiza with Bradyrhizobium japonicum, a highly germinating spore among 60 strains from 32 upland soils in southern part of Korea was obtained in Acaulospora sp., Gigaspora sp. and Glomus sp., respectively. As a result of dual inoculation of Glycin max cv. Dajangkong and Eunhakong both with $1{\times}10^8$cells of B. japonicum YCK 213 and 10 spores of each mycorrhiza in vermiculite pot, only Glomus sp. treatment together with the rhizobium showed significant increase ($P{\leqq}0.05$) both in shoot dry wt and nodule mass of not Eunhakong but Dajangkong. In red-yellow soils with pH 5.2($1:5H_2O$) and 203 mg of Lancaster P per kg of soil, in which $10^3$ cells of B. japonicum and $10{\pm}0.2$ spores of mycorrhizae per gram of soil were naturalized, grain yield of G. max cv. Dajangkong was increased to 3.9% by dual inoculation both of $4.8{\times}10^6$cells of B. japonicum and 10 spores of mycorrhizae per two seeds under condition applied with 30 kg $P_2O_5$ and 34 kg $K_2O$ per hectare compared to conventionally fertilized plot (2.75 MT $ha^{-1}$) added with 30 kg N $ha^{-1}$. However, there was not significant.

Effect of Chemotaxis on Nodulation in Bradyrhizobium-Soybean Symbiosis (근류균의 화학주성이 근류형성에 미치는 영향)

  • Kang, Sang-Jai;Park, Woo-Churl
    • Korean Journal of Soil Science and Fertilizer
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    • v.27 no.2
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    • pp.136-146
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    • 1994
  • To research the effect of chemotaxis of Rhizobia toward the root exudate on nitrogen fixing ability in soybean Rhizobia symbiosis system. Root exudate from seedlings of Glycine max. L was collected aseptic conditions. B. japonicum KCTC 2422 induced the formation of symbiotic nitrogen fixing nodules on the root of soybean plant and possessed motility and chemotaxis toward the 2mM proline. LPN-100 mutant was $Nod^-$, $Che^+$, and LPN-101 was $Che^-$, $Nod^+$ strains. Physiological properties of mutants were similar to parent strain. The crude root exudate was tested for its chemotactic ability using the capillary tube method. Chemotactic responses of RCR 3407 toward crude root exudate were 2.2, 2.6, 2.9, those of KCTC 2422 were 2.3, 2.9, 3.0, respectively. The crude root exudate was fractionated into neutral, cationic and anionic fractions. Chemotactic responses of KCTC 2422 was least with anionic fraction, most with neutral and intermediate with cationic fraction. B. japonicum KCTC 2422 was attracted by carbohydrates, amino acids and carboxylic acid. Carbohydrates and amino acids were good chemoattractants and carboxylic acids were intermediate chemoattractants. The peak concentration was $10^{-3}M$ for ribose, glucose, glutamine, aspartic acid and carboxylic acids, with exception of xylose, arabinose, tryptophan, which elicited maximum responses at $10^{-4}M$. The formation of nodules and nitrogenase activity of soybean inoculated with KCTC 2422 was determined in 7days after inoculation, and those of LPN-101 was detected in 15days after inoculation, but LPN-100 didn't form of nodules in soybean plants.

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