• Title/Summary/Keyword: inoculum potential

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Metallo-collagenase production by Arthrobacter creatinolyticus KP015744

  • Savita A. Kate;Madhuri Sahasrabudhe;Archana Pethe
    • Advances in environmental research
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    • v.11 no.1
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    • pp.1-16
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    • 2022
  • Amongst 27 isolates from deteriorated leather samples, Arthrobacter creatinolyticus KP015744 zzx28 was found to be an efficient collagenase producer. Collagenase production of 13.33 µmoles/min was shown at an optimum temperature at 37℃ after 72h and at pH 7.5 by using 2 ml/dL inoculum in 10 mg/ml collagen peptide type I as a substrate. In presence of Hg2+, EDTA and 𝛽-mercaptoethanol the collagenase production by the isolate was strongly inhibited however Fe2+, Ca2+and DMSO enhanced production of the enzyme. Specific activity was found to be 19.46×103 U/mg and molecular weight 66 kD by SDS PAGE. Isolate also has potential to hydrolyze keratin which is another important protein found in leather. Experimental results propose that collagenase can be effectively used as a tool for collagen and keratin rich solid waste treatment.

Conidial Disperse of the Pepper Anthracnose Fungus Colletotrichum acutatum and Its Density on Infected Fruits (고추 탄저병균(Colletotrichum acutatum)의 분생포자 비산과 과실병반에 형성된 전염원 밀도)

  • Jee, Hyeong-Jin;Shin, Shun-Shan;Lee, Ji-Hyun;Kim, Won-Il;Hong, Sung-Jun;Kim, Yong-Ki
    • Research in Plant Disease
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    • v.16 no.1
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    • pp.101-105
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    • 2010
  • This study was aimed to understand conidial disperse of the pepper anthracnose fungus Colletotrichum acutatum, elapse time for the disease development, and inoculum potentials on infected fruits. Most (99.2%) conidia of the fungus disseminated from inoculum source on the rainy day, while only 0.8% conidia dispersed on the sunny day. Among the conidia 93.3% were caught under 60 cm height at 30 cm distance; however, conidia were detected at 120 cm height at the distance. Relatively susceptible pepper fruits to anthracnose showed first visible symptoms at 4 days after infection under a mimic field condition. However, it seemed that over 10 days are required for the disease to develop on moderately resistant pepper fruits under unfavorable conditions. The number of conidia formed on a lesion was positively correlated with the lesion size ($R^{2}=0.88$). Over 10 millions of conidia were formed at a normal lesion size 1.5 cm in length. In some large coalesced lesions ca. 4cm in length produced over 100 millions of the fungal conidia. Results further confirmed that the rainfall is the key factor for the inoculum disperse of the pepper anthracnose pathogen, Colletotrichum acutatum, and a long distance dissemination is plausible according to rain and wind intensity. Consequently, rain-proof structures are ideal to avoid the disease, and removal of infected fruits and timely chemical spray are indispensible to reduce the inoculum potential in the field.

Isolation and characterization of a novel gossypol-degrading bacteria Bacillus subtilis strain Rumen Bacillus Subtilis

  • Zhang, Yunhua;Zhang, Zhengyou;Dai, Li;Liu, Ying;Cheng, Maoji;Chen, Lijuan
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.1
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    • pp.63-70
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    • 2018
  • Objective: The aim of the study was to isolate gossypol-degrading bacteria and to assess its potential for gossypol degradation. Methods: Rumen liquid was collected from fistulated cows grazing the experimental pasture. Approximately 1 mL of the rumen liquid was spread onto basal medium plates containing 2 g/L gossypol as the only source of carbon and was then cultured at $39^{\circ}C$ to isolate gossypol-degrading bacteria. The isolated colonies were cultured for 6 h and then their size and shape observed by microscope and scanning electron microscope. The 16S rRNA gene of isolated colonies was sequenced and aligned using National Center for Biotechnology Information-Basic Local Alignment Search Tool. The various fermentation conditions, initial pH, incubation temperature, inoculum level and fermentationperiod were analyzed in cottonseed meal (CSM). The crude protein (CP), total gossypol (TG), and free gossypol (FG) were determined in CSM after fermentation with isolated strain at $39^{\circ}C$ for 72 h. Results: Screening results showed that a single bacterial isolate, named Rumen Bacillus Subtilis (RBS), could use gossypol as a carbon source. The bacterium was identified by 16S rDNA sequencing as being 98% homologous to the sequence of Bacillus subtilis strain GH38. The optimum fermentation conditions were found to be 72 h, $39^{\circ}C$, pH 6.5, moisture 50%, inoculum level $10^7cell/g$. In the optimum fermentation conditions, the FG and TG content in fermented CSM decreased 78.86% and 49% relative to the control. The content of CP and the essential amino acids of the fermented CSM increased respectively, compared with the control. Conclusion: The isolation of a gossypol-degrading bacterium from the cow rumen is of great importance for gossypol biodegradation and may be a valuable potential source for gossypol-degradation of CSM.

Weed Management Technology with Host Specific of Biological Control Agents (기주특이성 잡초 활성 미생물을 이용한 잡초방제 기술)

  • Hong, Yeon-Kyu;Lee, Bong-Choon;Song, Seok-Bo;Park, Sung-Tae;Kim, Jeong-Nam;Geon, Min-Goo;Kim, In-Seob
    • Asian Journal of Turfgrass Science
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    • v.20 no.2
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    • pp.175-190
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    • 2006
  • The term mycoherbicide started in 1970, but its interest heightened due to increase costs of chemical herbicides. A classical biocontrol agent is expected to become a permanent part of its new environment and do no harm to it. Contemporary biological control agent(BCA) must be produced by artificial culture and could be applied like chemical herbicides over weeds. BCA is different from the classical approach in that it released through natural spread. To date 26 species of fungi are used as classical BCA against 26 species of weeds in seven countries. There are a number of examples of pathogens attacking non-target plants. But through risk assessments which include understanding the taxonomy, biology and ecology, the target and non-target species, it will be safe to introduce of exotic pathogens to control weeds. But pathogens have not been successfully used in practice. Many mycoherbicides show potential in laboratories, but are ineffective in the field and not consistent from year to year or field to field. There is also a lack of understanding humidity, dew formation and temperature and their effects on suppression of weeds by plant pathogens. Potential pathogen must be selected as a BCA. Previous studies suggest that these pathogens must (1) produce abundant and durable inoculum in artifical culture, (2) be genetically stable and weed specific and (3) kill weeds in control. A granular preparation of mycoherbicide into sodium alginate is lighter than liquids and less bulky than organic matter. Gel forms have also been used.

Effect of Substrate to Inoculum Ratio on Biochemical Methane Potential in the Thermal Pretreatment of Piggery Sludge (양돈분뇨의 열전처리에서 기질과 접종액의 비율이 메탄생산 퍼텐셜에 미치는 영향)

  • Kim, Seung-Hwan;Kim, Ho;Oh, Seong-Yong;Kim, Chang-Hyun;Yoon, Young-Man
    • Korean Journal of Soil Science and Fertilizer
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    • v.45 no.4
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    • pp.532-539
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    • 2012
  • This study was carried out to investigate the effect of substrate to inoculum ratio on ultimate methane potential ($B_u$) from piggery wastes. BMP(Biochemical methane potential) assays were executed for the three samples that have different organic characteristics (Filtrate of pig slurry, LF; Thermal hydrolysate of piggery sludge cake, TH; Mixture of LF and TH at the ratio of 4 to 1, Mix), and $B_u$ values obtained from BMP assays were compared with the theoretical methane potential ($B_{th}$) of each samples. While $B_u$ values (0.27, 0.44, and $0.46Nm^3\;Kg^{-1}-VS_{added}$) of TH sample that was pretreated with thermal hydrolysis were below the $B_{th}$ at all S/I ratios (0.1, 0.3, and 0.5), and $B_u$ values of LF (0.64 and $0.53Nm^3\;Kg^{-1}-VS_{added}$ for the S/I ratios of 0.1 and 0.3, respectively) at the lower S/I ratios of 0.1 and 0.3 exceeded the $B_{th}$ values ($0.418Nm^3\;Kg^{-1}-VS_{added}$). And also biodegradability ($B_u/B_{th}$) of LF sample were obtained as 152.07%, 122.67%, and 95.71% at the S/I ratios of 0.1, 0.3, and 0.5, respectively, and unreasonable $B_u/B_{th}$ values were presented at lower S/I ratios of 0.1 and 0.3. $B_u$ and $B_u/B_{th}$ of Mix sample showed a similar tendency with those of LF sample. Therefore, TH sample by thermal hydrolysis pretreatment showed lower anaerobic biodegradability than those of other samples (LF and Mix) and ultimate methane potentials of LF and Mix samples were overestimated in the lower S/I ratio of 0.1 and 0.3.

Detection, isolation, and characterization of the cucumber mosaic virus in Pseudostellaria heterophylla from Korea

  • Lee, Da Hyun;Kim, Jinki;Han, Jun Soo;Lee, Jae-Hyeon;Lee, ByulHaNa;Park, Chung Youl
    • Journal of Plant Biotechnology
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    • v.47 no.2
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    • pp.150-156
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    • 2020
  • Weeds play an important role in the survival of viruses and are potential inoculum sources of viral diseases for crop plants. In this study, specimens of Pseudostellaria heterophylla exhibiting symptoms of the cucumber mosaic virus (CMV) were collected in Bonghwa, Korea. The characteristics of the disease were described and leaf RNA was extracted and sequenced to identify the virus. Three CMV contigs were obtained and PCR was performed using specific primer pairs. RNA from positive samples exhibiting CMV leaf symptoms was amplified to determine the coat protein. A sequence comparison of the coat protein gene from the CMV BH isolate shared the highest nucleotide identity (99.2%) with the CMV ZM isolate. Phylogenetic analysis showed that CMV-BH belonged to subgroup IA and that the most closely-related isolate was CMV-ZM. All test plants used for the biological assay were successfully infected with CMV and exhibited CMV disease symptoms such as blistering, mosaic, and vein yellowing. To our knowledge, this is the first report of CMV infection in P. heterophylla from Korea.

Relationship between the Population of Ralstonia solanacearum in Soil and the Incidence of Bacterial Wilt in the Naturally Infested Tobacco Fields

  • Chung, Yun-Hwa;Yu, Yun-Hyun;Kang, Yue-Gyu
    • The Plant Pathology Journal
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    • v.20 no.4
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    • pp.289-292
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    • 2004
  • The population of Ralstonia solanacearum (Rs) in soil is very important as a primary inoculum source of bacterial wilt in tobacco fields. To investigate the population of Rs, physical properties and chemical components during the tobacco growing season, soil samples were taken from the fifteen fields which were located in the flue-cured tobacco growing area, Ansung, Kyunggi province and Wonju, Kangwon province. Two fields of the fifteen were bacterial wilt free. Six fields had less than 10% plants being diseased and seven over 10%. The Rs population level determined by using SMSA medium generally showed an up-and-down pattern being low in May, high in Jun and July and low in August. The soil population in May and June showed a positive correlation with the incidence of bacterial wilt (r=0.571$^*$, r=0.688$^{**}$), but P$_2O_5$, content of soil was negatively correlated with the disease incidence (r=-0.539$^*$). These results suggest that Rs population in soil examined in May or in June, and the P$_2O_5$ content in soil should be key factors to determine the bacterial wilt potential of tobacco fields.

Fusarium oxysporum Protects Douglas-fir (Pseudotsuga menziesii) Seedlings from Root Disease Caused by Fusarium commune

  • Dumroese, R. Kasten;Kim, Mee-Sook;James, Robert L.
    • The Plant Pathology Journal
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    • v.28 no.3
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    • pp.311-316
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    • 2012
  • Fusarium root disease can be a serious problem in forest and conservation nurseries in the western United States. Fusarium inoculum is commonly found in most container and bareroot nurseries on healthy and diseased seedlings, in nursery soils, and on conifer seeds. Fusarium spp. within the F. oxysporum species complex have been recognized as pathogens for more than a century, but attempts to distinguish virulence by correlating morphological characteristics with results of pathogenicity tests were unsuccessful. Recent molecular characterization and pathogenicity tests, however, revealed that selected isolates of F. oxysporum are benign on Douglas-fir (Pseudotsuga menziesii) seedlings. Other morphologically indistinguishable isolates, which can be virulent, were identified as F. commune, a recently described species. In a replicated greenhouse study, inoculating Douglas-fir seedlings with one isolate of F. oxysporum prevented expression of disease caused by a virulent isolate of F. commune. Moreover, seedling survival and growth was unaffected by the presence of the F. oxysporum isolate, and this isolate yielded better biological control than a commercial formulation of Bacillus subtilis. These results demonstrate that an isolate of nonpathogenic F. oxysporum can effectively reduce Fusarium root disease of Douglas-fir caused by F. commune under nursery settings, and this biological control approach has potential for further development.

Production of Cellulase and Xylanase by Aspergillus niger KKS

  • Kang, Seong-Woo;Kim, Seung-Wook
    • Journal of Microbiology and Biotechnology
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    • v.4 no.1
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    • pp.49-55
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    • 1994
  • A fungal strain capable of producing extracellular cellulase was isolated from farmland. It was identified as Aspergillus niger, and named Aspergillus niger KKS. Production of cellulase and xylanase by the A. niger KKS was studied through a shake-flask culture. The effects of culture conditions such as inoculum size, temperature, pH, and medium composition on the cellulase and xylanase production were examined. The optimum temperature and pH for the enzyme production were $30^{\circ}C$ and pH 7.0, respectively. The optimized medium was composed of 2.0% (w/v) rice straw, 0.5% (w/v) proteose peptone, 0.5% (w/v) $KH_2 PO_4$, 0.05% (w/v) yeast extract, 0.01% (w/v) $CoSO_4 \cdot 7H_2O$, and 0.05% (w/v) $CuSO$_4$\cdot 5H_2O$. When the strain was incubated with the optimized medium, it gave the activities of endoglucanase, $\beta$-glucosidase, $\beta$-xylosidase, xylanase were 3.80, 4.20, 4.00, 80.0 (IU/mL), respectively. Filter paper and cotton activities were 0.68 and 0.045 (IU/mL), respectively. The results of this study show that A. niger KKS is a potential organism with a wide spectrum of enzyme activities, such as those of $\beta$-glucosidase, $\beta$-xylosidase, and xylanase.

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Estimation of In vitro Digestibility of Barley Straw by Using a Homogenized Rumen Fluid and Artificial Saliva Mixed with Nitrogen and Energy Sources

  • Chaudhry, Abdul S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.11 no.1
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    • pp.13-16
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    • 1998
  • A $2{\times}2{\times}4$ factorial study was conducted to examine the possibility of improving estimates of in vitro digestibility, using untreated (UBS) and ammonia-treated (ABS) barley straw, through homogenization of rumen fluid (RF) and by additions of urea (U) and casein (C) as N sources and Xylose + Glucose (XG) as energy sources into artificial saliva. Digestibiltiy of ABS was significantly greater than that of UBS (p < 0.001). There was a significant decrease in digestibility when additions (U, UC, UCXG) were compared with the control (p < 0.001). A 2-way interaction between RF and straw type was significant (p < 0.05) for dry matter digestibility (DMD). Homogenization of RF increased DMD of ABS (p < 0.05) whereas it decreased DMD of UBS (p > 0.05). The study showed that addition of N alone or in combination with energy sources was not better than control, rather the reverse, digestion was inhibited by a combination of U and casein (UC). It was concluded that sufficient N and branched chain fatty acids were supplied in the inoculum from sheep fed high protein grass cubes to support the growth of cellulolytic microbes during in vitro incubation. Further studies are, however, required to explore the potential of homogenization in improving the in vitro method to estimate digestibility of cereal straws.