• Title/Summary/Keyword: inoculum

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Study on Low Temperature Tolerant Methane-Producing Bacteria for the Treatment of Agricultural and Livestock Wastes;III. Isolation of Low Temperature Tolerant Methanogens (농축산(農畜産) 폐기물(廢棄物) 처리(處理)를 위(爲)한 저온내성(低溫耐性) 메탄 생성균(生成菌)의 특성(特性)에 관(關)한 연구(硏究);III. 저온내성(低溫耐性) Methanogens의 분리(分離))

  • Kim, Kwang-Yong;Kim, Jai-Joung;Daniels, Lacy
    • Korean Journal of Environmental Agriculture
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    • v.15 no.3
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    • pp.362-371
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    • 1996
  • This study was conducted to investigate the biochemical properties of isolated bacteria, low temperature tolerant methanogens which were selected for use as inoculum for anaerobic fermentation of agricultural and livestock wasted at low temperature. The results, obtained were summarized as follows: Low temperature tolerant methanogens were isolated from the samples which showed the high methanogenesis rate by enrichment culture at low temperature in methanol medium. These methanogens, Methanobacterium M-251 and Methanobacterium M-253 were isolated from swampy sediment at latitude $56.9^{\circ}$, Methanosarcina mazei M-372 from lake sediment IV at latitude $55.0^{\circ}$ N, and Methanobacterium formicicum M-375 from tidal land soil at latitude $37.0^{\circ}N$, respectively. The isolated anaerobic bacteria could not use sugars as carbon sources. The optimum pH value for the growth of M-251 and M-375 was 6.8, but those for M-253 and M-372 6.5 and 7.0, respectively. The minimum growth temperature of isolated, M-251 and M-253 were $8^{\circ}C$ and the optimum temperature $30^{\circ}C$, while the minimum of M-392 and M-395 were $13^{\circ}C$ and the optimum $37^{\circ}C$. The growth rate of isolates at $17.5^{\circ}C$ were lower by 32-50% than that of $30^{\circ}C$. The isolated Methanobacterium strains such as M-251, M-253, and M-375 have lower cell yield, 0.38-1.21g/1M $CH_4$ than 1.14-1.51g/1M $CH_4$ of Methanosarcina mazei M-372.

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Anaerobic Biodegradability of Leachates Generated at Landfill Age (매립년한에 따른 침출수의 혐기성 생분해 특성)

  • Shin, Hang-Sik;Lee, Chae-young;Kang, Ki-hoon
    • Journal of the Korea Organic Resources Recycling Association
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    • v.8 no.1
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    • pp.90-96
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    • 2000
  • The composition of leachates varies depending on the waste characteristics, landfill age and landfilling method. Generally, leachates contain high dissolved organic substance and ammonia nitrogen whereas phosphorus concentration was very low. Leachate A produced from young landfill is characterized by high BOD5/COD ratio (0.8) whereas leachate C produced from old landfill has lower BOD5/COD ratio (0.1). Maximum biochemical methane potential of leachate A, B (from medium landfill) and C were 271,106 and 4 ml CH4/g-COD, respectively. On the other hand, the maximum biodegradability of leachate A, B, and C were 75,30, and 1%, respectively. These results indicated that anaerobic treatment of leachate from young landfill was effective in removing organic pollutants. In case of leachate C, carbon might reside in the form of large molecular weight organic compounds such as lignins, humic acids and other polymerized compounds of soils, which are resistant to biodegradation. The lag-phase period increased with the increasing organic concentration in leachate. In case of leachate A of concentration greater than 25%, the lag-phase period increased sharply. This implied that the start-up period of anaerobic process using an unacclimated inoculum could be extended due to the higher concentration of leachate. This relatively long lag-phase is probably related to the fact that most of the inhibitory compounds have been diluted beyond their inhibitory concentrations of less than 50%. Furthermore, the ultimate methane yield and methane production rate decreased as leachate concentration increased. It was anticipated the potential inhibition was related with the steady-state inhibition as well as the initial shock load.

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Controlling activity of Bion-M against bacterial spot caused by Xanthomonas euvesicatoria (고추 세균점무늬병에 대한 Bion-M의 방제 효과)

  • Kim, A-Hyeong;Yeon, Cho-Long;Kim, Joo-Hyung;Kim, Heung-Tae
    • The Korean Journal of Pesticide Science
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    • v.16 no.2
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    • pp.171-177
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    • 2012
  • Pepper seedlings of 6-leaf stage were inoculated with cell suspension of Xanthomonas euvesicatoria 1523 7 days after the application of Bion-M by soil-drenching. Disease severity in the treatment with 20 ${\mu}g\;mL^{-1}$ of acibenzolar-S-methyl, with which Bion-M was composed, was 19%, whereas that of the untreated control was 75%. The resulting control value of acibenzolar-S-methyl was calaulated as much as 74.7%. The control value of acibenzolar-S-methyl was dependent with the applied concentrations and ranged 29.3% to 49.3% at 4.0 and 0.8 ${\mu}g\;mL^{-1}$ of acibenzolar-S-methyl, respectively. Phytotoxicity was observed at 20.0 ${\mu}g\;mL^{-1}$, as lower leaves became to be yellowed and defoliated. The cell density of inoculum suspension of X. euvesicatoria 1523 affected the control value of acibenzolar-S-methyl. With optical density (O.D.) of 0.5 the control value of 4.0 ${\mu}g\;mL^{-1}$ of acibenzolar-S-methyl was 86.0%. However, the control value improved as high as 97.8% at the O.D. value of 0.1. The control value was 75.0% in adult plant of pepper, when acibenzolar-S-methyl was treated by soil-drenching 7 days before inoculation with cell suspension of X. euvesicatoria 1523. The control effect of acibenzolar-S-methyl on pepper bacterial spot was obtained in pepper field, showing that the control value at 10.0 ${\mu}g\;mL^{-1}$ was 71.2%.

Cross-Protection Effectiveness of Cucumber mosaic virus (CMV) Isolates Associated with Satellite RNA for Prevention of CMV Disease in Pepper Plants (Satellite RNA 보유 Cucumber mosaic virus(CMV)의 고추 CMV병에 대한 교차방어 효과)

  • 최장경;성미영;정혜진;홍진성;이상용
    • Research in Plant Disease
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    • v.7 no.3
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    • pp.155-163
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    • 2001
  • Two attenuated Cucumber mosaic virus (CMV) isolates, Paf-CMV and Rs2-CMV that had been selected from CMV isolates associated with satellite RNA (satRNA) were tested for cross-protection effect in pepper plants. The viruses selected as attenuated strains appeared to be identical serologically and physically to the challenge virus (Mf-CMV), but they were lower in the dilution end-point of infectivity of crude sap than Mf-CMV When symptoms were observed in several indicator plants after inoculation, Paf-CMV and Rs2-CMV were symptomless or showed mild mosaic symptoms while another satRNA isolate Ap-CMV developed severe mosaic symptoms on the leaves as Mf-CMV. The nucleotide sequences of the satRNAs were determined by sequencing full-length cDNA clones. Paf-, Rs2- and Ap-satRNAs were 386, 335, and 347 nucleotides long, respectively, The sequences were then compared with the other known Y-satRNA, revealing that nucleotide sequences of the satRNAs consisted of 5'- and 3'-terminal conserved regions. However variations occurred on the middle regions of the sequences, especially those related to symptom interference, showing significant differences between Paf-satRNA and other isolates. Infectious transcripts of Paf-satRNA and Rs2-satRNA induced mild mosaic symptoms in pepper plants when supported by genomic RNAs of Mf-CMV. Under greenhouse conditions, Paf-CMV and Rs2-CMV were tested for cross-protection effect in pepper and tobacco (Nicotiana tabacum cv, Xanthi nc) plants against Mf-CMV. No symptoms were developed on the plants vaccinated with Paf-CMV until 3 weeks after inoculation with the virulent strain; however another attenuated isolate, Rs2-CMV, showed less effectiveness in cross-protection. Depending on the concentration of the challenged virus, symptoms sometimes appeared later in the upper leaves. However, in plants challenged with low concentrations (below 0.2 mg/ml) of the challenge inoculum, symptoms caused by the virulent strain did not develop on the plants vaccinated with Paf-CMV. In the field experiments, the number of pepper plants with severe mosaic symptoms in the control plots was progressively increased after transplanting and reached approximately 50% after 50 days. On the other hand, the incidence of mosaic disease appeared very low on the plants that had received the protective inoculation with Paf-CMV.

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Development of an Efficient Simple Mass-Screening Method for Resistant Melon to Fusarium oxysporum f. sp. melonis (덩굴쪼김병 저항성 멜론을 위한 효율적이고 간편한 대량 검정법 개발)

  • Lee, Won Jeong;Jang, Kyoung Soo;Choi, Yong Ho;Kim, Heung Tae;Kim, Jin-Cheol;Choi, Gyung Ja
    • Research in Plant Disease
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    • v.21 no.3
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    • pp.201-207
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    • 2015
  • This study was conducted to establish a simple mass-screening method for resistant melon to Fusarium wilt caused by Fusarium oxysporum f. sp. melonis (FOM). Root-dipping inoculation method has been used to investigate resistance of melon plants to Fusarium wilt. However, the inoculation method requires a lot of labor and time because of complicate procedure. To develop a simple screening method on melon Fusarium wilt, occurrence of Fusarium wilt on susceptible and resistant cultivars of melon according to inoculation method including root-dipping, soil-drenching, tip, and scalpel methods was investigated. Scalpel and tip methods showed more clear resistant and susceptible responses in the melon cultivars than root-dipping inoculation method, but tip method represented slightly variable disease severity. In contrast, in the case of soil-drenching inoculation method, disease severity of the susceptible cultivars was very low. Thus we selected scalpel method as inoculation method of a simple screening method for melon Fusarium wilt. By using the scalpel inoculation method, resistance degrees of the cultivars according to incubation temperature after inoculation (25 and $30^{\circ}C$) and inoculum concentration ($1{\times}10^6$ and $1{\times}10^7conidia/ml$) were measured. The resistance or susceptibility of the cultivars was hardly affected by all the tested conditions. To look into the effectiveness of scalpel inoculation methods, resistance of 22 commercial melon cultivars to FOM was compare with root-dipping inoculation method. When the melon cultivars were inoculated by scalpel method, resistance responses of all the tested cultivars were clearly distinguished as by root-dipping method. Taken together, we suggest that an efficient simple mass-screening method for resistant melon plant to Fusarium wilt is to sow the seeds of melon in a pot (70 ml of soil) and to grow the seedlings in a greenhouse ($25{\pm}5^{\circ}C$) for 7 days, to cut the root of seedlings with a scalpel and then pour a 10 ml-aliquot of the spore suspension of $1{\times}10^6conidia/ml$ on soil. The infected plants were cultivated in a growth room at 25 to $30^{\circ}C$ for about 3 weeks with 12-hr light a day.

Growth inhibition of hydrotrope-combined copper against Microcystis aeruginosa and evaluation of its toxicity (Microcystis aeruginosa에 대한 hydrotrope-combined copper의 생장억제 및 독성 평가)

  • Park, Se-Keun;Ji, Jun-Gu;Jang, Hee Jung;Kim, Yeong-Kwan;Oh, Young-Sook;Choi, Sung-Chan
    • Korean Journal of Microbiology
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    • v.51 no.1
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    • pp.7-13
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    • 2015
  • Hydrotrope-combined copper (HCC) is a copper ($Cu^{2+}$)-based algicide, which is combined with a hydrotrope that keeps copper ion in solution to improve performance. This study assessed the growth inhibition effect of HCC against Microcystis aeruginosa which is one of the most common toxic cyanobacterium in eutrophic freshwater environment. Various HCC doses, ranging from 5.5 to $550{\mu}g/L$ as $Cu^{2+}$, were applied to either BG-11 or 1/4 diluted medium with low- or high-inoculum density of M. aeruginosa. Growth inhibition was monitored based on a decrease in chlorophyll-a content in culture medium during the incubation. Results showed that HCC significantly inhibited the growth of M. aeruginosa in a dose-dependent manner. In case of 1/4 diluted BG-11 medium, HCC dose as low as $5.5{\mu}g$ $Cu^{2+}/L$ completely inhibited the production of chlorophyll-a by M. aeruginosa. It was found that HCC did not induce any significant release of microcystin-LR from M. aeruginosa. Acute toxicity of HCC was tested using Daphnia magna, and the 24-h $EC_{50}$ value was 0.30 mg/L as $Cu^{2+}$ which was much higher than the actual inhibition dose. Ames test was performed using Salmonella enterica serovar Typhimurium TA100, and HCC showed no increase in the number of revertant colonies. The result suggested that HCC does not have any mutagenic potential in the aquatic environment. In addition, no genotoxic effect of HCC was also confirmed based on the SOS ChromoTest using Escherichia coli PQ37. Therefore, HCC could be used as a relatively safe and effective pre- and post-treatment agent to control hazardous algal blooming in aquatic environments.

Development of Efficient Screening Methods for Resistance of Tomato to Fusarium oxysporum f. sp. lycopersici (토마토 시들음병에 대한 효율적인 저항성 검정법 확립)

  • Park, Myung-Soo;Jeong, Bo-Ram;Jang, Kyoung-Soo;Choi, Yong-Ho;Kim, Jin-Cheol;Choi, Gyung-Ja
    • Horticultural Science & Technology
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    • v.30 no.4
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    • pp.426-431
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    • 2012
  • This study was conducted to establish an efficient screening method for resistant tomato to Fusarium wilt caused by Fusarium oxysporum f. sp. lycopersici (FOL). The resistance degrees of the six commercial cultivars of tomato to the pathogen were evaluated by dipping roots of the seedlings in spore suspension of five FOL isolates. On the basis of the results, two cultivars (Dotaerangmaster, resistant cultivar to FOL race 1; Supersunload, resistant cultivar to FOL race 2) and two isolates (KACC40043, FOL race 2; TF104, FOL race 3) were selected for system establishment. The disease development of the FOL isolates on the cultivars according to several conditions including root wounding, incubation temperature, inoculum concentration and dipping period of roots in spore suspension was investigated. The resistance of each cultivar to the disease was a race-specific response and hardly affected by the tested conditions except for incubation temperature of $20^{\circ}C$. The optimum temperature for disease development caused by FOL was 25 to $30^{\circ}C$. On the basis of the results, we suggest that an efficient screening method for resistant tomato cultivars to Fusarium wilt is to dip the non-cut roots of tomato seedlings at two-leaf stage in spore suspension of $1{\times}10^7\;conidia{\cdot}mL^{-1}$ for 0.5 hours and transplant the seedling to plastic pot with horticulture nursery media, and then to cultivate the plants in a growth room at $25^{\circ}C$ for 3 weeks with 12 hours light a day.

Effect of Omija(Schizandra chinensis) Extract on the Growth Inhibition of Food Borne Pathogens in Yoghurt (오미자 추출액 첨가 요구르트의 식중독균 증식 억제 효과)

  • 홍경현;남은숙;박신인
    • Food Science of Animal Resources
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    • v.23 no.4
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    • pp.342-349
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    • 2003
  • Escherichia coli O157:H7, Staphylococcus aureus and Salmonella enteritidis are food borne pathogens involved in food poisoning in numerous countries. This study aimed to obtain knowledges on the survival of Esc coli O157:H7, Sta aureus and Sal. enteritidis in the yoghurt added with water extract of Omija(Schizandra chinensis). The growth inhibition of Schizandra chinensis extract on the food borne pathogens were measured by total microbial count and effect of growth inhibition was correspondent to the concentration of Schizandra chinensis extract. The highest growth inhibition effect of Schizandra chinensis extract was shown on the Sta aureus followed by Sal. enteritidis and Esc. coli O157:H7. The number of surviving Esc. coli O157:H7 cell(3.55${\times}$10$\^$5/ CFU/mL) was decreased to 1.00${\times}$10$^1$∼3.00${\times}$10$^1$ CFU/mL after 24 hours incubation by the addition of 0.4∼l.0% of Schizandra chinensis extract in the yoghurt. And also the viable cell counts of surviving Sta. aureus cells (initial inoculum 1.24${\times}$10$\^$5/ CFU/mL) were decreased gradually to 4.00${\times}$10$^2$∼8.50${\times}$10$^2$ CFU/mL after 48 hours of incubation, but the viable cells of Sal. enteritidis were not detected after 24 hours of incubation. Growth of the food borne pathogens was strongly inhibited by the addition and incubation of Schizandra chinensis extract for 48 hours in the yoghurt.

Effects of Rennin and/or Starter Addition on the Manufacture and Ripening of Soybean Cheeses (Rennin 및 Starter의 첨가(添加)가 대두(大豆)치즈의 제조(製造) 및 숙성(熟成)에 미치는 영향(影響))

  • Jeong, Jae Hong;Choi, Woo Young
    • Korean Journal of Agricultural Science
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    • v.7 no.2
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    • pp.190-201
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    • 1980
  • This experiment was designed to optimize the process of manufacturing the soybean cheeses and to elucidate the chemical changes during ripening when the chemical changes during ripening when the milk components and enzyme preparations were added to the raw materials. Conditions for extracting soybean protein such as temperature, duration and amount of water added were determined; various coagulaters were compared by checking the curd texture and yield; starters from S. thermophilus, S. lactis MLB and S. cremoris EB-9 were tested as single- or multi-stain combinations; and the effects of skim milk and/or rennins-both microbial and calf origin-addition upon the process of manufacturing and ripening were studied. The results obtained were as follows. 1. optimal conditions for soybean extraction were found to be: temperature $100^{\circ}C$, duration 10 minutes, and amount of water added 9-fold, as considered the extraction rate of solids and proteins, and curd yield. 2. Sodium gluconate was the most effective among the coagulators tested, and 5% of single-strain starter from S. thermophilus was appered to be adequate inoculum for curd formation. 3. The effects of skim milk and/or rennins addition on the process of manufacturing and ripening of soybean cheeses were: 1) The addition of rennins resulted in fast formation of curd, especially with skim milk it was so. And Hansen rennet extracts brought better results in curd formation than Meito rennet extracts did. 2) No significant effect was observed on the changes in moisture content during ripening, however the levels of moisture contents in the products were higher in case of using Meito rennet extracts. 3) Effect on pH changes during ripening was also not significant in general, while levels of pH were decrease markedly during manufacturing and the initial stage of ripening. 4) The levels of bacterial counts were much higher in case of skim milk addition throughtout the ripening period. In general the numbers were reached to approximately $10^8cells/g$ during manufacturing, then decreased gradually to below $10^2cells/g$ in 8 weeks of ripening. 5) The addition of skim milk and/or rennin resulted in higher ripening index, and skim milk plus Meito rennet extracts was appeared to be best combination for the ripening index.

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Disinfection of Fusarium-infected Rice Seeds by Prochloraz and Gaseous Chlorine Dioxide

  • Jeon, Young-ah;Lee, Young-yi;Lee, Ho-sun;Sung, Jung-sook;Lee, Seokyoung
    • 한국균학회소식:학술대회논문집
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    • 2014.10a
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    • pp.25-25
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    • 2014
  • Three species of Fusarium, F. fujikuroi, F. verticillioides and F. proliferatum, are known to be associated with bakanae disease of rice [1, 2]. F. fujikuroi infects rice flowers and survive in endosperm and embryo of the seeds. Infected seed is an important source of primary inoculum of pathogens [3]. Seeds of rice (Oryza sativa cv. Boramchan) collected from bakanae-infected field were found to be 96% infected with Fusarium sp., 52% with F. fujikuroi, 42% with F. verticillioides, and 12% with F. proliferatum as determined by incubation method and species-specific PCR assays. F. fujikuroi was detected at lemma/palea, endosperm and embryo whereas F. verticillioides and F. proliferatum were recovered only from lemma/palea by means of component plating test. Seed disinfection methods have been developed to control bakanae disease and prochloraz has been most widely used for rice seeds. Two chemicals formulated with prochloraz (PC 1) and prochloraz + hexaconazole (PC 2) that inhibit biosynthesis of ergosterol strongly reduced the incidence of Fusarium spp. on selective media to 4.7% and 2.0%, respectively. Disease symptoms of rice seedlings in nursery soil were alleviated by chemical treatment; seedlings with elongated leaves or wide angle between leaf and stem were strikingly reduced from 15.6 to 3.2% (PC 1) and 0 (PC 2), stem rots were reduced from 56.9 to 26.2% (PC 1) and 32.1% (PC 2), and normal seedling increased from 0.4 to 13.3% (PC 2). Prochloraz has some disadvantages and risks such as the occurrence of tolerant pathogens [4] and effects on the sterol synthesis in animals and humans [5]. For these reasons, it is necessary to develop new disinfection method that do not induce fungal tolerance and are safe to humans and animals. Chlorine dioxide ($ClO_2$), that is less toxic, produces no harmful byproducts, and has high oxidizing power, has been reported to be effective at disinfection of several phytopathogenic fungi including Colletotrichum spp. and Alternaria spp. [6]. Gaseous $ClO_2$ applied to rice seeds at a concentration of 20 ppm strongly suppressed mycelial growth of Fusarium fujikuroi, F. verticillioides and F. proliferatum. The incidence of Fusarium spp. in dry seed with 8.7% seed moisture content (SMC) tended to decrease as the concentration of $ClO_2$ increased from 20 to 40 ppm. Applying 40 ppm $ClO_2$ at 90% relative humidity, incidence was reduced to 5.3% and resulted in significant reduction of disease symptoms on MS media. In nursery soil, stem rot was reduced from 56.9 to 15.4% and the number of normal seedlings increased from 0.4 to 25.5%. With water-soaked seeds (33.1% SMC) holding moisture in the endosperm and embryo, the effectiveness of disinfection using $ClO_2$ increased, even when treated with only 20 ppm for four hours. This suggests that moisture was a key element for action of $ClO_2$. Removal of the palea and lemma from seeds significantly decreased the incidence of Fusarium spp. to 3.0%. Seed germination appeared to decrease slightly by water-soaking at $30^{\circ}C$ because of increased SMC and by physical damage of embryos from hulling. These results indicate that the use of gaseous $ClO_2$ was effective as a means to disinfect rice seeds infected with Fusarium spp. and that moisture around the pathogens in the seed was an important factor for the action of $ClO_2$. Further investigations should be conducted to ascertain the best conditions for complete disinfection of Fusarium spp. that infect deep site of rice seeds.

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