• Title/Summary/Keyword: Spore production

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Optimal culture conditions for mass production of rock polypody (Polypodium vulgare L.)

  • Jang, Bo Kook;Park, Kyungtae;Han, Ahreum;Lee, Cheol Hee
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.04a
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    • pp.44-44
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    • 2019
  • This study aimed to develop a suitable method for inducing the proliferation of prothallus and producing sporophytes of rock polypody (Polypodium vulgare L.). The prothalli used in all experiments were obtained from spore germination and sub-cultured for 8-week intervals. The most appropriate media for prothallus propagation were investigated by culturing 300 mg of prothallus in MS ($1/4{\times}$, $1/2{\times}$, $1{\times}$, and $2{\times}$ strength) medium and in Knop medium for 8 weeks. Cultures were maintained at a temperature of $25{\pm}1^{\circ}C$, light intensity of $30{\pm}1.0{\mu}mol-m-2{\cdot}s-1$, and a photoperiod of 16/8 h (light/dark). Fresh weight of prothalli was 4.8 g on $1{\times}$ MS, 4.5 g on $1/2{\times}$ MS and 4.3 g on 1/4 MS medium. To select a suitable soil combination for sporophyte formation, 1.0 g of prothallus was ground with distilled water, spread in five combinations onto different soil substrates (decomposed granite, horticultural substrates, peat moss, and perlite), and then cultivated for 13 weeks. The sporophyte cultures were maintained at a temperature of $25{\pm}1^{\circ}C$, light intensity of $43{\pm}2.0{\mu}mol-m-2{\cdot}s-1$, humidity of $84{\pm}1.4%$, and a photoperiod of 16/8 h (light/dark). The results showed that a mixture containing a 2:1 (v:v) ratio of horticultural substrate and perlite, increased sporophyte formation to 462.5 sporophytes per pot (7.5 cm2). The other soil substrates produced from 314.5 to 405.3 sporophytes per pot. Therefore, our results will provide conditions suitable for mass production of Polypodium vulgare L.

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Production performances and antioxidant activities of laying hens fed Aspergillus oryzae and phytase co-fermented wheat bran

  • Huang, Chung Ming;Chuang, Wen Yang;Lin, Wei Chih;Lin, Li Jen;Chang, Sheng Chang;Lee, Tzu Tai
    • Animal Bioscience
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    • v.34 no.3_spc
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    • pp.371-384
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    • 2021
  • Objective: Wheat bran (WB) was co-fermented with Aspergillus oryzae and phytase (Phy) to determine whether co-fermentation improve WB phosphorus and fiber utilization in Isa-brown layers. Methods: A total of 112 Isa brown layer were randomly divided into 7 treatments with 8 replicates per a treatment and 2 hens per a replicate. The treatments included basal diet (control), basal diet supplemented with 250 unit/kg Phy (control+Phy), diet with 10% WB (10% WB), diet with 5% WB and 250 unit/kg Phy (5% WB+Phy) diet with 10% WB and 250 unit/kg Phy (10% WB+Phy), diet with 5% fermented WB supplemented with molasses and phy (PCFWH) and 125 unit/kg Phy (5% PCFWH), and diet with 10% PCFWH (10% PCFWH). The intestinal microbial population, intestinal morphology, serum antioxidant enzyme activities, and excreta phosphorus content were assessed. Results: In PCFWH, spore counts, protease activity, xylanase activity, and ferulic acid were 8.50 log/g dry matter (DM), 190 unit/g DM, 120 unit/g DM, and 127 ㎍/g, respectively. Xylobiose and xylotriose were released in PCFWH, while they were not detectable in WB. Antioxidant capacity was also enhanced in PCFWH compared to WB. The 10% WB+Phy and 10% PCFWH groups produced higher egg mass, but hens fed 5% WB+Phy had the lowest amount of feed intake. Eggs from 10% PCFWH had better eggshell weight, eggshell strength, and eggshell thickness. Birds fed with 10% PCFWH also had higher serum superoxide dismutase and catalase activities. Compare to control, 10% PCFWH significantly reduced excreta phosphorus content. Conclusion: Diet inclusion of 10% PCFWH improved egg quality, antioxidant status, and excreta phosphorus content of laying hens.

Characteristics of New Microsporidia S80 Isolated from Silkworm, Bombyx mori L. in Korea (가잠(家蠶)으로부터 분리(分離)된 새로운 Microsporidia S80의 특성(特性))

  • Lim, Jong Sung;Cho, Sae Yun
    • Current Research on Agriculture and Life Sciences
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    • v.1
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    • pp.67-83
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    • 1983
  • The new microsporidia S80 isolated from, Bombyx mori L. in Korea showed ovoid in the morphology of the spores and the size were measured $2.9{\pm}0.28{\mu}$ in length and $1.7{\pm}0.29{\mu}$ width. No other microsporidian spore like this has not been so far isolated from Silkworm. The length of the polar filament extruded in hydrogen peroxide ($H_2O_2$) at $30^{\circ}C$ was $26{\mu}$ of a round cytoplasm on the top. The spores were partly stained with Giemsa, Safranin-O and Gram as the same staining properties as Nosema bombycis, Microsporidia K 79 and other microsporidian spores. The fine structures were observed under scanning eleceron microscope through ultrathin sectioning. The spore wall was composed of three layers ; the thin exospore of an electron dense rippled layer, the thick electron lucent endospore which was thinning considerably at the polar filament insertion point, and the inner limiting membrane. Polar cap present at the sporeapex, with a long polar filament of 12-13 coils, subtending angle of $60^{\circ}$ to spore axis, which is tubular made up of a multilayered and are a benes core, light ring structure enclosing the dance core, the dark ring structure enclosing the inner light ring structure and the other than and light ring structure bounded from cytoplasm. Lamellate polaroplast occupied the anterior part of the spore, and the two neclei with dense nucleoplasm bounded by a double nuclear envelope were cited in the slight downer middle portion of spore. From the characteristics of the shape, size and fine structures, it is certain to reason the Microsporidia S80 belong to the phylum Microspora, class Microspora, order Microsporida, order Microsporida. The shape of two nuclei cited seems to be genus Nosema, but in the classification for the suborder it should be defined wheather pansporoblasts be formed or not and for the genis especial attempts have been made to define the characters which distinguish the disporous genera in the life cycle. Survey through the infection of the bad cocoons during 1980 to 1982 in South Korea the areas contaminated with new microsporidia were revealed 5 provinces of Kyung-Gi, Kang-Won, Chung-Nam and Chun-Nam. Pathological effects inoculated per os at second instar larvae of silkworm, the LD 50 was $7.1{\times}10^7/ml$ as lower pathogenecity than that of Nosema bombycis Naegeli of $1.2{\times}10_7/ml$. While on the other hand the inoculation of the microsporidia at fourth instar larvae lowerd the whole cocoon weight and cocoon shell weight and significant at 1% level. The microsporidia S80 defined it can not be transmitted transovarially from the result of predictive and collective examination of 21 egg batches from the infected female moth.

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Citric Acid Production from Glucose and Pumpkin by Using Immobilized Bead of Aspergillus niger (고정화 Aspergillus niger Bead를 이용한 포도당과 호박으로부터 구연산 생성)

  • 방병호
    • The Korean Journal of Food And Nutrition
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    • v.13 no.4
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    • pp.328-333
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    • 2000
  • The spore of Aspergillus niger KCTC-6144 was immobilized on alginate gel beads. When pumpkin powder was used with glucose for a medium of citric acid fermentation by Aspergillus niger beads, the beaded Aspergilus niger grew up inside the bead and mycelia penetrated through the pore of the bead membrane. The bead size became largely from 2.0∼2.5mm to 6∼8mm after growing at 30$\^{C}$ for 4 days. Studies of optimum culture conditions on citric acid fermentation using Aspergillus niger beads on pumpkin medium (pumpkin powder 1% +glucose 7%, pH 6.0) were carried out in submerged cultures on 250m1 Erlenmeyer flask. As a result, it was found that to reinforce 12% as carbon source was good for citric acid production and that 1% pumpkin powder was good as nitrogen and mineral source in orbital shaker (150rpm) at 30$\^{C}$ for 5 days. The optimum initial pH on citric acid production was pH 6.0 and it was found that 100 beads of immobilized Aspergillus niger was adequate for citric acid production in a 250ml Erlenmeyer flask containing 50m3 of pumpkin medium solution with orbital shaker at 30$\^{C}$ for 5 days. We also found that maximal production of citric acid was 23.5g/ℓ at optimal condition (at 30$\^{C}$ for 5 days, pH 6.0, and 100 beads and medium containing 1% pumpkin powder plus 12% glucose).

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Genetic Control of Asexual Sporulation in Fusarium graminearum

  • Son, Hokyoung;Kim, Myung-Gu;Chae, Suhn-Kee;Lee, Yin-Won
    • 한국균학회소식:학술대회논문집
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    • 2014.10a
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    • pp.15-15
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    • 2014
  • Fusarium graminearum (teleomorph Gibberella zeae) is an important plant pathogen that causes head blight of major cereal crops such as wheat, barley, and rice, as well as causing ear and stalk rot on maize worldwide. Plant diseases caused by this fungus lead to severe yield losses and accumulation of harmful mycotoxins in infected cereals [1]. Fungi utilize spore production as a mean to rapidly avoid unfavorable environmental conditions and to amplify their population. Spores are produced sexually and asexually and their production is precisely controlled. Upstream developmental activators consist of fluffy genes have been known to orchestrate early induction of condiogenesis in a model filamentous fungus Aspergillus nidulans. To understand the molecular mechanisms underlying conidiogenesis in F. graminearum, we characterized functions of the F. graminearum fluffy gene homologs [2]. We found that FlbD is conserved regulatory function for conidiogenesis in both A. nidulans and F. graminearum among five fluffy gene homologs. flbD deletion abolished conidia and perithecia production, suggesting that FlbD have global roles in hyphal differentiation processes in F. graminearum. We further identified and functionally characterized the ortholog of AbaA, which is involved in differentiation from vegetative hyphae to conidia and known to be absent in F. graminearum [3]. Deletion of abaA did not affect vegetative growth, sexual development, or virulence, but conidium production was completely abolished and thin hyphae grew from abnormally shaped phialides in abaA deletion mutants. Overexpression of abaA resulted in pleiotropic defects such as impaired sexual and asexual development, retarded conidium germination, and reduced trichothecene production. AbaA localized to the nuclei of phialides and terminal cells of mature conidia. Successful interspecies complementation using A. nidulans AbaA and the conserved AbaA-WetA pathway demonstrated that the molecular mechanisms responsible for AbaA activity are conserved in F. graminearum as they are in A. nidulans. F. graminearum ortholog of Aspergillus nidulans wetA has been shown to be involved in conidiogenesis and conidium maturation [4]. Deletion of F. graminearum wetA did not alter mycelial growth, sexual development, or virulence, but the wetA deletion mutants produced longer conidia with fewer septa, and the conidia were sensitive to acute stresses, such as oxidative stress and heat stress. Furthermore, the survival rate of aged conidia from the F. graminearum wetA deletion mutants was reduced. The wetA deletion resulted in vigorous generation of single-celled conidia through autophagy-dependent microcycle conidiation, indicating that WetA functions to maintain conidia dormancy by suppressing microcycle conidiation in F. graminearum. In A. nidulans, FlbB physically interacts with FlbD and FlbE, and the resulting FlbB/FlbE and FlbB/FlbD complexes induce the expression of flbD and brlA, respectively. BrlA is an activator of the AbaA-WetA pathway. AbaA and WetA are required for phialide formation and conidia maturation, respectively [5]. In F. graminearum, the AbaA-WetA pathway is similar to that of A. nidulans, except a brlA ortholog does not exist. Amongst the fluffy genes, only fgflbD has a conserved role for regulation of the AbaA-WetA pathway.

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Inhibition of growth and toxin production of ochratoxigenic Aspergillus spp. by isolated bacteria (분리세균에 의한 ochratoxin 생성 Aspergillus spp.의 생장 및 독소생성 저해)

  • Hwang, Ji-Seon;Choi, Ho-Yeong;Song, Hong-Gyu
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.226-233
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    • 2019
  • Ochratoxin A (OTA) that is one of mycotoxins produced mainly by Aspergillus spp. is a common contaminant of stored grains and poses health hazards to human and livestock. The aim of this study is to explore the ability of isolated bacteria Bacillus subtilis AF13 and Streptomyces shenzhenensis YR226 to inhibit growth and OTA production of 3 ochratoxigenic Aspergillus strains. The antifungal activity against mycelial growth and sporulation of Aspergillus strains was examined by coculture with AF13 and YR226 on potato dextrose agar plate. AF13 and YR226 reduced 77.58 and 78.48% of fungal colony radius, respectively, and both strains inhibited fungal sporulation up to 99% in 10 days of incubation. YR226 also reduced more than 91% of spore germination of 3 fungal strains. When Aspergillus strains were cocultured with AF13 or YR226 in yeast extract sucrose medium, mycelial growth and OTA production decreased in all three fungal strains. In particular, AF13 completely inhibited the mycelial growth of A. alutaceus and inhibited its OTA production by 99%, and YR226 also reduced mycelial growth and toxin production up to 99%, respectively. Antimicrobial substances produced by AF13 and YR226 included siderophore, chitinase, protease, ${\beta}$-1,3-glucanase and biosurfactant. These results suggest that AF13 and YR226 can be used in a biological method to prevent valuable crops against mycotoxigenic fungi, and therefore decrease economic damage in agriculture and feed industry.

Enhancement of Seaweed Rhizoid and Blade Formations by the Chlorophyte Codium fragile Extract (녹조류 청각 추출물에 의한 해조류 가근 및 유엽형성 촉진)

  • Getachew, Mehader;Getachew, Paulos;Cho, Ji-Young;Choi, Jae-Suk;Hong, Yong-Ki
    • Journal of Life Science
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    • v.26 no.4
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    • pp.484-489
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    • 2016
  • Living organisms can maintain or extend their territories by producing allelochemicals that influence the growth, survival, and reproduction of other organisms. To identify natural biostimulants of positive allelochemicals, we screened 18 common seaweed extracts for enhancement of rhizoid and blade production in a convenient Porphyra suborbiculata monospore assay. By addition of methanolic extract from the most potent green seaweed, Codium fragile, 100% and 50% enhancement doses reflecting the amount of C. fragile extract required to enhance rhizoid formation (in terms of number of spores with rhizoids per total spores tested) were approximately 100 and 50 μg/ml, respectively, in the P. suborbiculata monospore culture. The C. fragile extract quickly enhanced rhizoid formation, rhizoid numbers per rhizoid-holding spore, rhizoid length, blade formation (in number of spores with blade per total spores tested), and blade length from most monospores at early culture days. The extract enhanced rhizoid formation after 2 days of culture significantly, rhizoid numbers per rhizoid-holding spore after 3 days, rhizoid length after 3 days, blade formation after 2 days, and blade length after 1 day, respectively, from most monospores. The allelochemicals that enhanced favorite seaweed species may be efficacious for new seaweed management technologies, including the development of biostimulant agents based on natural products.

Enteritis Caused by Type A Clostridium perfringens Producing $\alpha$-Toxin in a Dog (개에서 $\alpha$-Toxin을 생성하는 Type A Clostridium perfringens에 의한 장염)

  • Han, Jae-Ik;Jang, Hye-Jin;Cheon, Haeng-Bok;Kim, Ji-Hye;Kim, Gon-Hyung;Chang, Dong-Woo;Na, Ki-Jeong
    • Journal of Veterinary Clinics
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    • v.27 no.2
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    • pp.190-193
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    • 2010
  • A 6-year-old, female, Siberian husky was referred with mucous diarrhea. On fecal examination, numerous clustered and individual large epithelial cells and rod-shaped, spore-forming bacteria were examined. By bacterial culture and molecular typing, the bacteria was identified as Clostridium perfringens (C. perfringens), and by toxin analysis of C. perfringens, production of $\alpha$-toxin was confirmed. Based on these results, the dog was diagnosed as enteritis caused by C. perfringens producing $\alpha$-toxin, and was treated with amoxicillin/clavulanate. After 1 week, the diarrhea was disappeared and no spore-forming bacteria were examined on fecal examination. This report shows that the rapid and exact diagnosis keeps a effective treatment for enteritis caused by C. perfringens producing $\alpha$-toxin in dogs.

Methods of Application and Beneficial Effects of Silicate-Coating Rice Seeds (볍씨의 규산코팅방법에 따른 이용특성과 육묘효과)

  • Kang, Yang-Soon;Kim, Wan Joong;Hwang, Duck Sang;Kim, Hee Kyu
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.65 no.1
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    • pp.30-39
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    • 2020
  • A new silicate coating technology was developed which reduces the impact of dust and loosening during seeding compared to existing silicate-coatings (Seed/Si/Zeolite), and therefore can lower the production costs of rice cultivation. In this method, 100 g of rice seed is coated with 18 mL of liquid silicic acid and then dressed with a mixture containing 80 g of dolomite and 5 g of iron. To determine the most effective method of application and ensure that seedlings developed healthily, a series of experiments were carried out. Infected seeds scattered in seedling boxes and pots (soil and hydroponic) were coated dry, without disinfection. In comparison to the seed which were not treated with the silicate-coating, the new seed (A) were 1.84 times heavier in weight, and were also improved in terms of coating strength and coating color. Compared to the seedlings grown from the non-coated seed, those grown from the new silicate-coated seed were of significantly higher quality (weight/length) and had erect, dark greenish leaves, which are ideal plant characteristics. This was most likely due to increased silicate uptake. The symptoms of bakanae disease in the non-coated seed peaked after 38 days to 54.2%, whereas the control value was 68.8% in the new silicate-coated seed (A). In the infected seedlings grown from the new silicate-coated rice seed, subnormal macro-conidia, namely, a sickle shape spore without a septum; a straight oblong shape spore without a septum and with a thick cell wall; and inter-septal necrosis of a normal spore were detected. It is believed that the strong alkalinity of silicic acid have acted as unfavorable conditions for pathogenicity. In seedlings grown from the new silicate coated rice seed under hydroponic conditions without nutrients, normal root activity and growth was maintained without leaf senescence. Therefore, it was possible to reduce the rate of fertilization. In the future, a new silicate-coated rice seed was required for the study of minimal nutrition for anti-aging of seedlings.

Effect of Physical Control Technology on Aspergillus ochraceus Reduction (물리적 제어기술이 Aspergillus ochraceus 저감화에 미치는 영향)

  • Lee, Eun-Seon;Kim, Jong-Hui;Kim, Bu-Min;Oh, Mi-Hwa
    • Journal of Food Hygiene and Safety
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    • v.36 no.5
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    • pp.447-453
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    • 2021
  • In this study, the effectiveness of physical control technology, a combined light sterilization (LED, UV) and hot water treatment in reducing Aspergillus ochraceus for food production environment was investigated. In brief, 1 mL aliquot of A. ochraceus spore suspension (107-8 spore/mL) was inoculated onto stainless steel chips, which was then dried at 37℃, and each was subjected to different physical treatment. Treatments were performed for 0.5, 1, 2, 5, 8, and 11 hours to reduce the strains using a light-emitting diode, but no significant difference was confirmed among the treatments. However, a significant reduction was observed on the chips treated with UV-C exposure and hot water immersion. After being treated solely with 360 kJ/m2 of UV-C on stainless steel chip, the fungi were significantly reduced to 1.27 log CFU/cm2. Concerning the hot water treatment, the initial inoculum amount of 6.49 log CFU/cm2 was entirely killed by immersion in 83℃ water for 5 minutes. Maintaining a high temperature for 5 minutes at the site is difficult. Thus, considering economic feasibility and usability, we attempted to confirm the appropriate A. ochraceus reduction conditions by combining a relatively low temperature of 60℃ and UV rays. With the combined treatments, even in lukewarm water, A. ochraceus decreased significantly through the increases in the immersion time and the amount of UV-C irradiation, and the yield was below the detection limit. Based on these results, if work tools are immersed in 60℃ lukewarm water for 3 minutes and then placed in a UV sterilization device for more than 10 minutes, the possibility of A. ochraceus cross-contamination during work is expected to be reduced.