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Occurrence of the Phytophthora Blight Caused by Phytophthora sansomeana in Atractylodes macrocephala Koidz. (Phytophthora sansomeana에 의한 큰꽃삽주 역병 발생 보고)

  • An, Tae Jin;Park, Myung Soo;Jeong, Jin Tae;Kim, Young Guk;Kim, Yong Il;Lee, Eun Song;Chang, Jae Ki
    • Korean Journal of Medicinal Crop Science
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    • v.27 no.6
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    • pp.404-411
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    • 2019
  • Background: In September 2017, wilting and rhizome rot symptoms were observed on Atractylodes macrocephala Koidz. in Jecheon-si and Eumseong-gun. This study was carried out to isolate hitherto unidentified pathogenic fungi from A. macrocephala and to test the pathogenicity of isolated fungi against Atractylodes spp. genus such as A. macrocephala, A. japonica, and their interspecific hybrids. Methods and Results: The diseased plants were washed with running tap water, and the boundary between the healthy area and the diseased area was cut while the pathogens were isolated by growing cultures from the diseased areas on Phytophthora semi-selective medium. The internal transcribed spacer (ITS) region of the isolates was used in this study for identification. Test plants were cultivated in the glasshouse at 20℃ - 30℃ for 4 months and then used for pathogenicity test. The pots with plants inoculated with mycelial plugs and zoospores were placed at 25℃ for 48 h in a dew chamber where relative humidity was above 95%, and then moved into the glasshouse at 20℃ - 30℃. The presence or absence of pathogenicity of the strains was determined by evaluating the symptom of plant wilting. The inoculation test was performed in three replicates with a non-treated control. Conclusions: On the basis of results of ITS sequence analysis, the strains isolated from the diseased plants was identified as Phytophthora sansomeana. Biological assay using test plants confirmed the pathogenicity of P. sansomeana against Atractylodes macrocephala. This is the first report of rhizome rot in A. macrocephala caused by P. sansomeana.

Effect of inclusion of different levels of Leucaena silage on rumen microbial population and microbial protein synthesis in dairy steers fed on rice straw

  • Nguyen, Thien Truong Giang;Wanapat, Metha;Phesatcha, Kampanat;Kang, Sungchhang
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.2
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    • pp.181-186
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    • 2017
  • Objective: Leucaena leucocephala (Leucaena) is a perennial tropical legume that can be directly grazed or harvested and offered to ruminants as hay, silage, or fresh. However, Leucaena contain phenolic compounds, which are considered anti-nutritional factors as these may reduce intake, digestibility and thus animal performance. Therefore, the objective of this experiment was to determine effects of Leucaena silage (LS) feeding levels on rumen microbial populations, N-balance and microbial protein synthesis in dairy steers. Methods: Four, rumen fistulated dairy steers with initial weight of $167{\pm}12kg$ were randomly assigned to receive dietary treatments according to a $4{\times}4$ Latin square design. Treatments were as followings: T1 = untreated rice straw (RS; Control), T2 = 70% RS+30% LS, T3 = 40% RS+60% LS, and T4 = 100% LS. Dairy steers were fed rice straw and LS ad libitum and supplemented with concentrate at 0.2% of body weight/d. Results: Results revealed that the rumen microbial population, especially cellulolytic, proteolytic bacteria and fungal zoospores were enhanced in steers that received 60% of LS (p<0.05), whereas the amylolytic bacteria population was not affected by treatments (p>0.05). Protozoal population was linearly decreased with increasing level of LS (p<0.05). Moreover, N-balance and microbial protein synthesis were enhanced by LS feeding (p<0.05) and were the highest in 60% LS group. Conclusion: Based on this study, it could be concluded that replacement of RS with 60% LS significantly improved microbial population and microbial protein synthesis in diary steers.

Paromomycin Derived from Streptomyces sp. AG-P 1441 Induces Resistance against Two Major Pathogens of Chili Pepper

  • Balaraju, Kotnala;Kim, Chang-Jin;Park, Dong-Jin;Nam, Ki-Woong;Zhang, Kecheng;Sang, Mee Kyung;Park, Kyungseok
    • Journal of Microbiology and Biotechnology
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    • v.26 no.9
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    • pp.1542-1550
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    • 2016
  • This is the first report that paromomycin, an antibiotic derived from Streptomyces sp. AG-P 1441 (AG-P 1441), controlled Phytophthora blight and soft rot diseases caused by Phytophthora capsici and Pectobacterium carotovorum, respectively, in chili pepper (Capsicum annum L.). Chili pepper plants treated with paromomycin by foliar spray or soil drenching 7 days prior to inoculation with P. capsici zoospores showed significant (p < 0.05) reduction in disease severity (%) when compared with untreated control plants. The disease severity of Phytophthora blight was recorded as 8% and 50% for foliar spray and soil drench, respectively, at 1.0 ppm of paromomycin, compared with untreated control, where disease severity was 83% and 100% by foliar spray and soil drench, respectively. A greater reduction of soft rot lesion areas per leaf disk was observed in treated plants using paromomycin (1.0 μg/ml) by infiltration or soil drench in comparison with untreated control plants. Paromomycin treatment did not negatively affect the growth of chili pepper. Furthermore, the treatment slightly promoted growth; this growth was supported by increased chlorophyll content in paromomycin-treated chili pepper plants. Additionally, paromomycin likely induced resistance as confirmed by the expression of pathogenesis-related (PR) genes: PR-1, β-1,3-glucanase, chitinase, PR-4, peroxidase, and PR-10, which enhanced plant defense against P. capsici in chili pepper. This finding indicates that AG-P 1441 plays a role in pathogen resistance upon the activation of defense genes, by secretion of the plant resistance elicitor, paromomycin.

Effects of Supplementation of Eucalyptus (E. Camaldulensis) Leaf Meal on Feed Intake and Rumen Fermentation Efficiency in Swamp Buffaloes

  • Thao, N.T.;Wanapat, M.;Kang, S.;Cherdthong, A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.7
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    • pp.951-957
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    • 2015
  • Four rumen fistulated swamp buffaloes were randomly assigned according to a $4{\times}4$ Latin square design to investigate the effects of Eucalyptus (E. Camaldulensis) leaf meal (ELM) supplementation as a rumen enhancer on feed intake and rumen fermentation characteristics. The dietary treatments were as follows: T1 = 0 g ELM/hd/d; T2 = 40 g ELM/hd/d; T3 = 80 g ELM/hd/d; T4 = 120 g ELM/hd/d, respectively. Experimental animals were kept in individual pens and concentrate was offered at 0.3% BW while rice straw was fed ad libitum. The results revealed that voluntary feed intake and digestion coefficients of nutrients were similar among treatments. Ruminal pH, temperature and blood urea nitrogen concentrations were not affected by ELM supplementation; however, ELM supplementation resulted in lower concentration of ruminal ammonia nitrogen. Total volatile fatty acids, propionate concentration increased with the increasing level of EML (p<0.05) while the proportion of acetate was decreased (p<0.05). Methane production was linearly decreased (p<0.05) with the increasing level of ELM supplementation. Protozoa count and proteolytic bacteria population were reduced (p<0.05) while fungal zoospores and total viable bacteria, amylolytic, cellulolytic bacteria were unchanged. In addition, nitrogen utilization and microbial protein synthesis tended to increase by the dietary treatments. Based on the present findings, it is suggested that ELM could modify the rumen fermentation and is potentially used as a rumen enhancer in methane mitigation and rumen fermentation efficiency.

Isolation, Identification and Characterization of Phytophthora katsurae, Causing Chestnut Ink Disease in Korea

  • Lee, Jong-Kyu;Jo, Jong-Won;Shin, Keum-Chul;Lee, Sang-Hyun;Lee, Sang-Yong
    • The Plant Pathology Journal
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    • v.25 no.2
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    • pp.121-127
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    • 2009
  • Since July 2005, survey of chestnut ink disease was carried out in chestnut stands located at southern parts of Korea. Dead chestnut trees showing inky ooze on necrotic trunks were found in two different locations. In order to isolate and identify the causal fungus, infected tissues and soil samples around dead or dying trees were collected and placed on Phytophthora-selective medium. Rhododendron and chestnut tree leaves were used as a bait to isolate the fungus from soil samples by attracting zoospores in soil suspensions. On V-8 culture medium, the isolates produced homothallic oogonia with protuberances ($34.0-46.2{\times}21.9-26.7{\mu}m$) abundantly, but did not produced sporangia. Mass production of sporangia was possible by immersing agar plugs with actively growing mycelium in the creek water at $18^{\circ}C$ for 3 days. Sporangia were papillate, and ovoid to obpyriform ($17.0-38.9{\times}14.6-29.2{\mu}m$) in shape. Comparison of the ITS sequences revealed that the isolates had 100% identity to the P. katsurae isolates from Japan and New Zealand and 99.6% identity to other P. katsurae isolates. All of the examined isolates from Korea were completely identical to each other in ITS sequence. Numerous sporangia were formed in filtered as well as unfiltered creek water, but no sporangia formed in sterilized distilled water. Light induced sporangia formation, but has no influence on oospore formation. Amendments of ${\beta}$-sitosterol in culture media have no significant effect on mycelial growth but significantly stimulate oospore and sporangia formation.

Identification of water mold from wild brook lamprey, Lethenterone reissneri (자연산 다묵장어, Lethenterone reissneri에서 발생한 물곰팡이병 원인체의 동정)

  • Kim, Hyoung Jun;Park, Jeong Su;Kim, Sung Yeon;Koo, Ja Geun;Bang, In-Chul;Kwon, Se Ryun
    • Journal of fish pathology
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    • v.26 no.1
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    • pp.39-44
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    • 2013
  • Saprolegnia isolate from wild brook lamprey was identified on the basis of its morphological and molecular characteristics. The isolates showed aseptic hyphae and clavate zoosporagium. Zoospores discharge was typically saprolegnoid. Neither oogomia nor antheridia was observed in this study. ITS sequence obtained from the isolate was compared with other Saprolegnia spp. to analyse their phylogenetic relationships. Results showed that the isolate belongs to clade I including Saprolegnia parasitica. Based on the asexual organs, zoospore discharge manner and ITS sequence analysis, the isolate was identified as S. parasitica.

Effects of Urea Level and Sodium DL-malate in Concentrate Containing High Cassava Chip on Ruminal Fermentation Efficiency, Microbial Protein Synthesis in Lactating Dairy Cows Raised under Tropical Condition

  • Khampa, S.;Wanapat, Metha;Wachirapakorn, C.;Nontaso, N.;Wattiaux, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.6
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    • pp.837-844
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    • 2006
  • Four, lactating dairy cows were randomly assigned according to a $2{\times}2$ Factorial arrangement in a $4{\times}4$ Latin square design to study supplementation of urea level (U) at 2 and 4% and sodium dl-malate (M) at 10 and 20 g/hd/d in concentrate. The treatments were as follows U2M10, U2M20, U4M10 and U4M20, respectively. The cows were offered the treatment concentrate at a ratio to milk yield at 1:2.5 and urea-treated rice straw was fed ad libitum. The results have revealed that rumen fermentation and blood metabolites were similar for all treatments. The populations of protozoa and fungal zoospores were significantly different as affected by urea level and sodium dl-malate. In addition, the viable bacteria were similar for amylolytic and proteolytic bacteria. Cellulolytic bacteria were significantly affected by level of sodium dl-malate especially Selenomonas ruminantium and Megasphaera elsdenii while Butyrivibrio fibrisolvens was significantly affected by level of urea supplementation. In conclusion, the combined use of concentrate containing high level of cassava chip at 75% DM with urea at 4% in concentrate and sodium dl-malate at 20 g/hd/d with UTS as a roughage could improv rumen ecology and microbial protein synthesis efficiency in lactating dairy cows.

Effect of Levels of Sodium DL-malate Supplementation on Ruminal Fermentation Efficiency of Concentrates Containing High Levels of Cassava Chip in Dairy Steers

  • Khampa, S.;Wanapat, Metha;Wachirapakorn, C.;Nontaso, N.;Wattiaux, M.A.;Rowlison, P.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.3
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    • pp.368-375
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    • 2006
  • Four rumen-fistulated dairy steers were randomly assigned according to a $4{\times}4$ Latin square design to investigate effects of supplementation levels of sodium dl-malate in concentrates on rumen ecology, ruminal fermentation, nitrogen balance, feed intake and digestibility of nutrients and ruminal microbial protein synthesis. The dietary treatments were cassava concentrate-based, containing sodium dl-malate supplementation at 0, 9, 18 and 27 g/hd/d with urea-treated rice straw (UTS) fed ad libitum. The experiment was conducted for four periods, each period lasting 21 days. Ruminal pH increased with incremental addition of malate (p<0.05). Additionally, molar proportions of propionate were higher in supplemented groups and was highest at 18 g/hd/d of malate supplement (p<0.05). Microbial protein synthesis tended to be higher in dairy steers receiving sodium dl-malate supplements and also was the highest at 18 g/hd/d. Variable bacterial populations, such as amylolytic, proteolytic and cellulolytic species were increased (p<0.05). Furthermore, protozoal populations were decreased significantly (p<0.05), while fungal zoospores were dramatically increased in dairy steers receiving sodium dl-malate supplement (p<0.05). These results suggested that supplementation of concentrate containing a high level of cassava chip at 18 g/hd/d with UTS in dairy steers could improve rumen fermentation efficiency and rumen microbial protein synthesis.

MICROBIAL COLONIZATION AND DIGESTION OF FEED MATERIALS IN CATTLE AND BUFFALOES I. GUINEA GRASS

  • Abdullah, N.;Ho, Y.W.;Mahyuddin, M.;Jalaludin, S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.5 no.2
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    • pp.323-327
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    • 1992
  • An experiment was conducted to determine whether there were any apparent differences in the microbial population, colonization pattern and digestion of guinea grass in situ, between cattle and swamp buffalo. Percentage losses in dry matter (DM), nitrogen (N) and neutral detergent fibre (NDF) of guinea grass were significantly (p<0.01) higher when incubated in the rumen of buffalo than in cattle. Buffalo also showed significantly (p<0.05) faster degradation rates than cattle for each grass component (DM, N, DNF). Light microscopy and SEM examination of the incubated grass materials showed that there were no apparent differences in the pattern of bacterial and fungal invasion and colonization of the grass materials between cattle and buffalo. Attachment of bacteria and fungal zoospores on the grass fragments occurred at 15 min after rumen incubation. After 3 h of rumen incubation, dense population of bacteria was observed in the thin-walled mesophyll and parenchyma tissues, whereas root-like fungal rhizoids were observed in both thin-walled and thick-walled cells. By 6 h, eroded zones were apparent in the thin-walled tissues and in thick-walled tissues with profuse rhizoids. After 24. 48 and 72 h of rumen incubation, most thin-walled tissues were degraded leaving mostly the thick-walled tissues. The predominant bacteria were the curved rods resembling Butyrivibrio sp., the thick rods resembling Fibrobacter sp., the diplococcoids resumbling Ruminococcus sp. And spirochetes. Fungi were predominantly those with spherical or oval sporangia. Fusiform sporangia with acuminate apices which resembled Ruminomyces sp. Were of lesser occurrence. Few protozoa were found on the grass fragments at all incubation times.

A Rapid Radicle Assay for Prescreening Antagonistic Bacteria Against Phytophthora capsici on Pepper

  • Chang, Sung-Hwan;Kwack, Min-Sun;Kim, Yun-Sung;Lee, Jung-Yeop;Kim, Ki-Deok
    • Mycobiology
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    • v.29 no.4
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    • pp.218-223
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    • 2001
  • A rapid radicle assay for prescreening antagonistic bacteria to Phytophthora capsic4 causal agent of Phytophthora blight of pepper was developed. Sixty-four bacterial strains with in vitro antifungal activity selected out of 1,400 strains isolated from soils of Ansung, Chunan, Koyang, and Paju, Korea in 1998 were used for development of the bioassay. Uniformly germinated pepper seeds dipped in bacterial cells for 3 hours were placed near the edges of growing mycelia of P. capsici on water agar containing 0.02% glucose. Five-week-old pepper plants(cv. Nockwang) were inoculated to compare with results of the radicle assay developed in this study. For plant inoculation, pepper seeds were sown in potting mixtures incorporated with the bacterial strains, then transplanted into steam-sterilized soils 3 weeks later. Plants were hole-inoculated with zoospores of P. capsici 2 weeks after transplanting. Disease incidence and severity were determined in radicle and plant assessments, respectively. In radicle assay, six strains, GK-B15, GK-B25, OA-B26, OA-B36, PK-B09, and VK-B14 consistently showed the significant(P=0.05) disease reduction against radicle infection by the fungus, four of which also did in plant assessments. Strains OA-B36 and GK-B15 consistently reduced the fungal infection in both the radicle assay and the plant assessment. Therefore, prescreening strains using the radicle assay developed in this study followed by plant assay could reduce time and labor, and improved the possibility of selecting antagonistic bacteria for control of Phytophthora blight of peppers.

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