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Analysis of Genetic Relationship of Cordyceps militaris in Korea by Random Amplified Polymorphic DNA (한국산 번데기동충하초의 RAPD 분석에 의한 종내 그룹의 유전적 유연관계 분석)

  • Sung, Jae-Mo;Kim, Sang-Hee;Yoon, Chul-Sik;Sung, Gi-Ho;Kim, Yong-Wook
    • The Korean Journal of Mycology
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    • v.27 no.4 s.91
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    • pp.256-273
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    • 1999
  • Seventy two isolates of Cordyceps militaris collected from 11 sites in Korea, including two isolates from ATCC, were used to assess genetic variation within Cordyceps militaris. The anamorph stage and cultural characteristics of C. militaris were observed through microscope and investigated on PDA respectively. The anamorphs of C. militaris were identified to be Verticillium. Isolates of C. militaris showed different growth rates, morphology and color. Fifty six isolates of single ascospore and seventy two isolates of mass ascospore from C. militaris were analysed using by Random Amplified Polymorphic DNA (RAPD) for genetic relationship analysis. Fifty six single ascospore isolates fell into two groups by phenogram constructed from distance values using the UPGMA method in NTSYS-pc software: group A from artificial fruit body of C18 except for isolate 51; group B from artificial fruit body of C738. The average genetic distance value within group A is 0.150 and group B is 0.163. The average genetic distance value between the two groups is 0.221. The average genetic distance value within 56 single ascospores is 0.207 and 72 mass ascospores is 0.330. Genetic relationships were not found among 72 mass ascospore isolates obtained from eleven geographically distant populations.

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Selection of Virulent Isolates of Fusarium circinatum and Investigation of Pitch Canker Severity of Pinus rigida and P. rigida × P. taeda Seed Orchards in Jeju Island (제주도(濟州道) 리기다 및 리기테다 채종원(採種園)에서 푸사리움가지마름병 피해도(被害度) 조사(調査) 및 병원성(病原性) 균주선발(菌株選拔))

  • Woo, Kwan-Soo;Kim, Young-Joung;Kim, Tae-Su;Lee, Seong-Kyu
    • Journal of Korean Society of Forest Science
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    • v.94 no.6
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    • pp.402-409
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    • 2005
  • This study was undertaken to compare and estimate the severity of pitch canker of individual trees of Pinus rigida and Pinus rigida ${\times}$ P. taeda in two seed orchards in Jeju island, in which the orchards have been damaged by the pitch canker for seven years. Wind-pollinated two-year-old seedlings of P. rigida and P. rigida ${\times}$ P. taeda, in which the seedlings of P. rigida ${\times}$ P. taeda were from seeds of phenotypically selected, uninfected(but untested) trees, were inoculated with the pathogenic fungus, Fusarium circinatum, isolated from P. rigida and P. thunbergii. The virulence of the isolates was also identified. Statistically significant difference was found in 'stem cankers'(SC; ${\chi}^2=7.76$, P=0.05) among 4 plantations of P. rigida ${\times}$ P. taeda of two seed orchards. P. rigida was higher in 'top kill' (TK) and 'branch tip symptoms' (BT) than those of P. rigida ${\times}$ P. taeda. In artificial inoculation tests, mortality of the seedlings from the resistant candidates was 14% higher than that of the seedlings from the susceptible candidates. This result may becaused by unknown pollen trees and/or candidate tree selection based only on phenotype. Two of five fungal isolates, C-6-L(9) and C-6-L(19), showed significantly higher mortality (68% and 60%, respectively) than others, suggesting that these isolates can be used as virulent isolates for a mass artificial inoculation. Resistance candidate seedlings that were selected from this study can be utilized as useful materials for fundamental studies of genetics and biochemistry to breed resistance varieties to pitch canker.

Sex Pheromone Trapping of Spodoptera frugiperda (Noctuidae: Lepidoptera) in Korea and the Distribution of Intraspecies-specific Single Nucleotide Polymorphisms in the Cytochrome c Oxidase Subunit 1 (CO1) (성페로몬을 이용한 열대거세미나방 포획과 시토크롬 옥시다제 1(CO1)에서 종내 변이군 특이적 단일염기다형성 분포)

  • Seo, Bo Yoon;Jung, Jin Kyo;Lee, Gwan-Seok;Yang, Chang Yeol;Cho, Jumrae;Kim, Yang Pyo
    • Korean journal of applied entomology
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    • v.59 no.3
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    • pp.217-231
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    • 2020
  • In 2019, the sex pheromones of Spodoptera frugiperda were used to examine moth trapping in cornfields in Gochang, Korea. Four types of traps were prepared, two funnel-types and two delta-types, each baited with 300 or 1000 ㎍ of a two-component (2C) blend of synthetic sex pheromones [100% (Z)-9-tetradecenyl acetate (Z9-14Ac) and 2% (Z)-7-dodecenyl acetate (Z7-12Ac)]. The greatest number of S. frugiperda were captured in the 300 ㎍ funnel-type trap (first catch: August 6). Large numbers of Mythimna loreyi (a non-target) were also caught in the funnel-type traps. Two wing-type traps were baited with 1000 ㎍ of the 2C blend or a four-component (4C) blend [100% (Z)-9-tetradecenyl acetate, 8% (Z)-11-hexadecenyl acetate (Z11-16Ac), 2% (Z)-7-dodecenyl acetate, and 1% (Z)-9-dodecenyl acetate (Z9-12Ac)] and the capture efficiency was assessed. Low numbers of S. frugiperda were captured regardless of the blend, and more M. loreyi were captured using the 4C blend. The two intraspecies groups clustered separately in a phylogenetic tree constructed using partial sequences (1004 bp) of cytochrome c oxidase subunit 1 (CO1). Of the 70 S. frugiperda captured in the pheromone traps, 66 belonged to CO1-RS (CO1 rice-strain) and 4 to CO1-CS (CO1 corn-strain). Twelve consistent single nucleotide polymorphisms (SNPs) were identified in CO1 between the CO1-RS and CO1-CS groups of S. frugiperda. Of the 73 S. frugiperda, 4 had the same SNP pattern as the CO1-CS group (including the corn strain) and 69 had the same SNP pattern as the CO1-RS group (including the rice strain).

Studies on the Manufacture of Concentrated Feed by the Use of Farm Product Waste Materials (농산물(農産物) 폐물(廢物)을 이용(利用)한 농후사료(濃厚飼料) 제조(製造)에 관(關)한 연구(硏究))

  • Kim, Sam-Soon;Lee, Ji-Yul;Park, Sung-Oh;Kim, Ki-Joo
    • The Korean Journal of Mycology
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    • v.1 no.2
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    • pp.15-23
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    • 1973
  • Mold producing cellulase were isolated from rotten woods, and identified as the three species: Aspergillus niger van Tieghem, Aspergillus schiemanni Thom and Trichoderma viride Pers. In this paper, culture conditions in the media and characteristics of these strains were investigated. Using these strains, we have conducted a research concerning the utilization of farm product waste materia's. 1. Optimum conditions for the cellulase formation were as follows. KM 10-1; pH 5.2-5.5, $35^{\circ}C$, incubation time 6 days. OL 11-1; pH 5.5, $30-35^{\circ}C$, incubation time 6 days. SH 9-2; pH 5.5, $30^{\circ}C$, incubatoin time 6 days. 2. Their cellulase activities in their optimum condition were as follows: KM 10-1; CMC-LP 78.5% CMC-SP 4.0 glucose mg/gm of the cultures/min. OL 11-1; CMC-LP 89.9%, CMC-SP 4.9 glucose mg/gm of the cultures/min. SH 9-2; CMC-I.P 77.4%, CMC-SP 3.9 glucose mg/gm of the cultures/min. 3. Hydrolysis of animal feed containing a large quantity (23-30%) of cellulose by means of the crude enzyme in the selected strains resolved 30% of the cellulose contained in the animal feed.

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Growth enhancement and cytotoxicity of Korean mistletoe fractions on human cell lines (한국산 겨우살이 분획물의 면역세포의 생육증진 및 세포독성)

  • Lee, So-Jin;Lee, Mi-Kyoung;Choi, Geun-Pyo;Yu, Chang-Yeon;Roh, Seong-Kyu;Kim, Jong-Dai;Lee, Hyeon-Yong;Lee, Jin-Ha
    • Korean Journal of Medicinal Crop Science
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    • v.11 no.1
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    • pp.62-70
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    • 2003
  • The biological activities on human immune and cancer cell lines of the four kinds of Korean mistletoes (Korean Viscum album, var. coloratum, : Korean Viscum sp. in Quercus acutissima Carr., Korean Viscum sp. in Castanea crenata, Korean Viscum sp. in Betula platyphylla, and Korean Viscum sp. in Salix koreensis) extracts were investigated. The extracts were preparated with ethanol, and fractionated with n-butanol, ethyl acetate, chloroform, hexane, and second distilled water. Cytotoxic potencies of the fractions on human normal lung cell line (HEL 299) showed under 28% in the concentration of 0.5 mg/ml. Growth inhibition effect of the Korean mistletoe extracts on the several human cancer cell lines depends on the concentration of the extracts, and extracting solvent. The hexane, chloroform, and ethyl acetate fractions indicated a strong anticancer activity, but not in aqueous and butanol fractions. Some mistletoe fractions have a different characteristic on the cancer cell lines. Stimulation on the growth of human immuno cell lines(B cell : Raji, T cell: Jurkat) of the extracts were confirmed in the ethyl acetate, chloroform, hexane fractions, but not in aqueous system.

Development of assay method for the activities of new compounds, and the effect of several fungicides against spore germination, adhesion, and myceial growth of Colletotrichum sp. causing red pepper anthracnose (고추 탄저병균의 포자 발아와 부착, 균사 생장에 미치는 화합물의 활성 검정법 확립 및 살균제의 효과)

  • Kim, Jae-Jeung;Kim, Joon-Tae;Park, Sung-Woo;Park, Eun-Suk;Kim, Heung-Tae
    • The Korean Journal of Pesticide Science
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    • v.7 no.3
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    • pp.159-168
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    • 2003
  • With microtiter plate, the assay method was developed for detecting the fungicidal activity of new compounds against spore germination, spore adhesion and mycelial growth of Colletotrichum sp. JC24 cal1Sing red pepper anthracnose. Also, the effects of some commercialized fungicides on fungal development like above mentioned were investigated by measuring the optical density of mycelia grown into wells of microtiter plate. For the standardization of assay method, some factors, such as the treatment of MTT and/or propanol, inodulum density and incubation period, affecting on mycelial optical density were investigated. For obtaining precise and consistent mycelial optical density, it was necessary the treatment of MTT for 12 hrs and propanol for 1 hr. inoculum density adjusted to $1\times10^5$ spores/mL and incubation period for 36 hrs at $25^{\circ}C$. For fungicidal activities, 6 protective fungicides, 6 ones inhibiting sterol biosynthesis, and one inhibiting respiration were used in this study. While mancozeb, chlorothalonil and dithianon among 6 protective fungicides inhibited strongly spore germination, adhesion, and mycelial growth at $6.25{\mu}g/mL$, propineb, iminoctadine and fluazinam inhibited intermediately spore germination and mycelial growth at $100{\mu}g/mL$. Washing above 3 fungicides with new PD broth, their activity against spore adhesion decreased. With hexaconazole, tebuconazole and myclobutanil, the tendency of the activity against fungal differentiation of the early infection stage was similar to the latter group of protective fungicides, showing the decrease of the inhibitory activity against spore adhesion by washing 2 hrs after incubation. However, kresoxim-methyl inhibited spore adhesion distinctly, depending on the applied concentrations. Based on these results, it might be able to assess the fungicidal activity of many compounds against spore germination, adhesion and mycelial growth by the use of microtiter plate in vitro. Using the assay developed in this report, it was possible to investigate the inhibitory activity of some commercialized fungicides, too.

Identification and Characterization of Three Isolates of Cucumber mosaic virus Isolated from Weed Hosts (잡초에서 분리한 3종 Cucumber mosaic virus의 동정과 특성)

  • Lee, Hyeok-Geun;Kim, Sung-Ryul;Jeon, Yong-Woon;Kwon, Soon-Bae;Ryu, Ki-Hyun;Choi, Jang-Kyung
    • Research in Plant Disease
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    • v.14 no.1
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    • pp.15-20
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    • 2008
  • Three isolates of Cucumber mosaic virus (CMV) were isolated from weed hosts showing typical mosaic symptoms, and some properties of the viruses were investigated. CMV isolates, designated as Is-CMV, Jd-CMV and Pla-CMV from Isodon inflexus, Jeffersonia dubia and Phryma leptostachya var. asiatica, respectively, were identified and characterized by biological reaction in several host plants, serological property, dsRNA analysis, reverse transcription-polymerase chain reaction (RT-PCR), restriction fragment-length polymorphism (RFLP). All isolates systemically infected in Nicotiana benthamiana, Cucurbita pepo cv. Black beauty and Cucumis sativus, and did not reveal any differences in these host plants between the isolates. However, remarkable difference in the symptoms was found between the CMVs in Capsicum annuum. Is-CMV induced an asymptomatic symptoms, while Jd-CMV and Pla-CMV produced severe mosaic symptoms in C. annuum plants. In dsRNA analysis, all isolates revealed four major bands with estimated molecular size of 3.4, 3.2, 2.1 and 1.0 kbp. The cDNAs of coat protein gene of the isolates were amplified by RT-PCR using a genus-specific single pair primers that designed to amplify a DNA fragment of approximately ranging from 938 to 966 bp. By restriction mapping analysis using RFLP of the RT-PCR products as well as by serological properties of gel diffusion test, the CMV isolates belong to a typical members of CMV subgroup IA. This is the first report on the occurrence of CMV in the three weed hosts.

Development of Efficient Screening Methods for Melon Plants Resistant to Fusarium oxysporum f. sp. melonis (멜론 덩굴쪼김병에 대한 효율적인 저항성 검정법 개발)

  • Lee, Won Jeong;Lee, Ji Hyun;Jang, Kyoung Soo;Choi, Yong Ho;Kim, Heung Tae;Choi, Gyung Ja
    • Horticultural Science & Technology
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    • v.33 no.1
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    • pp.70-82
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    • 2015
  • This study was conducted to establish an efficient screening system to identify melon resistant to Fusarium oxysporum f. sp. melonis. F. oyxsporum f. sp. melonis GR was isolated from infected melon plants collected at Goryeong and identified as F. oxysporum f. sp. melonis based on morphological characteristics, molecular analyses, and host-specificity tests on cucurbits including melon, oriental melon, cucumber, and watermelon. In addition, the GR isolate was determined as race 1 based on resistance responses of melon differentials to the fungus. To select optimized medium for mass production of inoculum of F. oxysporum f. sp. melonis GR, six media were tested. The fungus produced the most spores (microconidia) in V8-juice broth. Resistance degrees to the GR isolate of 22 commercial melon cultivars and 6 rootstocks for melon plants were investigated. All tested rootstocks showed no symptoms of Fusarium wilt. Among the tested melon cultivars, only three cultivars were susceptible and the other cultivars displayed moderate to high resistance to the GR isolate. For further study, six melon cultivars (Redqueen, Summercool, Superseji, Asiapapaya, Eolukpapaya, and Asiahwanggeum) showing different degrees of resistance to the fungus were selected. The development of Fusarium wilt on the cultivars was tested according to several conditions such as plant growth stage, root wounding, dipping period of roots in spore suspension, inoculum concentration, and incubation temperature to develop the disease. On the basis of the test results, we suggest that an efficient screening method for melon plants resistant to F. oxysporum f. sp. melonis is to remove soil from roots of seven-day-old melon seedlings, to dip the seedlings without cutting in s pore s uspension of $3{\times}10^5conidia/mL$ for 30 min, to transplant the inoculated seedlings to plastic pots with horticulture nursery media, and then to cultivate the plants in a growth room at 25 to $28^{\circ}C$ for about 3 weeks with 12-hour light per day.

Thermal Effects on the Development, Fecundity and Life Table Parameters of Aphis craccivora Koch (Hemiptera: Aphididae) on Yardlong Bean (Vigna unguiculata subsp. sesquipedalis (L.)) (갓끈동부콩에서 아카시아진딧물[Aphis craccivora Koch (Hemiptera: Aphididae)]의 온도발육, 성충 수명과 산란 및 생명표분석)

  • Cho, Jum Rae;Kim, Jeong-Hwan;Choi, Byeong-Ryeol;Seo, Bo-Yoon;Kim, Kwang-Ho;Ji, Chang Woo;Park, Chang-Gyu;Ahn, Jeong Joon
    • Korean journal of applied entomology
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    • v.57 no.4
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    • pp.261-269
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    • 2018
  • The cowpea aphid Aphis craccivora Koch (Hemiptera: Aphididae) is a polyphagous species with a worldwide distribution. We investigated the temperature effects on development periods of nymphs, and the longevity and fecundity of apterous female of A. craccivora. The study was conducted at six constant temperatures of 10.0, 15.0, 20.0, 25, 30.0, and $32.5^{\circ}C$. A. craccivora developed successfully from nymph to adult stage at all temperatures subjected. The developmental rate of A. craccivora increased as temperature increased. The lower developmental threshold (LT) and thermal constant (K) of A. craccivora nymph stage were estimated by linear regression as $5.3^{\circ}C$ and 128.4 degree-days (DD), respectively. Lower and higher threshold temperatures (TL, TH and TH-TL, respectively) were calculated by the Sharpe_Schoolfield_Ikemoto (SSI) model as $17.0^{\circ}C$, $34.6^{\circ}C$ and $17.5^{\circ}C$. Developmental completion of nymph stages was described using a three-parameter Weibull function. Life table parameters were estimated. The intrinsic rate of increase was highest at $25^{\circ}C$, while the net reproductive rate was highest at $20^{\circ}C$. Biological characteristics of A. craccivora populations from different geographic areas were discussed.

Development of cordycepin fortified milk production in Holstein cows I. Effects of various levels of Cordyceps militaris mycelia from grains supplement on cordycepin content in milk in dairy cows (Cordycepin 강화 우유 생산에 관한 연구 I. 동충하초 균사체의 적정사용량 결정을 위한 사양연구)

  • Yeo, J.M.;Lee, S.H.;Kim, D.H.;Hwang, J.H.;Kim, W.Y.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.11 no.1
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    • pp.103-111
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    • 2009
  • This study was carried out to determine effects of long-term supply of Cordyceps militaris mycelia on cordycepin content in milk in dairy cows. Ten Holstein cows in the early stages of lactation were divided into two groups. Control group received no supplement whereas treatment group received 6% of C. militaris mycelia of their dry matter intake for 6 months. Feed intake, milk urea nitrogen and somatic cell counts were not affected by long-term supply of C. militaris mycelia for the whole period. In addition, milk yield and milk composition were not affected by long-term supply of C. militaris mycelia at any time of the periods with the exception of milk protein content and yield. The average of milk protein content and yield from the whole period was higher for C. militaris mycelia supplement group than for the control group. As expected, cordycepin in whole blood and milk was not detected in the control group. The range of cordycepin content in the treatment was 0.31~0.38µ/ml and 0.18~0.26(µ/ml for whole blood and milk, respectively. Individual variation was found to be very high and, furthermore cordycepin was undetected in some milk samples. Thus, no clear pattern could be seen in cordycepin content in milk throughout the whole period. Overall, the results of the present study suggest that the transfer efficiency of cordycepin to milk by supplementing C. militaris mycelia in dairy cows was unpredictable and low.