• 한국균학회소식:학술대회논문집
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• 한국균학회 2014년도 추계학술대회 및 정기총회
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• pp.34-34
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• 2014

Filamentous fungi of the genus Colletotrichum (teleomorph, Glomerella) are considered major plant pathogens worldwide. Cereals, legumes, vegetables, and fruit trees may be seriously affected by this pathogen (1). Colletotrichum species cause typical disease symptoms known as anthracnoses, characterized by sunken necrotic tissue, where orange conidial masses are produced. Anthracnose appears in both developing and mature plant tissues (2). We investigated disease occurrence in apple orchards from 2013 to 2014 in northern Gyeongbuk province, Korea. Typical anthracnose with advanced symptoms was observed in all apple orchards studied. Of late, static fruit spot symptoms are being observed in apple orchards. A small lesion, which does not expand further and remains static until the harvesting season, is observed at the beginning of fruit growth period. In our study, static symptoms, together with the typical symptoms, were observed on apples. The isolated fungus was tested for pathogenicity on cv. 'Fuji apple' (fully ripe fruits, unripe fruits, and cross-section of fruits) by inoculating the fruits with a conidial suspension ($10^5$ conidia/ml). In apple inoculated with typical anthracnose fungus, the anthracnose symptoms progressed, and dark lesions with salmon-colored masses of conidia were observed on fruit, which were also soft and sunken. However, in apple inoculated with fungi causing static symptoms, the size of the spots did not increase. Interestingly, the shape and size of the conidia and the shape of the appressoria of both types of fungi were found to be similar. The conidia of the two types of fungi were straight and cylindrical, with an obtuse apex. The culture and morphological characteristics of the conidia were similar to those of C. gloeosporioides (5). The conidia of C. gloeosporioides germinate and form appressoria in response to chemical signals such as host surface wax and the fruitripening hormone ethylene (3). In this study, the spores started to germinate 4 h after incubation with an ethephon suspension. Then, the germ tubes began to swell, and subsequently, differentiation into appressoria with dark thick walls was completed by 8 h. In advanced symptoms, fungal spores of virtually all the appressoria formed primary hyphae within 16 h. However, in the static-symptom fungus spores, no primary hyphae formed by 16 h. The two types of isolates exhibited different growth rates on medium containing apple pectin, Na polypectate, or glucose as the sole carbon. Static-symptom fungi had a >10% reduction in growth (apple pectin, 14.9%; Na polypectate, 27.7%; glucose, 10.4%). The fungal isolates were also genetically characterized by sequencing. ITS regions of rDNA, chitin synthase 1 (CHS1), actin (ACT), and ${\beta}$-tubulin (${\beta}t$) were amplified from isolates using primer pairs ITS 1 and ITS 4 (4), CHS-79F and CHS-354R, ACT-512F and ACT-783R, and T1 and ${\beta}t2$ (5), respectively. The resulting sequences showed 100% identity with sequences of C. gloeosporioides at KC493156, and the sequence of the ${\beta}$t gene showed 100% identity with C. gloeosporioides at JX009557.1. Therefore, sequence data from the four loci studied proves that the isolated pathogen is C. gloeosporioides. We also performed random amplified polymorphic DNA-PCR, which showed clearly differentiated subgroups of C. gloeosporioides genotypes. The clustering of these groups was highly related to the symptom types of the individual strains.

• The Plant Pathology Journal
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• 제21권1호
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• pp.77-82
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• 2005

A summer annual weed of monochoria (Monochoria vaginalis) grows in the edges of rice paddies, ditches, and moist upland throughout Korea. It is very difficult to control with herbicide because of its sulfonylurea resistance. It is very competitive with fast growing pattern, that can cause reducing yields of rice. Brown leaf blight of monochoria (Monochoria vaginalis) occurred naturally in rice paddy, is first reported in Korea. The fungal isolate BWC01-54 was successfully isolated from the diseased leaves of monochoria. The fungus BWC 01-54 was grown well at $25-28^{\circ}C$, conidia of the greysh black brown mycelia were abundant produced on PDA at 15 days. The fungus was grown well in potato dextrose broth at $28^{\circ}C$ and fully grown within 10 days in 250 ml of flask. In host and pathogenicity test, conidia suspension of BWC01-54 was the most effective to control of monochoria compare to others isolates. Typical symptoms having pin point brown lesions were formed on stem and leaf and which severely affected the whole plants ware blighted within two weeks, respectively. Under paddies field condition, conidial suspension of the fungus BWC01-54 gave around 90% control. Therefore, we conclude that the fungus may have a potential as a biological control agent against sulfonylurea resistance weed in rice paddy.

• 식물병연구
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• 제9권3호
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• pp.166-169
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• 2003

고추재배에서 갈색점무늬병에 의한 반점 황화 조기낙엽 등의 피해로 수량감소를 일으키고 있어, 이 병해에 대한 저항성 육종을 위해서 병원균의 인위접종에 의한 발병조건을 확립하고자 한다. 본 실험 결과, 발병온도는 접종 후 야간 15~$20^{\circ}C$/주간 20~$25^{\circ}C$에서 습실을 경과하는 것이 발병이 가장 양호하였다. 접종묘령은 접종 후 병반 형성수가 많으면서 낙엽이 적게되는 30~40일 유묘가 발병이 가장 좋았다. 고추 유묘에 포자현탁액으로 접종할 경우 3.4 ${\times}$ $10^5$ conidia/ml 농도가 적당하며 접종 후 병반수로 저항성 정도를 조사할 때 20일까지 적당하였다.

• The Plant Pathology Journal
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• 제24권2호
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• pp.112-117
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• 2008

Pre-infection behavior of Fusarium circinatum on stems of pine species was investigated with scanning electron microscopy. Two-year-old stems of Pinus densiflora and p. rigida were inoculated with the fungal conidial suspension and subjected to $25^{\circ}C$ for up to 16 hr. Most microconidia germinated 12 hr after inoculation on pine stems. Conidia produced germ tubes from either one or both ends of microconidia. Germ tubes grew over the stem surface and appeared to enter host tissues through natural openings on pine stems. Surface cracks in the cork were entrance sites of germ tubes of F. circinatum. In addition, host cell wall cracks were often found at the tip of germ tubes. The cuticle appeared to be eroded either at the tip of germ tubes or around germlings. Germ tubes also produced appressoria-like structures, exhibiting swollen tips of germ tubes on the stem surface. There seems to be no significant differences in the pre-infection behavior of F. circinatum on stems between the two pine species.

• The Plant Pathology Journal
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• 제30권1호
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• pp.90-95
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• 2014

This study was conducted to evaluate the resistance of 212 accessions of lettuce germplasm to gray mold disease caused by Botrytis cinerea. The lettuce germplasm were composed of five species: Lactuca sativa (193 accessions), L. sativa var. longifolia (2 accessions), L. sativa var. crispa (2 accessions), L. saligna (2 accessions), and L. serriola (1 accession); majority of these originated from Korea, Netherlands, USA, Russia, and Bulgaria. After 35 days of spray inoculation with conidial suspension ($3{\times}10^7$ conidia/ml) of B. cinerea on the surface of lettuce leaves, tested lettuce germplasm showed severe symptoms of gray mold disease. There were 208 susceptible accessions to B. cinerea counted with 100% of disease incidence and four resistant accessions, IT908801, K000598, K000599, and K021055. Two moderately resistant accessions of L. sativa, K021055 and IT908801, showed 20% of disease incidence of gray mold disease at 45 days after inoculation; and two accessions of L. saligna, K000598 and K000599, which are wild relatives of lettuce germplasm with loose-leaf type, showed complete resistance to B. cinerea. These four accessions are candidates for breeding lettuce cultivars resistant to gray mold disease.

• 한국식물병리학회지
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• 제14권5호
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• pp.397-401
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• 1998

Asexual spores of Mycosphaerella nawae were profusely produced on PDA after a prolonged incubation at $25^{\circ}C$ for 90 days. When persimmon trees were artificially inoculated by the conidial suspension, typical symptoms of circular leaf spot of persimmon appeared on the leaves two month later. The imperfect stag of the fungus was identified as Ramularia sp. based on following morphological characteristics examined under a light microscope and a scanning electron microscope. Conidia were mostly ellipsoid, but occasionally cylindrical, elongated oval, taro, peanut or gourd shapes and measured as 12.2~32.6$\times$6.1~10.2 ${\mu}{\textrm}{m}$. erect, hyaline, colorless-light brown. Conidia were formed solitarily or in chains on a medium and infected leaves. Conidiophore was erect, hyaline, colorless-light brown. and the size was 20.4~102.0$\times$3.1~10.2 ${\mu}{\textrm}{m}$, respectively. In this paper, we firstly demonstratrated that asexual spores of M. nawae induced persimmon circular leaf spot in nature as well as sexual spores of the fungus. Therefore, it is hypothesized that the imperfect stage of the fungus plays an important role in nature for epidemics as secondary inoculum.

• 식물병연구
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• 제21권3호
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• pp.231-234
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• 2015

Elsinoe fawcettii and E. australis are two currently recognized scab pathogens of citrus. E. fawcettii has at least six pathotypes while E. australis has at least two pathotypes. Colonies of E. fawcettii and E. australis do not sporulate in artificial media including potato dextrose agar (PDA). Whiteside's method has been widely used for preparing conidial inoculum in vitro. This study was carried out to develop efficient method for conidia production from artificial media. We developed a shaking method which included the following steps: 1) Colony grown on PDA was mashed with a steel spatula; 2) Mycelia fragments were cultured in 50 ml sterilized rain water in a rotary shaker-incubator (180 rpm) at $25^{\circ}C$ for 24 h: 3) The conidia suspension was filtered through two layers of cheesecloth. Average conidia production of all isolates tested using this shaking method was approximately 13.1 times higher than that from Whiteside's method in this study.

• 한국식물병리학회지
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• 제13권3호
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• pp.167-171
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• 1997

올방개 지문무늬병균(E. mematosporus)의 접종농도, 처리회수, 올방개의 모령이 제초효과에 미치는 영향을 온실에서 검정하였다. 포자현택액의 농도를 $10^{5}$ conidia/ml로 조정하여 분무접종하였을 때 15~20일 이내에 올방개 지상부 줄기의 82.6%~92.1% 정도가 고사하였으나, 그 이하의 농도에서는 제초효과가 53.7%이하로 급격히 감소하였다. 또한 지상부 줄기의 고사는 지하부 괴경형성에서도 영향을 미쳐, $10^{5}$ conidia/ml의 농도에서는 무처리나 $10^[3},\;10^{4}$ 농도의 처리에 비해 약 6~21배의 괴격형성 억제력을 나타내었다. 올방개 지문무늬평균 분생포자현탁액($6.3\;{\times}\;10^{5}$ conidia/ml)은 여러번 처리할수록 제초효과가 높았는데 2회 이상 처리했을 때 지상부 줄기의 고사율(93.8%)이 급격히 증가하였고 고사하기까지의 시간도 단축되었으며 신초의 발생율은 1회 처리에 비해 2.5배 무처리에 비해 12배 감소시켰고 지하부 괴경형성도 1회처리에 비해 6배, 무처리에 비해 35배 억제하였다. 올방개의 모령에 따라서도 제초효율에 차이가 있었는데 신초 형성후 20일 되었을 때 처리하는 것이 줄기 고사율이 가장 높았고, 신초의 발생율이 현저히 감소하였으며 또한 지하부 괴경형성 억제력도 높게 나타났다. 이상의 결과로 보아 올방개 지문무늬병균의 최적처리의 포자량은 $10^[5}$ conidia/ml 이상, 1주일 간격으로 2~3회 처리하며 신초형성 20일 후 처리하는 것이 가장 효과적이라 생각된다.

• 식물병연구
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• 제20권2호
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• pp.71-78
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• 2014

감마선과 융복합 처리를 위한 은나노 입자(NA, NSS), 이염화이소시안산나트륨(NaDCC) 등의 화학대체제를 이용하여 백합잎마름병의 원인균인 Botrytis elliptica의 포자발아 및 균사생장 억제와 동시에 백합 잎 절편에서 발병 과정 중 방제를 연구하였다. 발병과정에서 감마선 융복합 처리효과를 알아보기 위한 실험은 곰팡이 침입을 인위적으로 도와주는 상처접종과 병원균 스스로 침입을 하는 분무접종 두 가지를 실시하였다. 병원균 곰팡이 포자 발아를 90%까지 억제시킬 수 있는 감마선 선량인 $D_{10}$은 526 Gy였다. 2000 Gy까지 감마선 선량을 병원균 균사에 조사한 후, 16, 19, 22, 25, $28^{\circ}C$에서 배양한 결과 곰팡이 균사는 가장 높았던 2000 Gy에서도 죽지 않고 생장이 약 1-2일 지연되었을 뿐이었다. 백합 잎마름병을 억제하기 위해 필요한 1-2 kGy의 감마선 처리는 절화백합에서 부작용이 보일만큼 너무 높기 때문에 감마선은 200 Gy로 조사하고 다른 화학대체제들과의 융복합 처리하는 방안을 강구하였다. 다양한 화학대체제들의 융복합 처리 농도를 $40{\mu}g/l$로 맞춘 이유는 화학제만으로 병원균을 완전히 죽이지 않으면서 감마선의 살균 효과를 거둘 수 있기 때문이었다. 병원균 포자 발아 융복합 실험에서 $40{\mu}g/l$의 NaDCC 처리는 무처리에 비해 포자발아를 1000배 이하로 줄일 수 있었다. 감마선만 처리한 실험에서 방제가는 상처접종에서 23%, 분무접종에서 19.5%로 기대만큼 높지 않았다. 상처접종에서 감마선 처리하지 않은 NaDCC 단독처리의 방제가는 89%였던 반면, NaDCC와 200 Gy 융복합 처리는 32%에 불과하였다. 자연상태에 더 가까운 분무접종 실험에서 NaDCC 단독처리의 방제가는 50%, NaDCC와 200 Gy의 감마선 융복합 처리 방제가는 24%였다.

• Mycobiology
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• 제46권2호
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• pp.92-100
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• 2018

The filamentous Ascomycota Colletotrichum gloeosporioides sensu lato is a fungus that has been reported worldwide as a causal agent of anthracnose disease in avocado and other crops. In Mexico, this species affects fruits from an early stage of development in the orchard until the post-harvest stage. Although fungicides are continuously applied to control Colletotrichum species, pericarp cankers and soft rot mesocarp in fruits are still frequently observed. Considering the lack of a precise description of the causative agent, the aim of the current study was to determine the pathogens involved in this symptomatology. Twenty-four isolates were consistently obtained from the pericarp of avocado fruits cv. "Hass" collected in the central avocado-producing area of Mexico. Morphological features such as colony growth, conidia size, and mycelial appressorium were assessed. Bayesian multilocus phylogenetic analyses were performed using amplified sequences of the internal transcribed spacer region of the nuclear ribosomal DNA; actin, chitin synthase, glyceraldehyde-3-phosphate dehydrogenase partial genes; and APn2-Mat1-2 intergenic spacer and mating type Mat1-2 partial gene from the nine selected isolates. In addition, fruits were inoculated with a conidial suspension and reproducible symptoms confirmed the presence of Colletotrichum fructicola in this area. This pathogenic species can now be added to those previously reported in the country, such as C. acutatum, C. boninense, C. godetiae, C. gloeosporioides, and C. karstii. Disease management programs to reduce the incidence of anthracnose should include C. fructicola to determine its response to fungicides that are routinely applied, considering that the appearance of new species is affecting the commercial quality of the fruits and shifting the original population structure.