• Title/Summary/Keyword: conidial suspension

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Morphological and Genetic Characteristics of Colletotrichum gloeosporioides Isolated from Newly Emerging Static-Symptom Anthracnose in Apple

  • Jeon, Yongho;Cheon, Wonsu
    • 한국균학회소식:학술대회논문집
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    • 2014.10a
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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.

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Weeding Efficacy of Sulfonylurea Resistance Weed, Monochoria (Monochoria vaginalis) with Brown Leaf Blight Caused by BWC01-54

  • Hong, Yeon-Kyu;Lee, Bong-Choon;Song, Seok-Bo;Hwang, Jae-Bok;Park, Sung-Tae
    • The Plant Pathology Journal
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    • v.21 no.1
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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.

Optimal Conditions for Resistance Screening of Cercospora leaf spot by Cercospora capsici on Pepper (저항성 검정을 위한 고추 갈색점무늬병(Cercospora capsici) 최적 발병조건)

  • Lim, Yang-Sook;Kim, Byung-Soo
    • Research in Plant Disease
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    • v.9 no.3
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    • pp.166-169
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    • 2003
  • Occurrence of cercospora leaf spot by Cercospora capsici is assamed to remarkbly cause the yield reduction through the leaf spot, yellowing of leaves and early defoliation in pepper cultivation. To develop resistant cultivar, optimal screening conditions of pepper against cercospora leaf spot were studied. Optimal screening conditions were 3.4 ${\times}$ $10^5$ conidia/ml conidial suspension were sprayed on to the leaves of 30~40 days old pepper seedlings, incubated at 15~$20^{\circ}C$/20~$25^{\circ}C$(day/night) and examined 20 days after inoculation.

Pre-Infection Behavior of the Pitch Canker Fungus Fusarium circinatum on Pine Stems

  • Thoungchaleun, Vilakon;Kim, Ki-Woo;Lee, Don-Koo;Kim, Chang-Soo;Park, Eun-Woo
    • The Plant Pathology Journal
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    • v.24 no.2
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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.

Evaluation of Lettuce Germplasm Resistance to Gray Mold Disease for Organic Cultivations

  • Shim, Chang Ki;Kim, Min Jeong;Kim, Yong Ki;Jee, Hyeong Jin
    • The Plant Pathology Journal
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    • v.30 no.1
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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.

Identification of the Imperfect Stage of Mycosphaerella nawae Causing Circular Leaf Spot of Persimmon in Korea (감나무 둥근무늬낙엽병균 Mycosphaerella nawae의 불완전 세대 동정)

  • 권진혁;강수웅;박창석;김희규
    • Korean Journal Plant Pathology
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    • v.14 no.5
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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.

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Improved Method to Increase Conidia Production from Isolates of Different Pathotypes of Citrus Scab Pathogen Elsinoe spp.

  • Hyun, Jae-Wook;Paudyal, Dilli Prasad;Hwang, Rok-Yeon
    • Research in Plant Disease
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    • v.21 no.3
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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.

Potential Appilication of Epicoccosorus nematosporus for the Control of Water chestnut (올방개 지문무늬병균의 효과적 처리방법에 의한 올방개 제초효과)

  • Hong, Yeon-Kyu;Cho, Jae-Min;Uhm, Jae-Youl;Ryu, Kil-Rim;,
    • Korean Journal Plant Pathology
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    • v.13 no.3
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    • pp.167-171
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    • 1997
  • To find optimum application methods of Epicoccosorus nematosporus for control of water chestnut, five different concentration of conidial suspensions ($10^[3}$ conidia/ml to $10^[7}$ conidia/ml) fo the fungus were applied 1 to 4 times on 10~40 days old seedlings of water chestnut in greenhouse. Inoculum levels equal to or greater than $10^[5}$ conidia/ml killed significantly more shoots (82.6%~92.1%) and suppressed significantly underground tuber formation compared to inoculum concentration less than $10^[4}$ conidia/ml. When the conidial suspension of ($6.3 {\times} 10^{5}$conidia/ml of E. nematosporus was sprayed 2 times in 7 days interval, percentage of the killed plants was up to 98.7%. Numbers of reshoots and tubers were alsosuppressed significantly compared to one time application. The percents of killed shoots were similar between 10-day-old and 20-day-old seedlings, and significantly higher than those treated on 30- or 40-day-old seedlings. The fungus treated on 20-day-old seedlings was the most effective because there was high number of reshoots from 10-day-old seedlings. Therefore, optimal application conditions for E. nematosporus is 2~3 times of application in 7 days interval with $10^[5}$ conidia/ml on 20-day-old seedling age.

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Effect of Gamma Irradiation and Its Convergent Treatments on Lily Leaf Blight Pathogen, Botrytis elliptica, and the Disease Development (감마선 및 화학 대체제 융복합 처리가 백합 잎마름병균 및 잎 절편 감염에 미치는 영향)

  • Kim, Ji-Hoon;Yun, Sung-Chul
    • Research in Plant Disease
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    • v.20 no.2
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    • pp.71-78
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    • 2014
  • Gamma irradiation and its convergence with nano-silver particles and sodium dichloroisocyanurate (NaDCC) were investigated to inhibit germination and mycelial growth of Botrytis elliptica, the pathogen of lily leaf blight. In addition, the same treatments were studied on the process of disease development with detached leaf of lily cv. Siberia. Spray inoculation, which is closer to natural infection than wound inoculation, can be a way to investigate infection ability of the treated pathogen. The irradiating dose required to reduce the population by 90%, $D_{10}$, was 526 Gy irradiating with 0-2000 Gy gamma ray on the conidial suspension as well as the growing mycelia. Even at 2000 Gy, the mycelium was not killed but just delayed its growth at 1-2 days behind. Convergent treatment with 40 mg/l of NaDCC just before 200 Gy gamma irradiation was the best way to decrease the conidial germination about 1/1000 times. The control values of gamma irradiation were 23% and 19.5% at wound inoculation and spray inoculation, respectively. On wound-inoculation, the control value of NaDCC only was 89%, and that of NaDCC convergent with 200 Gy gamma irradiation was 32%. On sprayinoculation, the highest control value was NaDCC at 50%, and that of NaDCC convergent with gamma irradiation was 24%.

Colletotrichum fructicola, a Member of Colletotrichum gloeosporioides sensu lato, is the Causal Agent of Anthracnose and Soft Rot in Avocado Fruits cv. "Hass"

  • Fuentes-Aragon, Dionicio;Juarez-Vazquez, Sandra Berenice;Vargas-Hernandez, Mateo;Silva-Rojas, Hilda Victoria
    • Mycobiology
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    • v.46 no.2
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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.