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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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Floristic study of Mt. Cheongtae area (Pyeongchang-gun, Gangwon-do) (청태산 지역(강원도 평창군)의 식물상)

  • Ji, Seong-Jin;Jung, Su-Young;Chang, Chin;Jang, Jeong-Won;Lee, Cheol-Ho;Yang, Jong-Cheol;Park, Myung-Soon;Chung, Gyu-Young
    • Korean Journal of Plant Taxonomy
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    • v.41 no.4
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    • pp.415-428
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    • 2011
  • The vascular flora in the Mt. Cheongtae area, located Pyeongchang-gun in Gangwon-do, were surveyed 14 times from April of 2008 to October of 2011. The result of this survey revealed 663 taxa in total, comprising 89 families, 342 genera, 577 species, 6 subspecies, 73 varieties and 7 forms. In the flora of this area, 19 taxa were Korean endemic plants and 24 taxa were rare and endangered plants of Korea. The floristic regional indicator plants found in this area were 136 taxa comprising 4 taxa of grade V, 16 taxa of grade IV, 31 taxa of grade III, 41 taxa of grade II and 44 taxa of grade I. Naturalized plants in Korea numbered37 taxa. In addition, 485 taxa were categorized by usage into 8 groups, as follows: 240 edible, 8 fiber, 177 medical, 60 ornamental, 245 pasturing, 2 industrial, 12 dyeing and 16 timer plants.

A Postharvest Control Technique of the Oriental Fruit Moth, Grapholita molesta, Infesting Apples Using CATTS (CATTS를 이용한 복숭아순나방 사과 수확 후 방제 기술)

  • Jung, Choongryul;Kwon, Kimyeon;Kim, Yonggyun
    • Korean journal of applied entomology
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    • v.53 no.1
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    • pp.73-80
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    • 2014
  • Postharvest insect pest control is necessary for agricultural industry including domestic consumer markets and exporting products to meet quarantine issue. Especially, the organic or environmentally friendly agricultural products do not fit to the traditional chemical postharvest treatments mostly using methyl bromide. As an alternative, a physical treatment called CATTS (controlled atmosphere and temperature treatment) has been developed to control various insect and mite pests on apple and several stone fruits. This study was designed to determine the CATTS conditions to control the oriental fruit moth, Grapholita molesta, which is restricted in exporting the infested apples. To apply CATTS on this insect pest, the most heat-tolerant stage was determined. Among the immature stages locating on the fruits, the fifth instar larvae were the most tolerant to $44^{\circ}C$ for 20 min. The ramping rate (the time to increase chamber temperature from $25^{\circ}C$ to $46^{\circ}C$) was positively correlated with the CATTS efficiency under 15% $CO_2$ and 1% $O_2$. After the ramping step, the duration of CATTS was positively correlated with CATTS efficiency. In addition, the CATTS efficiency was highly dependent on the fruit internal temperature at $44^{\circ}C$. From all these parameters, we developed a standard protocol yielding 100% control efficiency of CATTS against apples infested by 4,378 larvae including 2,104 fifth instar individuals.

Development of "Bt-Plus" Biopesticide Using Entomopathogenic Bacterial (Xenorhabdus nematophila, Photorhabdus temperata ssp. temperata) Metabolites (곤충병원세균(Xenorhabdus nematophila, Photorhabdus temperata ssp. temperata)의 대사물질을 이용한 "비티플러스" 생물농약 개발)

  • Seo, Sam-Yeol;Kim, Yong-Gyun
    • Korean journal of applied entomology
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    • v.50 no.3
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    • pp.171-178
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    • 2011
  • Bacillus thuringiensis (Bt) is a bacterial biopesticide against insect pests, mainly lepidopterans. Spodoptera exigua and Plutella xylostella exhibit significant decreases in Bt susceptibility in late larval instars. To enhance Bt pathogenicity, we used a mixture treatment of Bt and other bacterial metabolites which possessed significant immunosuppressive activities. Mixtures of Bt with culture broths of Xenorhabdus nematophila (Xn) or Photorhabdus temperata ssp. temperata (Ptt) significantly enhanced the Bt pathogenicity against late larval instars. Different ratios of Bt to bacterial culture broth had significant pathogenicities against last instar P. xylostella and S. exigua. Five compounds identified from the bacterial culture broth also enhanced Bt pathogenicity. After determining the optimal ratios, the mixture was applied to cabbage infested by late instar P. xylostella or S. exigua in greenhouse conditions. A mixture of Bt and Xn culture broth killed 100% of both insect pests when it was sprayed twice, while Bt alone killed less than 80% or 60% of P. xylostella and S. exigua, respectively. Other Bt mixtures, including Ptt culture broth or bacterial metabolites, also significantly increased pathogenicity in the semi-field assays. These results demonstrated that the Bt mixtures collectively names "Bt-Plus" can be developed into potent biopesticides to increase the efficacy of Bt.

Enhancement of Species-specific Attraction by Addition of a Minor Component of Sex Pheromone Gland of Grapholita dimorpha (복숭아순나방붙이 성페로몬샘의 미량성분 첨가에 따른 종특이적 유인력 증가 효과)

  • Jung, Chung Ryul;Kim, Seong-Ho;Kim, Yonggyun
    • Korean journal of applied entomology
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    • v.53 no.3
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    • pp.239-246
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    • 2014
  • Two congener species of Grapholita molesta and G. dimorpha have similar host ranges and adult occurrence patterns. In addition, the two species commonly use cis-8-dodecenyl acetate (Z8-12:Ac) and trans-8-dodecenyl acetate (E8-12:Ac) as their major sex pheromone components. The commercial lures made of the two components, therefore, suffer a mixed attraction of these two species. This study was conducted to assess any effect of their known minor sex pheromone components to reduce the mixed attraction. When cis-8-tetradecenyl acetate (Z8-14:Ac) was added to the commercial sex pheromone consisting of Z8-12:Ac/E8-12:Ac = 85/15, the attraction to G. dimorpha was not affected, but its attraction to G. molesta was significantly reduced. However, the addition of either dodecanyl acetate (12:Ac) or tetradedecanyl acetate (14:Ac) did not influence on the attraction of both species to the commercial lure. The addition of E8-14:Ac to 3-component sex pheromone, Z8-12:Ac/E8-12:Ac/Z8-14:Ac = 85/15/10, the attraction to G. dimorpha was rather significantly suppressed. From these results, the 3-component sex pheromone, Z8-12:Ac/E8-12:Ac/Z8-14:Ac = 85/15/10 was suggested as an improved composition of sex pheromone lure for G. dimorpha.

Toxicological Analysis of the Entomopathogenic Nematode, Steinernema carpocapsae, and the Symbiotic Bacteria, Xenorhabdus nematophilus on Beneficial Insects and Mammals (유용곤충과 포유류에 대한 곤충병원선충(Steinernema carpocapsae)과 공생세균(Xenorhabdus nematophilus)의 독성)

  • Park, Young-Jin;Kim, Mi-Kyung;Kim, Jin;Yang, Kyung-Hyung;Kim, Yong-Gyun
    • Korean journal of applied entomology
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    • v.40 no.3
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    • pp.259-264
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    • 2001
  • Toxicological studies of two potential biological control agents, the entomopathogenic nematode (Steinernema carpocapsae) and the symbiotic bacteria (Xenorhabdus nematophilus) were conducted against two beneficial insects and one mammal species. Two microbial agents varied in their toxicities between two insect species: an ant, Pristomyrmex pungens, and silkworm, Bombyx mori. In oral toxicity test, the symbiotic bacteria resulted in significant lethal [half lethal concentration of $1.4$\times$10^3$colony-forming units (cfu)/ml] on the ants, while they gave little lethal effect (half lethal concentration of more than $10^{8}$ cfu/ml) on the silkworms. The nematodes, however, gave significant lethal effect [half lethal concentration of 4 infected juveniles (IJs)/ml] on the silkworms, while they did little lethal effect (half lethal concentration of 150,000 IJs/ml) on the ants in topical assays. Both the nematodes and the bacteria did not give lethal effect to the albino rats, Rattus norvegicus, when they were fed orally into the rats. Also, any of these microbial agents were not detected in the internal organs of the treated rats.

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Variation in Development and DNA Methylation of Spodoptera exigua Fed with Different Diets (먹이에 따른 파밤나방 발육과 DNA 메틸화 변이)

  • Kim, Taehyung;Kumar, Sunil;Kim, Yonggyun
    • Korean journal of applied entomology
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    • v.54 no.4
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    • pp.359-367
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    • 2015
  • Physiological plasticity of insects can be closely related with their epigenetic change. This hypothesis was tested using a polyphagous lepidopteran insect, Spodoptera exigua, by assessing the effects of different diets on development and DNA methylation. Three different diets (Welsh onion (WO), Chinese cabbage (CC), artificial diet (AD)) were assessed by feeding a cohort of larvae from neonate to last instar. There were significant differences in larval developmental rate, pupal weight and adult emergence according to diet treatments. AD-fed larvae exhibited the fastest developmental rate along with the highest pupal weight and adult emergence. Among natural hosts, WO was more favorable for development of S. exigua than CC. Total hemolymph proteins and sugars in the last instar larvae were varied among different diets. Gene expression of an insulin-like peptide (SeILP1) presumably associated with development was also varied among diets. Cytosine methylation of genomic DNA was assessed using a monoclonal antibody. Genomic DNA of S. exigua larvae was methylated. DNA methylation was apparently varied among different diet-fed larvae. The facts that a cohort of S. exigua was differentiated in developmental rate and cytosine methylation by different diets suggest that epigenetic factor(s) may play a crucial role in the physiological plasticity.

Efficacy Test of Mating Disruptors Using Food Trap of Oriental Fruit Moth, Grapholita molesta (Busck) (먹이트랩을 이용한 복숭아순나방[Grapholita molesta (Busck)]의 교미교란제 효과 검정)

  • Kim, Yong-Gyun;Bae, Sung-Woo;Choi, Kyung-Hee;Lee, Dong-Hyuk;Lee, Soon-Won
    • Korean journal of applied entomology
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    • v.46 no.2
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    • pp.269-274
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    • 2007
  • This study was performed to estimate the efficacy of three commercial mating disruptors for the control of oriental fruit moth, Grapholita molesta (Busck), using a food-baited trap to collect field females. Mated female ratios estimated in the mating disruptor-treated areas were compared with the estimates of male trap captures, and both estimates were evaluated on the basis of crop damage measured by shoot tip damage caused by G. molesta. Both males and females were attracted to the food trap-baited with terpinyl acetate in apple orchard. Spermatophore of G. molesta was similar to female bursa copulatrix in size. Though there was more than 95% reduction in male trap captures indicating significant mating disruption, significant crop damage occurred on apple leaf buds, in which more than 35% of captured females were mated. This study demonstrates that assessment of the mated females would be more reasonable to represent the efficacy of mating disruptor(s) than the assessment of the male captures in G. molesta.

DNA Markers Applicable for Identification of Two Internal Apple Feeders, Grapholita molesta and Carposina sasakii (두 종의 사과 심식나방류 [복숭아순나방 (Grapholita molesta), 복숭아심식나방 (Carposina sasakii)] 동정용 DNA 분자지표)

  • Song, Seung-Baeck;Choi, Kyeung-Hee;Lee, Soon-Won;Kim, Yong-Gyun
    • Korean journal of applied entomology
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    • v.46 no.2
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    • pp.175-182
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    • 2007
  • Two fruit moths of the oriental fruit moth, Grapholita molesta (Busck), and the peach fruit moth, Carposina sasakii (Matsumura), infest apples in Korea by internally feeding behavior. C. sasakii is a quarantine insect pest from some other countries importing Korean apples. G. molesta is not a quarantine insect pest, but can be incorrectly identified as C. sasakii especially when it is found inside apple fruits at its larval stages because it is not easy to identify the two species by morphological characters alone. This incomplete identification results in massive economical loss by fruits needlessly destroyed or turned away at border inspection stations of the importing nations. This difficulty can be overcome by molecular DNA markers. Several polymorphic regions of mitochondrial DNA of both species were sequenced and used for developing specific striction sites and polymerase chain reaction (PCR) primers. Based on these sequences, three diagnostic PCR-restriction fragment length polymorphism (RFLP) sites were detected and validated for their practical uses. Also, species-specific PCR primers were devised to develop diagnostic PCR method for identifying the internal feeders.

A taxonomic study on the spikelet morphology of Korean Poa L. (Poaceae) (한국산 포아풀속(Poa L.)의 소수 형태에 의한 분류학적 연구)

  • Jung, Su Young;Chung, Gyu Young
    • Korean Journal of Plant Taxonomy
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    • v.38 no.4
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    • pp.477-502
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    • 2008
  • Morphological characters of inflorescence, spikelet, rachilla, glume, lemma and palea about 17 taxa of Korean Poa L. were investigated to evaluate their taxonomic significances. The bulbil present or not, inflorescence shape, callus shape of lemma and surface shape of palea were thought to be the subgenus characters. The keels shapes of palea, the number of lower glume, lateral nerve shape of lemma were thought to be the section characters. The number of floret, rachilla present or not, the number of lower glume, nerve shape of glume, surface shape of lemma were thought to be the species characters. Consequently, 17 taxa of the Korean Poa L. is classified into 3 subgenus (Ochlopoa, Stenopoa, Stenopoa), 7 section(Arenariae, Ochlopoa, Homalopoa, Poa, Pandemos, Tichopoa, Stenopoa). 6 taxa, P. ullungdoensis, P. takeshimana and P. matsumurae, P. nipponica, P. radula, P. viridula, not included in the existing classification system, were suggested to their taxonomic category in infragenic rank, and P. nemoralis, had been included in section Stenopoa of subgenus Stenopoa, was proposed to move to section Poa of subgenus Poa. P. ullungdoensis, reported as new species in 1955, was grasped its taxonomic identity.