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Identification and Characterization of Colletotrichum Species Associated with Bitter Rot Disease of Apple in South Korea

  • Oo, May Moe (Department of Applied Biology, College of Agriculture & Life Sciences, Chungnam National University) ;
  • Yoon, Ha-Yeon (Department of Applied Biology, College of Agriculture & Life Sciences, Chungnam National University) ;
  • Jang, Hyun A (Department of Applied Biology, College of Agriculture & Life Sciences, Chungnam National University) ;
  • Oh, Sang-Keun (Department of Applied Biology, College of Agriculture & Life Sciences, Chungnam National University)
  • Received : 2018.10.02
  • Accepted : 2018.11.22
  • Published : 2018.12.01

Abstract

Bitter rot caused by Colletotrichum species is a common fruit rotting disease of apple and one of the economically important disease in worldwide. In 2015 and 2016, distinct symptoms of bitter rot disease were observed in apple orchards in five regions of South Korea. In the present study, infected apples from these regions were utilized to obtain eighteen isolates of Colletotrichum spp. These isolates were identified and characterized according to their morphological characteristics and nucleotide sequence data of internal transcribed spacer regions and glyceraldehyde-3-phosphate-dehydrogenase. Molecular analyses suggested that the isolates of Colletotrichum causing the bitter rot disease in South Korea belong to 4 species: C. siamense; C. fructicola; C. fioriniae and C. nymphaeae. C. siamense and C. fructicola belonged to Musae Clade of C. gloeosporioides complex species while C. fioriniae and C. nymphaeae belonged to the Clade 3 and Clade 2 of C. acutatum complex species, respectively. Additionally, we also found that the isolates of C. gloeosporioides species-complex were more aggressive than those in the C. acutatum species complex via pathogenicity tests. Taken together, our results suggest that accurate identification of Colletotrichum spp. within each species complex is required for management of bitter rot disease on apple fruit in South Korea.

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Fig. 3. V-shaped necrotic tissues by cause of wound/drop inoculation of (A, E) Type 1, C. fioriniae; (B, F) Type 2, C. nymphaeae; (C, G) Type 3, C. siamense; (D, H) Type 4, C. fructicola on (A, B, C, D) Fuji cultivar and (E, F, G, H) Aori cultivar. Necrotic tissues symptoms were 14 days after inoculation (DAI).

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Fig. 4. Maximum parsimony analysis showing phylogenetic relationships among 18 isolates of Colletotrichum from apple and other related Colletotrichum spp. based on combined dataset of internal transcribed spacer (ITS) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene sequences. Numbers beside each branch represent bootstrap values obtained after a bootstrap test with 1,000 replications. Present isolates are shown in boldface. Bar indicates the number of nucleotide substitutions. The tree is rooted with Colletotrichum orbiculare.

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Fig. 1. (A) Colony morphology of 4 selected Colletotrichum species on PDA and V8 juice media. (B) Morphological characteristics of conidia and appressoria of Colletotrichum species in Type 1-4. I, Conidia; II-III, Appressoria. Bars = 10 μm.

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Fig. 2. (A) Symptoms on apple fruits after wound/drop inoculation with Colletotrichum spp. Symptoms produced by (I, IV) Type 1, C. fioriniae; (II, VII) Type 2, C. nymphaeae; (III, VIII) Type 3, C. siamense; (IV, IX) Type 4, C. fructicola; (V, X) control fruits. Fruits (I-V) on Fuji cultivar and (VI-X) on Aori cultivar. Lesions were 14 days after inoculation (DAI). (B) Comparison of lesion length among four types of Colletotrichum species on Fuji and Aori cultivars.

Table 1. Sources of isolates used in this study and GenBank accession number of Colletotrichum spp. isolated from apple fruits

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Table 2. Description of morphological characters of isolated Colletotrichum species based on colony character, conidial shape, and appressoria

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Table 3. Pathogenicity and fruits lesion diameter of apple (Fuji and Aori cultivars) fruits caused by four Colletotrichum species 14 days after inoculation with wound/drop method a

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Acknowledgement

Supported by : National Research Foundation of Korea

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