식물병연구 Research Article Open Access Res. Plant Dis. 23(4): 322-333 (2017) https://doi.org/10.5423/rpd.2017.23.4.322 Monilinia fructicola Characteristics of Brown Rot Caused by Monilinia fructicola on Stone Fruit in Korea 1,2 1 * 1 1 2 3 4 1, 2, 3, 4 *Corresponding author Tel: +82-63-290-6182 Fax: +82-63-290-6198 E-mail: choiiy21@korea.kr Hun-Tak Oh 1,2, In-Young Choi 1 *, Ju Kim 1, Young-Eun Na 1, Wang-Hyu Lee 2, Kui-Jae Lee 3, and Hyeon-Dong Shin 4 1 Jeollabuk-do Agricultural Research and Extension Services, Iksan 54591, Korea 2 Department of Agricultural Biology, Chonbuk National University, Jeonju 54896, Korea 3 Division of Biotechnology, College of Environmental and Bioresource Sciences, Chonbuk National University, Iksan 54596, Korea 4 Division of Environmental Science and Ecological Engineering, Korea University, Seoul 02841, Korea Received August 7, 2017 Revised September 10, 2017 Accepted September 10, 2017 In June and July 2015 and 2017, typical signs and symptoms of brown rot were observed on the fruit of Japanese apricot, peach, apricot, Japanese plum, and sweet cherry with incidence levels of 2-5% in Jeonju and Imsil, Korea. Early symptoms were small, circular, light brown spots that eventually destroyed entire fruit. Small sporodochia later appeared on the surface. Conidia isolated from each host were one-celled, hyaline, lemon-shaped and borne in branched monilioid chains. The optimal temperature range for hyphal growth of all the isolates was 20-25 o C. The growth of hyphae was faster on potato dextrose agar and oatmeal agar than others. Multiple alignments using the ITS sequences from different host showed that they matched each other (100%). The ITS sequences showed 100% identity to those of M. fructicola. Based on the morphological characteristics and phylogenetic analysis via internal transcribed spacer (ITS), all the isolate was identified as M. fructicola. Pathogenicity of representative isolates was proved by artificial inoculation, fulfilling Koch s postulates. This is the first confirmed report on brown rot caused by M. fructicola on stone fruit in Korea. Keywords: Brown rot, Monilinia fructicola, Stone fruit ( ), ( ), [Prunus mume Siebold & Zucc.], [P. persica (L.) Batsch], Research in Plant Disease pissn 1598-2262, eissn 2233-9191 www.online-rpd.org [P. ameniaca var. ansu Maxim.], [P. salicina Lindl.], [P. avium (L.) L.].,,,. 5-6, 6-7, 7, 6-7 5-7 6 (Rural Development Administration, 2007)., The Korean Society of Plant Pathology This is an open access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/ by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Research in Plant Disease Vol. 23 No. 4 323, 2000 1,114 ha 2015 12,082 ha 10 (Ministry of Agriculture, Food and Rural Affairs, 2017)., 1998 41 ton, 2008 3,383 ton (Ministry of Agriculture, Food and Rural Affairs, 2010).,. Alternaria sp., Botrytis cinerea (gray mold), Cercospora circumscissa (leaf spot), Cladosporium carpophilum (scab), Colletotrichum spp. (anthracnose), Monilinia fructicola (brown rot), Phomopsis vexans (brown rot), Phyllosticta circumscissa (leaf spot), Podosphaera tridactyla (powdery mildew), Septoria cerasina (leaf spot), Taphrina mume (leaf curl) (The Korean Society of Plant Pathology, 2009)., Monilinia, (Rural Development Administration, 2008)., Monilinia,,,,,,, M. fructicola (G. Winter) Honey, M. fructigena (Aderhold & Ruhland) Honey, M. laxa (Aderhold & Ruhland) Honey (Choi, 2016). M. fructicola,,, M. fructigena, M. laxa,,, (Fulton Brown, 1997; Hu, 2011; Shim, 2007)., M. fructicola M. laxa (blossom blight), brown rot. M. fructigena (Rungjindamai, 2014)., M. fructicola,,,,,, M. fructigena (The Korean Society of Plant Pathology, 2009). M. fructicola, (Shim, 2007), (Choi, 2014), (Choi, 2016).,.,,,,,.. 2015 2017 6-7,,,. 3,.. 1.5 ml eppendorf, 30. (Difco, MD, USA). 3, 25 o C 7. (Korea University Herbarium, KUS), (Korean Agricultural Culture Collection, KACC),,, (Table 1).. PDA 4 o C, 10 o C, 15 o C, 20 o C, 25 o C, 30 o C 7. PDA (Potato dextrose agar), OA (Oatmeal agar), CMA (Corn meal agar), WA (Water agar), RBA (Rose Bengal agar) 25 o C 7. 4, 1 mm, 35 mm.., 7 (scalpel) 1-3 10 5 conidia/ml. 5,. 100%,
324 Research in Plant Disease Vol. 23 No. 4 Table 1. Summary of Monilinia fructicola isolates investigated in this study Host plant/isolation source Japanese apricot [Prunus mume] Peach [P. persica] Apricot [P. ameniaca var. ansu] Japanese plum [P. salicina] Sweet cherry [P. avium] Geographic origin Date Incidence of brown rot (%) Herbarium voucher ITS GenBank Acc. No. Imsil, Korea Jun. 2015 KACC47998 KU513384 <2.0 Imsil, Korea Jul. 2015 JBARES-F33 MF597787 Imsil, Korea Jul. 2015 KACC47999 KU513385 <3.0 Jeoju, Korea Jul. 2015 JBARES-F35 MF597789 Jeonju, Korea Jun. 2015 KACC48000 KU513386 <5.0 Jeonju, Korea Jun. 2015 JBARES-F37 MF597788 Imsil, Korea Jun. 2015 JBARES-F41 MF591580 <2.0 Jeonju, Korea Jul. 2017 JBARES-F40 MF597786 Jeonju, Korea Jun. 2015 KACC48001 MF597791 <4.0 Jeonju, Korea Jun. 2017 JBARES-F39 MF597790 25 o C.. 7 Stereo (SteREO Discovery. V8, Zeiss, Munich, Germany) DIC (differential interference contrast) (AXIO, Zeiss) 200-1,000. (n=30) AxioVision LE Module Interactive Measurement (Zeiss).. 7 ribosomal DNA (rdna) internal transcribed spacer (ITS). Genomic DNA DNeasy Plant Mini kit (QIAGEN, Valencia, Canada), ITS1/ITS4 pirmer PCR (White, 1990). PCR. autosequencer (ABI 3030), GenBank database (National Centre for Biotechnology Information [NCBI] US National Institute of Health Bethesda, http://blast. ncbi.nlm.nih.gov/blast.cgi). GenBank ITS, MEGA 6.0 neighbor-joining. Sequence distance Tajima-Nei parameter model (Saitou Nei, 1987; Tamura, 2013)., Botrytis cinerea (GenBank accession No. KX229751) outgroup. [ ] (2009) Monilinia fructicola (G. Winter) Honey. M. fructicola (Japan), M. kusanoi (Japan), M. laxa (China, Japan), M. mume (Japan) (Farr Rossman, 2017).. 2015 6 7 (35 o 38'10"N; 127 o 19'02"E), <2.0% (Table 1)., (Fig. 1A).,, (Fig. 1B).,,.,.. 20-25 o C, 10-30 o C. 4 o C 30 o C. OA PDA, WA, RBA, CMA. -, (flat)/ (effuse), (lobate) (Table 2, Fig. 1E).. 4
Research in Plant Disease Vol. 23 No. 4 325 Fig. 1. Brown rot caused by Monilinia fructicola on Japanese apricot. (A) Occurrence of brown rot in a farm in June 2015, (B) A brown rotted fruit covered with the grayish spore mass, sporodochia, (C, D) Monilioid chains and lemon shaped conidia, (E) Five-day-old colony of M. fructicola on PDA., 6,... (monilioid chains) (Fig. 1C),,, - 13.9-18.3 8.1-10.9 m (Table 2, Fig. 1D).. KACC47998 JBARES-F33 ITS sequence 499 bp NCBI GenBank KU513384, MF597787. NCBI Blast search M. fructicola GenBank accession number KC544809, JN176564, FJ515894, KM279616 100%., M. fructicola (Table 1, Fig. 6)..,, Monilinia fructicola (G. Winter) Honey (Pellegrino, 2009). M. fructicola. M. kusanoi, M. laxa, M. mume (Farr Rossman, 2017). (2009). [ ] (2009) Monilinia fructicola (G. Winter) Honey. M. fructicola (Australia, Brazil, Canada, China, Croatia, Czech Republic, Ecuador, Greece, Hungary, Japan, Mexico, New Zealand, Norway, Slovenia, South Africa, Spain, Tasmania, United States, Venezuela), M. fructigena (Bulgaria, China, Cuba, Croatia, France, Hungary, Italy, Sicily, Netherlands, Spain), M. laxa (Australia, Brazil, Bulgaria, Chile, China, Croatia, Greece, Hungary, Iraq, Italy, Sicily, New Zealand, Norway, Serbia, Slovenia, South Africa, Spain, United States), M.
326 Research in Plant Disease Vol. 23 No. 4 Table 2. Comparison of morphological and cultural characteristics between the present isolates obtained from infected stone fruit and Monilinia fructicola species described previously Characters Present isolates a KACC47998 KACC47999 KACC48000 JBARES-F41 KACC48001 M. fructicola b Conidia size (mm ) 13.9-18.3 8.1-10.9 15.0-18.7 9.1-11.8 14.6-18.0 8.5-11.0 14.0-17.5 8.2-10.3 14.8-17.4 8.3-11.7 14.5-16.0 9.5-11.0 Conidia type Monilioid chains Monilioid chains Monilioid chains Monilioid chains Monilioid chains Monilioid chains Sporulation Abundant Abundant Abundant Abundant Abundant Abundant Colony morphology Flat/Effuse Lobate Flat/Effuse Lobate Flat/Effuse Lobate Raised Lobate Flat/Effuse Lobate Flat/Effuse Lobate a All structures were investigated on PDA plates incubated at 25 o C for 7 days. b Monilinia fructicola species described by EPPO/CABI (2010). Fig. 2. Brown rot caused by Monilinia fructicola on peach. (A) Occurrence of brown rot in a farm in July 2015, (B) A brown rotted fruit covered with the grayish spore mass, sporodochia, (C, D) Monilioid chains and lemon shaped conidia, (E) Five-day-old colony of M. fructicola on PDA. polystroma (Italy, Japan, Slovenia), M. seaveri (United States), Monilinia sp. (Brazil) (Farr Rossman, 2017).. 2015 7 (35 o 38'10"N; 127 o 19'02"E) (35 o 51'51"N; 127 o 02'49"E), <3.0% (Table 1)., (Fig. 2A). (Fig. 2B).,,.. 20-25 o C,
Research in Plant Disease Vol. 23 No. 4 327 10-25 o C. 4 o C 30 o C. OA PDA, WA, RBA, CMA. -, (flat)/ (effuse), (lobate) (Table 2, Fig. 2E).. 4, 5,... (monilioid chains) (Fig. 2C),,, - 15.0-18.7 9.1-11.8 m (Table 2, Fig. 2D).. KACC47999 JBARES-F35 ITS sequence 499 bp NCBI GenBank KU513385, MF597789. NCBI Blast search M. fructicola GenBank accession number KC544809, JN176564, FJ515894, KM279616 100%., M. fructicola (Table 1, Fig. 6)..,, Monilinia fructicola (G. Winter) Honey (Pellegrino, 2009). M. fructicola. M. fructigena, M. laxa, M. polystroma, M. seaveri (Farr Rossman, 2017). (2009). Fig. 3. Brown rot caused by Monilinia fructicola on Apricot, (A) Occurrence of brown rot in a farm in June 2015. The fruit was infected, dark black mummified the whole fruit, (B) A brown rotted fruit covered with the grayish spore mass, sporodochia, (C, D) Monilioid chains and lemon shaped conidia, (E) Five-day-old colony of M. fructicola on PDA.
328 Research in Plant Disease Vol. 23 No. 4 [ ] Research in Plant Disease (Choi, 2016) Monilinia fructicola (G. Winter) Honey. (Farr Rossman, 2017).. 2015 6 (35 o 52'18"N; 127 o 06'49"E), <5.0% (Table 1).., (Fig. 3A).,, (Fig. 3A). (Fig. 3B). CMA. -, (flat)/ (effuse), (lobate) (Table 2, Fig. 3E).. 3,, 5,... (monilioid chains) (Fig. 3C),,, - 14.6-18.0 8.5-11.0 m (Table 2, Fig. 3D).. 25 o C, 10-25 o C. 4 o C 30 o C. OA PDA, WA, RBA,. KACC48000 JBARES-F37 ITS sequence 499 bp NCBI GenBank KU513386, MF597788. NCBI Blast search M. fructicola Fig. 4. Brown rot caused by Monilinia fructicola on Japanese plum. (A) Occurrence of brown rot in a farm in July 2017, (B) A brown rotted fruit covered with the grayish spore mass, sporodochia, (C, D) Monilioid chains and lemon shaped conidia, (E) Five-day-old colony of M. fructicola on PDA.
Research in Plant Disease Vol. 23 No. 4 329 GenBank accession number KC544809, JN176564, FJ515894, KM279616 100%., M. fructicola (Table 1, Fig. 6)..,, Monilinia fructicola (G. Winter) Honey (Pellegrino, 2009). Choi (2016),. [ ] (2009) Monilinia fructicola (G. Winter) Honey. M. fructicola (Australia, Brazil, Chile, China, Japan, New Zealand, Spain, United States), M. fructigena (China), M. laxa (China, Japan, South Africa, United States) (Farr Rossman, 2017).. 2015 6 (35 o 35'18"N; 127 o 18'43"E) 2017 7 (35 o 52'42"N; 127 o 06'39"E), <2.0% (Table 1)., (Fig. 4A, 4B).,,. Fig. 4D).. JBARES-F40 JBARES-F41 ITS sequence 499 bp NCBI GenBank MF597786, MF591580. NCBI Blast search M. fructicola GenBank accession number KC544809, JN176564, FJ515894, KM279616 100%., M. fructicola (Table 1, Fig. 6)..,, Monilinia fructicola (G. Winter) Honey (Pellegrino, 2009). M. fructicola. M. fructigena, M. laxa (Farr Rossman, 2017). (2009). [ ] (Choi, 2014) Monilinia fructicola (G. Winter) Honey. M. fructicola (Australia, Canada, China, Greece, Japan, Maryland, United States), M. fructigena (Bulgaria, Serbia), M. kusanoi (Japan), M. laxa (Australia, Bulgaria, Canada, Greece, Japan, Norway, Serbia, Slovenia, United States), Monilinia sp. (Japan) (Farr Rossman, 2017).. 20-25 o C, 10-25 o C. 4 o C 30 o C. OA PDA, WA, RBA, CMA. -, (raised), (lobate) (Table 2, Fig. 4E).. 3, 4,... (monilioid chains) (Fig. 4C),,, - 14.0-17.5 8.2-10.3 m (Table 2,. 2015 2017 6 (35 o 52'42"N; 127 o 06'39"E), <4.0% (Table 1)..,, (Fig. 5A). (Fig. 5B).. 20-25 o C, 10-25 o C. 4 o C 30 o C. OA PDA, WA, RBA, CMA. -, (flat)/ (effuse), (lobate) (Table 2, Fig. 5E).
330 Research in Plant Disease Vol. 23 No. 4 Fig. 5. Brown rot caused by Monilinia fructicola on sweet cherry. (A) Occurrence of brown rot in a farm in June 2017, (B) A brown rotted fruit covered with the grayish spore mass, sporodochia, (C, D) Monilioid chains and lemon shaped conidia, (E) Five-day-old colony of M. fructicola on PDA.. 5, 7,... (monilioid chains) (Fig. 5C),,, - 14.8-17.4 8.3-11.7 m (Table 2, Fig. 5D).. KACC48001 JBARES-F39 ITS sequence 499 bp NCBI GenBank MF597791, MF597790. NCBI Blast search M. fructicola GenBank accession number KC544809, JN176564, FJ515894, KM279616 100%., M. fructicola (Table 1, Fig. 6)..,, Monilinia fructicola (G. Winter) Honey (Pellegrino, 2009). M. fructicola. M. fructigena, M. kusanoi, M. laxa (Farr Rossman, 2017). Choi (2014).,,, (Rural Development Administration, 2008). 2015 2017 6 7,,,,
Research in Plant Disease Vol. 23 No. 4 331 Fig. 6. Phylogenetic analysis by means of the neighbor-joining method comparing the internal transcribed spacer ribosomal DNA (rdna) region of Monilinia fructicola with that of other Monilinia spp. retrieved from GenBank. The numbers above the nodes are the bootstrap values obtained from 1,000 replicates. The isolates obtained in this study are shown in boldface.,.,,..,., 1-2. M. fructicola, (Gell, 2008). 1., (Sinclair Lyon, 2005)., 50-75% (Rural Development Administration, 2008)., M. fructicola (Pellegrino, 2009)., M. fructicola M. laxa,
332 Research in Plant Disease Vol. 23 No. 4 M. fructicola (Sinclair Lyon, 2005; Zhu, 2005). M. fructicola, Shim (2007) 30 o C. M. fructicola (Petroczy Palkovics, 2006; Pellegrino, 2009)., 10 o C 4 o C., OA PDA, WA, RBA, CMA. RBA WA, RBA, WA,. RBA, M. fructicola (Martin, 1950). OA, CMA (sporulation) PDA (Crous, 2009). M. fructicola, M. fructigena, M. laxa, M. yunnanensis ITS, ITS 100%, 4 (Hu, 2011). M. fructigena M. laxa M. fructicola small subunit (SSU) rdna 418 bp group-i intron M. fructigena M. laxa M. fructicola (Fulton Brown, 1997). M. fructicola M. fructicola, M. fructigena, M. laxa, M. polystroma, M. seaveri (Farr Rossman, 2017)., M. fructicola. 2015 2017 6-7,,,, 2-5%.,.,,,. (monilioid chains),,, -. 20-25 o C, OA PDA.,, ribosomal DNA (rdna) ITS Monilinia fructicola.. M. fructicola. Conflicts of Interest No potential conflict of interest relevant to this article was reported. Acknowledgement This study was carried out with the support of Cooperative Research Program for Agricultural Science & Technology Development (Project No. PJ011690) Rural Development Administration, Republic of Korea. References Choi, I. Y., Kim, J., Seo, K. W., Oh, H. T., Cho, J. H., Kim, J. H. and Song, Y. J. 2016. Occurrence of brown rot on apricot caused by Monilinia fructicola in Korea. Res. Plant Dis. 22: 122-126. (In Korean) Choi, H. W., Hong, S. K., Lee, Y. K., Nam, Y. J., Lee, J. G. and Shim, H. S. 2014. Characterization of Monilinia fructicola associated with brown rot of cherry fruit in Korea. Kor. J. Mycol. 42: 353-356. (In Korean) Crous, P. W., Verkley, G. J. M., Groenewald, J. Z. and Samon, R. A. 2009. Fungal Biodiversity, pp. 144-146. CBS-KNAW Fungal Biodiversity Centre Utrecht, The Netherlands. EPPO/CABI. 2010. Data sheets on quarantine pests: Monilinia fructicola. Farr, D. F. and Rossman, A. Y. Fungal Databases, Systematic Mycology & Microbiology Laboratory, ARS, USDA [Internet]. United States Department of Agriculture, Washington, DC. Cited 24th July 2017. URL http://nt.ars-grin.gov/fungaldatabases/ Fulton, C. E. and Brown, A. E. 1997. Use of SSU rdna group-i intron to distinguish Monilinia fructicola from M. laxa and M. fructigena. FEMS Microbiol. Lett. 157: 307-312.
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