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Potential Reasons for Prevalence of Fusarium Wilt in Oriental Melon in Korea

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Potential Reasons for Prevalence of Fusarium Wilt in Oriental Melon in Korea Plant Pathol. J. 33(3) : 249-263 (2017) https://doi.org/10.5423/PPJ.OA.02.2017.0026 The Plant Pathology Journal pISSN 1598-2254 eISSN 2093-9280 ©The Korean Society of Plant Pathology Research Article Open Access Potential Reasons for Prevalence of Fusarium Wilt in Oriental Melon in Korea Yunhee Seo and Young Ho Kim* Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul 08826, Korea (Received on February 2, 2017; Revised on March 8, 2017; Accepted on March 28, 2017) This study aims to examine the potential reasons for potentially from other Fusarium pathogen infection the current prevalence of the fusarium wilt in the ori- that produces seemingly wilting caused by severe stem ental melon. Twenty-seven Fusarium isolates obtained rotting. from oriental melon greenhouses in 2010–2011 were identified morphologically and by analysis of elonga- Keywords : Fusarium species, identification, pathogenic- tion factor-1 alpha gene (EF-1α) and internal tran- ity, root-knot nematode, virulence scribed spacer (ITS) rDNA sequences as 6 Fusarium species (8 isolates of F. oxysporum, 8 F. commune, 5 F. Handling Associate Editor : Jeon, Junhyun proliferatum, 3 F. equiseti, 2 F. delphinoides, and 1 F. andiyazi), which were classified as same into 6 EF-1α sequence-based phylogenetic clades. Pathogenicity of Plants belonging to Family Cucurbitaceae such as cu- the Fusarium isolates on the oriental melon was high- cumber, oriental melon and watermelon are important est in F. proliferatum, next in F. oxysporum and F. an- vegetable crops worldwide and major greenhouse-grown diyazi, and lowest in the other Fusarium species tested, fruit-bearing vegetables in Korea, of which the cultivation suggesting F. proliferatum and F. oxysporum were areas are 3,629 ha, 5,515 ha, and 12,299 ha with annual major pathogens of the oriental melon, inducing stem productions of 254,276 tons, 176,622 tons, and 672,914 rots and vascular wilts, respectively. Oriental melon tons, respectively (MAFRA, 2015). Particularly for the and watermelon were more susceptible to F. oxyspo- oriental melon (Cucumis melo var. makuwa Makino), the rum than shintosa and cucumber; and cucumber was greenhouse cultivation area comprises the highest ratio most, oriental melon and watermelon, medially, and (97.5%) of the total area among the other fruit-bearing shintosa was least susceptible to F. proliferatum, whose vegetables with the greenhouse ratios of around 80%, in- virulence varied among and within their phylogenetic dicating the growers’ preference to the continuous green- subclades. Severe root-knot galls were formed on all house-cropping of the oriental melon in limited areas, the crops infected with Meloidogyne incognita; how- which is probably because of the high benefit return from ever, little indication of vascular wilts or stem and/or the greenhouse farming. root rots was shown by the nematode infection. These The oriental melon has been suffered from a variety of results suggest the current fungal disease in the ori- diseases caused by 8 viruses, 4 bacteria, thirties of fungi ental melon may be rarely due to virulence changes and Oomycetes, and 3 root-knot nematodes (KSPP, 2009). of the fusarium wilt pathogen and the direct cause of Among all of these diseases, the most detrimental ones in the severe root-knot nematode infection, but may be the continuous greenhouse-cropping are caused by soil- borne pathogens such as Fusarium species and root-knot *Corresponding author. nematodes that accumulate their propagules during the Phone) +82-2-880-4675, FAX) +82-2-873-2317 successive cropping in greenhouse conditions that are E-mail) [email protected] favorable for their growth and reproduction. This facility cc This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http:// cultivation also ensures the survival of the pathogens in creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non- harshly cold weather conditions during the winter time in commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. Korea. Cultivation fields once infested with the fusarium wilt Articles can be freely viewed online at www.ppjonline.org. 250 Seo et al. pathogen (Fusarium oxysporum f. sp. melonis) face sig- Owen, 1955; Zhou and Everts, 2007). Fusarium spp. oth- nificant and continuous disease problems, consequently er than F. oxysporum such as F. equiseti, F. graminearum, resulting in the replant problem (Banihashemi and F. moniliforme, F. proliferatum, F. sambucinum, F. solani, deZeeuw, 1975). Also the root-knot nematodes reproduce and F. semitectum (F. incarnatum) were reported to be re- continuously without cessation in such warm-temperature lated with fusarium diseases in oriental melon areas (Kim conditions as in greenhouses and thereby the root-knot and Kim, 2004). These aspects make it difficult to find nematodes have been highly populated in oriental melon out the reasons for current fusarium wilt problems in the greenhouses because of the continuous cultivation for 20 oriental melon. Thus, this study aims to reveal the reasons years in Korea (Kim and Yeon, 2001; Kim et al., 2005). for the current prevalence of the fusarium wilts in the ori- This is evidenced by the population dynamics of the root- ental melon greenhouses firstly by examining the distribu- knot nematodes and their serious damages due to the suc- tion and pathogenicity of Fusarium species to find out the cessive cropping of the oriental melon in Korea (Byeon et major fusarium pathogens responsible for the fusarium al., 2014; Cho et al., 2000; Kim, 2001a, 2001b; Kim and wilt prevalence. This includes the pathological changes Yeon, 2001; Kwon et al., 1998; Park et al., 1995). of F. oxysporum and the occurrence of other Fusarium It is very difficult to control soil-borne diseases using spp. inducing the disease. Secondly it was examined on chemical pesticides in general (Agrios, 2004). For the fu- any direct relationships of the root-knot nematodes with sarium wilt of the oriental melon, no fungicides are com- the prevalence of the wilt symptoms when they have been mercially available to practice the disease control with a highly populated during the continuous greenhouse crop- full efficiency, but the use of resistant cultivars is suggest- ping so that their infections are severe (Agrios, 2004). ed as an efficient and environmentally friendly control strategy for the disease (Freeman et al., 2002). However, Materials and Methods this control strategy has rarely been implemented in the oriental melon until now in Korea as no oriental melon Isolation of Fusarium spp. Fusarium species were cultivar resistant to the Korean race of the pathogen has isolated from rhizosphere soils and stem tissues of the been developed yet (Lee et al., 2015; Matsumoto et al., greenhouse-grown oriental melon in two different lo- 2011). For the root-knot nematodes, the use of resistant cations, Seongju (Gyeongbuk province) and Yeoju cultivars is also an efficient and economical way of the (Gyeonggi province), Korea, during the disease survey nematode control, although there are several methods in 2010 and 2011. Soil samples taken from rhizospheres such as admixtures of soil, rotation and flooding reliable of the oriental melon with the presumed fusarium wilt for their control (Byeon et al., 2014; Kim and Choi, 2001; symptoms were dried for 24 h in a laminar flow hood. Kinloch and Hinson, 1972; Rhoades, 1976). However, no The soil samples were serially diluted to make soil sus- oriental melon cultivars resistant to the root-knot nema- pensions of the concentration of 10–3 g ml–1, and plated on todes have ever been commercialized in Korea yet. Fusarium-selective medium, Komada’s agar (Komada, In the cucurbitaceous crops, the vegetable production 1975). Stem samples from the presumably diseased ori- with grafted seedlings has been practiced for a long period ental melon plants were rinsed with tap water to remove of time from 1920, and currently it becomes a common adhering soils and debris, and cut with a flame-sterilized practice (constituting about 96% of the total plant propa- razor blade into c.a. 1-cm stem tissues. These stem tissues gation) to grow the oriental melon during the winter time were surface sterilized with 70% ethanol for 30 s and 1.0% to increase cold tolerance and to prevent the fusarium wilt sodium hypochlorite for 30 s, followed by rinsing with caused by F. oxysporum f. sp. melonis (Kim et al., 2005; sterile distilled water (SDW) two times, and then placed Lee, 1994; Lee and Oda, 2003). Several rootstocks for on Komada’s agar. Putative Fusarium colonies formed grafting the oriental melon have been developed for their on soil suspension-plated agar and mycelial pieces grown resistance to the fusarium wilt, among which shintosa out from the stem tissue samples were transferred and (Cucurbit maxima × Cu. moschata) has been widely used cultured on potato-dextrose agar (PDA; Difco, Detroit, as a rootstock plant since it was firstly used in 1920 (Lee, MI, USA) at 25°C for 10 days in an incubator. The my- 1994). However, the fusarium wilt still occurs prevalently celial plugs were transferred to carnation leaf agar (CLA) in oriental melon greenhouses with little indication of the and cultured at 25°C for three days, followed by storing disease subsided by the grafting rootstocks resistant to the at –80°C in a deep freezer until use. fungal disease. F. oxysporum with more than 80 forma speciales mor- Species identification of Fusarium isolates. Species phologically indistinguishable shows cross-pathogenicity of the Fusarium isolates were identified based on mi- among cucurbitaceous vegetables (Cafri et al., 2005; croscopic and macroscopic characteristics (totally mor- Prevalence of Fusarium Wilt in Oriental Melon 251 phological characteristics) of single-spored fusarium Pot experiments for pathogenicity (virulence) tests of Fu- isolates as described in other studies (Leslie and Sum- sarium isolates.
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