The Republic of Korea (South Korea) is the second largest chestnut producer in the world. Major cultivars planted in Korea, including cv. Daebo, Hyogo57, and Okkwang, are hybrids of Japanese chestnut (Castanea crenata) and Chinese chestnut (C. mollissima). Because of high perishability, most chestnuts harvested in September and October are preserved in cold rooms (0°C) for marketing. During a survey of postharvest diseases in April to August 2013, chestnut rots were continuously observed in cold rooms located in Buyeo County, Korea. Preliminary studies revealed that the most common agent of rot appeared to be a species of Mucor. When cut open, infected chestnuts showed partial interior discoloration varying from chalky white to dark brown. About 3 to 10% of chestnuts showed symptoms. Hyogo57 seemed to be the most susceptible variety with higher infection rates, up to 30% in some piles. Isolation was done by placing infected tissues on potato dextrose agar. A representative isolate was deposited in the Korean Agricultural Culture Collection (Accession No. KACC47727). Sporangiophores were mostly erect, branched sympodially, and hyaline. Sporangia were globose, pale yellow at first, then grayish brown at maturity. Columellae were obovoid to globose, subhyaline to pale brown, and usually with truncate base and collars. Sporangiospores were globose to irregular, and 4 to 10 μm in diameter. Chlamydospores were cylindrical to globose with oil drops. The fungus was identified as Mucor racemosus f. sphaerosporus (Hagem) Schipper based on the morphological characteristics and growth at low temperature (3). To conduct molecular analyses, genomic DNA was extracted with DNeasy Plant Mini Kits (Qiagen Inc., Valencia, CA). The primers ITS1/ITS4 and NL1/LR3 were used to amplify the internal transcribed spacer (ITS) region of rDNA and the D1/D2 region of the large subunit (4). The resulting 595-bp ITS sequences and 678 bp D1/D2 sequences were deposited in GenBank (Accession Nos. KJ769665 and KF769666). BLAST searches revealed that both the ITS sequences and D1/D2 sequences showed more than 99% similarity with those of M. racemosus f. sphaerosporus, respectively (JN939201 and AJ878775). To perform a pathogenicity test, a suspension of sporangiospores (1 × 105 spores/ml) was sprayed over 10 chestnuts cv. Hyogo57 wounded with a sewing needle and kept in plastic containers (0°C, 100% RH). Another 10 chestnuts wounded with a sewing needle and treated with sterile water served as controls. After 5 days, typical rots appeared on the inoculated chestnuts, whereas no symptoms were observed on controls. Koch's postulates were fulfilled with the re-isolation of M. racemosus from inoculated chestnuts. The pathogenicity test was carried out twice with similar results. M. hiemalis and M. mucedo have been recorded on chestnuts as postharvest pathogens in Switzerland (2) and Chile (1). To our knowledge, this is first report of postharvest rot of chestnut caused by M. racemosus f. sphaerosporus worldwide as well as in Korea. Further studies are necessary for control measures during cold storage of fresh chestnuts.
References: (1) D. F. Farr and A. Y. Rossman. Fungal Databases. Syst. Mycol. Microbiol. Lab., online publication, ARS, USDA, Retrieved May 23, 2014. (2) M. Jermini et al. J. Sci. Food Agric. 86:877, 2006. (3) M. A. A. Schipper. Stud. Mycol. 12:1, 1976. (4) G. Walther et al. Persoonia 30:11, 2013.
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