First Report of Gray Mold Caused by Botrytis cinerea on Red Raspberry (Rubus idaeus) in Korea

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February 2016
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February 2016, Volume 100, Number 2
Page 533
http://dx.doi.org/10.1094/PDIS-08-15-0869-PDN
DISEASE NOTES
<strong>First</strong> <strong>Report</strong> <strong>of</strong> <strong>Gray</strong> <strong>Mold</strong> <strong>Caused</strong> <strong>by</strong> <strong>Botrytis</strong>
<strong>cinerea</strong> on Red Raspberry (Rubus idaeus) in
Korea
J. Y. Kim, M. Aktaruzzaman, T. Afroz, and B. S. Kim, Department <strong>of</strong> Plant Science,
Gangneung-Wonju National University, Gangneung 25457, Korea; and H. D. Shin, Division
<strong>of</strong> Environmental Science and Ecological Engineering, Korea University, Seoul 02841, Korea.
Open Access.
ABSTRACT
Citation |
During May and June 2014, red raspberry ( Rubus idaeus L., Rosaceae) plants with symptoms
resembling gray mold were found in several plastic greenhouses with approximately 2 to
10% disease incidence (percentage <strong>of</strong> inflorescences infected) in Gangneung, Korea.
Symptoms included extensive growth <strong>of</strong> mycelia with gray conidia, mostly on blossoms and
fruits, rarely on leaves and twigs. Diseased fruit tissue was excised and surface sterilized <strong>by</strong>
immersion in 0.1% sodium hypochlorite (NaOCl) for 1 min, rinsed three times with sterilized
distilled water, placed on potato dextrose agar (PDA, Difco) plates, and incubated at 20 ±
2°C. Conidia (n = 50) were single-celled, ellipsoidal or ovoid, 6.3 to 8.2 × 5.7 to 9.7 μm on
naturally infected fruits and 5.9 to 10.8 × 4.9 to 7.3 μm on PDA. Monoconidial isolates were
obtained <strong>by</strong> plating conidial suspensions on PDA and selecting single colonies. After three
weeks, the fungus formed several black sclerotia ranging from 1.1 to 4.2 × 1.3 to 3.2 mm (n
= 20) near the edge <strong>of</strong> the Petri dish. A representative isolate (GRGM003) was deposited in
Gangneung-Wonju National University and used for further studies. Morphological
characteristics were consistent with those <strong>of</strong> <strong>Botrytis</strong> <strong>cinerea</strong> Pers.: Fr. (Ellis 1971). To
confirm the identity <strong>of</strong> the causal fungus, the internal transcribed spacer (ITS) region <strong>of</strong> rDNA
<strong>of</strong> the isolate was amplified with universal primers ITS1/ITS4 and sequenced. BLAST analysis
<strong>of</strong> the resulting 500-bp nucleotide ITS segment (GenBank Accession No. KP255842) showed
100% identity with the sequence <strong>of</strong> Botryotinia fuckeliana (teleomorph <strong>of</strong> <strong>Botrytis</strong> <strong>cinerea</strong>).
For further confirmation, three nuclear protein-coding genes were sequenced:
glyceraldehyde-3-phosphate dehydrogenase (G3PDH), heat-shock protein 60 (HSP60), and
DNA-dependent RNA polymerase subunit II (RPB2) (Staats et al. 2005). The G3PDH, HSP60,
and RPB2 sequences (KT025631, LC020016, and KT025632) <strong>of</strong> the isolate were 100%
identical to those <strong>of</strong> B. <strong>cinerea</strong> (e.g., KF015583, KJ018758, and KJ018756, respectively). For
the pathogenicity test, inoculum was prepared <strong>by</strong> harvesting conidia from 2-week-old
cultures on PDA. A suspension (1×10 6 conidia/ml) amended with 0.5% glucose and 0.25%
K 2 HPO 4 was sprayed onto three potted red raspberry plants with fruits. Three plants <strong>of</strong>
similar features were sprayed with sterilized water, serving as controls. Plants were covered
with plastic bags for 2 days after inoculation to maintain high relative humidity (90 ± 10%)
and were placed in a growth chamber at 20 ± 2°C. <strong>Gray</strong> mold symptoms on fruits and leaves
developed 3 to 5 days after inoculation, whereas control plants remained symptomless. The
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Article History
Print: 11 Feb 2016
Ahead <strong>of</strong> Print: 22 Dec 2015
<strong>First</strong> Look: 15 Oct 2015
Accepted: 27 Sep 2015

developed 3 to 5 days after inoculation, whereas control plants remained symptomless. The
pathogenicity test was performed twice with similar results. The pathogen was successfully
reisolated from inoculated leaves, fulfilling Koch’s postulates. Red raspberry gray mold has
been reported in many countries including the United States and China (Farr and Rossman
2015). To our knowledge, this is the first report <strong>of</strong> gray mold caused <strong>by</strong> B. <strong>cinerea</strong> on red
raspberry in Korea. According to our observations, fruit infections occurred under high
humidity <strong>of</strong> the greenhouse. We suppose that ventilation <strong>of</strong> the greenhouses may effectively
reduce severity <strong>of</strong> gray mold on raspberry fruits.
References:
Section:
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Ellis, M. B. 1971. Dematiaceous Hyphomycetes. CMI, Kew, Surrey, UK.
Farr, D. F., and Rossman, A. Y. Fungal Databases, Syst. Mycol. Microbiol. Lab., ARS,
USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/, August 5, 2015
Staats, M., et al. 2005. Mol. Biol. Evol. 22: 333. 10.1093/molbev/msi020 [CrossRef] [ISI]
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<strong>Gray</strong> mold caused <strong>by</strong> <strong>Botrytis</strong> <strong>cinerea</strong> on red raspberry (Rubus idaeus). A, Infected and
necrotic flowers. B and C, Infected fruits. D, Two-week-old colony <strong>of</strong> B. <strong>cinerea</strong> on potato
dextrose agar. E, Conidia. F and G, Scanning electron microscopy <strong>of</strong> B. <strong>cinerea</strong>.