First Report of Root and Collar Rot caused by Phytophthora nicotianae on ⨯Graptoveria 'Silver Star' in Korea.

⨯Graptoveria 'Silver Star' (a cross between Graptopetalum filiferum and Echeveria agavoides) from the Crassulaceae family, are an evergreen succulent with lotus constellation-shaped flowers, making it consumer favorite ornamental plant in Korea. In 2019, Korea's ornamental production was estimated at KRW 517.4 billion (EUR 382 million), from 4,244 ha of farming area according to the Ministry of Agriculture, Food and Rural Affairs of Korea. In July 2023, ⨯Graptoveria 'Silver Star' plants with chlorotic leaves, root and collar rot were observed in a greenhouse in Yongin (37°14'27.9"N, 127°10'39.19"E), Korea. To isolate the causal agent, small pieces (1 mm2) of symptomatic tissues were surface-sterilized using 1% NaOCl for 1 min, then put onto a water agar (WA) plate and incubated in the dark at 25℃ for five days. Two isolates (FD00202, FD00203) were obtained from diseased leaves, stem and roots by isolating single sporangium. To investigate the morphological characteristics of the isolates, the mycelium from potato dextrose agar (PDA) were transferred to V8 agar (V8A) followed by incubation at 25°C in the dark for 7 days. The isolates produced dense cottony mycelium, with slightly petaloid and light rossette pattern, with coralloid edges measuring 70 to 83 mm diameter. Sporangium were spheroid (30.0-48.0 µm long, 25.0-35.0 µm wide) with globose chlamydospores (17.0-50.0 µm long, 18.0-38.0 µm wide). Oogonia were not observed. Morphological and cultural characteristics of these isolates were phenotypically similar to that of Phytophthora nicotianae (Faedda et al. 2013; Abad et al. 2023). For molecular identification, genomic DNA was extracted from 5 days old cultures using the Maxwell® RSC PureFood GMO and Authentication Kit (Promega). Two gene regions, the rDNA-ITS, COX I were amplified and sequenced using primers ITS1/ITS4 and FM83/FM84, respectively (White et al. 1990; Martin and Tooley 2003). The resulting sequences were deposited in GenBank with accession no. LC783858 to LC783861. A BLASTn search of the DNA sequences from ITS, COX I showed 99.81 and 98.94% identity to P. nicotianae isolate IMI 398853, respectively. Maximum likelihood phylogenetic analyses were performed for the combined data set with ITS, COX I using MEGA7 under the Tamura-Nei model (Kumar et al. 2016). The isolates formed a monophyletic group with P. nicotianae isolate IMI 398853, CPHST BL162, and CPHST BL 44. Based on morphological characteristics and molecular analysis, the isolates were identified as P. nicotianae. T confirm their pathogenicity, inoculum was prepared in accordance with Ann (2000). Artificially wounded healthy plant roots were dipped in zoospore suspension (3.0 × 106 zoospore/ml) for 24 hours, with mock-treated plants (control) dipped in sterile distilled water (Ann. 2000). Thereafter, the plants were transplanted into new medium and kept under high humidity. Symptoms were observed after 10 days of incubation. The plants inoculated with P. nicotianae showed similar symptoms of chlorotic leaves with root and collar rot, while control remained symptomless. The pathogen was re-isolated from all inoculated plants and confirmed as P. nicotianae by morphological and molecular analysis. but not from controls, fulfilling Koch's postulates. Phytophthora nicotianae was previously report on Echeveria derenbergii and Kalanchoe blossfeldiana causing brown spot on stems and roots in California and Korea, respectively (French 1989; Oh and Son 2008). To best of our knowledge, this is the first report of P. nicotianae causing root and collar rot on ⨯Graptoveria 'Silver Star' plants in the Korea.

Occurence of leaf spot on Narrow-head ragwort (Ligularia stenocephala) caused by Alternaria cinerariae in Korea.

Narrow-head ragwort (Ligularia stenocephala (Maxim.) Matsum. & Koidz.) has been used for medicinal purposes and a leafy vegetable in South Korea (Choi et al., 2007; Debnath et al., 2017). In May 2022, brown spots and blight were observed on the leaves in a farmer's field in Namwon (35°26'58.9"N, 127°28'55.9"E), Korea. About 80% of the plants in a 3000 m2 cultivation area were infected. To isolate the causal agent, small pieces (1 mm2) surface-sterilized (1% NaOCl for 1 min) symptomatic leaf tissues were put onto a water agar (WA) plate and incubated in the dark at 25℃. After five days of incubation, two isolates (FD00021, FD00022) were obtained from diseased leaves using a single spore isolation technique. Morphological characteristics were examined after seven days of incubation at 25℃ in the dark. Colonies were 51.6 to 65.3 mm in diameter, gray-green in the center, and ivory at the edge. Conidiophores were straight or curved and 10 - 34 × 4 - 5 µm. Conidia were solitary or two to four in a chain, long ellipsoid to obclavate, one to thirteen transverse septa, 52 - 169 ×14 - 34 µm, blunt-tapered beak variable in size 4 - 56 × 3 - 10 µm (n=75). The morphological and cultural characteristics of the isolates were consistent with that of Alternaria cinerariae (Nishikawa and Nakashima, 2015; Simmons, 2007). For molecular identification, genomic DNA was extracted from 5-day-old cultures using the Maxwell® RSC PureFood GMO and Authentication Kit (Promega). Five gene regions, including rDNA ITS, GPD, Alt a, RPB2, and EF1-α were amplified and sequenced using ITS1/ITS4, gpd1/gpd2, Alt-a1-for/ Alt-a1-rev, RPB2-5F2/RPB2-7cR, and EF1-728F/EF1-986R primer sets respectively (Wang et al., 2022; Garibaldi et al., 2022). The resulting sequences were deposited in GenBank with accession no. OP785152, OP785153 and OP832000 to OP832007). The concatenated genes (rDNA ITS, GPD, Alt a, RPB2, and EF1-α) sequence identity of the FD00021 and FD00022 against the reference strain A. cinerariae CBS 116495 is 99.92% (2572/2574) and 99.84% (2570/2574), respectively. Maximum Likelihood tree was inferred based on the concatenated sequences of the five gene regions using the Kimura 2-parameter model with 1,000 bootstrap replications. The phylogenetic tree showed that the present strains and A. cinerariae CBS 116495 fell into the same clade with high bootstrap support (100%). Based on morphological characteristics and molecular analysis, the isolates were identified as A. cinerariae. To confirm their pathogenicity, drops (70 µl) of conidial suspension (1×104 spores/ml) were applied on intact healthy leaves (3 leaves/plant) of plants (3 plants/isolate) that had been cultivated for one month after transplantation as seedlings. Controls were treated with sterile distilled water. The treated plants were covered with plastic boxes to maintain humidity around 90% and were maintained in an incubator at 25℃ in a 12-hour light-dark cycle. Symptoms appeared only on inoculated leaves after four days of inoculation, while controls remained asymptomatic. The A. cinerariae isolates were re-isolated from infected tissue of the inoculated leaves, thus fulfilling Koch's postulates. Alternaria cinerariae is an important plant pathogen that can cause leaf spot and blight on a variety of host plants including Cineraria spp. and Tussilago farfara (He et al. 2020). This is the first report on leaf spot on Narrow-head ragwort caused by A. cinerariae in the world. Leaf spot disease caused by Alternaria cinerariae is a significant threat to narrow-head ragwort agriculture in South Korea. Therefore, its control strategies are necessary for increasing productivity.

First report of Choanephora rot on Lettuce (Lactuca sativa L.) caused by Choanephora cucurbitarum in Korea.

Lettuce (Lactuca sativa L.) is a vegetable belonging to the family Asteraceae, and one of the major leafy vegetables in Korea. Lettuce has long been consumed as a fresh vegetable. In 2020, 96,774 tons were produced in about 3,800 ha according to Statistics Korea, and various types of lettuce have been bred for year-round production in line with consumer needs (Jang 2007). In August 2020, symptoms of a disease on young leaves of green lettuce (cv. Cheongpungyeoreumchima) were observed in the commercial greenhouse, Jeongeup (35°37'17.5"N, 126°53'40.4"E), Korea. About 60% of plants in 500 m2 of the cultivation area were infected. The tips of infected leaves appeared water-soaked, with brown to black lesions and were covered with dark masses of sporangiophores. Two fungal isolates NC20860 and NC20861 were isolated from monosporous sporangiola in infected leaves. Two isolates produced fast-growing colonies on potato dextrose agar medium (PDA) with abundant white mycelium at the top and bright yellow pigmentation at the underside in only 2 days. Morphological characteristics of the two fungal isolates were investigated from 30 days after inoculation on PDA. Sporangiophores bearing sporangia were erect, non-septate, unbranched, hyaline, and straight. Sporangia were hyaline to brown, globose to subglobose, 28.4 to 33.6 µm diameter. Sporangiospores from sporangia were brown to dark brown, ellipsoid to ovoid, distinctly longitudinally striate, with hyaline appendages at each pole, 6.3 to 7.5 µm wide and 13.0 to 16.0 µm long. Sporangiophores bearing sporangiola were erect, unbranched, hyaline and sporangiola were fusiform, ellipsoid to broadly ellipsoid, 7.6 to 9.8 µm wide, 13.3 to 17.8 µm long. Based on the morphological and cultural characteristics, this fungus was identified as Choanephora cucurbitarum (Berk.&Ravenel) Thaxt (Kirk 1984). To identify the species of the two isolates, DNA was extracted by using the Maxwell® RSC PureFood GMO and Authentication Kit (Promega). The internal transcribed spacer (ITS) region of ribosomal DNA was amplified using nTaq-Tenuto polymerase chain reaction (PCR) kit and therein specified protocol (Enzynomics). The primers ITS1F and ITS4 were used for PCR and sequencing (Gardes 1991). The ITS sequences of the isolates NC20860 and NC20861 from Korea (NCBI GenBank accession no.MZ960299, MZ960300) and the sequences of C. cucurbitarum strains CBS 150.51 (MH856791.1), and KUS-F29113 (KU316934.1) were compared. The sequences of the isolates were identical to those of the reference strains of C. cucurbitarum CBS 150.51 (reference/isolates base pairs, 566/566) and KUS-F29113 (572/572) at the 100% of sequence similarity. Pathogenicity test was conducted using seedlings of 8-week-old green lettuce (cv. Cheongsimchima) by spraying spore suspension (2⨉104 spores/ml). The plants were covered with plastic bags and incubated at 25 ℃ and 70 to 90% RH with a 12-h photoperiod for 20 days. The symptoms were observed only on the inoculated plants within 20 days after inoculation. The inner leaves of the inoculated lettuces had water-soaked lesions. No symptoms were observed in controls. C. cucurbitarum was reisolated from inoculated lettuce, fulfilling Koch's postulates. This fungus has been reported as the causal pathogen of fruit and blossom rot in cucurbits and other plants (Akwaji 2014). However, to our knowledge, occurrence of this pathogen on lettuce has not been reported as yet. This disease is crucial for producers because infected leaves directly results in yield loss of lettuce. This is the first report of Choanephora rot on lettuce caused by C. cucurbitarum in Korea and worldwide.

First Report of Fusarium ipomoeae causing Fusarium wilt on Glycine max in South Korea.

Soybean (Glycine max L.) is one of the most important crops worldwide. In South Korea, three species of Fusarium have been reported as causal pathogens of Fusarium wilt of soybean (KSPP, 2021). From 2017 to 2018, wilted soybeans were observed in two soybean fields in Daegu (36.62°126.91°) and Yesan (35.89°128.44°), South Korea. The incidence rate was about 2 to 5% of the total 0.1ha, respectively. The diseased soybeans were yellowed from the lower leaves or dried up, and the inside of the root and stem were turned brown. Fragments (each 5 mm × 5 mm) of the symptomatic vascular tissue were surface-sterilized with 1% NaOCl for 1 min, and then rinsed twice in sterilized distilled water. The seven pieces each from two diseased plants were placed on water agar and incubated at 25°C for 5 days. Two single spore isolates were cultured on carnation leaf agar at 25°C for 14 days under near ultra violet/dark conditions for 12 hours. Macroconidia of two isolates were mostly 3- to 5-septate, dorsiventral curvature, hyaline, apical cell hooked to tapering, basal cell foot-shaped, and measured 51.3 - 62.2 × 3.7 - 4.7 µm (DG43821) and 63.8-74.8×3.1-4.4 µm (YS37232). Microconidia were not observed. Chlamydospores were produced in chains or pairs, subglobose and thick walled. The color of the aerial mycelium was pinkish white and the reverse of the colony was brownish orange on potato dextrose agar. Based on morphological and cultural characteristics, the two isolates were identified as belonging to Fusarium incarnatum-equiseti species complex (Leslie and Summerell 2006). To confirm the accurate species identification of the two isolates, DNA sequencing of the internal transcribed spacers and intervening 5.8S (ITS), partial translation elongation factor 1-alpha (TEF) and RNA polymerase II largest subunit (RPB2) genes was carried out using primer sets of ITS1/ITS4, EF1 / EF2 and 7cf / 11ar, respectively (O'Donnell et al. 2010). The nucleotide sequences obtained of two isolates were deposited in GenBank with accession numbers of MW375694, MW375695, MW382963, MW382964, MZ364324 and MZ364325. Identities of the ITS region, TEF and RPB2 gene sequences of the two isolates were 490/492, 482/483, 632/633, 631/632, 870/870 and 931/931 with those of ex-type strain F. ipomoeae LC12165 (MK280832, MK289599 and MK289752) in GenBank, respectively. Thus, based on molecular characteristics, the two isolates were confirmed as F. ipomoeae. A pathogenicity test of the two isolates was conducted using root-dip inoculation on seedlings of one soybean cultivars, Pyeongwon. A spore suspension was prepared by flooding 10-day-old cultures on PDA with sterilized distilled water. Fifteen soybean seedlings at the VC stage per each isolate were inoculated by dipping the roots in the spore suspension (1 × 106 conidia/mL) for 2 hours. Inoculated plants were transplanted into pots containing sterilized soil and maintained in the greenhouse at 28±3°C with 14 h/10 h light/dark. An equal number of plants inoculated with sterilized distilled water served as controls. Five days after inoculation, withered symptoms were observed on two or four of the inoculated seedlings, and by 10 days after inoculation, all inoculated plants had withered and died. No symptoms were observed in the non-inoculated control soybeans. The pathogen was consistently re-isolated from only inoculated plants, thus fulfilling Koch's postulates. To our knowledge, this is the first report of F. ipomoeae causing Fusarium wilt on soybean in South Korea, as well as worldwide. This pathogen has been reported on peanut in China as a causal agent of leaf spot (Xu et al., 2021). Understanding the host range of this pathogen and the distribution of F. ipomoeae affecting legume crops in South Korea is important, to ensure an effective management of Fusarium wilt on soybeans.

First Report of Colletotrichum sublineola causing Anthracnose on Sorghum bicolor in South Korea.

Sorghum (Sorghum bicolor (L.) Moench) is one of the top five cereal crops in the world, but the cultivation area in Korea is estimated to be about 3,000 ha (MIFFAF, 2012). In August 2014, anthracnose symptoms on sorghum leaves were observed in two fields in Yecheon (36.62°, 128.41°) and Youngwol (37.20°, 128.49°), South Korea. Symptoms on leaves were brownish red irregular lesions with yellow and tan borders. Some darkened conidiomata and setae were observed on the lesions of infected leaves. Approximately 20% of sorghum plants (cv. Hwanggeumchal) were affected in each field with an area of about 0.1 ha. Fragments of diseased infected leaves were surface sterilized with 1% NaOCl for 30sec. The pieces were placed on water agar and incubated at 25°C for 7days. Two isolates were obtained through single sporing and cultured on synthetic nutrient poor agar at 25°C for 14days. Conidia (n=30) of YN1458 isolate were falcate and measured 22.0 to 32.7 × 4.2 to 6.4 µm. Brown to black setae (n=20) had 1-3 septa, with tapering acute apices and 53.7 to 95.2 × 4.7 to 7.8 µm in size. Appressoria (n=30) were dark brown, usually irregular and 10.5 to 16.9 × 8.6 to 13.6 µm in size. Colonies on PDA produced salmon spore masses in the center of the colony, and whitish grey to dark color in reverse. The morphological characteristics of two isolates were similar. Based on morphology, two isolates were tentatively identified as Colletotrichum graminicola species complex (Cannon et al. 2012; Crouch and Tomaso-Peterson 2012). To clarify taxonomic placement, DNA extracted from mycelia of the two isolates was PCR amplified and sequenced targeting internal transcribed spacer (ITS) regions of rDNA, actin (ACT), chitin synthase 1(CHS-1), and beta-tubulin (TUB) genes (Weir et al. 2012). The sequences of the above four loci of YN1458 and YN1728 were deposited in GenBank with accession numbers KT351801, KT351802 (ITS); KY769869, KY69870 (ACT); KY769871, KY769872 (CHS-1); and KY769873, KY769874 (TUB), respectively. The sequencing results of two isolates showed 99.6% (ITS), 99.6% (ACT of YN1458), 100% (ACT of YN1728), 100% (CHS-1), 100% (TUB of YN1458) and 99.8% (TUB of YN1728) similarity with C. sublineola CBS 131301 (JQ005771, JQ005834, JQ005792, and JQ005855) by BLASTn. Based on the morphological characteristics and multigene sequence analysis, the two isolates were identified as C. sublineola. Pathogenicity of two isolates was confirmed by spraying conidial suspensions (106 conidia/mL) on leaves of 3-week-old sorghum seedlings (cv. Hwanggeumchal) using a pot assay (5 plants per isolate). The same number of seedlings were sprayed with sterile distilled water and served as controls. All plants were maintained in a greenhouse at 25/32°C with natural light. After one week, symptoms similar to those in the field were observed on the leaves inoculated with the pathogen, but not on the control leaves. Colletotrichum sublineola was consistently re-isolated from the inoculated leaves showing anthracnose symptoms and the pathogen identity was confirmed by observing morphological characteristics. So far, C. graminicola was known as the only causal agent pathogen of sorghum anthracnose in South Korea (KSPP, 2009). To our knowledge, this is the first report of C. sublineola causing anthracnose on sorghum in South Korea. Although sorghum is a small-scale crop in South Korea, it is necessary to study the biological and pathogenic characteristics of C. sublineola for effective control of sorghum anthracnose.

Species composition of and fumonisin production by the Fusarium fujikuroi species complex isolated from Korean cereals.

To assess the risk of fumonisin contamination in Korean cereals, we isolated colonies of the Fusarium fujikuroi species complex (FFSC) from barley, maize, rice and soybean samples from 2011 to 2015. A total of 878 FFSC strains were isolated mostly from maize and rice, and species identity of the isolates were determined using the DNA sequence of the translation elongation factor 1-α (TEF-1α) and RNA polymerase II (RPB2) genes. Fusaria recovered from Korean cereals included F. fujikuroi (317 isolates and a frequency of 36%), F. proliferatum (212 isolates and 24.1%), F. verticillioides (170 isolates and 19.4%), F. concentricum (86 strains and 9.8%), F. andiyazi (56 isolates and 6.4%), F. subglutinans (28 isolates and 3.2%), F. thapsinum (5 isolates and 0.6%), and F. circinatum (2 isolates and 0.2%). The rice samples were dominated by F. fujikuroi (47.4%), F. proliferatum (27.3%), and F. concentricum (15.1%), whereas maize samples were dominated by F. verticillioides (33.9%), F. fujikuroi (25.3%), and F. proliferatum (21.1%). A phylogenetic analysis of 70 representative isolates demonstrated that each species was resolved as genealogically exclusive in the ML tree. Fumonisin production potential was evaluated using a PCR assay for the fumonisin biosynthesis gene, FUM1 in all of the isolates. Most of the isolates tested (94%) were positive for FUM1. All of the isolates assigned to F. fujikuroi, F. proliferatum, F. verticillioides and F. thapsinum were positive for FUM1 irrespective of their host origin. Seventy-seven representative isolates positive for FUM1 were examined for fumonisin production in rice medium. The majority of F. proliferatum (26/27, 96.3%), F. verticillioides (16/17, 94.1%) and F. fujikuroi (19/25, 76.0%) produced both FB1 and FB2. Notably, 16 of 19 fumonisin-producing F. fujikuroi produced >1000µg/g of fumonisins (FB1+FB2) in rice medium, which is higher than that in previous reports. These results suggest that F. fujikuroi can produce high levels of fumonisins similar to F. verticillioides and F. proliferatum.

Characterization of Bacillus amyloliquefaciens DA12 Showing Potent Antifungal Activity against Mycotoxigenic Fusarium Species.

In an attempt to develop a biological control agent against mycotoxigenic Fusarium species, we isolated Bacillus amyloliquefaciens strain DA12 from soil and explored its antimicrobial activities. DA12 was active against the growth of mycotoxigenic F. asiaticum, F. graminearum, F. proliferatum, and F. verticillioides both in vitro and in planta (maize). Further screening using dual culture extended the activity range of strain DA12 against other fungal pathogens including Botrytis cinerea, Colletotrichum coccodes, Endothia parasitica, Fusarium oxysporum, Raffaelea quercus-mongolicae, and Rhizoctonia solani. The butanol extract of the culture filtrate of B. amyloliquefaciens DA12 highly inhibited the germination of F. graminearum macroconidia with inhibition rate 83% at a concentration of 31.3 µg/ml and 100% at a concentration of 250 µg/ml. The antifungal metabolite from the butanol extract was identified as iturin A by thin layer chromatography-bioautography. In addition, volatile organic compounds produced by DA12 were able to inhibit mycelial growth of various phytopathogenic fungi. The volatile compounds were identified as 2-heptanone, 5-methyl heptanone and 6-methyl heptanone by gas chromatography-mass spectrometry (GC-MS) analysis. These results indicate that the antagonistic activity of Bacillus amyloliquefaciens DA12 was attributable to iturin A and volatile heptanones, and the strain could be used as a biocontrol agent to reduce the development of Fusarium diseases and mycotoxin contamination of crops.

Mycobiota of ground red pepper and their aflatoxigenic potential.

To investigate contamination of ground red pepper with fungi and mycotoxin, we obtained 30 ground red pepper samples from 15 manufacturers in the main chili-pepper-producing areas in Korea. Fungal contamination was evaluated by spreading diluted samples on potato dextrose agar plates. The total fungi counts ranged from 0 to 7.3 × 103 CFU/g. In the samples, the genus Aspergillus had the highest incidence, while Paecilomyces was isolated most frequently. The next most frequent genera were Rhizopus, Penicillium, Cladosporium, and Alternaria. Within Aspergillus, A. ruber was predominant, followed by A. niger, A. amstelodami, A. ochraceus, A. terreus, A. versicolor, A. flavus, and A. fumigatus. The samples were analyzed for aflatoxins, ochratoxin A, and citrinin by ultra-perfomance liquid chromatography (UPLC) with a fluorescence detector. Ochratoxin A was detected from three samples at 1.03‒2.08 µg/kg, whereas no aflatoxins or citrinin were detected. To test the potential of fungal isolates to produce aflatoxin, we performed a PCR assay that screened for the norB-cypA gene for 64 Aspergillus isolates. As a result, a single 800-bp band was amplified from 10 A. flavus isolates, and one Aspergillus sp. isolate. UPLC analyses confirmed aflatoxin production by nine A. flavus isolates and one Aspergillus sp. isolate, which produced total aflatoxins at 146.88‒909.53 µg/kg. This indicates that continuous monitoring of ground red pepper for toxigenic fungi is necessary to minimize mycotoxin contamination.

Occurrence of Toxigenic Fusarium vorosii among Small Grain Cereals in Korea.

Fusarium graminearum species complex (FGSC) causes Fusarium head blight in small grain cereals. To date, four species (F. graminearum, F. asiaticum, F. boothii, and F. meridionale ) belonging to FGSC frequently occur in Korean cereals. In addition, we first reported the occurrence of additional species (F. vorosii ) within FGSC, which was isolated from barley, corn, and rice in Korea. Phylogenetic analysis of the Fusarium isolates of this group using combined multi-gene sequences confirmed species identification. Moreover, the macroconidia produced by these isolates were morphologically similar to those of the F. vorosii holotype. Chemical analysis indicated that the F. vorosii isolates produced various trichothecenes such as nivalenol and deoxynivalenol with their acetyl derivatives along with zearalenone. Pathogenicity tests demonstrated that all of the F. vorosii isolates examined were pathogenic on barley, corn, and rice with variation in aggressiveness. This study is the first report of F. vorosii in Korean cereals, their pathogenicity towards barley and corn, and their ability to produce trichothecenes and zearalenone.

Occurrence of Soybean Sleeping Blight Caused by Septogloeum sojae in Korea.

Sleeping blight was observed on soybean plants grown in Yanggu, Suwon and Geumsan from 2005 to 2011. Symptoms developed on stems and pods of affected soybean plants. Five fungal isolates were obtained from the diseased plants and identified as Septogloeum sojae based on their morphological, cultural and molecular characteristics. Pathogenicity of the fungus was confirmed on soybean plants by artificial inoculation. This is the first report of S. sojae causing sleeping blight in soybean plants in Korea.

Leaf Spot and Stem Rot on Wilford Swallowwort Caused by Stemphylium lycopersici in Korea.

In June 2012, leaf spot and stem rot were observed on Wilford Swallowwort plants grown in Cheonan, Korea. Three fungal isolates obtained from the diseased leaves and stems were identified as Stemphylium lycopersici, based on morphological, cultural, and molecular characteristics and pathogenicity. This is the first report of leaf spot and stem rot on Wilford Swallowwort caused by S. lycopersici.

Occurrence of Clubroot on Shepherd's-purse Caused by Plasmodiophora brassicae.

Clubroot symptoms were frequently observed on roots of shepherd's-purse (Capsella bursa-pastoris) grown in a field in Nonsan, Chungnam province, Korea in March, 2009. Many resting spores were found in the cells of the root gall tissues collected from the field. The clubroot pathogen was identified as Plasmodiophora brassicae based on its morphological and pathological characteristics. This is the first report that P. brassicae causes clubroot of shepherd's-purse in Korea.

Occurrence of Violet Root Rot on Membranous Milk Vetch Caused by Helicobasidium mompa in Korea.

Severe violet root rot occurred in a field of membranous milk vetch in Bonghwa, Korea, in October 2010. Two fungal isolates from the diseased plants were identified as Helicobasidium mompa based on their morphological, cultural, and molecular characteristics. This is the first report that H. mompa causes violet root rot on membranous milk vetch in Korea.

Leaf Spot of Yam Caused by Pseudophloeosporella dioscoreae in Korea.

Leaf spot symptoms were frequently observed on yam plants grown in the Yeoju area in Korea during a disease survey in 2008. A total of five isolates of Pseudophloeosporella sp. were obtained from the infected leaves of yam plants. All of the isolates were identified as Pseudophloeosporella dioscoreae based on their morphological and cultural characteristics. A phylogenetic tree derived from the internal transcribed spacer sequences of the fungal isolates showed that the fungus is distinctly separated from species in other related genera. P. dioscoreae isolates caused very tiny spots on leaves of yam plants two weeks after artificial inoculation which were similar to those observed in the field. This is the first report that Pseudophloeosporella dioscoreae causes leaf spot in yams in Korea.

Identification of Calonectria colhounii Associated with Basal Stem Rot on Blueberry Seedlings Imported from the United States of America.

Basal stem rot symptoms were found on blueberry seedlings imported from the United States of America in 2008. The fungus obtained from the diseased seedlings was identified as Calonectria colhounii based on morphological and molecular characteristics. The consignments of the blueberry seedlings infected with C. colhounii were destroyed to prevent introduction of the fungus to Korea.

Occurrence of Clubroot on Pak-Choi Caused by Plasmodiophora brassicae.

Clubroot symptoms occurred severely on roots of Pak-Choi (Brassica campestris ssp. chinensis) grown in greenhouses in Gwangju city, Gyeonggi province, Korea in September, 2008. The incidence of the disease symptoms reached as high as 90% in three greenhouses investigated. The root galls collected from the greenhouses were sectioned using a scalpel and observed by light microscope. Many resting spores were found in the cells of the root gall tissues. Suspension of resting spores was prepared from the root galls and inoculated to roots of healthy Pak-Choi plants. Each of five resting spore suspensions caused clubroot symptoms on the roots, which were similar to those observed during the greenhouse survey. Resting spores of the pathogen were observed in the cells of the affected roots. The clubroot pathogen was identified as Plasmodiophora brassicae based on its morphological and pathological characteristics. This is the first report that Plasmodiophora brassicae causes clubroot of Pak-Choi.

Occurrence of Dumontinia tuberosa in Korea.

Thirty-seven single spore isolates were obtained from specimens of ascomycetous fruiting bodies collected from Mt. Suri, Anyang in Korea. The fungal specimens and isolates were identified as Dumontinia tuberosa based on their morphological and cultural characteristics. This is the first record of this fungus occurring in Korea.

Occurrence of Fuligo gyrosa Causing Slime Mold of Oriental Melon.

Recently, a severe slime mold infestation affected oriental melon plants in fields in Chilgok county, Gyeongbuk province, Korea. Specimens were collected from the fields and examined for identification. A species of Myxomycetes, Fuligo gyrosa, was identified based on its morphological characteristics. This is the first report that F. gyrosa causes slime mold of oriental melon.

Mating Types and Optimum Culture Conditions for Sexual State Formation of Fusarium fujikuroi Isolates.

Twenty-five isolates of Fusarium fujikuroi acquired from rice seeds and rice plants evidencing symptoms of Bakanae disease were evaluated to determine their mating types and characterize the formation of their sexual state. The mating types of the isolates were evaluated via multiplex PCR with the diagnostic primers of the mating-type (MAT) region: GFmat1a, GFmat1b, GFmat2c, and GFmat2d. Among the 25 isolates, 11 were identified as MAT-1 (male), and 14 as MAT-2 (female). Four MAT-1 isolates and three MAT-2 isolates were mated and cultured to evaluate the optimal culture conditions for the production of their sexual states. Among four tested media, 10% V8 juice agar proved optimal for the perithecial production of the isolates. The isolates also generated the largest numbers of perithecia when incubated at 23℃ in alternating cycles of 12 hr fluorescent light and NUV fluorescent light and 12 hr darkness.

Occurrence of anthracnose on highbush blueberry caused by colletotrichum species in Korea.

A total of 82 isolates of Colletotrichum species were obtained from anthracnose symptoms of highbush blueberry trees grown in the Gochang area of Korea during a disease survey in 2008. Out of the isolates, 75 were identified as Colletotrichum gloeosporioides and the others as C. acutatum based on their morphological and cultural characteristics. Twenty six of C. gloeosporioides isolates produced their teleomorph Glomerella cingulata in PDA culture. Three isolates of each C. gloeosporioides and C. acutatum caused anthracnose symptoms on the leaves by artificial inoculation, which were similar to what was observed in the orchards. Previously in Korea, only C. gloeosporioides has been reported as causing anthracnose in blueberries. This is the first report that C. acutatum causes anthracnose in the highbush blueberry in Korea.