First Report of Crown Rot on Gypsophila (Gypsophila paniculata) Caused by Fusarium proliferatum in Korea

Gypsophilas commonly cultivated are Gypsophila elegans B. and G. paniculata L. In September of 2009 and 2010, a severe wilt symptom due to crown rot was observed on G. paniculata (cv. Bristol Fairy) in greenhouses in Yeosu, South Korea. The area of cultivation (~8 ha) in Yeosu covers 90% of production in the Jeonnam Province. Disease outbreak was 20 to 30% in affected greenhouses. Early symptoms included brown discoloration surrounding basal stems and slight wilting. Late symptoms included a sunken stem rot next to the roots, root rot, severe wilting, and dying plants. The causal fungus appeared to invade plants through the basal stem, causing a crown rot that prevented the plant from taking up water and nutrients. Crown rot occurred on young and mature plants. Ten fungal isolates were recovered from basal stems and roots of wilted plants. Microconidia were abundantly produced on potato dextrose agar (PDA), V8 juice agar (VA), carnation leaf agar (CLA), and oatmeal agar (OA). Microconidia were single celled, variable, oval-ellipsoid cylindrical, straight to curved, club-to-kidney shaped or spindle shaped on OA, more slender on VA. Macroconidia were not found on any media used. Microconidia on PDA were 5.9 to 15.1 (9.9) × 2.7 to 4.3 (3.5) μm. Germinated conidia (or false conidia) were often formed on CLA. Conidiophores as phialides were singly formed but often branched. Length of conidiophores was up to 31.1 μm on CLA. Small-sized chlamydospores were rarely found. Fusarium isolates (EML-GYP1, 2, and 3) were selected and identified. From extracted genomic DNA, the internal transcribed spacer (ITS) region including 5.8S rDNA was amplified using ITS1F (5′-CTTGGTCATTTAGAGGAAGT-3′) and LR5F (5′-GCTATCCTGAGGGAAAC-3′) primers. Sequence analyses by BLAST indicated that the isolates (GenBank HM560019, HM560020, and HM560021) were most similar to F. proliferatum (EF4534150) with sequence identity values of 99.3, 99.4, and 99.1%, respectively. The causal fungus was determined to be F. proliferatum based on morphological data and ITS rDNA sequences. Pathogenicity tests with the three isolates were performed on 10 plants of G. paniculata using the dipping method. Healthy roots and basal stems were soaked in a conidial suspension adjusted to ~1.2 × 106 conidia/ml (distilled water) for 15 min. Plants were potted in sterile soil, kept in a humid chamber for 72 h, and moved to a greenhouse. The experiment was carried out in duplicate and repeated two times. Similar symptoms to those observed in the greenhouses were seen 7 days after inoculation. The causal fungus was reisolated from the artificially inoculated basal stems, fulfilling Koch's postulates. Control plants whose basal stems and roots were dipped in sterile water showed no crown rot and wilt symptoms. EML-GYP2 was determined to be the most pathogenic. Ten records of disease caused by three Fusarium species (Fusarium sp., F. oxysporum, and F. udum) have been found on gypsophilas (1), but only F. oxysporum has been reported to cause wilt on G. elgans in Korea (2). To our knowledge, this is the first report of crown rot on gypsophila caused by F. proliferatum in Korea as well as the world. References: (1) D. F. Farr and A. Y. Rossman. Fungal Databases, Systematic Mycology and Microbiology Laboratory, ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ , October 27, 2010, (2) W.-G. Kim and H.-M. Koo. Page 381 in: List of Plant Diseases in Korea. The Korean Society of Plant Pathology, 2009.

Download Full-text

First Report of Leaf Blight Caused by Septoria allii on Garlic Chives in Korea

Plant Disease ◽

10.1094/pdis-08-12-0809-pdn ◽

2013 ◽

Vol 97 (1) ◽

pp. 147-147

Author(s):

J. H. Park ◽

S. E. Cho ◽

K. S. Han ◽

H. D. Shin

Keyword(s):

Disease Incidence ◽

Morphological Characteristics ◽

Its Region ◽

Leaf Blight ◽

Plant Diseases ◽

Cultivated Plants ◽

Sequence Information ◽

Conidial Suspension ◽

Korean Society ◽

First Report

Garlic chives, Allium tuberosum Roth., are widely cultivated in Asia and are the fourth most important Allium crop in Korea. In June 2011, a leaf blight of garlic chives associated with a Septoria spp. was observed on an organic farm in Hongcheon County, Korea. Similar symptoms were also found in fields within Samcheok City and Yangku County of Korea during the 2011 and 2012 seasons. Disease incidence (percentage of plants affected) was 5 to 10% in organic farms surveyed. Diseased voucher specimens (n = 5) were deposited at the Korea University Herbarium (KUS). The disease first appeared as yellowish specks on leaves, expanding to cause a leaf tip dieback. Half of the leaves may be diseased within a week, especially during wet weather. Pycnidia were directly observed in leaf lesions. Pycnidia were amphigenous, but mostly epigenous, scattered, dark brown to rusty brown, globose, embedded in host tissue or partly erumpent, separate, unilocular, 50 to 150 μm in diameter, with ostioles of 20 to 40 μm in diameter. Conidia were acicular, straight to sub-straight, truncate at the base, obtuse at the apex, hyaline, aguttulate, 22 to 44 × 1.8 to 3 μm, mostly 3-septate, occasionally 1- or 2-septate. These morphological characteristics matched those of Septoria allii Moesz, which is differentiated from S. alliacea on conidial dimensions (50 to 60 μm long) (1,2). A monoconidial isolate was cultured on potato dextrose agar (PDA). Two isolates have been deposited in the Korean Agricultural Culture Collection (Accession Nos. KACC46119 and 46688). Genomic DNA was extracted using the DNeasy Plant Mini DNA Extraction Kit (Qiagen Inc., Valencia, CA). The internal transcribed spacer (ITS) region of rDNA was amplified using the ITS1/ITS4 primers and sequenced. The resulting sequence of 482-bp was deposited in GenBank (JX531648 and JX531649). ITS sequence information was at least 99% similar to those of many Septoria species, however no information was available for S. allii. Pathogenicity was tested by spraying leaves of three potted young plants with a conidial suspension (2 × 105 conidia/ml), which was harvested from a 4-week-old culture on PDA. Control leaves were sprayed with sterile water. The plants were placed in humid chambers (relative humidity 100%) for the first 48 h. After 7 days, typical leaf blight symptoms started to develop on the leaves of inoculated plants. S. allii was reisolated from the lesions of inoculated plants, confirming Koch's postulates. No symptoms were observed on control plants. The host-parasite association of A. tuberosum and S. allii has been known only from China (1). S. alliacea has been recorded on several species of Allium, e.g. A. cepa, A. chinense, A. fistulosum, and A. tuberosum from Japan (4) and A. cepa from Korea (3). To the best of our knowledge, this is the first report of S. allii on garlic chives. No diseased plants were observed in commercial fields of garlic chives which involved regular application of fungicides. The disease therefore seems to be limited to organic garlic chive production. References: (1) P. K. Chi et al. Fungous Diseases on Cultivated Plants of Jilin Province, Science Press, Beijing, China, 1966. (2) P. A. Saccardo. Sylloge Fungorum Omnium Hucusque Congnitorum. XXV. Berlin, 1931. (3) The Korean Society of Plant Pathology. List of Plant Diseases in Korea, Suwon, Korea, 2009. (4) The Phytopathological Society of Japan. Common Names of Plant Diseases in Japan, Tokyo, Japan, 2000.

Download Full-text

First Report of Powdery Mildew on Trident Maple Caused by Sawadaea nankinensis in Korea

Plant Disease ◽

10.1094/pdis-93-12-1348c ◽

2009 ◽

Vol 93 (12) ◽

pp. 1348-1348

Author(s):

H. B. Lee ◽

C. J. Kim ◽

H. Y. Mun ◽

J. P. Hong ◽

D. A. Glawe

Keyword(s):

Powdery Mildew ◽

Its Region ◽

Agricultural Science ◽

Morphological Data ◽

First Report ◽

5.8S Rdna ◽

Rdna Sequences ◽

National University ◽

Causal Fungus ◽

Pathogenicity Tests

Trident maple (Acer buergerianum Miq.) is widely grown in Korea as an ornamental tree as well as for the art of bonsai. During 2008 and 2009, a powdery mildew was observed on trident maple plants at the campus of Chonnam National University, Gwangju, Korea. Further surveys revealed the disease to be widespread on this species in other areas including Jeonbuk and Chungnam provinces in Korea. White, superficial mycelia were observed on young shoots and leaves early in spring. Both macroconidia and microconidia were produced beginning in May and conidial production continued through the summer into September and October. Production of chasmothecia was observed starting in September and continued into October. Macroconidia were produced in chains that were sinuate in outline. Individual macroconidia were barrel shaped and 23.4 to 30.0 (26.6) × 15.6 to 21.1 (18.1) μm. Foot cells of macroconidial conidiophores were 26.7 to 110.7 (48) × 7.1 to 11.2 (8.8) μm with one to five following cells. Microconidia were broadly ellipsoidal to subglobose and 8.9 to 12.5 (10.5) × 4.3 to 5.8 (5.1) μm. Chasmothecia typically were formed on adaxial leaf surfaces and 193.2 to 238.1 (216.8) μm in diameter. Appendages bore uncinate to circinate apices and were 176.8 to 267.7 (211.5) × 4.3 to 8.0 (6.2) μm. From extracted genomic DNA, internal transcribed spacer (ITS) region inclusive of 5.8S rDNA was amplified with ITS1F (5′-CTTGGTCATTTAGAGGAAGT-3′) and LR5F (5′-GCTATCCTGAGGGAAAC-3′) primers. The causal fungus was determined to be Sawadaea nankinensis (F.L. Tai) S. Takam. & U. Braun (2) on the basis of morphological data and ITS rDNA sequences. A BLAST search of GenBank with an ITS sequence from this fungus determined that the five sequences exhibiting the highest max score values (1,811 to 2,004) were from S. nankinensis; these sequences produced max ident values from 94% to 99%. In contrast, max score and max ident values from sequences of other Sawadaea spp. were lower, including scores of 1,063 and 98% similarity for S. polyfida var. japonica, 915 and 97% for S. tulasnei, and 913 and 97% for S. bicornis. Pathogenicity tests were conducted on field-grown plants in two replicates. These plants were inoculated with a paintbrush to apply conidia (~5 × 106/ml) collected from powdery-mildew-infected leaves. Inoculated plants developed powdery mildew symptoms within 5 days of inoculation and resembled those observed on naturally infected plants. S. nankinensis (synonym Uncinula nankinensis) was first reported on A. buergerianum from China in 1930 (2). Recently, S. nankinensis (F.L. Tai) S. Takam & U. Braun was reported to occur on A. buergerianum in Japan (3). Until now, three Sawadaea spp. (S. bicornis (Wallr.) Homma, S. negundinis Homma, and S. tulasnei (Fuckel) Homma) have been reported to cause powdery mildew on A. ginnala, but only S. bicornis (= U. circinata Cooke & Peck) has been reported to cause powdery mildew on A. ginnala in Korea (1). However, no Sawadaea sp. previously was reported to cause powdery mildew on A. buergerianum. To our knowledge, this is the first report of powdery mildew on trident maple (A. buergerianum) caused by S. nankinensis in Korea. References: (1) H. D. Shin. Erysiphaceae of Korea. National Institute of Agricultural Science and Technology, 2000. (2) F. L. Tai. Page 1517 in: Sylloge Fungorum Sinicorum. Science Press, Academia Sinica, Peking, 1979. (3) S. Takamatsu et al. Mycoscience 49:161, 2008.

Download Full-text

First report of Fusarium commune causing root and crown rot on maize in Italy

Plant Disease ◽

10.1094/pdis-01-21-0075-pdn ◽

2021 ◽

Author(s):

Monica Mezzalama ◽

Vladimiro Guarnaccia ◽

Ilaria Martino ◽

Giulia Tabome ◽

Maria Lodovica GULLINO

Keyword(s):

Plant Disease ◽

Tap Water ◽

Morphological Characteristics ◽

Plant Diseases ◽

Crown Rot ◽

Conidial Suspension ◽

Fusarium Spp ◽

First Report ◽

Root And Crown Rot ◽

Pathogenicity Tests

Maize (Zea mays L.) is a cereal crop of great economic importance in Italy; production is currently of 62,587,469 t, with an area that covers 628,801 ha, concentrated in northern Italy (ISTAT 2020). Fusarium species are associated with root and crown rot causing failures in crop establishment under high soil moisture. In 2019 maize seedlings collected in a farm located in San Zenone degli Ezzelini (VI, Italy) showed root and crown rot symptoms with browning of the stem tissues, wilting of the seedling, and collapsing due to the rotting tissues at the base of the stem. The incidence of diseased plants was approximately 15%. Seedlings were cleaned thoroughly from soil residues under tap water. Portions (about 3-5 mm) of tissue from roots and crowns of the diseased plants were cut and surface disinfected with a water solution of NaClO at 0.5% for 2 minutes and rinsed in sterile H20. The tissue fragments were plated on Potato Dextrose Agar (PDA) amended with 50 mg/l of streptomycin sulfate and incubated for 48-72 hours at 25oC. Over the 80 tissue fragments plated, 5% were identified as Fusarium verticillioides, 60% as Fusarium spp., 35% developed saprophytes. Fusarium spp. isolates that showed morphological characteristics not belonging to known pathogenic species on maize were selected and used for further investigation while species belonging to F. oxysporum were discarded. Single conidia of the Fusarium spp. colonies were cultured on PDA and Carnation Leaf Agar (CLA) for pathogenicity tests, morphological and molecular identification. The colonies showed white to pink, abundant, densely floccose to fluffy aerial mycelium. Colony reverse showed light violet pigmentation, in rings on PDA. On CLA the isolates produced slightly curved macronidia with 3 septa 28.1 - 65.5 µm long and 2.8-6.3 µm wide (n=50). Microconidia were cylindrical, aseptate, 4.5 -14.0 µm long and 1.5-3.9 µm wide (n=50). Spherical clamydospores were 8.8 ± 2.5 µm size (n=30), produced singly or in pairs on the mycelium, according to the description by Skovgaard et al. (2003) for F. commune. The identity of two single-conidia strains was confirmed by sequence comparison of the translation elongation factor-1α (tef-1α), and RNA polymerase II subunit (rpb2) gene fragments (O’Donnell et al. 2010). BLASTn searches of GenBank, and Fusarium-ID database, using the partial tef-1α (MW419921, MW419922) and rpb2 (MW419923, MW419924) sequences of representative isolate DB19lug07 and DB19lug20, revealed 99% identity for tef-1α and 100% identity to F. commune NRRL 28387(AF246832, AF250560). Pathogenicity tests were carried out by suspending conidia from a 10-days old culture on PDA in sterile H2O to 5×104 CFU/ml. Fifty seeds were immersed in 50 ml of the conidial suspension of each isolate for 24 hours and in sterile water (Koch et al. 2020). The seeds were drained, dried at room temperature, and sown in trays filled with a steamed mix of white peat and perlite, 80:20 v/v, and maintained at 25°C and RH of 80-85% for 14 days with 12 hours photoperiod. Seedlings were extracted from the substrate, washed under tap water, and observed for the presence of root and crown rots like the symptoms observed on the seedlings collected in the field. Control seedlings were healthy and F. commune was reisolated from the symptomatic ones and identified by resequencing of tef-1α gene. F. commune has been already reported on maize (Xi et al. 2019) and other plant species, like soybean (Ellis et al. 2013), sugarcane (Wang et al. 2018), potato (Osawa et al. 2020), indicating that some attention must be paid in crop rotation and residue management strategies. To our knowledge this is the first report of F. commune as a pathogen of maize in Italy. References Ellis M L et al. 2013. Plant Disease, 97, doi: 10.1094/PDIS-07-12-0644-PDN. ISTAT. 2020. http://dati.istat.it/Index.aspx?QueryId=33702. Accessed December 28, 2020. Koch, E. et al. 2020. Journal of Plant Diseases and Protection. 127, 883–893 doi: 10.1007/s41348-020-00350-w O’Donnell K et al. 2010. J. Clin. Microbiol. 48:3708. https://doi.org/10.1128/JCM.00989-10 Osawa H et al. 2020. Journal of General Plant Pathology, doi.org/10.1007/s10327-020-00969-5. Skovgaard K 2003. Mycologia, 95:4, 630-636, DOI: 10.1080/15572536.2004.11833067. Wang J et al. 2018. Plant Disease, 102, doi/10.1094/PDIS-07-17-1011-PDN Xi K et al. 2019. Plant Disease, 103, doi/10.1094/PDIS-09-18-1674-PDN

Download Full-text

First Report of Anthracnose Caused by Colletotrichum gloeosporioides on Malus prunifolia in Korea

Plant Disease ◽

10.1094/pdis-01-12-0063-pdn ◽

2012 ◽

Vol 96 (5) ◽

pp. 766-766 ◽

Cited By ~ 1

Author(s):

W. Cheon ◽

Y. S. Kim ◽

Y. H. Jeon

Keyword(s):

Colletotrichum Gloeosporioides ◽

Sequence Similarity ◽

Morphological Characteristics ◽

Its Rdna ◽

Plant Diseases ◽

Conidial Suspension ◽

Korean Society ◽

First Report ◽

Malus Prunifolia ◽

Crab Apple

In 2010 and 2011, crab apples in Andong Province, Korea were found with dark brown spots on the fruit and mummified fruit on a tree. The fruit surface had red, circular spots that contained smaller, white spots; the color of the inner spots later changed to brown or black. Eventually, the rotten fruit dried and became mummified. Microscopic examination revealed the presence of acervuli and dark brown-to-black, needle-shaped setae. To isolate potential pathogens from infected fruit, small sections (5 to 10 mm2) were excised from the margins of lesions. These sections were surface sterilized with 70% ethanol and 1% NaOCl for 1 min and then rinsed three times with sterile distilled water. The fungus that was isolated produced whitish mycelia when grown on potato dextrose agar (PDA); the mycelia later became gray to dark gray with aerial mycelia in tufts and numerous conidia were produced. The conidia were straight, cylindrical with an obtuse apex and a truncated base, and measured 11.4 to 17.5 × 4.2 to 7.1 μm. The measurements and taxonomic characteristics coincide with those of Colletotrichum gloeosporioides (Penz.) (1). The isolated fungus was tested for pathogenicity on crab apples and cv. Fuji apples by inoculation with a conidial suspension (105 conidia/ml) prepared from 20-day-old PDA cultures. A 20-μl drop of the conidial suspension was placed onto crab apple and apple fruits that had been wounded by piercing them 1 to 2 mm deep with a pin. Small, dark lesions were observed on the artificially inoculated fruit 3 days after inoculation. Nine days after inoculation, dark lesions with salmon-colored masses of conidia were observed on fruit, which were also soft and sunken. Abundant masses of conidia were produced in the decayed tissues. The fungus was reisolated from the parts of the fruits showing the symptoms. The internal transcribed spacer (ITS) rDNA of the isolated fungus was amplified and sequenced by PCR as described by White et al. (2). The resulting 582-bp of ITS rDNA sequence was deposited in GenBank (Accession No. JQ405742). A BLAST analysis for sequence similarity of the ITS region revealed 100% identity with nucleotide sequences for C. gloeosporioides isolates (Accession Nos. HQ645080 and AB458667). The results obtained on morphological characteristics, pathogenicity, and molecular data corresponded with those of C. gloeosporioides described by Sutton (1). To our knowledge, this is the first report of the presence of C. gloeosporioides on crab apple in Korea (3). Crab apple is used as a pollinator for single-cultivar apple orchards and may become a possible source of inoculum for cultivated apple. References: (1) T. B. Sutton. Compendium of Apple and Pear Diseases. The American Phytopathological Society, St. Paul, MN, 1990. (2) T. J. White et al. PCR Protocols: A Guide to Methods and Applications, Academic Press, Inc., New York, 1990. (3) S. H. Yu. List of Plant Diseases in Korea. 5th ed. (in Korean). The Korean Society of Plant Pathology, 2009.

Download Full-text

First Report of Fusarium tricinctum Causing Stem and Root Rot on Lanzhou Lily (Lilium davidii var. unicolor) in China

Plant Disease ◽

10.1094/pdis-11-13-1146-pdn ◽

2014 ◽

Vol 98 (7) ◽

pp. 999-999 ◽

Cited By ~ 8

Author(s):

Q. H. Shang ◽

X. Zhao ◽

Y. Y. Li ◽

Z. K. Xie ◽

R. Y. Wang

Keyword(s):

Root Rot ◽

Gansu Province ◽

Plant Diseases ◽

Pathogenicity Test ◽

Conidial Suspension ◽

Sterile Water ◽

First Report ◽

5.8S Rdna ◽

Lanzhou Lily ◽

Lilium Davidii

Lanzhou lily (Lilium davidii var. unicolor Cotton) is an important bulb edible crop which mostly distributes in middle area of Gansu Province in China (2). Recently, plants of Lanzhou lily developed symptoms of severe wilting. In early autumn of 2012 to 2013, a survey of Lanzhou lily disease was carried out in Yuanjiawan, Caoyuan, Xiguoyuan, and Hutan villages of Lanzhou City and Xuding and Guanshan villages of Linxia Prefecture. Disease symptoms included stem and root rot, vessels showed a brown to dark brown discoloration, plus a progressive yellowing and wilting of leaves from the base. Small pieces of symptomatic leaves, stems, and roots were surface disinfected with 75% ethanol for 30 s, 3% sodium hypochlorite for 5 min, and then washed three times in sterile distilled water. The tissues were placed on Martin Agar at 25°C for 7 days. Three isolates were consistently isolated from diseased tissues and all isolates with morphology similar to Fusarium spp. Isolates were transferred to potato dextrose agar (PDA) and carnation leaf agar (CLA) and incubated at 25°C in darkness. These isolates grew rapidly on PDA and formed abundant dense aerial mycelium, initially white, that became deep pink with age and formed red pigments in the medium. On CLA, macroconidia with 3 to 5 septa were abundant, relatively slender, and curved to lunate. Microconidia were abundant, oval and 0 to 1 septa. Chlamydospores were globose with a smooth outer wall in chains. The rDNA internal transcribed spacer (ITS) region comprising ITS1, ITS2, and 5.8S rDNA was amplified using primers ITS-1 and ITS-4 (3) and sequenced. On the basis of a comparison of 563 bp, all the three isolates had the identical sequence (GenBank Accession No. KF728675). BLASTn analysis of the sequence showed 100% match with the ITS sequences of those F. tricinctum sequences in GenBank (Accession Nos. FJ233196, AY188923, and JF776663). Pathogenicity test was performed by transplanting 2-month-old tissue culture seedlings to plastic pots in a sterile mixture of vermiculite and torf substrate at 1:3 (v/v). Seedlings were inoculated with 6 ml of the conidial suspension (104 conidia/ml) on the roots of plant in each pot, three plants per pot, and three replicates for each treatment. Seedlings treated with sterile water served as controls. The seedlings were placed in a plant growth chamber maintained at 22 ± 3°C, relative humidity >70%, 16 h light per day, and irrigated with sterile water. After 4 weeks, inoculated plants exhibited wilting foliage that with symptoms similar to those observed in the field, while the control plants remained healthy. F. tricinctum was re-isolated from all inoculated plants. The disease has been reported previously in ornamental lily in China (1). However, to the best of our knowledge, this is the first report of F. tricinctum causing wilt on edible Lanzhou lily in China and the disease must be taken into consideration of current disease management. This work supported by NSFC No. 31370447 and Hundred Talents Program of CAS “Molecular mechanism of biological control on plant diseases.” References: (1) Y. Y. Li et al. Plant Dis. 97:993, 2013. (2) R. Y. Wang et al. Virol. J. 7:34, 2010. (3) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.

Download Full-text

First Report of Powdery Mildew Caused by Erysiphe alphitoides on Japanese Snailseed (Cocculus trilobus) in Korea

Plant Disease ◽

10.1094/pdis-01-13-0031-pdn ◽

2014 ◽

Vol 98 (5) ◽

pp. 685-685 ◽

Cited By ~ 2

Author(s):

W. Cheon ◽

S. G. Lee ◽

Y. H. Jeon

Keyword(s):

Powdery Mildew ◽

Morphological Characteristics ◽

Its Region ◽

Plant Diseases ◽

Its Sequences ◽

Korean Society ◽

First Report ◽

Erysiphe Alphitoides ◽

Oak Tree ◽

Mother Cells

Japanese snailseed (Cocculus trilobus DC.) has been known as a medicinal herb to treat dieresis, rheumatoid arthritis, and dropsy. In September 2011, severe powdery signs were found on several Japanese snailseed plants near Andong, Korea. Diseased leaves showed chlorotic or necrotic lesions, along with leaf distortion and senescence. Diseased leaves were associated with a fungus that resulted in what appeared to be white colonies, predominately associated with the upper leaf surfaces, and rarely on the lower surfaces. The colonies increased in size and coalesced, subsequently covering the entire surface. The fungus-produced chasmothecia were 92 to 123 μm in diameter, blackish brown, and had a depressed, globose shape. Each chasmothecium had approximately 8 to 12 appendages that were straight to mildly bent, and were four to six times dichotomously branched and often entwined. There were three to six asci per chasmothecium, 38 to 57 × 32 to 43 μm in size, each of which held six to eight ascospores. Conidiophores were single or sometimes two on a hyphal cell, arising from the upper part of mother cells, mostly positioned central, 6.5 to 8 μm with width. Conidiophores were erect and up to 150.5 μm long. Conidia were ellipsoidal or sometimes lemon-shaped. The conidial size was 31.5 to 40 × 19 to 24.5 μm with length/width. These morphological characteristics were identified as being similar to Erysiphe alphitoides (1). DNA was extracted from collected hyphae of infected leaves using the NucleoSpin Tissue Kit (Macherey-Nagel, Duren, Germany). The ITS region of rDNA was amplified using primers ITS4/ITS5 and sequenced (GenBank Accession No. KF734882). The isolate (APEC-F1203) was 99% homologous to other E. alphitoides isolates from oak trees in Japan (AB292704, AB292699, AB292697, and AB292701) and Europe (EF672350, AJ417497). In Korea, this fungus is an oak tree pathogen (2). As proof of pathogenicity, infected leaves having abundant sporulation were pressed onto leaves of five healthy plants. Inoculated and non-inoculated plants were incubated in a moist chamber for 48 h and then maintained in a greenhouse at 15 to 22°C. After 10 to 12 days, powdery mildew colonies developed on inoculated plants. Uninoculated control plants did not show powdery mildew. Microscopic observation of the pathogen growing on the inoculated plants revealed that it was the same as the original fungus. We also observed powdery mildews on oak tree leaves around Japanese snailseed and analyzed their ITS sequences with the above-mentioned methods. As a result, the ITS sequences of powdery mildew pathogens obtained from Japanese snailseed and oak tree were identical. To our knowledge, this is the first report of the presence of E. alphitoides on Japanese snailseed in Korea. This fungus has been reported in association with numerous oak (Quercus spp.) species in Korea, showing that it may be a potential source of inoculum in Japanese snailseed. References: (1) S. Takamatsu et al. British Mycol. Res. 111:809. 2007. (2) S. H. Yu. List of Plant Diseases in Korea, 5th ed. The Korean Society of Plant Pathology, 2009.

Download Full-text

First Report of Zucchini Collapse by Fusarium solani f. sp. cucurbitae Race 1 and Plectosporium tabacinum in Italy

Plant Disease ◽

10.1094/pdis-91-3-0325a ◽

2007 ◽

Vol 91 (3) ◽

pp. 325-325 ◽

Cited By ~ 7

Author(s):

S. Vitale ◽

M. Maccaroni ◽

A. Belisario

Keyword(s):

Fusarium Solani ◽

Fusarium Species ◽

Morphological Characteristics ◽

Its Region ◽

Conidial Suspension ◽

Single Spore ◽

Running Water ◽

First Report ◽

Race 1 ◽

Rot Disease

Zucchini plant collapse has been often associated with Fusarium solani f. sp. cucurbitae race 1, which is the causal agent of Fusarium crown and foot rot disease of cucurbits. In Italy, F. solani f. sp. cucurbitae race 1 has been reported on zucchini (Cucurbita pepo) in a greenhouse in the Tuscany Region (4). In spring 2005, a severe outbreak was observed on zucchini in a vast area of cultivation in the province of Venice. Isolations from necrotic vessels gave more than 20 single-spore cultures. On the basis of morphological characteristics, they were identified as F. solani (2) and Plectosporium tabacinum (3). The internal transcribed spacer (ITS) region of rDNA was amplified and sequenced. A fragment of 454 and 531 bp was 99% homologous with sequence PSU66732 and AF150472 of F. solani f. sp. cucurbitae race 1 and P. tabacinum, respectively, in the NCBI database. The nucleotide sequences have been assigned Accession No. AM408782 for F. solani f. sp. cucurbitae race 1 and AM408781 for P. tabacinum. Pathogenicity tests were conducted with four isolates of each species on 15-day-old zucchini plants and on fruit. Plants were inoculated by dipping the roots in a conidial suspension of 106 spores ml-1 for 10 min. Control plants were dipped in sterile water. Five replicates for the inoculated and control plants were used. All plants were maintained in a greenhouse at approximately 24°C. After 14 days, inoculations with F. solani f. sp. cucurbitae race 1 gave symptoms of a cortical rot at the base of the stem with a progressive yellows and wilting of leaves, while plants inoculated with P. tabacinum displayed a moderate wilting. Fruit were washed under running water, disinfected with a solution of 3% sodium hypochlorite and 5% ethanol for 1 min, and inoculated with 6-mm-diameter mycelial plugs cut from the margin of 10-day-old cultures grown on PDA. Plugs were inserted into holes (approximately 2 mm deep) made with a sterile 7-mm-diameter cork borer. Five replicates per isolate were used. Fruit were kept at room temperature (22 to 24°C) in a moist chamber. All isolates induced symptoms of fruit rotting 10 days after inoculation. All controls remained healthy. The colonies reisolated from the inoculated plants and fruit were morphologically identical to the original isolates. The results obtained proved that F. solani f. sp. cucurbitae race 1 can be considered the major pathogen in zucchini collapse, at the same time P. tabacinum may play a role in this syndrome as reported for other cucurbits (1). To our knowledge, this is the first report of zucchini plant collapse caused by F. solani f. sp. cucurbitae race 1 and P. tabacinum, and the first report of P. tabacinum on zucchini in Italy. References: (1) V. J. Garcia-Jimenez et al. EPPO Bull. 30:169, 2000. (2) P. E. Nelson et al. Fusarium Species: An Illustrated Manual for Identification. Pennsylvania State University, University Park, 1983. (3) M. E. Palm et al. Mycologia 87:397, 1995. (4) G. Vannacci and P. Gambogi. Phytopathol. Mediterr. 19:103, 1980.

Download Full-text

First Report of Pepper Fruit Rot Caused by Fusarium concentricum in China

Plant Disease ◽

10.1094/pdis-03-13-0325-pdn ◽

2013 ◽

Vol 97 (12) ◽

pp. 1657-1657 ◽

Cited By ~ 6

Author(s):

J. H. Wang ◽

Z. H. Feng ◽

Z. Han ◽

S. Q. Song ◽

S. H. Lin ◽

...

Keyword(s):

Fusarium Species ◽

Fruit Rot ◽

Post Inoculation ◽

Distilled Water ◽

Conidial Suspension ◽

Average Growth ◽

First Report ◽

Control Fruit ◽

Pepper Fruit ◽

Fruit Samples

Pepper (Capsicum annuum L.) is an important vegetable crop worldwide. Some Fusarium species can cause pepper fruit rot, leading to significant yield losses of pepper production and, for some Fusarium species, potential risk of mycotoxin contamination. A total of 106 diseased pepper fruit samples were collected from various pepper cultivars from seven provinces (Gansu, Hainan, Heilongjiang, Hunan, Shandong, Shanghai, and Zhejiang) in China during the 2012 growing season, where pepper production occurs on approximately 25,000 ha. Pepper fruit rot symptom incidence ranged from 5 to 20% in individual fields. Symptomatic fruit tissue was surface-sterilized in 0.1% HgCl2 for 1 min, dipped in 70% ethanol for 30 s, then rinsed in sterilized distilled water three times, dried, and plated in 90 mm diameter petri dishes containing potato dextrose agar (PDA). After incubation for 5 days at 28°C in the dark, putative Fusarium colonies were purified by single-sporing. Forty-three Fusarium strains were isolated and identified to species as described previously (1,2). Morphological characteristics of one strain were identical to those of F. concentricum. Aerial mycelium was reddish-white with an average growth rate of 4.2 to 4.3 mm/day at 25°C in the dark on PDA. Pigments in the agar were formed in alternating red and orange concentric rings. Microconidia were 0- to 1-septate, mostly 0-septate, and oval, obovoid to allantoid. Macroconidia were relatively slender with no significant curvature, 3- to 5-septate, with a beaked apical cell and a foot-shaped basal cell. To confirm the species identity, the partial TEF gene sequence (646 bp) was amplified and sequenced (GenBank Accession No. KC816735). A BLASTn search with TEF gene sequences in NCBI and the Fusarium ID databases revealed 99.7 and 100% sequence identity, respectively, to known TEF sequences of F. concentricum. Thus, both morphological and molecular criteria supported identification of the strain as F. concentricum. This strain was deposited as Accession MUCL 54697 (http://bccm.belspo.be/about/mucl.php). Pathogenicity of the strain was confirmed by inoculating 10 wounded, mature pepper fruits that had been harvested 70 days after planting the cultivar Zhongjiao-5 with a conidial suspension (1 × 106 spores/ml), as described previously (3). A control treatment consisted of inoculating 10 pepper fruits of the same cultivar with sterilized distilled water. The fruit were incubated at 25°C in a moist chamber, and the experiment was repeated independently in triplicate. Initially, green to dark brown lesions were observed on the outer surface of inoculated fruit. Typical soft-rot symptoms and lesions were observed on the inner wall when the fruit were cut open 10 days post-inoculation. Some infected seeds in the fruits were grayish-black and covered by mycelium, similar to the original fruit symptoms observed at the sampling sites. The control fruit remained healthy after 10 days of incubation. The same fungus was isolated from the inoculated infected fruit using the method described above, but no fungal growth was observed from the control fruit. To our knowledge, this is the first report of F. concentricum causing a pepper fruit rot. References: (1) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Ames, IA, 2006. (2) K. O'Donnell et al. Proc. Nat. Acad. Sci. USA 95:2044, 1998. (3) Y. Yang et al. 2011. Int. J. Food Microbiol. 151:150, 2011.

Download Full-text

First Report of Anthracnose of Onion Caused by Colletotrichum coccodes in Ohio

Plant Disease ◽

10.1094/pdis-01-14-0032-pdn ◽

2014 ◽

Vol 98 (9) ◽

pp. 1271-1271 ◽

Cited By ~ 2

Author(s):

F. Baysal-Gurel ◽

N. Subedi ◽

D. P. Mamiro ◽

S. A. Miller

Keyword(s):

Leaf Tissue ◽

Its Region ◽

The United States ◽

Tween 20 ◽

Colletotrichum Coccodes ◽

Academic Press ◽

Conidial Suspension ◽

First Report ◽

Allium Cepa L ◽

Total Dna

Dry bulb onion (Allium cepa L. cvs. Pulsar, Bradley, and Livingston) plants with symptoms of anthracnose were observed in three commercial fields totaling 76.5 ha in Huron Co., Ohio, in July 2013. Symptoms were oval leaf lesions and yellowing, curling, twisting, chlorosis, and death of leaves. Nearly half of the plants in a 32.8-ha field of the cv. Pulsar were symptomatic. Concentric rings of acervuli with salmon-colored conidial masses were observed in the lesions. Conidia were straight with tapered ends and 16 to 23 × 3 to 6 μm (2). Colletotrichum coccodes (Wallr.) S. Hughes was regularly isolated from infected plants (2). Culturing diseased leaf tissue on potato dextrose agar (PDA) amended with 30 ppm rifampicin and 100 ppm ampicillin at room temperature yielded white aerial mycelia and salmon-colored conidial masses in acervuli. Numerous spherical, black microsclerotia were produced on the surface of colonies after 10 to 14 days. To confirm pathogen identity, total DNA was extracted directly from a 7-day-old culture of isolate SAM30-13 grown on PDA, using the Wizard SV Genomic DNA Purification System (Promega, Madison, WI) following the manufacturer's instructions. The ribosomal DNA internal transcribed spacer (ITS) region was amplified by PCR using the primer pair ITS1 and ITS4 (2), and sequenced. The sequence, deposited in GenBank (KF894404), was 99% identical to that of a C. coccodes isolate from Michigan (JQ682644) (1). Ten onion seedlings cv. Ebenezer White at the two- to three-leaf stage of growth were spray-inoculated with a conidial suspension (1 × 105 conidia/ml containing 0.01% Tween 20, with 10 ml applied/plant). Plants were maintained in a greenhouse (21 to 23°C) until symptoms appeared. Control plants were sprayed with sterilized water containing 0.01% Tween 20, and maintained in the same environment. After 30 days, sunken, oval lesions each with a salmon-colored center developed on the inoculated plants, and microscopic examination revealed the same pathogen morphology as the original isolates. C. coccodes was re-isolated consistently from leaf lesions. All non-inoculated control plants remained disease-free, and C. coccodes was not re-isolated from leaves of control plants. C. coccodes was reported infecting onions in the United States for the first time in Michigan in 2012 (1). This is the first report of anthracnose of onion caused by C. coccodes in Ohio. Unusually wet, warm conditions in Ohio in 2013 likely contributed to the outbreak of this disease. Timely fungicide applications will be necessary to manage this disease in affected areas. References: (1) A. K. Lees and A. J. Hilton. Plant Pathol. 52:3. 2003. (2) L. M. Rodriguez-Salamanca et al. Plant Dis. 96:769. 2012. (3) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.

Download Full-text

First Report of Anthracnose Caused by Colletotrichum fragariae on Cyclamen in North Carolina

Plant Disease ◽

10.1094/pdis-06-11-0475 ◽

2011 ◽

Vol 95 (11) ◽

pp. 1480-1480 ◽

Cited By ~ 3

Author(s):

B. Liu ◽

M. Munster ◽

C. Johnson ◽

F. J. Louws

Keyword(s):

North Carolina ◽

Sequence Analysis ◽

Room Temperature ◽

Plant Diseases ◽

Pathogenicity Test ◽

Conidial Suspension ◽

Disease Symptoms ◽

First Report ◽

Leaf Spots ◽

Colletotrichum Fragariae

In November 2009, cyclamen (Cyclamen persicum) plants with disease symptoms from a commercial greenhouse operation in the western part of North Carolina were sent to the Plant Diseases and Insect Clinic at North Carolina State University. Symptoms consisted of coalescing reddish and tan necrotic leaf spots with concentric circles. Other symptoms included darkened vascular tissue and decay of the corm, large roots, and petioles. Diseased leaves and stems were surface sterilized in 0.5% sodium hypochlorite for 3 min, air dried, and placed in petri dishes containing alkaline water agar. After 3 days of incubation at room temperature, fungal colonies were transferred to acidified potato dextrose agar. Isolation frequency after 5 days was 33% (three of nine pieces) and 16% (one of six pieces) from small leaf spots and petioles, respectively. Pure cultures of isolates were gray and black with abundant, aerial, gray whitish mycelia. Diseased plants were also incubated in a moist chamber at room temperature and sporulation was observed within 7 days. Conidia were tapered with rounded ends and produced in the acervulus and on the tips of setae, which is consistent with the morphology of described isolates of Colletotrichum fragariae. Similar setae were also observed directly on the fine roots of the original sample. The pathogenicity of single-spore cultures was tested by spraying four 2-month-old cyclamen plants with a conidial suspension (106 conidia/ml) and the plants were kept in a humid chamber for 24 h. Noninoculated controls (four plants) were sprayed with distilled water and subjected to the same conditions. The pathogenicity test was also repeated. Inoculated plants and controls were placed in a greenhouse with a temperature range from 22 to 25°C. After 7 to 10 days, symptomatic leaves and stems were observed on all the inoculated plants but not on the control plants. Fungi reisolated from 10 symptomatic leaf tissues had identical morphological features as the original isolates. Fungal DNA was extracted with DNeasy Plant Mini DNA Extraction Kits following the manufacturer's protocol (Qiagen Inc., Valencia, CA). Sequence analysis of the rRNA internal transcribed spacer (ITS) region of the cyclamen isolate (GenBank Accession No. HQ188923), based on the fragment amplified with ITS1 and ITS4 primers, showed 100% similarity to isolates of C. fragariae deposited in GenBank (Accession Nos. FJ172290 [ATCC MYA-4443 from cyclamen] and FJ810510 [ATCC MYA-4442 from silver date palm]) and Florida isolate C16 isolated from strawberry (1). In addition, the morphology and ITS sequences of the cyclamen isolate were identical to those of the C. fragariae voucher isolate from strawberry (GU174546). Results from disease symptoms, colony and spore morphology, pathogenicity tests, and ITS sequence analysis suggest that C. fragariae was the pathogen responsible for the disease symptoms on cyclamens. To our knowledge, this is the first report of a disease caused by C. fragariae on cyclamen in North Carolina and complements an earlier report from Florida (1). Reference: (1) S. J. MacKenzie et al. Plant Dis. 92:1432, 2008.

Download Full-text