First Report of Gray Mold (Amphobotrys ricini) on Copperleaf (Acalypha wilkesiana) in Brazil

Acalypha wilkesiana (Euphorbiaceae), common names copperleaf or Jacob's coat (in Brazil, crista-de-peru), is a popular ornamental native from the Pacific islands. It is widely used in gardens in Brazil (4). In January 2012, a group of diseased A. wilkesiana was found in a nursery at the municipality of Itaboraí (state of Rio de Janeiro, Brazil). Later, another group of individuals of the same plant species bearing identical disease symptoms were found in a botanic garden in the city of Rio de Janeiro (Jardim Botânico do Rio de Janeiro). Diseased plants had intense leaf blight. Such leaves dropped over healthy leaves of the same or other plants and necrosis was hence initiated on such leaves. Inflorescences were also affected by blight and after becoming necrotic a dieback of supporting stems also resulted. Abundant grayish sporulation was easily observed over necrotic tissues. Samples were collected, dried in a plant press, and representative specimens were deposited in the herbarium at the Universidade Federal de Viçosa. These were from Itaboraí (VIC 31822) and from Rio de Janeiro (VIC 31931). Structures were mounted in lactophenol for observation under a microscope and isolated in pure culture on PCA plates. Isolates were deposited in the culture collection of the Universidade Federal de Viçosa with accession numbers of COAD 1112 and COAD 1108, respectively. The fungus had the following morphology: conidiophores cylindrical, up to 1,200 μm branching dicotomously at mid-length in broad angles and then branching secondarily, light brown; conidiogenous cells ampulliform, terminal, denticulate; conidia globose, 6 to 11 μm diam, subhyaline to pale brown, smooth. This combination of features is typical of Amphobotrys ricini (2), a common pathogen of castor bean (1) and several other members of the Euphorbiaceae. DNA was extracted from each isolate growing in pure culture and ITS sequences were generated and deposited in GenBank under the accession numbers JX961613 (COAD 1108) and JX961614 (COAD 1112). These were compared by BLASTn with other entries in GenBank, and the closest match for both isolates was A. ricini (JF433374) with 97% nucleotide homology (over 97% query coverage) for COAD 1112 and 98% nucleotide homology (over 98% query coverage) for COAD 1112. Pathogenicity of the isolate from A. wilkesiana was demonstrated through brush inoculation of a conidial suspension (3 × 106 conidia. mL–1) onto healthy leaves of a A. wilkesiana individual followed by its transfer to a humid chamber for 48 h. Symptoms appeared after 3 days of inoculation and sporulation appeared over necrotic tissues after 10 days. Despite the importance of A. ricini as a plant pathogen, little has been investigated on its taxonomy with molecular tools. Although morphology and host-association are the basis for the delimitation of A. ricini, our preliminary results for ITS sequences suggest that this species may include cryptic taxa that are not properly discriminated on a morphological and pathological basis. This report follows other novel reports of A. ricini on ornamental Euphorbiaceae in Brazil (3) and, to our knowledge, represents the first report of A. ricini on A. wilkesiana worldwide. References: (1) G. H. Godfrey. J. Agric. Res. 23:679, 1923. (2) G. L. Hennebert. Persoonia 7:183, 1973. (3) B. V. Lima et al. Australas. Plant Dis. Notes 3:5, 2008. (4) H. Lorenzi and H. M. Souza. Plantas Ornamentais no Brasil - Arbustivas, Herbáceas e Trepadeiras. Nova Odessa: Instituto Plantarum, 1999.

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First Report of Stigmina palmivora Causing Leaf Spots on Phoenix roebelenii in Brazil

Plant Disease ◽

10.1094/pdis-10-13-1030-pdn ◽

2014 ◽

Vol 98 (6) ◽

pp. 849-849 ◽

Cited By ~ 1

Author(s):

A. Colmán ◽

R. A. da Silva ◽

R. Alves ◽

M. Silva ◽

R. W. Barreto

Keyword(s):

Pure Culture ◽

Leaf Surface ◽

The United States ◽

Culture Collection ◽

Commonwealth Mycological Institute ◽

First Report ◽

Leaf Spots ◽

Nucleotide Homology ◽

Query Coverage ◽

Representative Samples

Phoenix roebelenii (Arecaceae), known as dwarf date (tamareira-anã in Brazil), is a palm native to Southeast Asia and widely cultivated worldwide because of its ornamental value and ease of adaptation to a broad range of climates and soil types (4). In June 2012, some individuals were observed in a private garden in the municipality of Viçosa (state of Minas Gerais, Brazil) bearing numerous necrotic lesions on its leaves. Representative samples were taken, dried in a plant press, and brought to the laboratory for examination. A fungus was regularly associated with the leaf spots. Fungal structures were mounted in lactophenol and slides were examined under a microscope (Olympus BX 51). Spores were taken from sporulating colonies with a sterile fine needle and plated on PDA for isolation. A pure culture was deposited in the culture collection of the Universidade Federal de Viçosa (accession COAD1338). A dried herbarium sample was deposited in the local herbarium (VIC39741). The fungus had the following morphology: conidiophores grouped on sporodochia, cylindrical, 12 to 29 × 5 to 6 μm, dark brown; conidiogenous cells, terminal, proliferating percurrently (annellidic), 8 to 20 × 5 to 6 μm, pale to dark brown; conidia obclavate to subcylindrical, straight, 58 to 147 × 5 to 6 μm, 6 to 16 septate, hila thickened and darkened with a thin-walled projecting papilla, dark brown, and verrucose. The morphology of the Brazilian collections agrees well with the description of Stigmina palmivora (2), a species known to cause leaf spots on P. roebelenii in the United States (Florida) and Japan (3). Pathogenicity was demonstrated through inoculation of leaves of healthy plants by placing 6 mm diameter cuture disks of COAD1338 on the leaf surface followed by incubation in a moist chamber for 48 h and then transferred to a greenhouse bench at 21 ± 3°C. Typical leaf spots were observed 15 days after inoculation. DNA was extracted from the isolate growing in pure culture and ITS and LSU sequences were generated and deposited in GenBank under the accession numbers KF656785 and KF656786, respectively. These were compared by BLASTn with other entries in GenBank, and the closest match for each region were Mycosphaerella colombiensis strain X215 and M. irregulariamosa strain CPC 1362 (EU514231, GU2114441) with 93% of nucleotide homology (over 100% query coverage) for ITS and 98% of nucleotide homology (over 100% query coverage) for LSU. There are no sequences for S. palmivora deposited in public databases for comparison, but for Stigmina platani, the type species in this genus, 86% and 96% nucleotide homology for ITS and LSU with S. palmivora were found. The genus Stigmina is regarded as being polyphyletic (1) and this is probably reflected by these low homology levels found in the BLASTn search. To our knowledge, this is the first report of Stigmina palmivora in Brazil. References: (1) P. W. Crous et al. Stud. Mycol. 75:37, 2012. (2) M. B. Ellis. Dematiaceous Hyphomycetes. Commonwealth Mycological Institute, Kew, UK, 1971. (3) D. F. Farr and A. Y. Rossman. Fungal Databases. Syst. Mycol. Microbiol. Lab. ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ , 2013. (4) H. Lorenzi et al. Palmeira no Brasil: Exóticas e Nativas, 2nd ed. Editora Plantarum, Nova Odessa, Brazil, 2005.

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First Report of Leaf Blight on Duying Caused by Phyllosticta anacardiacearum in China

Plant Disease ◽

10.1094/pdis-93-5-0546b ◽

2009 ◽

Vol 93 (5) ◽

pp. 546-546 ◽

Cited By ~ 2

Author(s):

B. G. Lou ◽

Y. D. Xu ◽

C. Sun ◽

X. M. Lou

Keyword(s):

Southern China ◽

Disease Incidence ◽

Its Region ◽

Culture Collection ◽

Leaf Blight ◽

Its Sequences ◽

Dead Leaf ◽

Conidial Suspension ◽

First Report ◽

Branch Dieback

Duying (Elaeocarpus glabripetalus Merr.; Elaeocarpaceae) is widely cultivated as an ornamental tree of commercial importance in southern China. From 2003 to 2008, severe outbreaks of Duying leaf blight occurred in the Hangzhou area, Zhejiang Province. Disease incidence was greater than 20% and mainly infected young leaves and shoots in the spring and autumn. Severely infected leaves and shoots died and eventually led to branch dieback. The overall growth decline of affected trees occurs over 4 to 6 years before tree death. Infection symptoms are characterized by grayish, round, semicircular- or irregular-shaped spots (5 mm to 5 cm long) with dark brown borders and the appearance of black, granular pycnidia within the dead leaf tissues. The primary infection zones are commonly observed on the leaf margins and apices, are brown, up to 2 mm in diameter, and often surrounded by a yellow zone. Pycnidia were globose and 122 to 127 μm (average 123.5 μm) in diameter. A fungus was consistently isolated from symptomatic tissues on potato dextrose agar (PDA). Ash-black pycnidia appeared on PDA after 10 days. Ascospores developed on modified PDA (1 liter of PDA + 20 g of Duying leaves) after 18 days. Conidiogenous cells were cylindrical to obpyriform. The hyaline conidia were obovoid and guttulate, 10 to 13 × 6 to 8 μm (average 11.5 × 7.5 μm), and usually surrounded by a mucilaginous sheath with a hyaline apical appendage that was 5 to 8 μm long. Pseudothecia were solitary and subglobose with long necks. Asci were 45 to 70 × 7.5 to 12 μm (average 62.5 × 10.8 μm). Ascospores were 12 to 13 × 4 to 5 μm with rounded apices and hyaline, mucilaginous, apical caps. The fungus was morphologically identified as Phyllosticta anacardiacearum van der Aa (teleomorph Guignardia mangiferae A. J. Roy). This identification was also confirmed by the China General Microbiological Culture Collection Center (CGMCC). Six representative fungal isolates were identified by sequencing the internal transcribed spacer (ITS) region of the rDNA and comparing the sequences with those in GenBank using BLAST searches. The ITS sequences of six cultures (GenBank Accession Nos. EU821356–EU821361) showed 100% identity with the ITS sequences of an isolate of a Phyllosticta sp. (GenBank Accession No. AF532314) (2) and G. mangiferae (GenBank Accession No. AY277717) (1). To fulfill Koch's postulates, a conidial suspension (106 conidia per ml) collected from PDA cultures (isolate phy01) was used to spray inoculate leaves of potted 3-year-old Duying trees. Inoculated trees were kept for 48 h under a polyethylene sheet cover and grown at 10 to 15°C in a greenhouse. A total of 30 leaves of five healthy trees were inoculated with the pathogen. In addition, five 3-year-old trees were sprayed with sterile water to serve as uninoculated controls. After 10 to 14 days, inoculated leaves showed infection symptoms resembling those observed on Duying trees naturally infected with P. anacardiacearum. The pathogen was reisolated from the margins of necrotic tissues, but not from controls. To our knowledge, this is the first report of leaf blight on E. glabripetalus caused by P. anacardiacearum in China. Reference: (1) F. R. Katia et al. Mycol. Res. 108:45, 2004. (2) A. K. Pandey et al. Mycol. Res. 107:439, 2003.

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First Report of Leaf Spot Caused by Ascochyta marginata on Aralia elata in Korea

Plant Disease ◽

10.1094/pdis-08-11-0658 ◽

2012 ◽

Vol 96 (1) ◽

pp. 147-147

Author(s):

S. H. Lee ◽

C. K. Lee ◽

M. J. Park ◽

H. D. Shin

Keyword(s):

East Asia ◽

Leaf Spot ◽

Morphological Characteristics ◽

Culture Collection ◽

Research Report ◽

Conidial Suspension ◽

First Report ◽

Commercial Cultivation ◽

Aralia Elata ◽

Initial Symptoms

Aralia elata (Miq.) Seem., known as Japanese angelica tree, is a deciduous shrub belonging to the Araliaceae, which is native to East Asia. The young shoots have long been used in various dishes in East Asia. Commercial cultivation of this shrub, especially in polytunnels, is expanding in Korea. Several diseases including Sclerotinia rot have been known to be present on this plant (1,2). In early September 2007, leaf spot symptoms were first observed on several trees in Hongcheon, Korea. Microscopic observations revealed that the leaf spots were associated with an Ascochyta sp. Further surveys of the Ascochyta leaf spot showed the occurrence of the disease in approximately 5 to 10% of the trees in the 3 ha of commercial fields surveyed in Chuncheon, Gapyeong, Inje, and Jinju, Korea. Initial symptoms on leaves were circular to irregular, brown to dark brown, becoming zonate, and finally fading to grayish brown in the center with a yellow halo. Representative samples were deposited in the herbarium of Korea University. Conidiomata on leaf lesions were pycnidial, amphigenous, but mostly epiphyllous, immersed or semi-immersed in host tissue, light brown to olive brown, and 60 to 200 μm in diameter. Ostioles were papillate, 20 to 35 μm wide, and surrounded by a ring of darker cells. Conidia were hyaline, smooth, cylindrical to clavate, straight to mildly curved, slightly constricted at the septa, medianly one-septate, sometimes aseptate, 8 to 16 × 2.5 to 3.5 μm, and contained small oil drops. These morphological characteristics were consistent with the previous reports of Ascochyta marginata J.J. Davis (3,4). A monoconidial isolate was cultured on potato dextrose agar (PDA) plates and accessioned in the Korea Agricultural Culture Collection (Accession KACC43082). The conidia were readily formed on PDA. Inoculum for the pathogenicity tests was prepared by harvesting conidia from 30-day-old cultures of KACC43082 and a conidial suspension (approximately 2 × 106 conidia/ml) was sprayed onto leaves of three healthy seedlings. Three noninoculated seedlings served as controls. Inoculated and noninoculated plants were covered with plastic bags for 48 h in a glasshouse. After 7 days, typical leaf spot symptoms started to develop on the leaves of the inoculated plants. The fungus, A. marginata, was reisolated from those lesions, confirming Koch's postulates. No symptoms were observed on control plants. Previously, the disease was reported in Japan (4) and China (3). To our knowledge, this is the first report of A. marginata on Japanese angelica trees in Korea. According to our field observations in Korea, the Ascochyta leaf spot mostly occurred on plants growing in a humid environment, especially during the rainy season. The seedlings as well as the trees growing in sunny, well-ventilated plots were nearly free from this disease. Therefore, the growing conditions seemed to be the most important factor for the development and severity of the disease. References: (1) C. K. Lee et al. Plant Pathol. J. 26:426, 2010. (2) S. H. Lee et al. Diseases of Japanese Angelica Tree and Their Control. Research Report 08-10. Korea Forest Research Institute. Seoul, Korea, 2008. (3) J. Sun et al. Acta Mycol. Sin. 14:107, 1995. (4) M. Yoshikawa and T. Yokoyama. Mycoscience 36:67, 1995.

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First Report of Colletotrichum boninense Causing Anthracnose on Pepper in China

Plant Disease ◽

10.1094/pdis-04-12-0403-pdn ◽

2013 ◽

Vol 97 (1) ◽

pp. 138-138 ◽

Cited By ~ 7

Author(s):

Y. Z. Diao ◽

J. R. Fan ◽

Z. W. Wang ◽

X. L. Liu

Keyword(s):

Internal Transcribed Spacer ◽

Severe Disease ◽

Causal Agent ◽

Its Sequences ◽

Universal Primers ◽

Pathogenicity Test ◽

Conidial Suspension ◽

Single Spore ◽

First Report ◽

Species Specific

Anthracnose, caused by Colletotrichum spp., is a severe disease and results in large losses in pepper (Capsicum frutescens) production in China (4). Colletotrichum boninense is one of the Colletotrichum species in pepper in China. In August 2011, anthracnose symptoms (circular, sunken lesions with orange to black spore masses) were observed on pepper fruits in De-Yang, Sichuan Province, China. Three single-spore isolates (SC-6-1, SC-6-2, SC-6-3) were obtained from the infected fruits. A 5-mm diameter plug was transferred to potato dextrose agar (PDA); the isolates formed colonies with white margins and circular, dull orange centers. The conidia were cylindrical, obtuse at both ends, and 10.5 to 12.6 × 4.1 to 5.0 μm. The colonies grew rapidly at 25 to 28°C, and the average colony diameter was 51 to 52 mm after 5 days on PDA at 25°C. Based upon these characters, the causal agent was identified as C. boninense. To confirm the identity of the isolates, the internal transcribed spacer (ITS) regions were amplified with the ITS1/ITS4 universal primers (1). The internal transcribed spacer (ITS) sequences (Accession No. JQ926743) of the causal fungus shared 99 to 100% homology with ITS sequences of C. boninense in GenBank (Accession Nos. FN566865 and EU822801). The identity of the causal agent as C. boninense was also confirmed by species-specific primers (Col1/ITS4) (2). In a pathogenicity test, five detached ripe pepper fruits were inoculated with 1 μl of a conidial suspension (106 conidia/mL) or five fruits with 1 μl of sterile water were kept as control. After 7 days in a moist chamber at 25°C, typical anthracnose symptoms had developed on the five inoculated fruits but not on control fruits. C. boninense was reisolated from the lesions, and which was confirmed by morphology and molecular methods as before. There have reports of C. boninense infecting many species of plants, including pepper (3). To our knowledge, this is the first report of C. boninense causing anthracnose on pepper in China. References: (1) A. K. Lucia et al. Phytopathology 93:581, 2002. (2) S. A. Pileggi et al. Can. J. Microbiol. 55:1081, 2009. (3) H. J. Tozze et al. Plant Dis. 93:106, 2009. (4) M. L. Zhang. J. Anhui Agri. Sci. 2:21, 2000.

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First Report of Anthracnose of Tricyrtis macropoda Caused by Colletotrichum gloeosporioides in Korea

Plant Disease ◽

10.1094/pdis-03-12-0277-pdn ◽

2012 ◽

Vol 96 (7) ◽

pp. 1070-1070 ◽

Cited By ~ 1

Author(s):

J. H. Park ◽

K. S. Han ◽

Y. D. Kwon ◽

H. D. Shin

Keyword(s):

Colletotrichum Gloeosporioides ◽

Morphological Characteristics ◽

Its Region ◽

Culture Collection ◽

Perennial Herb ◽

Pathogenicity Test ◽

Conidial Suspension ◽

Natural Habitats ◽

First Report ◽

Small Water

Tricyrtis macropoda Miq. (syn. T. dilatata Nakai), known as speckled toadlily, is a perennial herb native to China, Japan, and Korea. The plant has been highly praised for its beautiful flowers and rare populations in natural habitats. In September 2006, several dozen plants were heavily damaged by leaf spots and blight in cultivated plantings in the city of Pocheon, Korea. The infections with the same symptoms were repeated every year. In July 2011, the same symptoms were found on T. macropoda in the cities of Gapyeong and Osan, Korea. The leaf lesions began as small, water-soaked, pale greenish to grayish spots, which enlarged to form concentric rings and ultimately coalesced. A number of blackish acervuli were formed in the lesions. Acervuli were mostly epiphyllous, circular to ellipsoid, and 40 to 200 μm in diameter. Setae were two- to three-septate, dark brown at the base, paler upwards, acicular, and up to 100 μm long. Conidia (n = 30) were long obclavate to oblong-elliptical, sometimes fusiform-elliptical, guttulate, hyaline, and 12 to 20 × 4 to 6.5 μm (mean 15.4 × 5.2 μm). These morphological characteristics of the fungus were consistent with the description of Colletotrichum gloeosporioides (Penz.) Penz. & Sacc. (2). Voucher specimens (n = 7) were deposited in the Korea University herbarium (KUS). Two isolates, KACC46374 (ex KUS-F25916) and KACC46405 (ex KUS-F26063), were deposited in the Korean Agricultural Culture Collection. Fungal DNA was extracted and the complete internal transcribed spacer (ITS) region of rDNA was amplified with the primers ITS1/ITS4 and sequenced. The resulting sequences of 549 bp were deposited in Genbank (Accession Nos. JQ619480 and JQ619481). They showed 100% similarity with a sequence of C. gloeosporioides (EU32619). Isolate KACC46374 was used in a pathogenicity test. Inoculum was prepared by harvesting conidia from 3-week-old cultures on potato dextrose agar. A conidial suspension (2 × 106 conidia/ml) was sprayed onto 15 leaves of three plants. Three noninoculated plants served as controls. Plants were covered with plastic bags to maintain 100% relative humidity for 24 h and then kept in a greenhouse (22 to 28°C and 70 to 80% RH). After 5 days, typical leaf spot symptoms, identical to the ones observed in the field, started to develop on the leaves of inoculated plants. No symptoms were observed on control plants. C. gloeosporioides was reisolated from the lesions of inoculated plants, thus fulfilling Koch's postulates. An anthracnose associated with C. tricyrtii (Teng) Teng was recorded on T. formosana and T. latifolia in China (3) and on T. formosana in Taiwan (1), respectively, without etiological studies. The morphological features of C. tricyrtii are within the variation of C. gloeosporioides (2). To our knowledge, this is the first report of anthracnose of T. macropoda. This report has significance to indigenous plant resource conservation managers and scientists because T. macropoda has been listed as one of the 126 “Rare and Endangered Plants” by the Korea Forest Service since 1991. References: (1) K. Sawada. Rep. Dept. Agric. Gov. Res. Inst. Formosa 87: 1, 1944. (2) B. C. Sutton. Pages 1–27 in: Colletotrichum Biology, Pathology and Control. J. A. Bailey and M. J. Jeger, eds. CAB International, Wallingford, U.K. 1992. (3) S. C. Teng. Contrib. Biol. Lab. Sci. Soc. China 8:36, 1932.

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First Report of Volutella Blight on Pachysandra Caused by Volutella pachysandricola in China

Plant Disease ◽

10.1094/pdis-11-11-0997 ◽

2012 ◽

Vol 96 (4) ◽

pp. 584-584

Author(s):

Q. Bai ◽

Y. Xie ◽

R. Dong ◽

J. Gao ◽

Y. Li

Keyword(s):

Morphological Characteristics ◽

Stem Canker ◽

Botanical Garden ◽

The United States ◽

Culture Collection ◽

Leaf Blight ◽

Conidial Suspension ◽

First Report ◽

Leaf Spots ◽

Plastic Bags

Pachysandra (Pachysandra terminalis, Buxaceae) and Japanese Pachysandra, also called Japanese Spurge, is a woody ornamental groundcover plant distributed mostly in Zhejiang, Guizhou, Henan, Hubei, Sichuan, Shanxi, and Gansu provinces in China. In April 2010, P. terminalis asymptomatic plants were shipped from Beijing Botanical Garden Institute of Botany Chinese Academy of Science to the garden nursery of Jilin Agricultural University (43°48′N, 125°23′E), Jilin Province. In June 2011, Volutella blight (sometimes called leaf blight and stem canker) of P. terminalis was observed on these plants. Infected leaves showed circular or irregular, tan-to-brown spots often with concentric rings and dark margins. The spots eventually grew and coalesced until the entire leaf died. Cankers appeared as greenish brown and water-soaked diseased areas, subsequently turning brown or black, and shriveled and often girdled the stems and stolons. During wet, humid weather in autumn, reddish orange, cushion-like fruiting structures of the fungus appeared on the stem cankers and undersides of leaf spots. Symptoms of the disease were consistent with previous descriptions (2–4). Five isolates were obtained from necrotic tissue of leaf spots and cankers of stems and stolons and cultured on potato dextrose agar. The colony surface was salmon colored and slimy. Conidia were hyaline, one celled, spindle shaped, and 12.57 to 22.23 × 3.33 to 4.15 μm with rounded ends. Morphological characteristics of the fungus were consistent with the description by Dodge (2), and the fungus was identified as Volutella pachysandricola (telemorph Pseudonectria pachysandricola). The internal transcribed spacer (ITS) regions of the nuclear rDNA were amplified using primers ITS4/ITS5 (1). The ITS sequences were 553 bp long and identical among these five isolates (GenBank Accession No. HE612114). They were 100% identical to Pseudonectria pachysandricola voucher KUS-F25663 (Accession No. JN797821) and 99% identical to P. pachysandricola culture-collection DAOM (Accession No. HQ897807). Pathogenicity was confirmed by spraying leaves of clonally propagated cuttings of P. terminalis with a conidial suspension (1 × 106 conidia/ml) of the isolated V. pachysandricola. Control leaves were sprayed with sterile water. Plants were covered with plastic bags and kept in a greenhouse at 20 to 25°C for 72 h. After 5 to 8 days, typical disease symptoms appeared on leaves, while the control plants remained healthy. V. pachysandricola was reisolated from the leaf spots of inoculated plants. Pachysandra leaf blight and stem canker also called Volutella blight, is the most destructive disease of P. terminalis and previously reported in the northern humid areas of the United States (Illinois, Connecticut, Ohio, Indiana, Iowa, Massachusetts, Missouri, Kentucky, and Wisconsin), northern Europe (Britain, Germany, and Poland), and the Czech Republic. To our knowledge, this is the first report of the disease caused by V. pachysandricola in China. The disease may become a more significant problem in P. terminalis cultivation areas if the disease spreads on P. terminalis in nursery beds. References: (1) D. E. L. Cooke et al. Mycol. Res. 101:667, 1997. (2) B. O. Dodge. Mycologia 36:532, 1944. (3) S. M. Douglas. Online publication. Volutella Blight of Pachysandra. The Connecticut Agricultural Experiment Station, 2008. (4) I. Safrankova. Plant Protect. Sci.43:10, 2007.

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First Report of Leaf Spot Caused by Phoma dictamnicola on Dictamnus dasycarpus in Korea

Plant Disease ◽

10.1094/pdis-06-14-0658-pdn ◽

2014 ◽

Vol 98 (10) ◽

pp. 1443-1443

Author(s):

J. H. Park ◽

S. E. Cho ◽

C. K. Lee ◽

S. H. Lee ◽

H. D. Shin

Keyword(s):

Leaf Spot ◽

Its Region ◽

Molecular Data ◽

Culture Collection ◽

Morphological Identification ◽

Conidial Suspension ◽

Aesthetic Value ◽

Transparent Plastic ◽

First Report ◽

Plastic Bags

Dictamnus dasycarpus Turcz, known as densefruit pittany, is a perennial herbal plant belonging to the Rutaceae. In Oriental medicine, this plant is used for treatment of various ailments (4). Since the white and purple striped flowers and glossy leaves are of aesthetic value, the plant is popular in gardens throughout Korea. In July 2012, a leaf spot was observed on hundreds of D. dasycarpus with nearly 100% incidence in a garden in Gapyeong County, Korea. Lesions on leaves reaching up to 20 mm in diameter were circular to irregular, brown to dark brown, then becoming zonate with age, and finally fading to grayish brown in the center with a reddish brown margin. The disease caused premature defoliation and reduced plant vigor as well as aesthetic value. In June 2014, the same symptoms were found on D. dasycarpus in a nursery in Jinju City, Korea. Representative samples were deposited in the Korea University Herbarium (KUS). Pycnidia on lesions were epiphyllous, immersed or semi-immersed in host tissue, light brown to olive brown, and 90 to 210 μm in diameter. Ostioles were 15 to 30 μm wide and surrounded by a ring of darker cells. Conidia were hyaline, smooth, ellipsoidal to nearly reniform, straight to mildly curved, aseptate or rarely medianly 1-septate with age, 5.5 to 9.6 × 1.8 to 3.6 μm, and contained small oil drops. These characteristics were consistent with the previous descriptions of Phoma dictamnicola Boerema, Gruyter & Noordel. (1,2). A monoconidial isolate was cultured on potato dextrose agar plates and deposited in the Korea Agricultural Culture Collection (Accession No. KACC46948). Morphological identification of the fungus was confirmed by molecular data. Genomic DNA was extracted using a DNeasy Plant Mini 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 505 bp was deposited in GenBank (Accession No. KM047023). A BLAST search showed that the ITS sequence shared >99% similarity with that of P. dictamnicola (GU237877). For the pathogenicity tests, inoculum was prepared by harvesting conidia from 30-day-old cultures of KACC46948 and a conidial suspension (2 × 106 conidia/ml) was sprayed onto leaves of five healthy seedlings. Five seedlings were sprayed with sterile distilled water, serving as controls. The plants were covered with transparent plastic bags for 48 h in a 25°C glasshouse with a 12-h photoperiod. After 10 days, typical leaf spot symptoms started to develop on the leaves of the inoculated plants. The fungus, P. dictamnicola, was re-isolated from those lesions, confirming Koch's postulates. No symptoms were observed on control plants. Previously, Phoma leaf spot on Dictamnus spp. has been reported in the Netherlands and North America (3) and recently in China (1). To our knowledge, this is the first report of leaf spot on D. dasycarpus caused by P. dictamnicola in Korea. Our observations suggest that low humidity with good ventilation as well as removal of infected leaves and plant debris might be main strategies for preventing this disease. References: (1) Q. Bai et al. Plant Dis. 95:771, 2011. (2) G. H. Boerema et al. Phoma Identification Manual: Differentiation of Specific and Infra-Specific Taxa in Culture. CABI Publishing. Wallingford, UK, 2004. (3) D. F. Farr and A. Y. Rossman. Fungal Databases. Syst. Mycol. Microbiol. Lab., Online publication, USDA ARS, Retrieved June 19, 2014. (4) J. L. Yang et al. Planta Med. 77:271, 2011.

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First Report of Anthracnose Caused by Colletotrichum gloeosporioides on Soybean (Glycine max) in Malaysia

Plant Disease ◽

10.1094/pdis-10-12-0944-pdn ◽

2013 ◽

Vol 97 (6) ◽

pp. 841-841 ◽

Cited By ~ 1

Author(s):

F. Mahmodi ◽

J. B. Kadir ◽

M. Y. Wong ◽

A. Nasehi ◽

A. Puteh ◽

...

Keyword(s):

Glycine Max ◽

Morphological Characterization ◽

Negative Control ◽

Culture Collection ◽

Conidial Suspension ◽

First Report ◽

Representative Isolate ◽

Detached Leaves ◽

Soybean Plants ◽

Management Recommendations

Soybean (Glycine max L.) is one of the most economically important crops in the world, and anthracnose is known to infect soybean in most countries. Colletotrichum truncatum is the common pathogen causing anthracnose of soybean. However, at least five species of Colletotrichum have been reported on soybean worldwide (2). In July 2010, anthracnose symptoms were observed on soybean in the experimental fields of the agriculture station in Ladang Dua, University Putra Malaysia located in Selangor state of Malaysia. Symptoms were initially observed on a few plants randomly within one field, but after 4 weeks, the disease was found in two additional fields scattered across an area of 1 km2. Pinkish-brown lesions were observed on the pods, and the formation of dark lesions on the leaves and stems was sometimes followed by stem girdling, dieback, and distorted growth. At later stages, numerous epidermal acervuli developed in the lesions, and mucilaginous conidial masses appeared during periods of high relative humidity. Conidia produced in acervuli were straight, cylindric, hyaline, and aseptate, with both ends rounded. Conidia measured (mean ± SD) 14.2 ± 0.6 × 3.6 ± 0.7 μm, and the L/W ratio was 3.95 μm. Six isolates of the fungus were obtained and identified as C. gloeosporioides on the basis of morphological characterization (3). The isolates were deposited in the University Putra of Malaysia Culture Collection (UPMCC). PDA cultures were white at first and subsequently became grayish to pink to reddish-brown. Amplification and sequence analysis of coding and none-coding regions of the ITS-rDNA (GenBank JX669450), actin (JX827430), β-tubulin (JX827454), histone (JX827448), chitin synthase (JX827436), and glyceraldehyde-3-phosphate dehydrogenase (JX827442) obtained from the representative isolate, CGM50, aligned with deposited sequences from GenBank and revealed 99 to 100% sequence identity with C. gloeosporioides strains (JX258757, JX009790, GQ849434, HM575301, JQ005413, and JX00948 from GenBank). One representative isolate, CGM50, was used for pathogenicity testing. Four non-infected detached leaves and pods of 24-day-old G. max var. Palmetto were surface-sterilized and inoculated by placing 10 μl of a conidial suspension (106 conidia ml–1) using either the wound/drop or non-wound/drop method (4), with 10 μl distilled water as a negative control. Leaves and pods were incubated at 25°C, 98% RH. The experiment was repeated twice. Five days after inoculation, the development of typical field symptoms, including acervuli formation, occurred on the leaves and pods of inoculated plants, but not on the negative controls. A fungus with the same colony and conidial morphology as CGM50 was recovered from the lesions on the inoculated leaves and pods. Anthracnose caused by C. gloeosporioides on soybean plants has been reported previously in different countries, but not in Malaysia (3). Geographically, the climate of Malaysia is highly conducive to maintain and cause outbreaks of anthracnose all year round; thus, the development of management recommendations will be inevitable for anthracnose control. To our knowledge, this is the first report of C. gloeosporioides causing anthracnose on soybean in Malaysia. References: (1) U. Damm et al. Fungal Diversity 39:45, 2009. (2) S. L. Chen et al. J. Phytopathol. 154:654, 2006. (3) B. C. Sutton. The Genus Glomerella and its Anamorph Colletotrichum. CAB International, Wallingford, UK, 1992. (4) P. P. Than et al. Plant Pathol. 57:562, 2008. ERRATUM: A correction was made to this Disease Note on May 19, 2014. The author N. Soleimani was added.

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First Report of Leptosphaeria biglobosa Causing Black Leg on Brassica campestris ssp. chinensis var. purpurea in Central China

Plant Disease ◽

10.1094/pdis-01-14-0098-pdn ◽

2014 ◽

Vol 98 (8) ◽

pp. 1156-1156 ◽

Cited By ~ 1

Author(s):

X. Cai ◽

L. Yang ◽

J. Zhang ◽

G. Q. Li

Keyword(s):

Gene Sequence ◽

Brassica Campestris ◽

Its Sequences ◽

Central China ◽

Colony Morphology ◽

Conidial Suspension ◽

First Report ◽

Black Leg ◽

Species Specific ◽

Β Tubulin

Purple cai-tai (Brassica campestris ssp. chinensis var. purpurea) is a traditional vegetable widely grown in southern China. In 2012 and 2013, black leg disease was observed on purple cai-tai in three surveyed cities (Jingzhou, Qianjiang, and Huanggang) in Hubei Province of China. Disease incidence ranged from 5 to 88% in eight surveyed fields. White cankers occurred on basal stems and numerous black pycnidia and pink conidia were present on the stem surface. Surface-sterilized (5% NaOCl for 90 s, rinsed in sterilized water three times) stem pieces were plated on potato dextrose agar (PDA) and incubated at 20°C and 12 h light/12 h dark for 7 days. A total of 22 isolates were obtained. All of the isolates appeared similar in colony morphology on PDA (20°C, 7 to 10 days), producing yellow pigment and black-brown, globose pycnidia containing cylindrical hyaline conidia (4 to 5 × 2 μm). These characteristics matched the description for Phoma lingam, the anamorph Leptosphaeria maculans and L. biglobosa (2). Species-specific primers LbigF, LmacF, and LmacR (1) were used in PCR-based identification of the isolates. A 444-bp DNA fragment characteristic of L. biglobosa was amplified from DNA extracted from all of the collected isolates. DNA amplification from the isolate UK-1 of L. maculans from B. napus in Hertfordshire of the United Kingdom yielded a 331-bp fragment. Two isolates, HGHCT2-1 and HGHCT2-2, were further identified by cloning and analysis of the ITS sequences and the partial sequences encoding β-tubulin and actin (3,4). The ITS sequences (586 bp, GenBank Accession. Nos. KF371660 and KF371661) were 100% identical to L. biglobosa ‘brassicae’ strain UK28 (DQ133893). The DNA sequences for β-tubulin (479 bp, KF307760 and KF307761) and actin (899 bp, KF307758 and KF307759) were 99 and 100% identical to the partial β-tubulin gene sequence (AY748997) and the partial actin gene sequence (AY748949) of the L. biglobosa ‘brassicae’ strain 2379-4, respectively. Pathogenicity of six randomly selected isolates was determined on two purple cai-tai cvs. Wanzi Qianhong and Jiu Yue Xian. Cotyledons of 10-day-old seedlings grown in potting mix in pots were pricked with a sterilized needle, and each wound was inoculated with 10 μl of conidial suspension (1 × 107 conidia/ml) of an isolate or 10 μl sterilized water (control). There were 12 cotyledons for each isolate and control. The experiment was repeated once. The treated seedlings were incubated at 20°C in an incubator under 12 h light/12 h dark for 12 days. The control cotyledons were healthy, but necrotic lesions were developed on the cotyledons that were inoculated with L. biglobosa and formation of pycnidia was observed on some lesions. Fungi re-isolated from the lesions were similar to the original L. biglobosa isolates both in colony morphology on PDA and in species-specific PCR testing. No fungi were isolated from the control cotyledons. This is the first report of L. biglobosa causing black leg on B. campestris ssp. chinensis var. purpurea in central China. The finding will be useful for understanding of the epidemiology of black leg on cruciferous crops and for management of this disease. References: (1) S. Y. Liu et al. Plant Pathol. 55:401, 2006. (2) R. A. Shoemaker and H. Burn. Can J Bot. 79:412, 2001. (3) L. Vincenot et al. Phytopathology 98:321, 2008. (4) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.

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First Report of Oak Anthracnose Caused by Apiognomonia errabunda on Oriental White Oak in Korea

Plant Disease ◽

10.1094/pdis-02-13-0181-pdn ◽

2013 ◽

Vol 97 (8) ◽

pp. 1121-1121

Author(s):

C. K. Lee ◽

S. H. Lee ◽

J. H. Park ◽

S. E. Cho ◽

H. D. Shin

Keyword(s):

North America ◽

Morphological Characteristics ◽

Its Region ◽

Culture Collection ◽

Deciduous Tree ◽

Pathogenicity Test ◽

Conidial Suspension ◽

Natural Forests ◽

White Oak ◽

First Report

Oriental white oak, Quercus aliena Blume, is native to East Asia including Korea. It is one of the major deciduous tree species in natural forests in Korea. In May 2012, several hundred trees were found to be heavily damaged by a previously unknown leaf disease in a forest near Songjiho Lake in Goseong County of central Korea. Leaf symptoms began as small, water-soaked, pale greenish to grayish lesions, which enlarged to follow the veins or midribs and to be bounded by them, often killing part of the leaf. Leaf distortion and blight resulted in the later stage of disease development. A number of grayish brown to nearly black acervuli were formed on the lesions, especially on the midribs and veins. Acervuli were mostly hypophyllous, intraepidermal, erumpent, circular to ellipsoid in outline, cushion-like, and 70 to 220 μm in diameter. Conidia (n = 30) were elliptical to fusiform-elliptical, occasionally obclavate, aguttulate or guttulate, hyaline, aseptate, and 7.5 to 20 × 5 to 7.5 μm (mean 14.6 × 6.1 μm). These morphological characteristics of the fungus were consistent with the description of conidial state of Apiognomonia errabunda (Roberge ex Desm.) Höhn. (3,4). Voucher specimens were deposited in the Korea University Herbarium (KUS). An isolate obtained from KUS-F26690 was deposited in the Korean Agricultural Culture Collection (Accession No. KACC46842). Fungal DNA was extracted with DNeasy Plant Mini DNA Extraction Kits (Qiagen Inc., Valencia, CA). The complete internal transcribed spacer (ITS) region of rDNA was amplified with the primers ITS1/ITS4 and sequenced. The resulting 549-bp sequence was deposited in GenBank (KC426947). This showed >99% similarity with sequences of A. errabunda (AJ888475 to 888477). For pathogenicity test, inoculum was prepared by harvesting conidia from 4-week-old cultures on potato dextrose agar. A conidial suspension (1 × 106 conidia/ml) was sprayed onto young leaves of three potted seedlings. Three seedlings treated with sterile distilled water served as controls. Plants were covered with plastic bags to maintain 100% relative humidity for 24 h and then kept in a greenhouse (20 to 26°C and 60 to 80% RH). After 26 days, typical leaf spot symptoms, identical to the ones observed in the field, developed on the inoculated leaves. No symptoms were observed on controls. A. errabunda was reisolated from the lesions of inoculated plants, fulfilling Koch's postulates. Oak anthracnose associated with A. errabunda (including A. quercina) has been recorded in Europe and North America (1,4). Oak anthracnose of evergreen Quercus glauca Thunb. (syn. Cyclobalanopsis glauca (Thunb.) Oerst.) associated with A. supraseptata in Japan is not related to this disease (2). To our knowledge, this is the first report of oak anthracnose of Q. aliena globally and also the first finding of A. errabunda in Asia as well as in Korea. This pathogen is known as one of the major forest pathogens in oak stand in Europe and North America (3). Pending further studies, including a risk assessment, A. errabunda may be considered as a potentially new and serious threat in native and planted ranges of Q. aliena in Korea. References: (1) D. F. Farr and A. Y. Rossman. Fungal Databases. Syst. Mycol. Microbiol. Lab., Online publication, ARS, USDA, retrieved February 18, 2013. (2) S. Kaneko and T. Kobayashi. Trans. Mycol. Soc. Japan 25:11, 1984. (3) A. Ragazzi et al. Phytopathol. Mediterr. 46:295, 2007. (4) M. V. Sogonov et al. Mycol. Res. 111:693, 2007.

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