First Report of Zonate Leaf Spot of Artocarpus altilis Caused by Cristulariella moricola in Taiwan

Breadfruit (Artocarpus altilis (Parkinson) Fosberg) is an important landscape and garden tree in Taiwan. During the spring of 2002, zonate leaf spots of breadfruit were observed at a Taipei nursery in northern Taiwan. Initially, several small, brown, zonate lesions developed on leaves. As lesions enlarged, they coalesced, leading to blighting of leaves and premature defoliation. Sporophores on the host were generally hypophyllous but sometimes amphigenous, solitary, erect, easily detachable, and as much as 850 μm long. The upper portion of the sporophore is considered an individual conidium and consisted of a pyramidal head that was fusiform to ventricose and cristulate, 495 to 534 μm long and 210 to 290 μm wide at the broadest point. Branches within the pyramidal head were short and compact, and dichotomously or trichotomously branched. The conidia were hyaline, broad, septate, tapering toward an acute apex, and sometimes constricted at the basal septum. Conidiophores were 400 to 680 × 20 to 100 μm. The fungus was isolated from infected tissue and maintained on potato dextrose agar (PDA). Sclerotia were produced on PDA after 4 to 5 weeks at 20°C without light, but conidia were not observed in culture. The fungus was identified as Cristulariella moricola (Hino) Redhead based on morphological characteristics (1,2). To complete Koch's postulates, three sporophores from infected leaves or three sclerotia from cultures were placed individually on each of 10 breadfruit leaves. The plants were placed in plastic bags and incubated at 16 to 20°C. Symptoms were observed after 2 to 3 days on 100% of plants inoculated with sporophores and after 6 days on 50% of plants inoculated with sclerotia. The pathogen was reisolated from lesions on plants inoculated with sporophores and sclerotia. No symptoms were observed on the control plants. C. moricola has been known to cause a bull's eye or zonate leaf spot and defoliation on woody and annual plants, including at least 51 host species and 36 families distributed in the central and eastern United States (1). To our knowledge, this is the first report of zonate leaf spot and defoliation of breadfruit caused by C. moricola. References: (1) T. T. Chang. Bull. Taiwan For. Res. Inst. New Ser. 10 (2):235, 1995. (2) S. A. Redhead. Can. J. Bot. 53:700, 1975.

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First Report of Zonate Leaf Spot of Vigna vexillata var. tsusimensis Caused by Cristulariella moricola

Plant Disease ◽

10.1094/pdis.2002.86.4.440c ◽

2002 ◽

Vol 86 (4) ◽

pp. 440-440 ◽

Cited By ~ 3

Author(s):

H. B. Lee ◽

C.-J. Kim

Keyword(s):

Leaf Spot ◽

Morphological Characteristics ◽

Leaf Spot Disease ◽

Eastern United States ◽

Leaf Spots ◽

Vigna Vexillata ◽

Initial Symptoms ◽

Pathogenicity Tests ◽

Basal Septum ◽

Bull's Eye

A zonate leaf spot disease on a wild bean variety, Vigna vexillata L. var. tsusimensis Mat., occurred in the patch fields and foothills of Chungnam and Kyonggi districts in Korea during late September, October, and early November of 1999 to 2001. The zonate lesions were particularly prevalent in October following periods of heavy dew accumulation. Initial symptoms were small, circular lesions with darkbrown marginal rings that later developed into large spots with characteristic target-shaped rings. The spots were gray to bright or blackish brown, progressed rapidly, and sometimes fused together to form lesions of up to 20 mm in diameter. Sporophores on the natural host were generally hypophyllous but sometimes amphigenous, abundant on large spots, fewer on small spots, solitary, erect, easily detachable, and up to 864 μm long. The upper portion of the sporophore is considered an individual conidium and consisted of a pyramidal head that was fusiform to ventricose and cristulate, 495 to 534 μm long, and 210 to 290 μm wide at the broadest point. Branches within the pyramidal head were short and compact, and dichotomously or trichotomously branched. The central axis within the conidium was hyaline, broad, septate, tapering toward an acute apex, and sometimes constricted at the basal septum. Conidiophores were 272 to 330 μm long and up to 24 μm wide. The fungus was identified as Cristulariella moricola (Hino) Redhead based on morphological characteristics (1,2). The fungus was isolated from Vigna leaf spots, placed on 2% water agar or potato dextrose agar (PDA), and maintained on PDA amended with 2% Vigna leaf extract. For pathogenicity tests, 4- to 5-week-old leaves of V. vexillata var. tsusimensis were surface-sterilized in 1% NaOCl. Agar disks (approximately 10 mm diameter) containing mycelia of the fungus were placed on the upper leaf surface. The inoculated plants (two leaflets per plant × 2) were then sprayed with distilled water, covered with premoistened polyethylene bags, and incubated at 15 to 25°C. Within 5 days, small leaf spots appeared that were similar to those originally observed on all inoculated leaflets. Uninoculated control leaves exposed to the same environmental conditions remained healthy. C. moricola was consistently reisolated from the infected leaves. The hyphomycete fungus C. moricola has been known to cause a bull's eye or zonate leaf spot and defoliation on woody and annual plants, including at least 73 host species and 36 families distributed in the central and eastern United States and Japan (1). In Asia, the occurrence of Cristulariella spp. on several hosts has been reported only in Taiwan and Japan (3,4). No species in the genus has ever been reported from Korea. To our knowledge, V. vexillata var. tsusimensis represents a previously unreported host for C. moricola. References: (1) M. C. Niedbalski et al. Mycologia 75:988, 1983. (2) S. A. Redhead. Mycologia 71:1248, 1979. (3). H. J. Su and S. C. Leu. Plant Dis. 67:915, 1983. (4) T. Yokoyama and K. Tubaki. Trans. Mycol. Soc. Jpn. 15:189, 1974.

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First Report of Zonate Leaf Spot of Cinnamomum kanehirae Caused by Hinomyces moricola in Taiwan

Plant Disease ◽

10.1094/pdis-03-12-0252-pdn ◽

2012 ◽

Vol 96 (8) ◽

pp. 1226-1226

Author(s):

C. H. Fu ◽

F. Y. Lin

Keyword(s):

Morphological Characteristics ◽

Pathogenicity Test ◽

Annual Plants ◽

Eastern United States ◽

Transparent Plastic ◽

Leaf Spots ◽

Plastic Bags ◽

Initial Symptoms ◽

Medicinal Fungi ◽

Basal Septum

Cinnamomum kanehirae, a native tree of Taiwan, is an important tree that hosts popular medicinal fungi. In the winter of 2011, zonate leaf spots were observed at a nursery garden in Wu-Lai, Taiwan. Initial symptoms included small brown lesions on leaves that became larger leaf spots after expanding or fusing together, causing a leaf blight and eventually defoliation. Sporophores on the host were generally hypophyllous but sometimes amphigenous, solitary, erect, easily detachable. The upper portion of the sporophore was considered an individual conidium and consisted of a pyramidal head that was fusiform to ventricose, 320 to 580 μm long and 100 to 130 μm wide at the broadest point. Branches within the pyramidal head were short and compact, and dichotomously or trichotomously branched. The sporophore initials were hyaline, broad, septate, tapering toward an acute apex, and sometimes constricted at the basal septum. Sclerotia were observed in older lesions, grey or black, spherical, and 1 to 2.5 mm in diameter. The fungus was isolated from infected tissue and sporophores, maintained on potato dextrose agar (PDA) at 20°C in darkness. Sclerotia were produced on PDA after 4 to 5 weeks and were irregular or spherical, but sporophores didn't develop on agar medium. The fungus was identified as Hinomyces moricola on the basis of morphological characteristics (1). Koch's postulates were performed by inoculating four 1-year-old, asymptomatic, potted C. kanehirae plants; every plant was inoculated with sporophores from infected leaves on each of five leaves. Four noninoculated plants were kept in separate pots and served as controls. All plants were covered with transparent plastic bags individually and incubated in a growth chamber at 18 to 20°C. Symptoms were observed after 2 to 4 days on every inoculated plant but not on uninoculated plants. The leaf spots were similar to those originally observed. The pathogen was reisolated from spots of inoculated plants. The pathogenicity test was repeated once. H. moricola is known to cause severe defoliation on woody and annual plants, including at least 73 host species and 36 families distributed in the eastern United States and Japan (2). References: (1) N.-S. Tomoko et al. Mycoscience. 47:351, 2006. (2) J. C. Trolinger et al. Plant Dis. Reptr. 62:710, 1978.

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First Report of Pestalotiopsis microspora Causing Leaf Spot of Hidcote (Hypericum patulum) in Japan

Plant Disease ◽

10.1094/pdis-94-8-1064b ◽

2010 ◽

Vol 94 (8) ◽

pp. 1064-1064 ◽

Cited By ~ 3

Author(s):

M. Zhang ◽

H. Y. Wu ◽

T. Tsukiboshi ◽

I. Okabe

Keyword(s):

Leaf Spot ◽

Morphological Characteristics ◽

Its Region ◽

Basal Cells ◽

Single Spore ◽

Management Options ◽

First Report ◽

Leaf Spots ◽

Plastic Bags ◽

Pestalotiopsis Microspora

Hidcote, Hypericum patulum Thunb. ex Murray, is a deciduous shrub that is cultivated as an ornamental in landscape gardens and courtyards in Japan. In early August 2008, severe leaf spotting was observed on plants growing in a courtyard in Nasushiobara, Tochigi, Japan. More than 30% of the leaves on five shrubs exhibited leaf spot symptoms. Small, round, pale brown lesions were initially observed. Later, they expanded to 5 to 12 mm in diameter, round to irregular-shaped with pale brown centers and dark brown margins. Under continuously wet or humid conditions, black acervuli developed on the leaf lesions. Conidia were straight or slightly curved, fusiform to clavate, and five-celled with constrictions at the septa. Conidia ranged from 17 to 21 × 5 to 8 μm with hyaline apical and basal cells. Fifteen percent of apical cells had two and the rest had three appendages (setulae) ranging from 10 to 21 μm long. The basal hyaline cell tapered into a 2 to 4 μm pedicel. The three median cells ranged from light or dark brown to olive green. These morphological characteristics matched those of Pestalotiopsis microspora (Speg.) G.C. Zhao & N. Li (1,2). The identity of the fungus was confirmed by DNA sequencing of the internal transcribed spacer (ITS) region (GenBank Accession No. GU908473) from single-spore isolates, which revealed 100% homology with those of other P. microspora isolates (e.g., GenBank Accession Nos. FJ459950 and DQ456865). Koch's postulates were confirmed using leaves of three detached branches of a field-grown asymptomatic plant of H. patulum. Thirty leaves of each branch were inoculated by placing mycelial plugs obtained from the periphery of 7-day-old single-spore cultures on the leaf surface. Potato dextrose agar plugs without mycelium served as controls. Leaves on branches were covered with plastic bags for 24 h to maintain high relative humidity in a greenhouse (approximately 24 to 28°C). After 5 days, all inoculated leaves showed symptoms identical to those described above, whereas control leaves remained symptom free. Reisolation of the fungus from lesions on inoculated leaves confirmed that the causal agent was P. microspora. To our knowledge, this is the first report of leaf spots on H. patulum caused by P. microspora in Japan. Management options may have to be developed and implemented to protect Hidcote plants in areas where leaf spot cannot be tolerated. References: (1) P. A. Saccardo. Sylloge Fungorum III:789, 1884. (2) G. C. Zhao and N. Li. J. Northeast For. Univ. 23(4):21, 1995.

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First Report of Bipolaris peregianensis Causing Leaf Spot of Cynodon dactylon in China

Plant Disease ◽

10.1094/pdis-12-11-1066-pdn ◽

2012 ◽

Vol 96 (6) ◽

pp. 917-917 ◽

Cited By ~ 2

Author(s):

Z. Y. Wang ◽

S. N. Xie ◽

Y. Wang ◽

H. Y. Wu ◽

M. Zhang

Keyword(s):

Leaf Spot ◽

Cynodon Dactylon ◽

Control Strategies ◽

Tap Water ◽

Morphological Characteristics ◽

Its Region ◽

Basal Cells ◽

First Report ◽

Leaf Spots ◽

Plastic Bags

Cynodon dactylon is widely cultivated as a sod crop in warm climates worldwide. In early September 2009, heavy leaf spot infection was observed on C. dactylon from Zhengzhou, Henan, China. Early symptoms appeared as small, elliptical, pale brown lesions on the leaves. Lesions later expanded to 5 to 10 mm long and 1 to 2 mm wide, becoming brown to dark brown. A fungus was consistently isolated from leaf spots on potato dextrose agar (PDA), but with poor sporulation. Morphological characteristics were observed from single-conidium cultures on tap water agar + wheat straw (TWA+W) after 5 to 7 days. Conidiophores were light to medium brown, cylindrical, solitary or clustered, unbranched, usually with basal cells enlarged, and 94.5 to 147.0 × 4.0 to 9.0 μm. Conidia were fusoid, strongly curved, end cells broadly hemiellipsoidal, brown, 58.5 to 84.5 × 13.5 to 18.5 μm, with 6 to 10 septa. These morphological characteristics are similar to those of Bipolaris peregianensis (2). The identity of our fungus was confirmed to be B. peregianensis by DNA sequencing of the internal transcribed spacer (ITS) region (GenBank Accession No. JQ316121), which was 99% homologous to those of other B. peregianensis isolates (= Cochliobolus peregianensis; Accession Nos AF071328 and AF158111) (1). Koch's postulates were performed with the leaves of three pots of C. dactylon. Leaves were sprayed with 1 × 105 conidia/ml of B. peregianensis; an equal number of leaves in the pots of the same plant sprayed with sterile distilled water served as the control. All test plants were covered with plastic bags for 24 h to maintain high relative humidity at 23 to 25°C. After 7 days, more than 50% of inoculated leaves showed symptoms identical to those observed in natural condition, whereas controls remained symptom free. Reisolation of the fungus from lesions on inoculated leaves confirmed that the causal agent was B. peregianensis. To our knowledge, this is the first report of leaf spots caused by B. peregianensis on C. dactylon in China. The disease cycle and the control strategies in the regions are being further studied. References: (1) M. L. Berbee et al. Mycologia 91:964, 1999. (2) A. Sivanesan. Mycol. Pap. 158:1, 1987.

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First Report of Alternaria alternata Causing Black Leaf Spot of Rubber Tree in China

Plant Disease ◽

10.1094/pdis-09-14-0954-pdn ◽

2015 ◽

Vol 99 (2) ◽

pp. 290-290 ◽

Cited By ~ 11

Author(s):

Z. Y. Cai ◽

Y. X. Liu ◽

G. H. Li ◽

Y. F. Wang ◽

M. Zhou

Keyword(s):

Alternaria Alternata ◽

Leaf Spot ◽

Rubber Tree ◽

Morphological Characteristics ◽

Correct Identification ◽

Post Inoculation ◽

First Report ◽

Leaf Spots ◽

Plastic Bags ◽

Black Leaf Spot

We first reported Alternaria heveae (E.G. Simmons ) to be the pathogen that caused black leaf spot of rubber tree (Hevea brasiliensis Muell. Arg) in Heikou county in July 2014 (1). Black leaf spots that resembled the symptoms caused by A. heveae were observed on the leaves of rubber trees of the whole propagule collection nursery in Jingping County (22°68′ N and 103°05′ E) of Yunnan Province. Black foliar spots (0.1 to 2 mm in diameter) surrounded by a yellow halo with lesions slightly sunken on the leaf surface were observed. To confirm whether the disease was caused by the same pathogen, 5-mm2 sections were removed from the leading edge of the lesion and were surface-sterilized in 75% ethanol, air-dried, plated on potato carrot agar (PCA), and incubated at 28°C in the dark. Colonies of the fungus on PCA had round margins and little aerial mycelia with gray-black coloration after 6 days of growth on PCA (2). Medium brown conidia were found to be in short chains of two to eight spores, ovoid, obclavate, and obpyriform, with or without a short conical or cylindrical-shaped apical beak. Conidia ranged from 22.5 to 67.5 μm long (mean 39.9 μm) × 10 to 15 μm wide (mean 12.5 μm; 100 colodia were measured), with three to six transverse septa and zero to three longitudinal or oblique septa. Morphological characteristics matched the descriptions of A. alternata [(Fries) Keissler] (4).The ITS1-5.8S-ITS2 region of one single-spore isolate, Ah02JP1, was amplified with primers ITS1 and ITS4. The PCR product was sequenced directly and deposited in GenBank (Accession No. KM111289). A BLAST search of the GenBank database revealed 100% similarity with A. alternata isolates KJ829535.1, KJ677246.1, and KF813070.1. Therefore, the pathogen was identified as A. alternata on the basis of its morphological characteristics and ITS sequence. Pathogenicity of a representative isolate, Ah02JP1 was confirmed using a field rubber tree inoculation method. Three rubber plants (the clone of rubber tree Yunyan77-4) were grown to the copper-colored leaf stage. Leaves were spray-inoculated (104 conidia per milliliter spore suspension) until drops were equally distributed using a manual pressure sprayer. Three rubber plants sprayed with sterile distilled water were used as controls. After inoculation, the plants were covered with plastic bags to maintain high relative humidity. The plastic bags were removed 2 days post-inoculation (dpi), and the plants were monitored daily for symptom development. Five days post-inoculation, spots similar to the original ones seen on the field trees developed on all inoculated leaves, while control leaves remained symptomless. A. alternata was re-isolated from spray-inoculated leaves, confirming Koch's postulates. A. alternata has been reported as the causal agent of leaf blight of rubber tree in India, which initially appeared as minute spots on leaves and enlarged with the growth of the leaves (3). However, in the present study, the symptoms (black leaf spots) remained small over time after inoculation. To our knowledge, this is the first report of A. alternata on rubber tree in China. Correct identification of pathogens is essential for disease management strategies. This report will establish a foundation for the further study of Alternaria alternata to address the disease effectively. References: (1) Z. Y. Cai et al. Plant Dis. 98:1011, 2014. (2) E. Mirkova. J. Phytopathol. 151:323, 2003. (3) C. B. Roy et al. J. Plantation Crops 34:499, 2006. (4) T. Y. Zhang. Page 32 in: Flora Fungorum Sinicorum, Vol. 16: Alternaria. Science Press, Beijing, 2003.

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First Report of Leaf Spot Caused by Cercospora fukushiana on New Guinea Impatiens in Korea

Plant Disease ◽

10.1094/pdis-04-14-0363-pdn ◽

2014 ◽

Vol 98 (9) ◽

pp. 1280-1280

Author(s):

B. S. Kim ◽

K. S. Baek ◽

C. H. Pak ◽

J. H. Park ◽

H. D. Shin

Keyword(s):

Leaf Spot ◽

Morphological Characteristics ◽

Its Region ◽

New Guinea ◽

Culture Collection ◽

Distilled Water ◽

First Report ◽

Leaf Spots ◽

Plastic Bags ◽

New Guinea Impatiens

New Guinea impatiens, Impatiens hawkeri W. Bull, is widely cultivated as a potted plant and garden plant. In July 2013, hundreds of young plants (cv. Fanfare) showing symptoms of leaf spot with approximately 50% incidence were found in polyethylene tunnels in Yongin City, Korea. Leaf spots were circular to oblong, reaching 6 mm or more in diameter. The spots were initially uniformly brown to reddish brown, turning gray with reddish brown margin. Diseased plants defoliated prematurely and were abandoned without marketing due to signs of discoloration and yellowing on leaves. A cercosporoid fungus was consistently observed in association with disease symptoms. Stromata were brown, small, and composed of a few swollen hyphal cells. Conidiophores were emerging through the cuticle, fasciculate (n = 2 to 20), olivaceous to brown, paler toward the apex, straight to mildly curved, geniculate, 30 to 260 μm long, 3.5 to 5 μm wide, 1- to 6-septate, and with conspicuous conidial scars. Conidia were hyaline and acicular. Smaller conidia were straight and longer conidia were mildly curved. Conidia were subacute to obtuse at the apex, truncate to obconically truncate at the base, 2- to 18-septate, 30 to 320 × 3.5 to 5.5 μm, and with thickened, darkened hila at the base. Morphological characteristics of the fungus were consistent with the previous reports of Cercospora fukushiana (Matsuura) W. Yamam. (1). Voucher specimens were housed in the Korea University herbarium (KUS). An isolate from KUS-F27438 was deposited in the Korean Agricultural Culture Collection (Accession No. KACC47640). Fungal DNA was extracted with DNeasy Plant Mini Kits (Qiagen Inc., Valencia, CA). The complete internal transcribed spacer (ITS) region of rDNA was amplified with the primers ITS1/ITS4 (4) and sequenced. The resulting sequence of 497 bp was deposited in GenBank (Accession No. KJ620981). This showed >99% similarity with sequence of C. fukushiana (EF600954) on I. balsamina from Korea. Isolate of KACC47640 was used in the pathogenicity tests. Hyphal suspensions were prepared by grinding 3-week-old colonies grown on PDA with distilled water using a mortar and pestle. Five plants were inoculated with hyphal suspensions and five plants were sprayed with sterile distilled water. The plants were covered with plastic bags to maintain a relative humidity of 100% for 24 h and then transferred to a 25 ± 2°C greenhouse with a 12-h photoperiod. Typical symptoms of necrotic spots appeared on the inoculated leaves 10 days after inoculation, and were identical to the symptoms observed in the field. C. fukushiana was re-isolated from symptomatic leaf tissues, confirming Koch's postulates. No symptoms were observed on water-inoculated control plants. Previously, leaf spots of Impatiens spp. associated with C. apii, C. balsaminae, and C. fukushiana have been reported (1,2,3). To our knowledge, this is the first report of C. fukushiana on I. hawkeri in Korea. Our observations in the nurseries of I. hawkeri suggest that low humidity with good ventilation as well as plant hygiene in greenhouses might be main strategies for preventing this disease. References: (1) C. Chupp. A Monograph of the Fungus Genus Cercospora. Ithaca, NY, 1953. (2) D. F. Farr and A. Y. Rossman. Fungal Databases. Syst. Mycol. Microbiol. Lab., online publication, ARS, USDA, retrieved March 25, 2014. (3) J. M. Soares et al. Plant Dis. 93:1214, 2009. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.

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First Report of Alternaria Leaf Spot of Banana Caused by Alternaria alternata in the United States

Plant Disease ◽

10.1094/pdis-01-13-0007-pdn ◽

2013 ◽

Vol 97 (8) ◽

pp. 1116-1116 ◽

Cited By ~ 3

Author(s):

V. Parkunan ◽

S. Li ◽

E. G. Fonsah ◽

P. Ji

Keyword(s):

United States ◽

Alternaria Alternata ◽

Leaf Spot ◽

Morphological Characteristics ◽

The United States ◽

Its Sequencing ◽

Single Spore ◽

First Report ◽

Alternaria Leaf Spot ◽

Leaf Spots

Research efforts were initiated in 2003 to identify and introduce banana (Musa spp.) cultivars suitable for production in Georgia (1). Selected cultivars have been evaluated since 2009 in Tifton Banana Garden, Tifton, GA, comprising of cold hardy, short cycle, and ornamental types. In spring and summer of 2012, 7 out of 13 cultivars (African Red, Blue Torres Island, Cacambou, Chinese Cavendish, Novaria, Raja Puri, and Veinte Cohol) showed tiny, oval (0.5 to 1.0 mm long and 0.3 to 0.9 mm wide), light to dark brown spots on the adaxial surface of the leaves. Spots were more concentrated along the midrib than the rest of the leaf and occurred on all except the newly emerged leaves. Leaf spots did not expand much in size, but the numbers approximately doubled during the season. Disease incidences on the seven cultivars ranged from 10 to 63% (10% on Blue Torres Island and 63% on Novaria), with an average of 35% when a total of 52 plants were evaluated. Six cultivars including Belle, Ice Cream, Dwarf Namwah, Kandarian, Praying Hands, and Saba did not show any spots. Tissue from infected leaves of the seven cultivars were surface sterilized with 0.5% NaOCl, plated onto potato dextrose agar (PDA) media and incubated at 25°C in the dark for 5 days. The plates were then incubated at room temperature (23 ± 2°C) under a 12-hour photoperiod for 3 days. Grayish black colonies developed from all the samples, which were further identified as Alternaria spp. based on the dark, brown, obclavate to obpyriform catenulate conidia with longitudinal and transverse septa tapering to a prominent beak attached in chains on a simple and short conidiophore (2). Conidia were 23 to 73 μm long and 15 to 35 μm wide, with a beak length of 5 to 10 μm, and had 3 to 6 transverse and 0 to 5 longitudinal septa. Single spore cultures of four isolates from four different cultivars were obtained and genomic DNA was extracted and the internal transcribed spacer (ITS1-5.8S-ITS2) regions of rDNA (562 bp) were amplified and sequenced with primers ITS1 and ITS4. MegaBLAST analysis of the four sequences showed that they were 100% identical to two Alternaria alternata isolates (GQ916545 and GQ169766). ITS sequence of a representative isolate VCT1FT1 from cv. Veinte Cohol was submitted to GenBank (JX985742). Pathogenicity assay was conducted using 1-month-old banana plants (cv. Veinte Cohol) grown in pots under greenhouse conditions (25 to 27°C). Three plants were spray inoculated with the isolate VCT1FT1 (100 ml suspension per plant containing 105 spores per ml) and incubated under 100% humidity for 2 days and then kept in the greenhouse. Three plants sprayed with water were used as a control. Leaf spots identical to those observed in the field were developed in a week on the inoculated plants but not on the non-inoculated control. The fungus was reisolated from the inoculated plants and the identity was confirmed by morphological characteristics and ITS sequencing. To our knowledge, this is the first report of Alternaria leaf spot caused by A. alternata on banana in the United States. Occurrence of the disease on some banana cultivars in Georgia provides useful information to potential producers, and the cultivars that were observed to be resistant to the disease may be more suitable for production. References: (1) E. G. Fonsah et al. J. Food Distrib. Res. 37:2, 2006. (2) E. G. Simmons. Alternaria: An identification manual. CBS Fungal Biodiversity Center, Utrecht, Netherlands, 2007.

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First Report of Leaf Spot of Sweet Basil Caused by Cercospora guatemalensis in Korea

Plant Disease ◽

10.1094/pdis-05-12-0436-pdn ◽

2012 ◽

Vol 96 (10) ◽

pp. 1580-1580

Author(s):

J. H. Park ◽

K. S. Han ◽

J. Y. Kim ◽

H. D. Shin

Keyword(s):

Leaf Spot ◽

Morphological Characteristics ◽

Its Region ◽

Culture Collection ◽

Distilled Water ◽

First Report ◽

Sweet Basil ◽

Organic Farm ◽

Leaf Spots ◽

Close Phylogenetic Relationship

Sweet basil, Ocimum basilicum L., is a fragrant herb belonging to the family Lamiaceae. Originated in India 5,000 years ago, sweet basil plays a significant role in diverse cuisines across the world, especially in Asian and Italian cooking. In October 2008, hundreds of plants showing symptoms of leaf spot with nearly 100% incidence were found in polyethylene tunnels at an organic farm in Icheon, Korea. Leaf spots were circular to subcircular, water-soaked, dark brown with grayish center, and reached 10 mm or more in diameter. Diseased leaves defoliated prematurely. The damage purportedly due to this disease has reappeared every year with confirmation of the causal agent made again in 2011. A cercosporoid fungus was consistently associated with disease symptoms. Stromata were brown, consisting of brown cells, and 10 to 40 μm in width. Conidiophores were fasciculate (n = 2 to 10), olivaceous brown, paler upwards, straight to mildly curved, not geniculate in shorter ones or one to two times geniculate in longer ones, 40 to 200 μm long, occasionally reaching up to 350 μm long, 3.5 to 6 μm wide, and two- to six-septate. Conidia were hyaline, acicular to cylindric, straight in shorter ones, flexuous to curved in longer ones, truncate to obconically truncate at the base, three- to 16-septate, and 50 to 300 × 3.5 to 4.5 μm. Morphological characteristics of the fungus were consistent with the previous reports of Cercospora guatemalensis A.S. Mull. & Chupp (1,3). Voucher specimens were housed at Korea University herbarium (KUS). An isolate from KUS-F23757 was deposited in the Korean Agricultural Culture Collection (Accession No. KACC43980). 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 sequence of 548 bp was deposited in GenBank (Accession No. JQ995781). This showed >99% similarity with sequences of many Cercospora species, indicating their close phylogenetic relationship. Isolate of KACC43980 was used in the pathogenicity tests. Hyphal suspensions were prepared by grinding 3-week-old colonies grown on PDA with distilled water using a mortar and pestle. Five plants were inoculated with hyphal suspensions and five plants were sprayed with sterile distilled water. The plants were covered with plastic bags to maintain a relative humidity of 100% for 24 h and then transferred to a 25 ± 2°C greenhouse with a 12-h photoperiod. Typical symptoms of necrotic spots appeared on the inoculated leaves 6 days after inoculation, and were identical to the ones observed in the field. C. guatemalensis was reisolated from symptomatic leaf tissues, confirming Koch's postulates. No symptoms were observed on control plants. Previously, the disease was reported in Malawi, India, China, and Japan (2,3), but not in Korea. To our knowledge, this is the first report of C. guatemalensis on sweet basil in Korea. Since farming of sweet basil has recently started on a commercial scale in Korea, the disease poses a serious threat to safe production of this herb, especially in organic farming. References: (1) C. Chupp. A Monograph of the Fungus Genus Cercospora. Ithaca, NY, 1953. (2) D. F. Farr and A. Y. Rossman. Fungal Databases. Systematic Mycology & Microbiology Laboratory, ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ , May 5, 2012. (3) J. Nishikawa et al. J. Gen. Plant Pathol. 68:46, 2002.

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First Report of Leaf Spot Disease on Walnut Caused by Colletotrichum fioriniae in China

Plant Disease ◽

10.1094/pdis-09-14-0938-pdn ◽

2015 ◽

Vol 99 (2) ◽

pp. 289-289 ◽

Cited By ~ 9

Author(s):

Y. Z. Zhu ◽

W. J. Liao ◽

D. X. Zou ◽

Y. J. Wu ◽

Y. Zhou

Keyword(s):

Dna Sequences ◽

Leaf Spot ◽

Juglans Regia ◽

Morphological Characteristics ◽

Leaf Spot Disease ◽

Koch’S Postulates ◽

First Report ◽

Spot Disease ◽

Leaf Spots ◽

Koch's Postulates

In May 2014, a severe leaf spot disease was observed on walnut tree (Juglans regia L.) in Hechi, Guangxi, China. Leaf spots were circular to semicircular in shape, water-soaked, later becoming grayish white in the center with a dark brown margin and bordered by a tan halo. Necrotic lesions were approximately 3 to 4 mm in diameter. Diseased leaves were collected from 10 trees in each of five commercial orchards. The diseased leaves were cut into 5 × 5 mm slices, dipped in 75% ethanol for 30 s, washed three times in sterilized water, sterilized with 0.1% (w/v) HgCl2 for 3 min, and then rinsed five times with sterile distilled water. These slices were placed on potato dextrose agar (PDA), followed by incubating at 28°C for about 3 to 4 days. Fungal isolates were obtained from these diseased tissues, transferred onto PDA plates, and incubated at 28°C. These isolates produced gray aerial mycelium and then became pinkish gray with age. Moreover, the reverse of the colony was pink. The growth rate was 8.21 to 8.41 mm per day (average = 8.29 ± 0.11, n = 3) at 28°C. The colonies produced pale orange conidial masses and were fusiform with acute ends, hyaline, sometimes guttulate, 4.02 to 5.25 × 13.71 to 15.72 μm (average = 4.56 ± 0.31 × 14.87 ± 1.14 μm, n = 25). The morphological characteristics and measurements of this fungal isolate matched the previous descriptions of Colletotrichum fioriniae (Marcelino & Gouli) R.G. Shivas & Y.P. Tan (2). Meanwhile, these characterizations were further confirmed by analysis of the partial sequence of five genes: the internal transcribed spacer (ITS) of the ribosomal DNA, beta-tubulin (β-tub) gene, glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene, chitin synthase 3(CHS-1) gene, and actin (ACT) gene, with universal primers ITS4/ITS5, T1/βt2b, GDF1/GDR1, CHS1-79F/CHS1-354R, and ACT-512F/ACT-783R, respectively (1). BLAST of these DNA sequences using the nucleotide database of GenBank showed a high identify (ITS, 99%; β-tub, 99%; GAPDH, 99%; CHS-1, 99%; and ACT, 100%) with the previously deposited sequences of C. fioriniae (ITS, KF278459.1, NR111747.1; β-tub, AB744079.1, AB690809.1; GAPDH, KF944355.1, KF944354.1; CHS-1, JQ948987.1, JQ949005.1; and ACT, JQ949625.1, JQ949626.1). Koch's postulates were fulfilled by inoculating six healthy 1-year-old walnut trees in July 2014 with maximum and minimum temperatures of 33 and 26°C. The 6-mm mycelial plug, which was cut from the margin of a 5-day-old colony of the fungus on PDA, was placed onto each pin-wounded leaf, ensuring good contact between the mycelium and the wound. Non-colonized PDA plugs were placed onto pin-wounds as negative controls. Following inoculation, both inoculated and control plants were covered with plastic bags. Leaf spots, similar to those on naturally infected plants, were observed on the leaves inoculated with C. fioriniae within 5 days. No symptoms were observed on the negative control leaves. Finally, C. fioriniae was re-isolated from symptomatic leaves; in contrast, no fungus was isolated from the control, which confirmed Koch's postulates. To our knowledge, this is the first report of leaf disease on walnut caused by C. fioriniae. References: (1) L. Cai et al. Fungal Divers. 39:183, 2009. (2) R. G. Shivas and Y. P. Tan. Fungal Divers. 39:111, 2009.

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First Report of Alternaria heveae Causing Black Leaf Spot of Rubber Tree in China

Plant Disease ◽

10.1094/pdis-01-14-0065-pdn ◽

2014 ◽

Vol 98 (7) ◽

pp. 1011-1011 ◽

Cited By ~ 1

Author(s):

Z. Y. Cai ◽

Y. X. Liu ◽

G. X. Huang ◽

M. Zhou ◽

G. Z. Jiang ◽

...

Keyword(s):

Rubber Tree ◽

Morphological Characteristics ◽

Black Spot ◽

Its Region ◽

Post Inoculation ◽

First Report ◽

Leaf Spots ◽

Plastic Bags ◽

Manual Pressure ◽

Tree Leaf

Rubber tree (Hevea brasiliensis Muell. Arg.) is an important industrial crop of tropical areas for natural rubber production. In October 2013, foliar spots (0.1 to 0.4 mm in diameter), black surrounded by a yellow halo, and with lesions slightly sunken were observed on the rubber tree leaf in a growing area in Heikou County of Yunnan Province. Lesion tissues removed from the border between symptomatic and healthy tissue were surface sterilized in 75% ethanol and air-dried, plated on PDA plates, and incubated at 28°C with alternating day/night cycles of light. The pathogen was observed growing out of many of the leaf pieces, and produced abundant conidia. Colonies 6.1 cm in diameter developed on potato carrot agar (PCA) after 7 days, with well-defined concentric rings of growth. Colonies on PCA were composed of fine, dark, radiating, surface and subsurface hyphae. Conidia produced in PCA culture were mostly solitary or in short chains of 2 to 5 spores, long ovoid to clavate, and light brown, 40 to 81.25 × 8 to 20 μm (200 colonies were measured), with 3 to 6 transverse septa and 0 to 2 longitudinal or oblique septa. Morphological characteristics were similar to those described for Alternaria heveae (3,4). A disease of rubber tree caused by Alternaria sp. had been reported in Mexico in 1947 (2). DNA of Ah01HK13 isolate was extracted for PCR and sequencing of the ITS region with ITS1 and ITS4 primers was completed. From the BLAST analysis, the sequence of Ah01HK13 (GenBank Accession No. KF953884), had 97% similarity to A. dauci, 96% identical to A. macrospora (AY154701.1 and DQ156342.1, respectively), indicating the pathogen belonged to Alternaria genus. According to morphological characteristics, this pathogen was identified as A. heveae. Pathogenicity of representative isolate, Ah01HK13 was confirmed using a field rubber tree inoculation method. Three rubber plants (the clone of rubber tree Yunyan77-4) were grown to the copper-colored leaf stage and inoculated by spraying spore suspension (concentration = 104 conidia/ml) to the copper-colored leaves until drops were equally distributed on it using manual pressure sprayer. Three rubber plants sprayed with sterile distilled water were used as controls. After inoculation, the plants were covered with plastic bags. The plastic bags were removed after 2 days post-inoculation (dpi) and monitored daily for symptom development (1). The experiment was repeated three times. The typical 0.1 to 0.4 mm black leaf spots were observed 7 dpi. No symptoms were observed on control plants. A fungus with the same colony and conidial morphology as A. heveae were re-isolated from leaf lesions on inoculated rubber plants, but not from asymptomatic leaves of control plants, fulfilling Koch's postulates. Based on these results, the disease was identified as black spot of rubber tree caused by A. heveae. To our knowledge, this is the first report of A. heveae on rubber tree in China. References: (1) Z. Y. Cai et al. Microbiol Res. 168:340, 2013. (2) W. J. Martin. Plant Dis. Rep. 31:155, 1947. (3) E. G. Simmons. Mycotaxon 50:262, 1994. (4) T. Y. Zhang. Page 111 in: Flora Fungorum Sinicorum: Alternaria, Science Press, Beijing, 2003.

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