First Report of Leaf Spot Caused by Alternaria alternata on Kiwifruit in Italy

A leaf spot of kiwifruit (Actinidia deliciosa (A. Chev.) C. F. Liang & A. R. Ferg.) leaves was recently observed on plants of the cultivar Hayward in an orchard near Salerno, in southern Italy. The affected plants showed early severe defoliation. The fungus isolated from the infected leaves was identified as Alternaria alternata (Fr.:Fr.) Keissl., based on conidial morphological characteristics. Pathogenicity tests were made by inoculating detached leaves of male pollinator cultivar Tomuri and the female cultivars Hayward and Bruno with a 7-mm disk taken from actively growing cultures of the fungus on potato dextrose agar (PDA). After 14 days, necrotic leaf spots developed and A. alternata was consistently isolated from the inoculated leaves. A. alternata has been observed as a pathogen on leaves and fruits in New Zealand. In the Mediterranean, it has been reported in Israel (2) and in the island of Crete (1). This is the first report of Alternaria leaf spot on kiwifruit in Italy. References: (1) V. A. Bourbos and M. T. Skoudridakis. Petria 7:111, 1997. (2) A. Sive and D. Resnizky. Alon Hanotea 41:409, 1987.

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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 on Japanese Plum Caused by an Alternaria sp. in Korea

Plant Disease ◽

10.1094/pd-89-0343a ◽

2005 ◽

Vol 89 (3) ◽

pp. 343-343 ◽

Cited By ~ 3

Author(s):

Youngjun Kim ◽

Hyang Burm Lee ◽

Seung Hun Yu

Keyword(s):

Leaf Spot ◽

Morphological Characteristics ◽

The United States ◽

Conidial Suspension ◽

First Report ◽

Japanese Plum ◽

Leaf Spots ◽

Causal Fungus ◽

Pathogenicity Tests ◽

Alternaria Sp

Japanese plum (Prunus salicina Lindley) is a deciduous tree in the family Rosaceae. In Korea, this plant is widely distributed in orchards as an important stone fruit as well as in gardens as an ornamental tree because of their abundant white blossoms. Every September to November since 2001, leaf spots were observed on Japanese plum in a garden in Cheongyang, Chungnam District, Korea. Early symptoms consisted of small, brown spots that were 2 to 5 mm in diameter. Later, the leaf lesions became circular or irregular, dark brown, expanded to 15 mm in diameter, and resulted in discoloration with necrosis on twisted leaves that was followed by defoliation. In November, older lesions sometimes appeared blackish brown as sporulation occurred on the lesions. The causal fungus was isolated from diseased leaves and cultured on potato dextrose agar. A culture has been placed in the CABI Herbarium (IMI Accession No. 387139). Conidial dimension averaged 34 × 12 μm. On the basis of morphological characteristics of conidia and conidiophores, the causal fungus was identified as a small-spored species of Alternaria as described by E. G. Simmons (1). Pathogenicity tests were conducted by inoculating slightly wounded and nonwounded leaves with a conidial suspension adjusted to 1 × 106 conidia/ml. Four leaves per each experiment were either wounded or not and inoculated with a spore suspension. The eight leaves were placed in a moist chamber at 25°C. After 6 to 10 days, small brown spots appeared on 87% of the wounded and nonwounded leaves. Control leaves sprayed with distilled water did not develop any symptoms. The causal fungus was consistently reisolated from the leaf spots. Results from pathogenicity tests were similar in a repeated test. It is possible that small-spored Alternaria spp. isolates are host specific (2). Eight Alternaria spp., including A. alternata, A. tenuis, A. tenuissima, and A. citri, have been found to cause black spot on fifteen Prunus spp. in China, Japan, Hong Kong, Libya, Mexico, Australia, and the United States (2). Further studies on the host-specific toxin production, geographical distribution, and host ranges for the species of Alternaria isolated from Japanese plum are in progress. To our knowledge, this is the first report of leaf spot on Japanese plum (P. salicina) caused by a small-spored Alternaria sp. in Korea. References: (1) E. G. Simmons. Mycotaxon 55:79, 1995. (2) K. Inoue and H. Nasu. J. Gen. Plant Pathol. 66:18, 2002.

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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 Alternaria alternata as a Dieback Pathogen of Kiwifruit

Plant Disease ◽

10.1094/pdis.2000.84.3.371c ◽

2000 ◽

Vol 84 (3) ◽

pp. 371-371 ◽

Cited By ~ 4

Author(s):

P. C. Tsahouridou ◽

C. C. Thanassoulopoulos

Keyword(s):

Alternaria Alternata ◽

Morphological Characteristics ◽

Potato Dextrose Agar ◽

Conidial Suspension ◽

Actinidia Chinensis ◽

Northern Greece ◽

Disease Symptoms ◽

First Report ◽

Surface Sterilization ◽

Pathogenicity Tests

During a survey of diseases on kiwifruit (Actinidia chinensis) cv. Hayward during spring 1998 in Northern Greece, leaves of kiwifruit trees were found covered with small, necrotic brown spots. Intense spotting was associated with defoliation. Furthermore, small, sunken, dark brown cankers appeared a few centimeters below the twig tip, and twigs died distal to the canker a few days later, while the twig remained healthy below the canker, often producing a new shoot below the canker. Isolations from symptomatic leaves and internal tissues of twigs on potato dextrose agar (PDA), after surface-sterilization with 0.5% NaOCl and 90% alcohol, respectively, consistently yielded a fungus that, based on conidial morphological characteristics, was identified as Alternaria alternata (Nees:Fr.) Keissl. (2). Pathogenicity tests were performed on 3-year-old kiwi plants. Tests on twigs were conducted by inoculating 35 twigs with 5-mm disks from 7-day-old cultures on PDA at 25°C. Each twig was inoculated with three disks. Tests on leaves were conducted by spraying a conidial suspension (5 × 106 conidia per ml) of the cultures on leaves. Plants were kept in a glasshouse at 22 to 25°C. Symptoms, identical to those observed in the field, developed on leaves and twigs 6 and 18 days, respectively, after inoculation. All inoculated leaves exhibited symptoms of the disease, whereas more than 95% of inoculated twigs exhibited disease symptoms. A. alternata was consistently reisolated from inoculated tissues. A. alternata is widely known to be a leaf spot pathogen of kiwifruit (1). This is the first report of A. alternata causing twig dieback. References: (1) L. Corazza and L. Luongo. Plant Dis. 83:487, 1999. (2) E. G. Simmons. Mycotaxon 37:79, 1990.

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First Report of Alternaria alternata Causing Leaf Spot on English Walnut

Plant Disease ◽

10.1094/pdis.1999.83.7.696a ◽

1999 ◽

Vol 83 (7) ◽

pp. 696-696 ◽

Cited By ~ 5

Author(s):

A. Belisario ◽

E. Forti ◽

L. Corazza ◽

H. A. van Kesteren

Keyword(s):

Alternaria Alternata ◽

Leaf Spot ◽

Juglans Regia ◽

Deciduous Trees ◽

Sterile Water ◽

First Report ◽

Leaf Spots ◽

Pathogenicity Tests ◽

Necrotic Spots ◽

English Walnut

A foliar disease of English walnut (Juglans regia L.) was observed in 1998 in an orchard in northern Italy. Symptoms consisted of circular, necrotic spots bordered with concentric zones of darker tissue. Average lesion diameter was 20 mm, although lesions could extend to half of the leaflet lamina. A fungus identified as Alternaria alternata (Fr.:Fr.) Keissl. (1) was observed on the necrotic tissue and was consistently isolated from the margins of the necrosis. Conidia from leaves were brown, ellipsoid to ovoid (primary conidia 28 to 60 × 8 to 15 μm; secondary conidia 10 to 30 × 7 to 13 μm), with walls often ornamented, 1 to 6 transversely septate and 0 to 3 longitudinally septate. For pathogenicity tests, three isolates were grown on potato dextrose agar for 2 weeks. Inoculations were performed on detached, surface-sterilized, healthy J. regia leaflets. Four drops (5 μl each) of a sterile water suspension of 1 × 105 conidia per ml were placed on each leaflet; three leaves per isolate were used. Leaves were incubated in a moist chamber. After 10 days, leaf spots similar to the original symptoms developed on all the inoculated points for all three isolates, and the pathogen was reisolated. Control leaflets inoculated with sterile, distilled water remained symptomless. The experiment was performed three times and the results were similar. Alternaria alternata is a well-known pathogen on many crops but a few records report this fungus as a causal agent of leaf spot on deciduous trees. This is the first report of A. alternata on English walnut. Reference: (1) E. G. Simmons. Mycotaxon 37:79, 1990.

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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 Occurrence of Eggplant Wilt Caused by Verticillium dahliae in Korea

Plant Disease ◽

10.1094/pdis.2000.84.10.1152a ◽

2000 ◽

Vol 84 (10) ◽

pp. 1152-1152

Author(s):

S. K. Kim ◽

S. S. Hong ◽

K. W. Kim ◽

E. W. Park

Keyword(s):

Verticillium Dahliae ◽

Solanum Melongena ◽

Morphological Characteristics ◽

Wilt Disease ◽

Potato Dextrose Agar ◽

First Report ◽

Vascular Tissues ◽

Fungal Isolates ◽

Pathogenicity Tests ◽

Root Cutting

A wilt disease occurred on greenhouse-grown eggplants (Solanum melongena L.) at Hanam and Yeojoo, Korea, in 1997. Lower leaves on the 2-month-old wilted eggplants exhibited gradual yellowing, interveinal necrosis, and marginal crinkling and dropped prematurely. Vascular tissues of diseased stems were discolored and turned black. Vertical sections of the stems revealed that the pith had been colonized by the fungus. The disease progressed from lower parts of the plants upward. Incidence of diseased eggplants in greenhouses was 5% on 23 May 1997. Although the incidence increased to 10% on 13 June, it remained constant through early July. Fungal isolates from discolored vascular tissues were initially whitish to cream color on potato-dextrose agar, which turned black due to the formation of microsclerotia. The fungus also produced abundant verticillate conidiophores with phialides and conidia. Based on these cultural and morphological characteristics, the fungus was identified as Verticillium dahliae Klebahn. Pathogenicity tests by root cutting, root dipping, or soil drenching resulted in similar symptoms observed in the naturally infected eggplants. Symptoms were first observed on lower leaves of each eggplant 3 weeks after inoculation. Isolation from symptomatic leaves of the inoculated eggplants yielded V. dahliae. This is the first report of occurrence of Verticillium wilt of eggplant in Korea.

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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 Leaf Spot of Houttuynia cordata Caused by Alternaria alternata in China

Plant Disease ◽

10.1094/pdis-10-10-0724 ◽

2011 ◽

Vol 95 (3) ◽

pp. 359-359

Author(s):

L. Zheng ◽

R. Lv ◽

Q. Li ◽

J. Huang ◽

Y. Wang ◽

...

Keyword(s):

Alternaria Alternata ◽

Leaf Spot ◽

Southern China ◽

Sclerotium Rolfsii ◽

Tween 20 ◽

Herbaceous Plant ◽

Houttuynia Cordata ◽

First Report ◽

Perennial Herbaceous Plant ◽

Pathogenicity Tests

Houttuynia cordata is a perennial herbaceous plant (family Saururaceae) that is native to southern China, Japan, Korea, and Southeast Asia where it grows well in moist to wet soils. It is commonly used as a Chinese herbal medicine and as a vegetable. In North America and Europe it is also used as an ornamental. From September 2007 to November 2009, symptoms of leaf spot were found on H. cordata leaves in Dangyang County, Hubei, China, with the crop area affected estimated to be over 600 ha per year. Rhizome yield was reduced by 20% on average, with up to 70% yield losses in some fields during the autumn growing season. Lesions were initially small, brown, and oval or circular that developed into dark spots and sometimes formed target spots with white centers. These spots enlarged and overlapped, extending until the leaves withered entirely usually within 2 months. A fungus was consistently recovered from symptomatic leaf samples collected in October 2008 or 2009 with an average 90% isolation rate from ~60 leaf pieces that were surface sterilized with 0.1% mercuric chloride solution. Three isolates, HCDY-2, HCDY-3, and HCDY-4, were used to further evaluate characteristics of the pathogen. On potato dextrose agar, all cultures initially developed white colonies and the centers turned gray or brown after 4 days of incubation. Conidiophores were single or fasciculate, straight or knee curved, gray-brown with regular septa, and 42 to 61 × 4 to 5 μm. Conidia were obclavate or ovate, brown, and 26 to 38 × 12 to 20 μm with three to five transverse and one to three longitudinal or oblique septa. The tops of some conidia developed into secondary conidiophores, which were cylindrical, beige, and 5 to 17 × 3 to 5 μm. The pathogen was identified as Alternaria alternata based on descriptions in Simmons (3). Genomic DNA of HCDY-2 was extracted, and the rDNA-internal transcribed spacer sequence showed 99.6% identity to A. alternata (GenBank No. AY513941). Pathogenicity tests were performed with the three isolates by spraying conidial suspensions (1 × 106 conidia/ml) containing 0.1% Tween 20 onto upper and lower surfaces of leaves of 40-day-old 15-cm high plants. There were 20 leaves from five replicate plants for each isolate. Control plants were treated with sterilized water containing 0.1% Tween 20 only. All plants were incubated with a 16-h photoperiod at 25°C and 90% relative humidity in an artificial climate chamber. Five days after inoculation, typical brown spots were observed on all inoculated leaves but no symptoms were seen on water-treated control plants. Koch's postulates were fulfilled by reisolation of A. alternata from diseased leaves. The pathogenicity tests were carried out twice. A survey of the literature revealed only a few fungal diseases associated with H. cordata (1,2,4), including Phyllosticta houttuyniae, Pseudocercospora houttuyniae, Rhizoctonia solani, and Sclerotium rolfsii. Although A. alternata is a cosmopolitan plant pathogen, it has not been reported on any species in the four genera in Saururaceae (Anemopsis, Gymnotheca, Houttuynia, and Saururus) (3). To our knowledge, this is the first report of A. alternata infecting H. cordata worldwide. References: (1) Y. L. Guo and W. X. Zhao. Acta Mycol. Sin. 8:118, 1989. (2) K. Sawada. Spec. Publ. Taiwan Univ. 8:138, 1959. (3) E. G. Simmons. Alternaria: An Identification Manual. The American Phytopathological Society, St. Paul, MN, 2007. (4) Y. Wu et al. J. Changjiang Vegetables (In Chinese) 2:19, 2007.

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First Report of Myrothecium roridum Causing Leaf Spot on Garden Hydrangea in the United States

Plant Disease ◽

10.1094/pdis-04-10-0264 ◽

2010 ◽

Vol 94 (10) ◽

pp. 1266-1266 ◽

Cited By ~ 7

Author(s):

M. T. Mmbaga ◽

Y. Li ◽

M.-S. Kim

Keyword(s):

United States ◽

Leaf Spot ◽

Incubation Temperature ◽

Morphological Characteristics ◽

The United States ◽

Control Treatment ◽

Medium Size ◽

First Report ◽

Detached Leaves ◽

Myrothecium Roridum

Garden hydrangea (Hydrangea macrophylla) is a popular flowering shrub that grows well in Tennessee but foliar diseases impact their appearance, health, and market value. Leaves of garden hydrangea showed necrotic lesions with concentric rings of brown and dark brown at the Tennessee State University Research Center in McMinnville. A fungus was recovered from June and July leaf samples with 20% frequency of isolation from approximately 40 leaf pieces that were surface sterilized and plated in potato dextrose agar (PDA). Isolates developed white colonies and dark gray-to-black, spore-bearing mycelial cushions (sporodochia) that formed on older colonies (30 to 45 days old) at 25 ± 2°C. Conidia were hyaline to slightly dark, one-celled, ovoid to elongate with rounded ends, and 2.0 to 2.5 × 5.5 to 6.5 μm. These morphological characteristics were consistent with those described for Myrothecium roridum Tode ex Fr. (1). DNA sequence for three isolates of this fungus showed identical internal transcribed spacer (ITS) region sequences (GenBank Accession No. HM215150) with 99% maximum sequence identity to M. roridum isolates (GenBank Accession Nos. AJ301994.1 and AJ608978). Another close match (97%) was with M. gramineum (GenBank Accession No. FJ235084) and M. tongaense (GenBank Accession No. AY254157). Pathogenicity of M. roridum was evaluated on detached leaves from three hydrangea cultivars, Nikko Blue, All Summer Beauty, and Blue bird. Four, medium-size, detached leaves were placed in moist chambers and inoculated with 5-mm mycelial plugs from 14-day-old cultures; sterile PDA was used as the control treatment. A randomized, complete-block experimental design was used with a replication of four leaves per cultivar. Incubation temperature was 26 ± 2°C. Necrotic lesions started 4 to 5 days after inoculation in all inoculated leaves; lesions expanded to cover 10 to 25% of the leaf surface and formed concentric rings; sterile PDA plugs did not produce leaf lesions. This experiment was repeated twice and similar symptoms were produced; M. roridum was reisolated from all inoculated leaves. Spray inoculation of detached leaves of hydrangea cv. Pretty Maiden with 5 × 104 spores/ml produced similar symptoms; leaves sprayed with water remained symptom free. M. roridum has a wide host range and similar symptoms have been reported on other ornamentals including salvia (2), begonia ( http://mrec.ifas.ufl.edu/foliage/folnotes/begonias.htm ), gardenia ( http://cfextension.ifas.ufl.edu/agriculture/ nursery_production/ documents/Gardenia.pdf ), and cotton (3). To our knowledge, this is the first report of M. roridum causing leaf spot on H. macrophylla in the United States. References: (1) M. B. Ellis. Page 465 in: More Damatacous Hyphomycetes. CABI, Wallingford, UK. 1993. (2) J. A. Mangandi et al. Plant Dis. 91:772, 2007. (3) R. L. Munjal. Indian Phytopathol. New Delhi, 13:150, 1960.

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