First Report of Colletotrichum dracaenophilum on Dracaena sanderiana in Bulgaria

In the winter of 2007, severe damage was observed on numerous indoor potted plants of Dracaena sanderiana hort. Sander ex Mast. (“lucky bamboo”) in Plovdiv, Bulgaria, which were imported from a country of unknown origin. These plants were already in the retail distribution stream. Initially, the internodes of infected stems appeared pale green with yellowish lesions. An upward spreading necrosis led to a weakness of the stems with wilt and death of the plants occurring within 2 weeks. Eventually, entire stems were covered with numerous, black, globose-to-ellipsoid acervuli with sparse, black setae. The fungus was aseptically isolated from stem lesions on potato dextrose agar (PDA) on which it produced fast-growing, pale pink colonies. On the plant and in culture, the conidia were hyaline, broadly clavate to cylindrical, occasionally slightly curved, and measured 20 to 34 × 6.7 to 10.0 μm (average 28 × 8.5 μm). On the basis of the symptoms on the plant and morphological, cultural, and molecular characteristics, the fungus was identified as Colletotrichum dracaenophilum D.F. Farr & M.E. Palm (1). Pathogenicity of the fungus was confirmed by artificial inoculation of healthy plants of D. sanderiana (three replicates). Stems were inoculated by inserting small mycelial plugs from 7-day-old PDA cultures into wounds that were subsequently covered with Parafilm strips. After 2 weeks, pale green lesions started developing on all inoculated plants and the fungus was successfully reisolated. No symptoms were found around the pure agar control wounds. The specimen from Bulgaria was deposited in the U.S. National Fungus Collections (BPI 877337) with the derived culture deposited as CBS 121453. In addition, the internal transcribed spacer region of the nrDNA of this isolate was sequenced and deposited as GenBank Accession No. EU003533. To our knowledge, this is the first report of C. dracaenophilum on potted plants outside of China and is the first report of this species in Bulgaria. Reference: (1) D. F. Farr et al. Mycol. Res. 110:1395, 2006.

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First Report of Stem Blight on Perilla (Perilla frutescens) Caused by Corynespora cassiicola in Korea

Plant Disease ◽

10.1094/pdis-93-5-0550a ◽

2009 ◽

Vol 93 (5) ◽

pp. 550-550 ◽

Cited By ~ 2

Author(s):

H. B. Lee ◽

C. J. Kim ◽

H. Y. Mun

Keyword(s):

Perilla Frutescens ◽

Morphological Data ◽

Conidial Suspension ◽

Corynespora Cassiicola ◽

Cucumis Sativus L ◽

First Report ◽

Potted Plants ◽

Rdna Sequences ◽

Pathogenicity Tests ◽

Stem Lesions

Perilla or kkaennip (Perilla frutescens (L.) Britton), an annual herb of the mint family, Lamiaceae, is used in salads and kimchi and for wrapping sliced raw fish. In September 2007, a disease occurred on greenhouse-produced perilla (cv. Manchu) in Gwangyang and Jeonnam provinces, Korea. Symptoms included leaf blight and irregularly shaped stem lesions approximately 1 to 3 cm long. Plants eventually died. In some greenhouses, 10 to 30%, and occasionally as much as 70%, of the plants were affected. Isolations on potato dextrose agar yielded a fungus with single conidiophores (439 to 656 [average 524] μm long × 6.2 to 11.6 [average 9.2] μm wide) with three to eight septa. Conidia were fusiform, obclavate to subcylindrical, straight or curved, and 30.4 to 180.1 (average 98.2) μm long × 6.7 to 18.1 (average 10.5) μm wide with 5 to 16 (commonly 13) distosepta. On the basis of morphological data and ITS rDNA sequences, the fungus was identified as Corynespora cassiicola (Berk. & Curt.) Wei. (1,2). Sequences of one isolate, EML-COR1, were more than 99% identical to sequences of C. cassiicola ATCC64204 (GenBank Accession No. AY238606) and C. cassiicola (GenBank Accession No. EF490450). In pathogenicity tests, the stems and leaves of two 2-month-old wounded and nonwounded potted plants (cv. Manchu) were sprayed until runoff with a conidial suspension of 5 × 104 conidia per ml. The plants were maintained for 48 h in a humid chamber and then moved to a greenhouse. Symptoms similar to those observed in the commercial greenhouse developed on wounded stems within 10 days. On nonwounded plants, symptoms developed 3 to 4 weeks after inoculation. C. cassiicola was reisolated from these lesions. Control plants (sprayed with distilled water) remained symptomless. The experiment was repeated with similar results. Although C. cassiicola causes blight of cucumber (Cucumis sativus L.), sesame (Sesamum indicum L.), and other crops, to our knowledge, this is the first report of C. cassiicola on perilla. References: (1) M. B. Ellis. Page 372 in: Dematiaceous Hyphomycetes. 1971. (2) J. L. D. Silva et al. Plant Pathol. 55:580, 2006.

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First Report of Colletotrichum boninense Causing Anthracnose on Rosa chinensis in China

Plant Disease ◽

10.1094/pdis-01-21-0102-pdn ◽

2021 ◽

Author(s):

Haixia Ding ◽

Wan Peng Dong ◽

Wei Di Mo ◽

Lijuan Peng ◽

Zuo-Yi Liu

Keyword(s):

Botanical Garden ◽

Guizhou Province ◽

Likelihood Method ◽

Leaf Spot Disease ◽

Molecular Characteristics ◽

Pathogenicity Test ◽

Single Spore ◽

Internal Transcribed Spacer Region ◽

First Report ◽

Rosa Chinensis

Chinese rose (Rosa chinensis Jacq.) is cultivated for edible flowers in southwestern China (Zhang et al. 2014). In March 2020, a leaf spot disease was observed on about 3-5% leaves of Chinese rose cultivar ‘Mohong’ in Guizhou Botanical Garden (26°37' 45'' N, 106°43' 10'' E), Guiyang, Guizhou province, China. The symptomatic plants displayed circular, dark brown lesions with black conidiomata in grey centers on leaves, and leaf samples were collected. After surface sterilization (0.5 min in 75% ethanol and 2 min in 3% NaOCl, washed 3 times with sterilized distilled water) (Fang 2007), small pieces of symptomatic leaf tissue (0.3 × 0.3 cm) were plated on potato dextrose agar (PDA) and incubated at 28oC for about 7 days. Two single-spore isolates, GZUMH01 and GZUMH02, were obtained, which were identical in morphology and molecular analysis. Therefore, the representative isolate GZUMH01 was used for further study. The pathogenicity of GZUMH01 was tested through a pot assay. Ten healthy plants were scratched with a sterilized needle on the leaves. Plants were inoculated by spraying a spore suspension (106 spores ml-1) onto leaves until runoff, and the control leaves sprayed with sterile water. The plants were maintained at 25°C with high relative humidity (90 to 95%) in a growth chamber. The pathogenicity test was carried out three times using the method described in Fang (2007). The symptoms developed on all inoculated leaves but not on the control leaves. The lesions were first visible 48 h after inoculation, and typical lesions similar to those observed on field plants after 7 days. The same fungus was re-isolated from the infected leaves but not from the non-inoculated leaves, fulfilling Koch’s postulates. Fungal colonies on PDA were villiform and greyish. The conidia were abundant, oval-ellipsoid, aseptate, 15.8 (13.7 to 18.8) × 5.7 (4.3 to 6.8) µm. The fungal colonies, hyphae, and conidia were consistent with the descriptions of Colletotrichum boninense Moriwaki, Toy. Sato & Tsukib. (Damm et al. 2012; Moriwaki et al. 2003). The pathogen was confirmed to be C. boninense by amplification and sequencing of the internal transcribed spacer region (ITS), the glyceraldehyde-3-phosphate dehydrogenase (GADPH), actin (ACT), and chitin synthase 1 (CHS-1) genes using primers ITS1/ITS4, GDF1/GDR1, ACT512F/ACT783R, and CHS-79F/CHS-345R, respectively (Damm et al. 2012; Moriwaki et al. 2003). The sequences of the PCR products were deposited in GenBank with accession numbers MT845879 (ITS), MT861006 (GADPH), MT861007 (ACT), and MT861008 (CHS-1). BLAST searches of the obtained sequences of the ITS, GADPH, ACT, and CHS-1 genes revealed 100% (554/554 nucleotides), 100% (245/245 nucleotides), 97.43% (265/272 nucleotides), and 99.64% (279/280 nucleotides) homology with those of C. boninense in GenBank (JQ005160, JQ005247, JQ005508, and JQ005334, respectively). Phylogenetic analysis (MEGA 6.0) using the maximum likelihood method placed the isolate GZUMH01 in a well-supported cluster with C. boninense. The pathogen was thus identified as C. boninense based on its morphological and molecular characteristics. To our knowledge, this is the first report of the anthracnose disease on R. chinensis caused by C. boninense in the world.

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First Report of Sea buckthorn Stem Wilt Caused by Fusarium sporotrichioides in Gansu, China

Plant Disease ◽

10.1094/pdis-03-21-0627-pdn ◽

2021 ◽

Author(s):

Bo Xia ◽

Yue Liang ◽

Jianzhong Hu ◽

Xiaoling Yan ◽

Liqiang Yin ◽

...

Keyword(s):

Elongation Factor ◽

Morphological Characteristics ◽

Gansu Province ◽

Sea Buckthorn ◽

Molecular Characteristics ◽

Ear Rot ◽

Fusarium Sporotrichioides ◽

Tree Roots ◽

Internal Transcribed Spacer Region ◽

First Report

Sea buckthorn (Hippophae rhamnoides) is an important deciduous shrub for fruit and ecological restoration in arid and semi-arid regions of China. Twelve Chinese and Russian cultivars (cv. Shenqiuhong, eshi01, ... eshi11) were planted about 1.6 acre area in a seedling nursery, located in Qingyang City of Gansu province in northwest China, where high mortality (more than 70%) of sea buckthorn was observed in late July 2019. Symptoms consisted of massive chlorosis, drooping leaves and dried-up stems on 5-year-old trees. Pieces of tree roots and stems with irregular light-brown discoloration in the xylem vessels were selected. Small pieces of discolored tissue were surface disinfested (1 min in 1% sodium hypochlorite, followed by three rinses with sterile distilled water), air-dried, and placed on potato dextrose agar (PDA) medium for 5 days at 25°C in the dark. A fungus was consistently isolated from both diseased roots and stems tissues. Colonies on PDA grew rapidly. Dense mycelia were pinky-white initially, and became carmine red color with age on the undersurface of the plate. Macroconidia were moderately curved, 3 to 5 marked septa, hyaline, thick walled, and measuring 27.8± 3.6 µm × 4.8 ± 0.5 µm (n = 30). Microconidia were abundant, pear-shaped, ellipsoid to fusoid, often with a papilla at the base, and 8.4 ± 2.2 µm ×3.1 ± 0.3 µm (n = 30). Genomic DNA was extracted for amplification and sequencing of the internal transcribed spacer region (ITS1 and ITS4 primers) (White et al. 1990) of the ribosomal DNA (Accession Nos. MN160235 to MN160238) and translation elongation factor-1 alpha (EF1 and EF2 primers, accession Nos. MN429075 to MN429078) (O’Donnell et al. 1998). The sequences revealed 99% similarity to the sequences of the ITS (AY188917), and 100% identity with EF1-α (JF740808) regions of Fusarium sporotrichioides. Based on morphological and molecular characteristics, the fungus was identified as F. sporotrichioides (Leslie and Summerell 2006). Koch’s postulates were fulfilled on healthy, potted 1-year-old sea buckthorn seedings using two isolates in a greenhouse at 25 °C, 90% relative humidity, and 12-hour light/dark photoperiod. Ten potted seedings were inoculated on the stems by placing a 5-mm-diameter mycelial plug (5-day-old PDA cultures for each isolate) into the surface of a wound created with a needle, and the inoculation sites were covered with Parafilm to maintain moisture. Ten seedings were inoculated with PDA plugs as controls. Seven to ten days after inoculation, typical symptoms of dark-brown necrotic lesions on chlorotic leaf margins were observed. About 2 weeks after inoculation, the inoculated stems were gradually dry up, accompanied by withering and fallen leaves. Control plants remained asymptomatic. Pathogens were successfully isolated from the inoculated stems again, exhibiting morphological characteristics identical to those of F. sporotrichioides. Previous papers reported F. sporotrichioides as a common pathogen caused lavender wilt (Cosic et al. 2012), foliar spots on forage corn (Moya-Elizondo et al. 2013) and maize ear rot (Wang et al. 2019). To our knowledge, this is the first report of sea buckthorn stem wilt caused by F. sporotrichioides on several Chinese and Russian cultivars in Gansu province of China. In Heilongjiang province, the same disease was reported in 2010 (Song et al. 2010), nearly 30 longitudes away from Gansu province. Therefore, this disease appears to be a serious risk for future sea buckthorn production.

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First Report of Stem and Leaf Anthracnose on Blueberry Caused by Colletotrichum gloeosporioides in China

Plant Disease ◽

10.1094/pdis-11-12-1056-pdn ◽

2013 ◽

Vol 97 (6) ◽

pp. 845-845 ◽

Cited By ~ 1

Author(s):

C. N. Xu ◽

Z. S. Zhou ◽

Y. X. Wu ◽

F. M. Chi ◽

Z. R. Ji ◽

...

Keyword(s):

Colletotrichum Gloeosporioides ◽

Morphological Characteristics ◽

Its Region ◽

Pathogenic Fungi ◽

Molecular Characteristics ◽

First Report ◽

Potted Plants ◽

Leaf Spots ◽

Pathogenicity Tests ◽

Highbush Blueberries

Blueberry (Vaccinium spp.) is becoming increasingly popular in China as a nutritional berry crop. With the expansion of blueberry production, many diseases have become widespread in different regions of China. In August of 2012, stem and leaf spots symptomatic of anthracnose were sporadically observed on highbush blueberries in a field located in Liaoning, China, where approximately 15% of plants were diseased. Symptoms first appeared as yellow to reddish, irregularly-shaped lesions on leaves and stems. The lesions then expanded, becoming dark brown in the center and surrounded by a reddish halo. Leaf and stem tissues (5 × 5 mm) were cut from the lesion margins and surface-disinfected in 70% ethanol for 30 s, followed by three rinses with sterile water before placing on potato dextrose agar (PDA). Plates were incubated at 28°C. Colonies were initially white, becoming grayish-white to gray with yellow spore masses. Conidia were one-celled, hyaline, and cylindrical with rounded ends, measuring 15.0 to 25.0 × 4.0 to 7.5 μm. No teleomorph was observed. The fungus was tentatively identified as Colletotrichum gloeosporioides (PenZ.) PenZ & Sacc. (teleomorph Glomerella cingulata (Stoneman) Spauld. & H. Schrenk) based on morphological characteristics of the colony and conidia (1). Genomic DNA was extracted from isolate XCG1 and the internal transcribed spacer (ITS) region of the ribosomal DNA (ITS1–5.8S-ITS2) was amplified with primer pairs ITS1 and ITS4. BLAST searches showed 99% identity with C. gloeosporioides isolates in GenBank (Accession No. AF272779). The sequence of isolate XCG1 (C. gloeosporioides) was deposited into GenBank (JX878503). Pathogenicity tests were conducted on 2-year-old potted blueberries, cv. Berkeley. Stems and leaves of 10 potted blueberry plants were wounded with a sterilized needle and sprayed with a suspension of 105 conidia per ml of sterilized water. Five healthy potted plants were inoculated with sterilized water as control. Dark brown lesions surrounded by reddish halos developed on all inoculated leaves and stems after 7 days, and the pathogen was reisolated from lesions of 50% of inoculated plants as described above. The colony and conidial morphology were identical to the original isolate XCG1. No symptoms developed on the control plants. The causal agent of anthracnose on blueberry was identified as C. gloeosporioides on the basis of morphological and molecular characteristics, and its pathogenicity was confirmed with Koch's postulates. Worldwide, it has been reported that blueberry anthracnose might be caused by C. acutatum and C. gloeosporioides (2). However, we did not isolate C. acutatum during this study. To our knowledge, this is the first report of stem and leaf anthracnose of blueberry caused by C. gloeosporioides in China. References: (1) J. M. E. Mourde. No 315. CMI Descriptions of Pathogenic Fungi and Bacteria. Kew, Surrey, UK, 1971. (2) N. Verma, et al. Plant Pathol. 55:442, 2006.

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First Report of Neofusicoccum parvum Associated with Dieback of Mango Trees in Peru

Plant Disease ◽

10.1094/pdis-93-4-0426b ◽

2009 ◽

Vol 93 (4) ◽

pp. 426-426 ◽

Cited By ~ 12

Author(s):

J. Javier-Alva ◽

D. Gramaje ◽

L. A. Alvarez ◽

J. Armengol

Keyword(s):

Dna Sequences ◽

Mangifera Indica ◽

Elongation Factor ◽

Tree Canopy ◽

Internal Transcribed Spacer Region ◽

Neofusicoccum Parvum ◽

First Report ◽

Early Maturation ◽

Mycelial Plug ◽

Stem Lesions

Mango (Mangifera indica L) is one of the most important cash crops of northern Peru. Since 2003, adult mango trees (cvs. Criollo and Kent) located in Piura Province developed symptoms of dieback characterized by the death of twigs and branches in the tree canopy. Additional disease symptoms involved darkened, elongated lesions on the peduncle, causing an early maturation of the fruit, and in advanced symptoms, stem-end rot of fruits. Symptoms were frequent in the spring months (September to November) when the lesions expand rapidly. Diseased tissues from branches and fruits were collected and small pieces of necrotic tissues were surface disinfected and plated onto potato dextrose agar (PDA) with 0.5 g L–1 streptomycin sulfate. Plates were incubated at 25°C in the dark. All affected tissues consistently developed colonies with a white mycelium, moderately dense, and becoming olivaceous gray after 5 to 6 days. Pycnidia were produced on sterile mango twigs placed on the surface of potato carrot agar (PCA) after 10 days. Conidia were hyaline, guttulate, aseptate, measuring (15-) 18.5 (-22.5) × (4-) 5.2 (-7.5) μm. Conidia became olivaceous and developed one or two septa before germination. Isolates were identified as Neofusicoccum parvum (Pennycook & Samuels) Crous, Slippers, & A.J.L. Phillips (1). DNA sequences of the rDNA internal transcribed spacer region (ITS) and part of the translation elongation factor 1-alpha (EF1-α) genes were used to confirm the identification through BLAST searches in GenBank (ITS: 99% identity to Accession No. EU080928; EF1-α: 98% identity to Accession No. AY343367). Representative sequences of the studied DNA regions were deposited at GenBank (ITS: Accession No. FJ528596; EF1-α: Accession No. FJ528597). Pathogenicity tests were conducted on 18-month-old potted mango plants cv. Kent with two N. parvum strains (A4 and A5). A mycelial plug (3 cm in diameter) taken from the margin of an actively growing colony of each isolate was put in a wound made with a cork borer of the same diameter on the stem of each plant. Inoculation wounds were wrapped with Parafilm. Controls were inoculated with sterile PDA plugs. Ten replicates for each isolate were used with an equal number of control plants. Plants were maintained in a greenhouse with a temperature range of 22 to 28°C. After 4 weeks, mango plants showed necrotic stem lesions originating from the inoculation point affecting also the branches of the inoculated plants. No differences in lesion area between strains were obtained. No lesions developed in the control plants. Reisolations from necrotic tissues were successful and both isolates were morphologically identical to those used for inoculations. N. parvum was isolated from all symptomatic trees in all surveyed areas. This pathogen has already been reported on mango (2) and currently represents a serious problem in the mango-producing areas of Peru. To our knowledge, this is the first report of N. parvum affecting mango in Peru. References: (1) P. W. Crous et al. Stud. Mycol. 55:235, 2006. (2) B. Slippers et al. Mycologia 97:99, 2005.

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First Report of Alternaria alternata Causing a Leaf Spot on Philodendron ‘con-go’ in China

Plant Disease ◽

10.1094/pdis-07-14-0726-pdn ◽

2014 ◽

Vol 98 (11) ◽

pp. 1588-1588 ◽

Cited By ~ 3

Author(s):

Z. Zhou ◽

Y. L. Li ◽

C. Y. Yuan ◽

P. L. Duan

Keyword(s):

Alternaria Alternata ◽

Leaf Spot ◽

Its Region ◽

Control Treatment ◽

Distilled Water ◽

Internal Transcribed Spacer Region ◽

First Report ◽

Potted Plants ◽

Similar Disease ◽

Residential District

Philodendron ‘con-go’ is widely cultivated indoors in China as an evergreen potted plant. In October 2013, a leaf spot on Philodendron ‘con-go’ was observed in the residential district of Luoyang (112.46° E, 34.62° N), Henan Province, China. The disease was characterized by oval-shaped, 10 to 20 × 25 to 55 mm, yellow to brown lesions with darker brown borders. Fifty potted plants were surveyed, and less than 2% of the leaves were infected. Lesions appeared mostly in old leaves. The symptomatic leaves affected on the plants' ornamental value, but had little impact on their health. Some lesions merged to form a large irregular lesion that could cover a whole leaf. Two infected leaves from one plant were selected randomly for the isolation of the pathogen. Lesions were cut into 1 cm2 pieces, soaked in 70% ethanol for 30 s, sterilized with 1% sodium hypochlorite for 5 min, then washed three times in sterilized distilled water. The pieces were incubated at 25°C on potato dextrose agar (PDA) for 4 to 5 days. A fungus was consistently isolated. Colonies of the fungus were deep green with white mycelium borders. Conidiophores were light brown with 2 to 4 septa. Conidia were obclavate, 14.6 to 49.1 × 8.3 to 16.4 μm, with a short beak, and with 1 to 5 transverse septa and 0 to 3 longitudinal septa, light brown to olive-brown. Based on morphology, the pathogen was identified as Alternaria alternata. Three isolates were selected randomly for further identification. To confirm pathogenicity, eight leaves of potted Philodendron ‘con-go’ plants were wounded with a sterile pin after wiping each leaf surface with 70% ethanol and washing each leaf with sterilized distilled water three times. The isolates were grown on PDA for 7 days and suspended in sterile distilled water to produce a final concentration of 2 × 105 spores/ml. A 5-μl drop of spore suspension was placed on each pin-wounded leaf. Each of three fungal isolates was inoculated on two leaves, and the control treatment (water inoculated) was done similarly on two leaves. The plants were placed in a growth chamber at 28°C with 80% relative humidity, 50 to 60 klx/m2 light intensity, and a 10-h photoperiod. After 7 days, lesions appeared on inoculated leaves, but the control leaves remained symptomless. Pathogenicity tests were repeated three times. Similar disease symptoms and re-isolation of A. alternata fulfilled Koch's postulates. To confirm the fungal identification, the rDNA of the internal transcribed spacer region in three isolates were amplified using primers ITS1 and ITS4 (1) and sequenced. The nucleotide sequence of the ITS region was submitted to GenBank under accession KJ829535 and showed 100% sequence identity with the strain A. alternata LPSC 1187 (KF753947.1). To our knowledge, this is the first report of a leaf spot of Philodendron ‘con-go’ by A. alternate in China. Reference: (1) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, CA, 1990.

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First Report of Bipolaris cactivora Causing Flower Rot of Pitaya (Hylocereus costaricensis) in China

Plant Disease ◽

10.1094/pdis-09-20-2055-pdn ◽

2020 ◽

Author(s):

Fang Qiu ◽

Jinsong Yang ◽

Changping Xie ◽

Xi Li ◽

Jing Li ◽

...

Keyword(s):

Large Subunit ◽

South Florida ◽

Fungal Isolate ◽

Fruit Rot ◽

Bootstrap Support ◽

Molecular Characteristics ◽

Conidial Suspension ◽

Internal Transcribed Spacer Region ◽

First Report ◽

Dark Green

Pitaya (Hylocereus costaricensis), belonging to the Cactaceae family, has rich functional substances, such as a variety of amino acids, which are popular with consumers (Wichienchot et al. 2010). In May 2019, flowers showed symptoms of rot, with an incidence of 15% in a plantation (233.3 ha) in Changjiang (19°46′N; 108°93′E) (Hainan province), China. The initial disease symptoms of flower were small scattered purple-red spot (1~2 mm), including circular, long oval or irregular in shape. The spots were gradually expanded and coalesced, forming abundant reddish-brown lesions. Later, this disease resulted in rotting and blackening of the whole flower. Many black mildew layers (conidiophores and conidia) on the surface of the lesions were observed under compound microscopy. Symptomatic flower tissue (4 cm2) from collecting samples was disinfected in 75% ethanol for 25 s, followed by 1 min in 5% sodium hypochlorite, rinsed 3 times with sterile water, plated on potato dextrose agar (PDA) for 3 days, and incubated at 28ºC. A fungus was consistently isolated from symptomatic flower samples with 90% isolation rate. Resultant colony of the fungus was circular, dark green, velvety, hairy, after 7 days, incubated at 28ºC. Hyphae were septate, 6.2-8.9 μm (average 7.6±0.5) in diameter. Conidia were straight, obclavate, pale to mid brown, 2-6 septate, 23.0 to 42.2 μm (average 31.0±3.2) × 6.5 to 9.8 μm (average 8.0±0.6) (n = 100). The conidia were normally produced germ tubes from one end or both ends. The width of conidiophore was 5.1 to 6.6 μm (average 5.8±0.4) (n = 50). Sequences were generated from the isolate using primers for the internal transcribed spacer region (ITS) (ITS1/ITS4) (White et al. 1990), ribosomal large subunit (LSU) (LROR/LR5) (Vilgalys et al. 1990), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (GPD1/GPD2) (Berbee et al. 1999) loci. The resulting sequences were deposited in GenBank with accession numbers MN960109, MN966852, and MT542865. BLAST analysis demonstrated that these sequences were 99% similar to ITS (HM193535), LSU (MH869295), and GAPDH (HM598681) of Bipolaris cactivora. A maximum likelihood phylogenetic analysis based on combined dataset of ITS, LSU, and GAPDH sequences using MEGA7.0 revealed that the isolate was placed in the same clade as B. cactivora with 100% bootstrap support. A conidial suspension (1 × 105 conidia/ml) of the fungal isolate was prepared by harvesting conidia from pure culture of the fungus grown on PDA 25 days. The 10 mL suspension was sprayed onto ten flowers with no wounding. Ten additional flowers sprayed with sterile distilled water were served as controls. All flowers were covered with plastic bags to maintain high humidity and incubated under natural condition. Typical symptoms of purple-red spot were observed on all the inoculated flowers on the third day. Abundant dark-brown to dark lesions were observed on the surface of flowers and were similar to those observed on the naturally infected flowers after 5 days. The control flowers remained asymptomatic. The fungal isolate of B. cactivora was reisolated from lesion of the flowers and reidentified by morphological and molecular characteristics, thus fulfilled Koch’s postulates. Pathogenicity tests were repeated thrice with the same results. B. cactivora had been reported causing flowers and fruit rot of pitaya in South Florida (Tarnowski et al. 2010). This is the first report of B. cactivora causing flower rot of pitaya (H. costaricensis) in China. The flower rot may provide inoculum for the fruit rot, which will cause reduction of pitaya yield.

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First Report of Downy Mildew on Nursery-Grown Hellebore Caused by Peronospora pulveracea in California

Plant Disease ◽

10.1094/pdis-93-3-0319c ◽

2009 ◽

Vol 93 (3) ◽

pp. 319-319 ◽

Cited By ~ 1

Author(s):

C. Y. Warfield ◽

C. L. Blomquist ◽

E. E. Lovig

Keyword(s):

Downy Mildew ◽

The United States ◽

Early Spring ◽

Economic Losses ◽

Late Winter ◽

Internal Transcribed Spacer Region ◽

First Report ◽

Potted Plants ◽

Rdna Sequences ◽

Young Leaves

Hellebore or Lenten rose (Helleborus × hybridus) is an evergreen, herbaceous perennial in the family Ranunculaceae. Hellebores are sold as decorative, potted plants and as shade-loving landscape plants favored for their attractive and prolonged blooms in late winter or early spring. In April of 2008, downy mildew-like growth was observed on the foliage of approximately 60 containerized plants of Helleborus ‘Blue Lady’, ‘Pink Lady’, ‘White Lady’, and ‘Royal Heritage’ grown outdoors in a retail nursery in coastal San Mateo County, California. Infected foliage had angular, vein-delimited, dark brown-to-black speckled lesions on adaxial leaf surfaces turning dry and necrotic with age. Young leaves were small and distorted. Affected flowers were spotted and brown. The abaxial sides of affected leaves had light brown-to-purplish downy mildew-like growth. Subhyaline conidia, globose to ellipsoid in shape, ranged from 25 to 31 × 17 to 24 μm (average 28 × 21 μm). Conidiophores ranged from 265 to 375 × 5 to 11.5 μm (average 333 × 8.9 μm), branching dichotomously four to five times in the upper half. Morphological measurements fell within the range previously described for Peronospora pulveracea and P. alpicola, which were reported on Helleborus spp. and Ranunculus aconitifolius, respectively (1,2). DNA sequence of the internal transcribed spacer region of rDNA of our isolate (Genbank Accession No. FJ384778) matched sequences of P. pulveracea (Genbank Accession No. AY198270) and P. alpicola (Genbank Accession No. AY198271) with 100% identity. These two organisms are taxonomically indistinguishable by rDNA sequences and are likely to be the same species (3). To our knowledge, this is the first report of P. pulveracea on Helleborus × hybridus in California and the United States. Lenten rose is commercially propagated by seed, which is a potential pathway for introduction of this pathogen. Mature plants are sold and shipped intra- and interstate as decorative flowering plants or nursery stock. The importance and economic impact of this disease is limited, but significant economic losses could occur during production. References: (1) E. A. Gäumann. Beitr. Kryptogamenflora Schweiz 5:113, 1923. (2) G. Hall. Mycopathologia 126:57, 1994. (3) H. Voglmayr. Mycol. Res. 107:1132, 2003.

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First Report of Colletotrichum boninense Causing Anthracnose on Pepper in Brazil

Plant Disease ◽

10.1094/pdis-93-1-0106a ◽

2009 ◽

Vol 93 (1) ◽

pp. 106-106 ◽

Cited By ~ 16

Author(s):

H. J. Tozze ◽

N. M. Massola ◽

M. P. S. Câmara ◽

R. Gioria ◽

O. Suzuki ◽

...

Keyword(s):

Morphological Characteristics ◽

Hypodermic Needle ◽

Molecular Characteristics ◽

Width Ratio ◽

Conidial Suspension ◽

Rio Grande Do Sul ◽

First Report ◽

Potted Plants ◽

Length Width ◽

Orange Color

Colletotrichum boninense was isolated from pepper (Capsicum annuum) fruits (cv. Amanda) with preharvest anthracnose symptoms collected in the Brazilian states of Rio Grande do Sul and São Paulo in July of 2005. In the field, the disease affected mature fruits and leaves with an incidence near 25%. Typical symptoms in fruits were circular, sunken lesions with orange spore masses in a dark center. Three single conidia isolates were obtained from infected fruits. When grown on potato dextrose agar at 25°C with a 12-h photoperiod, these isolates produced white colonies with a cream-to-orange color in the opposite side, but no sclerotia. Conidia were cylindrical, had obtuse ends and a hilum-like low protuberance at the base, and measured 13.5 to 15.5 × 4.6 to 5.1 μm. Conidial length/width ratio was 2.8 to 3.0. These morphological characteristics are consistent with the description of C. boninense (1). To confirm pathogen identity, the internal transcribed spacer rRNA region was sequenced (GenBank Accession Nos. FJ010199, FJ010200, and FJ010201) and compared with the same region of C. boninense (GenBank Accession No. DQ286160.1). Similarity between these sequences was 98 to 99%. The pathogenicity of the three isolates was determined on pepper fruits cv. Amanda. Attached as well as detached fruits from potted plants were inoculated. Inoculation was performed by depositing 40-μl droplets of a suspension (105 conidia per ml) on the surfaces of nonwounded (detached n = 5; attached n = 5) and wounded (detached n = 5; attached n = 5) fruits with a sterilized hypodermic needle. Incubation took place in a moist chamber for 12 days at 25°C with a 12-h photoperiod. Inoculation of control fruits was similar in procedure and number to that of test fruits, except sterile distilled water was used instead of the conidial suspension. Symptoms, observed in wounded and nonwounded test fruits 3 to 5 days after inoculation, were characterized by necrotic, sunken zones containing acervuli, black setae, and orange spore masses. Control fruits presented no symptoms. Pathogens reisolated from infected fruits showed the same morphological and molecular characteristics of the isolates previously inoculated. To our knowledge, this is the first report of C. boninense infecting pepper in Brazil. Reference: (1) J. Moriwaki et al. Mycoscience 44:47, 2003.

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First Report of Leaf Spot, Blight, and Stem Lesions Caused by Cylindrocladium pauciramosum on Callistemon in Spain

Plant Disease ◽

10.1094/pdis-91-8-1057c ◽

2007 ◽

Vol 91 (8) ◽

pp. 1057-1057 ◽

Cited By ~ 6

Author(s):

A. Pérez-Sierra ◽

L. A. Álvarez ◽

M. León ◽

P. Abad-Campos ◽

J. Armengol ◽

...

Keyword(s):

Uv Light ◽

Single Spore ◽

First Report ◽

Potted Plants ◽

Leaf Spots ◽

Plastic Bags ◽

The Family ◽

Stem Lesions ◽

The Uk ◽

Β Tubulin

Callistemons (Callistemon citrinus (Curtis) Skeels), evergreen plants of the family Myrtaceae, are commonly grown in Mediterranean gardens for their red bottlebrush-like flowers. During November of 2006, 1-year-old potted plants of callistemon showed leaf spots and blight in commercial nurseries in Valencia, Spain. Symptoms consisted initially of minute brown spots on the leaves, developing into black-gray blotches that finally coalesced. Diseased plants also showed stem lesions and blight of young shoots. Approximately 30% of the plants were affected. A Cylindrocladium sp. was isolated consistently from the infected tissues. Six single conidial isolates were grown on carnation leaf agar (CLA) under near-UV light at 25°C for 7 days (1). The macroconidiophores comprised of a stipe, a sterile elongation, and a penicillated arrangement of primary, secondary, and tertiary branches. The stipes were septate, 110 to 175 (138) μm long, with a terminal obpyriform vesicle measuring 3.75 to 7.5 (5.8) μm wide. Phialides (12.5 × 3.6 μm) were hyaline, doliiform to reniform, with conidia 40 to 55 × 3.7 to 5 μm, cylindrical with rounded ends, aseptate or one septate. Chlamydospores were brown and formed microsclerotia. These features conformed to the description of Cylindrocladium pauciramosum (3). Further confirmation was obtained by sequence analysis. The 5′ end of the β-tubulin gene was amplified using primers T1 and βt2b (2). Comparison with other sequences in GenBank revealed that the isolates described here were identical with C. pauciramosum (Accession No. AY880064) isolated from Ceanothus in the UK. To confirm pathogenicity, 1-year-old plants of callistemon were inoculated with two isolates by spraying with a spore suspension of the fungus (1 × 105 conidia per ml) obtained from 14-day-old single spore colonies on CLA. Control plants were treated with sterile distilled water. After inoculation, all plants were maintained in plastic bags and kept at 22 ± 2°C. Four days after inoculation, the plants developed symptoms similar to those observed in natural infections, and C. pauciramosum was reisolated, successfully completing Koch's postulates. No symptoms were observed on the control plants. C. pauciramosum has been recorded on several hosts, including Callistemon citrinus, in Italy (4). To our knowledge, this is the first report of C. pauciramosum on callistemon in Spain. References: (1) P. W. Crous and M. J. Wingfield. Mycotaxon 51:341, 1994. (2) B. Henricot and A. Culham. Mycologia 94:980, 2002. (3) C. L. Schoch et al. Mycologia 91:286, 1999. (4) C. L. Schoch et al. Plant Dis. 85:941, 2001.

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