First Report of Colletotrichum boninense Causing Anthracnose on Pepper in Brazil

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 Anthracnose Caused by Colletotrichum theobromicola on Barbados Cherry (Malpighia emarginata) in Brazil

Plant Disease ◽

10.1094/pdis-01-14-0099-pdn ◽

2014 ◽

Vol 98 (9) ◽

pp. 1272-1272 ◽

Cited By ~ 2

Author(s):

C. A. D. Bragança ◽

A. F. Nogueira Junior ◽

F. Rogério ◽

N. S. Massola

Keyword(s):

Nutritional Value ◽

Morphological Characteristics ◽

Fluorescent Light ◽

Width Ratio ◽

Conidial Suspension ◽

First Report ◽

Colletotrichum Species ◽

Malpighia Emarginata ◽

Tropical America ◽

Length Width

Barbados cherry, also called acerola, is a fruit originated from tropical America that is well-known for its high content of vitamin C and nutritional value. Anthracnose is one of the most common diseases on Barbados cherry. In Brazil, this disease is associated with Colletotrichum gloeosporioides sensu lato (2). In 2012, necrotic and sunken spots were observed on Barbados cherry fruit (cv. Rubra) in Sao Paulo State, Brazil, from which a Colletotrichum species was isolated on potato dextrose agar (PDA). The isolate was grown on PDA at 25°C and 12-h photoperiod under fluorescent light. The colony was gray on the upper surface and the reverse part was dark gray. Conidia (n = 50) were cylindrical to subcylindrical, hyaline, and 12 to 15 (avg. 12.7) × 3.8 to 5.9 (avg. 4.3) μm. Conidia length/width ratio was 2 to 3.6. Pathogenicity was confirmed on Barbados cherry fruit. Inoculation was carried out by depositing 40-μl droplets of a conidial suspension (1 × 105 conidia ml−1) on fruit wounded with a sterilized needle and on non-wounded fruit. Fruit were incubated in a moist chamber at 25°C. First symptoms appeared 3 and 5 days after inoculation on wounded and non-wounded fruit, respectively. No symptoms were observed on control fruit inoculated with water. Six isolates recovered from inoculated fruit showed the same morphological characteristics of the previous isolate. The DNA of the fungus was extracted by a CTAB protocol (1) and the sequences of ITS, GAPDH, ACT, CHS-1, TUB, and CAL genes (4) were generated. Sequences were used in BLAST searches in GenBank and were 100% similar to C. theobromicola, except for GAPDH. The ITS (KC566724) and CAL (KC566437) sequences matched strain ICMP 17099 (JX010285 and JX009588, respectively) with 100% identity. The BTUB (KC566148), GAPDH (KC566578), ACT (KC566870), and CHS-1(KC566292) sequences matched with the strains ICMP 18649 (JX010447, 100% identity), ICMP 17099 (JX009957, 99% identity, 1 pb), ICMP 18567 (JX009457, 100% identity), and ICMP 18613 (JX009771, 100% identity), respectively. The sequences were also compared with authentic culture of C. gloeosporioides (IMI 356878) and the identities were: ITS 99% (JX010148), CAL 91% (JX009729), BTUB 90% (JX010445), GAPDH 83% (GU174561), ACT 93% (JX009494), and CHS-1 98% (JX009747). Based on the multi-gene sequencing, the isolate was identified as C. theobromicola. C. theobromicola was described in 2010 (3) and it is considered as a widely distributed species occurring on different hosts in tropical and subtropical regions (4). This report shows the necessity of the identification of Colletotrichum species from tropical fruits to elucidate the etiology of anthracnose diseases of which C. gloeosporioides sensu lato is considered to be the causal agent. To our knowledge, this is the first report of C. theobromicola on Barbados cherry. References: (1) M. G. Murray and W. F. Thompson. Nucleic Acids Res. 8:4321, 1980. (2) R. Ritzinger et al. Acerola em Foco 13:1, 2007. (3) E. I. Rojas et al. Mycologia 102:1318, 2010. (4) B. S. Weir et al. Stud. Mycol. 73:115, 2012.

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The First Report of Downy Mildew Caused by Peronospora belbahrii on Sweet Basil in Greenhouses in the Czech Republic

Plant Disease ◽

10.1094/pdis-07-13-0747-pdn ◽

2014 ◽

Vol 98 (11) ◽

pp. 1579-1579 ◽

Cited By ~ 3

Author(s):

I. Šafránková ◽

L. Holková

Keyword(s):

Czech Republic ◽

Downy Mildew ◽

Width Ratio ◽

Specific Sequence ◽

The Czech Republic ◽

First Report ◽

Sweet Basil ◽

Potted Plants ◽

Length Width ◽

Necrotic Spots

Sweet basil (Ocimum basilicum L.) is an aromatic plant that is cultivated as a pot plant in greenhouses or in fields in the Czech Republic. The plants are intended for direct consumption or for drying. In April of 2012, the first large chlorotic from the middle necrotic spots occurred gradually on leaves of pot plants O. basilicum cv. Genovese in greenhouses in Central Bohemia. The characteristic gray to brown furry growth of downy mildew appeared on abaxial surfaces of leaves in the place of chlorotic spots within 3 to 4 days. The infested leaves fell off in the late stages of pathogenesis. The infestation gradually manifested itself in ever-younger plants and in July, cotyledons and possibly the first true leaves were already heavily infected and damaged and these plants rapidly died. The plant damage reached 80 to 100%, so it was necessary to stop growing the plants in the greenhouse at the end of July. The causal agent was isolated and identified as Peronospora belbahrii Thines by means of morphological and molecular characters (2,3). Conidiophores were hyaline, straight, monopodial, 280 to 460 μm, branched three to five times, ended with two slightly curved branchlets with a single conidia on each branchled tip. The longer branchlets measured 13 to 24 μm (average 18.2 μm), the shorter one 4 to 15 μm (average 9.7 μm). Conidia were rounded or slightly ovoid, from brownish to dark brownish, measured 22 to 31 × 20 to 28 μm (length/width ratio 1.2). A pathogen-specific sequence was detected with the help of the pathogen ITS rDNA specific primers in symptomatic leaves (1). DNA from plant tissues was isolated using the DNeasy plant Mini Kit (Qiagen, Germany) following the standard protocol. PCR was performed using KAPA2G Robust HotStar kit (Kapa Biosystems, United States) according to the conditions recommended in Belbahri et al. (1). The specific products were visualized by electrophoresis through 1.5% agarose gels. Leaves of 20-day-old potted plants O. basilicum ‘Genovese’ were inoculated by spraying with 5 × 105 conidia/ml of the pathogen. Each pot contained 10 plants. Sterilized distilled water was applied to control plants. Plants were covered with polyethylene bags during the entire incubation period to maintain high humidity, and kept at a temperature of 22 to 24°C. Typical disease symptoms appeared on leaves 5 to 9 days after inoculation. Control plants were symptomless. P. belbahrii was re-isolated from the lesions of inoculated plants, thus fulfilling Koch's postulates. Downy mildew on sweet basil was reported in countries in Africa, Europe, and South and North America (4). To our knowledge, this is the first report of downy mildew on sweet basil in the Czech Republic. References: (1) L. Belbahri et al. Mycol. Res. 109:1276, 2005. (2) Y.-J. Choi et al. Mycol. Res. 113:1340, 2009. (3) M. Thines et al. Mycol. Res. 113:532, 2009. (4) C. A. Wyenandt et al. HortScience 45:1416, 2010.

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First report of the cactus cyst nematode, Cactodera cacti, infecting Cereus jamacaru (Cactaceae) in Brazil

Plant Disease ◽

10.1094/pdis-09-21-1924-pdn ◽

2021 ◽

Author(s):

Francisco Bruno da Silva Café ◽

Rhannaldy Benício Rebouças ◽

Juvenil H. Cares ◽

Cristiano Souza Lima ◽

Francisco de Assis Câmara Rabelo Filho ◽

...

Keyword(s):

Body Length ◽

Morphological Characteristics ◽

Its Region ◽

The Body ◽

Control Treatment ◽

Molecular Characteristics ◽

Width Ratio ◽

Morphometric Characteristics ◽

First Report ◽

Second Stage

During a survey in 2018 for plant nematodes associated with roots and soil in cactus cultivation areas in Ceará State (3°44'48"S, 38°34'29"W), cysts were found on roots of mandacaru, Cereus jamacaru DC. This cactus is native to Brazil, can grow to 6-10 meters in height, and is widely distributed in the Northeast region (Romeiro-Brito et al. 2016) where it is used in construction, in disease remedies, as forage, and as an ornamental (Sales et al. 2014). Several cysts, second-stage juveniles (J2) and eggs extracted from the soil and roots, using sucrose centrifugation, were examined by scanning electron microscopy (SEM) and light microscopy (LM) to determine morphological and morphometric characteristics. Molecular characteristics were determined by DNA extraction from J2 and embryonated eggs using a protocol specific for Heteroderidae (Subbotin et al., 2018). The internal transcribed spacer sequence (ITS) region of the rDNA and D2-D3 regions of the 28S rDNA were amplified using the universal primers TW81 (5′-GTTTCCGTAGGTGAACCTGC-3′) and AB28 (5′-ATATGCTTAAGTTCAGCGGGT-3′), D2A(5′-ACAAGTACCGTGAGGGAAAGTTG-3′) and D3B(5′-TCGGAAGGAACCAGCTACTA-3′), respectively. To confirm that mandacaru is a host for C. cacti, six plantlets of mandacaru were inoculated with 1,800 eggs of the nematode, and kept in a greenhouse at 31 ± 3 ºC and irrigated daily. Six non inoculated mandacaru plantlets served as control treatment. Morphometric characteristics of cysts (n=35) were body length, excluding neck, 555.8 ± 87.8 (354,9 - 727,6) μm, body width 392.1 ± 63.4 (297.9 - 553.7) μm, neck length 63.5 ± 25.8 (49.8-105.0) μm, length to width ratio 1.4 ± 0.2 (1.0-1.8) μm and vulval cone length 48.4 ± 15.2 (40.7 –53.6) μm. Cysts had a rough surface, were lemon-shaped to rounded and had a zigzag cuticular pattern with a protruding vulval cone. They were circumfenestrate without underbridge and bullae, but with the presence of vulval denticles. Measurements of second-stage juveniles (n = 13) included the body length 511.2 ± 33.7 (452.7 - 551.5) μm, stylet length 28.0 ± 2.8 (25.4 - 34.0) μm, tail length 50.7 ± 5.1 (40.6 - 57.4) μm, tail hyaline region 22.7 ± 2.2 (18.9 – 27.1), with a = 20.9 ± 2.2 (17.7-24.3) μm, b = 5.4 ± 0.4 (5.1-5.8) μm, b'= 3.4 ± 0.4 (3.1-3.9) μm, c = 10.2 ± 1.3 (8.9-13.3) μm and c' = 3.8 ± 0.4 (3.0-4.5) μm. The observations of essential morphological characteristics for identification indicated that the species found on C. jamacaru was Cactodera cacti (Filipjev & Schuurmans-Stekhoven, 1941) Krall & Krall, 1978. The sequences of the studied rDNA regions were submitted to GenBank (ITS: MW562829 and D2–D3 regions of 28S: MW562830). The samples used for molecular analysis showed a high degree of sequence identity (99.59%) with C. cacti, from China, Iran and USA for the ITS region. The identity of the D2-D3 regions of 28S sequence was 99.54% with C. cacti isolates from Germany and 99.41% with isolates from USA. Phylogenetic analyses were performed using Maximum likelihood (ML) method for both individual loci, confirming the species as Cactodera cacti. All inoculated mandacaru plantlets showed C. cacti cysts on the roots after 60 days, confirming that mandacaru is a host for C. cacti. This species was reported in São Paulo State, in 2001, associated with ornamental cactus cultivated in pots, but plant species were not identified (Santos et al., 2001). The second report in Brazil was to Schlumbergera sp., an ornamental plant (Oliveira et al. 2007). In both studies, the nematode was not morphologically nor molecularly characterized. Cactodera cacti has been commonly associated with cactus worldwide (Esser, 1992). It has been reported in association with C. jamacaru was first reported in 2011 in China (Duan et al. 2012). This is the first report of the occurrence of C. cacti on C. jamacaru in field conditions in Brazil, and its presence in cactus cultivation areas with agricultural importance represents a threat to cactus production in the country.

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First Report of Colletotrichum tropicale Causing Anthracnose on Common Fig in Taiwan

Plant Disease ◽

10.1094/pdis-06-21-1329-pdn ◽

2021 ◽

Author(s):

Chung-hang Duan ◽

Guan-ying Chen

Keyword(s):

Manganese Superoxide Dismutase ◽

Morphological Characteristics ◽

Width Ratio ◽

Conidial Suspension ◽

Gene Sequences ◽

First Report ◽

Fungal Isolates ◽

Common Fig ◽

Colletotrichum Tropicale ◽

Central Taiwan

Ficus carica L. known as common fig is one of the most profitable fruit crops in Taiwan. Their fruit are harvested for high-priced market. Common fig can be eaten fresh or dried and processed to make different food products. In September 2015, an anthracnose-like disease was widely observed on common fig fruit planted in an orchard in Lukang township (24°04'36" N, 120°27'15" E) in Changhua County, central Taiwan. Symptoms were sunken, water-soaked lesions covered with salmon-colored spore masses and were observed on all stages of fruit, especially when fruit was ripe. Four fungal isolates were collected from four diseased fruit of different plants in the same orchard. Conidia were spread on 2% water agar, and a single conidium was separated by a handmade glass needle. Fungal isolates were grown on potato dextrose agar (PDA) at 24 to 28°C with diffused light. All four strains produced white, aerial, and cottony mycelia covered with abundant salmon-colored conidial masses on PDA. The conidia were hyaline, single celled, round cylindrical on both ends, thin walled, and the contents guttulate. The sizes of conidia were 15.4 (18.5 to 13.1) × 4.73 (5.8 to 3.6) μm [average (max. to min.); length/width ratio = 3.25, n = 40]. DNA was isolated from the representative isolate FC1 and used for amplification of partial sequences of the internal transcribed spacer (ITS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), actin (ACT), β-tubulin 2 (TUB2), manganese-superoxide dismutase (SOD2), calmodulin (CAL), chitin synthase 1 (CHS-1) (Weir et al. 2012) and the intergenic region of apn2 and MAT1-2-1 gene (ApMat) genes (Sharma et al. 2013). A BLAST search against the NCBI database revealed that FC1 gene sequences [GenBank accession nos. MT192648 (ITS), MT155819 (GAPDH), MT199873 (ACT), MT199874 (TUB2), MT815916 (SOD2), MT815917 (CAL), MW684717 (CHS-1) and MT221652 (ApMat)] displayed 99.1, 98.2, 99.3, 99.6, 99.5, 100.0, 92.8 and 100.0% nucleotide identity to the respective gene sequences of Colletotrichum tropicale CBS 124949 (ICMP18653) (JX010264, JX010007, JX009489, JX010407, JX010329, JX009719, JX009870 and KC790728). Multilocus phylogenetic analysis performed with reference sequences showed that the isolate FC1 clustered with C. tropicale in accordance with BLAST results. A conidial suspension (1 × 106 conidia/mL) prepared from FC1 isolate was inoculated by spraying onto detached, ripe, healthy, non-wounded and surface-disinfected common fig fruit (cv. China, n = 4). Fruit sprayed with sterile water were used as control. Fruit were kept in a moist chamber (greater than 90% relative humidity, 24 to 28°C) for 24 h and then maintained in the lab for additional 5 days. The inoculated fruit developed lesions similar to the disease symptoms in the orchard. No symptom was observed on fruit treated with water. C. tropicale was re-isolated from symptomatic fruits and had similar morphological characteristics to FC1 isolate, thus fulfilling Koch’s postulates. The experiment was repeated once showing similar results. The FC1 isolate of C. tropicale with the identification number BCRC FU31436 has been deposited at Taiwan Bioresource Collection and Research Center. This fungus had previously been found on lotus and mango in Taiwan (Chen and Kirschner 2018; Wu et al. 2020), while the pathogenicity among the isolates from different origins is not yet known. To our knowledge, this is the first report of C. tropicale causing anthracnose on common fig fruit in Taiwan.

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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 Powdery Mildew Caused by Erysiphe heraclei on Parsley in Korea

Plant Disease ◽

10.1094/pdis-09-13-0972-pdn ◽

2014 ◽

Vol 98 (6) ◽

pp. 847-847

Author(s):

S. E. Cho ◽

M. J. Park ◽

J. H. Park ◽

J. Y. Kim ◽

H. D. Shin

Keyword(s):

Powdery Mildew ◽

Morphological Characteristics ◽

Its Region ◽

Petroselinum Crispum ◽

Width Ratio ◽

Anethum Graveolens ◽

First Report ◽

Organic Farm ◽

Length Width ◽

A Minor

Parsley, Petroselinum crispum (Mill.) Nyman, is a minor but important leaf crop in Korea. In June 2010, parsley plants (cv. Paramount) showing typical symptoms of powdery mildew were found with approximately 90% incidence (percentage of plants showing symptoms) in polyethylene-film-covered greenhouses in an organic farm in Icheon County of Korea. Symptoms first appeared as thin white colonies, which subsequently showed abundant growth on the leaves with chlorosis and crinkling. Most diseased plantings were unmarketable and shriveled without being harvested. The damage due to powdery mildew infections on parsley has reappeared in Icheon County and Gangneung City with confirmation of the causal agent made again in 2011 and 2012. Voucher specimens were deposited in the Korea University Herbarium (KUS). Appressoria on the mycelium were multilobed or moderately lobed. Conidiophores were cylindrical, 75 to 125 × 8 to 10 μm, straight in foot-cells, and produced conidia singly, followed by 2 to 3 cells. Conidia were oblong-elliptical to oblong, 32 to 55 × 14 to 20 μm with a length/width ratio of 1.7 to 2.9, lacked fibrosin bodies, and produced germ tubes on the perihilar position, with angular/rectangular wrinkling of the outer walls. First-formed conidia were apically conical, basally subtruncate to rounded, and generally smaller than the secondary conidia. Chasmothecia were not found. These structures are typical of the powdery mildew Pseudoidium anamorph of the genus Erysiphe. The specific measurements and morphological characteristics were consistent with those of E. heraclei DC. (1). To confirm the identity of the causal fungus, the complete ITS region of rDNA from isolate KUS-F25037 was amplified with primers ITS5 and P3 (3) and sequenced directly. The resulting 606-bp sequence was deposited in GenBank (Accession No. KF680162). A GenBank BLAST search of this sequence revealed 100% identity with that of E. heraclei on Anethum graveolens from Korea (JN603995) and >99% similarity with those of E. heraclei on Daucus carota from Mexico (GU252368), Pimpinella affinis from Iran (AB104513), Anthriscus cerefolium from Korea (KF111807), and many other parsley family (Apiaceae) plants. Pathogenicity was verified through inoculation by gently pressing diseased leaves onto leaves of five healthy potted parsley plants. Five non-inoculated plants served as negative controls. Inoculated plants developed symptoms after 7 days, whereas the control plants remained symptomless. The fungus present on the inoculated plants was morphologically identical to that originally observed on diseased plants. Parsley powdery mildew caused by E. heraclei has been known in Europe, North America, Brazil, and Japan (2,4). To our knowledge, this is the first report of powdery mildew infections by E. heraclei on parsley in Korea. Since cultivation of parsley was only recently started on a commercial scale in Korea, powdery mildew infections pose a serious threat to safe production of this herb, especially those grown in organic farming where chemical options are limited. References: (1) U. Braun and R. T. A. Cook. Taxonomic Manual of the Erysiphales (Powdery Mildews), CBS Biodiversity Series No. 11. CBS, Utrecht, 2012. (2) D. F. Farr and A. Y. Rossman. Fungal Databases. Syst. Mycol. Microbiol. Lab., Online publication, ARS, USDA, retrieved September 17, 2013. (3) S. Takamatsu et al. Mycol. Res. 113:117, 2009. (4) Y. Tsuzaki and K. Sogou. Proc. Assoc. Plant Prot. Shikoku 24:47, 1989.

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First Report of Fusarium commune causing Stem Rot of Tobacco (Nicotiana tabacum) in Hunan Province, China

Plant Disease ◽

10.1094/pdis-07-20-1466-pdn ◽

2020 ◽

Author(s):

Quan Zhong ◽

Yan song Xiao ◽

Bin He ◽

Zhi Hui Cao ◽

Zhi Guo Shou ◽

...

Keyword(s):

Nicotiana Tabacum ◽

Pathogenic Fungus ◽

Morphological Characteristics ◽

Disease Diagnosis ◽

Hunan Province ◽

Stem Rot ◽

Molecular Characteristics ◽

Conidial Suspension ◽

First Report ◽

Representative Isolate

Tobacco (Nicotiana tabacum L.) is a leafy, annual, solanaceous plant grown commercially for its leaves. It is one of the most important cash crops in China. In April of 2020, tobacco stems in commercial tobacco fields developed a brown to dark brown rot, in the Hunan Province of China. Almost 20% of the plants were infected. Symptoms appeared as round water-soaked spots, then turned dark black and developed into brown necrotic lesions leading to the stem becoming girdled and rotted. Diseased stem tissue was cut and sterilized with 70% ethanol for 10 s, 0.1% HgCl2 for 2 min, rinsed with sterile distilled water three times, and then plated on potato dextrose agar (PDA) and incubated at 26°C in the dark. Six isolates with similar morphology were obtained. Colonies cultured on PDA have morphological characteristics of Fusarium spp. producing white to orange-white, densely aerial mycelium with magenta to dark violet pigmentation. Macroconidia were produced on carnation leaf agar plates (Xi et al. 2019), which were slightly curved, with apical and basal cells curved, and usually contained three or five septa, 25.50 to 41.50×3.55 to 5.80 μm (n=50). Microconidia were cylindrical, ovate-oblong, straight to slightly curved, aseptate and 5.80 to 13.75 × 3.10 to 4.10 μm (n=50). For molecular identification, the translation elongation factor 1-alpha (EF1-α), the largest subunit of RNA polymerase II gene sequences (RPB2) and the mitochondrial small subunit rDNA (mtSSU) of a representative isolate CZ3-5-6 were amplified using the primer pairs ef1/ef2 (O’Donnell et al. 1998), 5F2/7Cr (O’Donnell et al. 2010) and NMS1/ NMS2 (Li et al. 1994). The obtained EF1-α, RPB2 and mtSSU sequences (GenBank accession nos. MT708482, MT708483 and MW260121, respectively) were 99.70 %, 100% and 100% identical to strains of F. commune (HM057338.1 for EF1-α, KU171700.1 for RPB2 and MG846025 for mtSSU). Moreover, Fusarium-ID database searches revealed that the EF1-α and RPB2 were 100% identical to F. commune strains (FD_01140_EF-1a and FD_02411_RPB2). Based on the morphological and molecular characteristics of the representative isolate, the fungal species was identified as F. commune. Pathogenicity testing of a representative isolate was performed by inoculating tobacco plants, which were grown for 2.5 months in a sterile pot with autoclaved soil. Each tobacco stem was injected with 20 μl of conidial suspension (105 spores/ml). Plants inoculated with sterilized water served as control. The pathogenicity tests were performed twice using three replicate plants, and all plants were kept in humid chambers (80 × 50 × 80 cm) at 26°C with a 12-h photoperiod. After 10 days, dark brown necrotic symptoms around the inoculated site, similar to those observed in natural field, were developed in all inoculated plants, whereas no symptoms were observed on the control plants. The pathogenic fungus was re-isolated from symptomatic tissue and identified as F. commune but was not recovered from the control plants. Fusarium commune has been reported to cause root rot or stalk and stem rot on some plants, such as sugarcane (Wang et al. 2018), Gentiana scabra (Guan et al. 2016) and maize (Xi et al. 2019). However, to our knowledge, this is the first report of F. commune causing stem rot on tobacco in China. Identification of F. commune as a stem rot causing pathogen might provide important insights for disease diagnosis on tobacco caused by different Fusarium species. Overall, this disease might bring a threat to tobacco production, and appropriate control measures should be adopted to reduce losses in tobacco fields.

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Occurrence of leaf spot caused by Neodeightonia phoenicum on pygmy date plam (Phoenix roebelenii) in China

Plant Disease ◽

10.1094/pdis-10-21-2190-pdn ◽

2021 ◽

Author(s):

Wu Zhang ◽

Xiu Li Song

Keyword(s):

Leaf Spot ◽

Elongation Factor ◽

Morphological Characteristics ◽

Aerial Mycelium ◽

Likelihood Method ◽

Molecular Characteristics ◽

Infected Leaf ◽

Conidial Suspension ◽

Tropical Regions ◽

Potted Plants

The pygmy date palm (Phoenix roebelenii) is a popular ornamental plant widely cultivated in tropical regions as well as in China. In June 2018, a new leaf spot symptoms were observed on P. roebelenii in several different parks in Zhanjiang City of China. The early symptoms of infected leaves were presented with small, round, pale brown spots. As the size of these spots increased, they coalesced to form larger irregular necrotic lesions surrounded by dark brown edges, which eventually led to leaf wilted and defoliation. A filamentous fungus was consistently isolated from infected leaf samples. Colonies on PDA at 25°C (12 h light/dark) were initially white with abundant aerial mycelium, which turned fluffy and dark olivaceous after one-week culture. Pycnidial conidiomata were black and globose and formed on pine needles in water agar at 25°C (12 h light/dark) after 21 days. Conidiogenous cells were hyaline, cylindrical, holoblastic. The conidia was ovoid to ellipsoid, thick-walled, which was initially hyaline and aseptate, later turned into dark brown and 1-septate with a striate appearance to conidia, 11.6~25.0 μm×9.6~12.0 μm (av. 20.4 μm×10.1 μm). For molecular identification, the partial sequences of internal transcribed spacer (ITS) regions, translation elongation factor (EF-1α) and β-tubulin (TUB) genes of two representative isolates RYCK-1, RYCK-2 were amplified and sequenced using primer pairs ITS/ITS4 (White et al. 1990), EF-688F/EF-986R (Carbone and Kohn 1999), and Bt2a/Bt2b (Glass and Donaldson 1995), respectively. The sequences of the above three loci of the two isolates (accession nos. ITS, OK329968 and OK329969; EF-1α, OK338067 and OK338068; TUB, OK338069 and OK338070) showed 98.4-100.0 % identity with the existing sequences of ex-type culture CBS 122528 of N. phoenicum. A multilocus phylogenetic analysis of the three loci concatenated sequences using the maximum likelihood method showed the isolates that belongs to N. phoenicum. Based on the morphological characteristics and molecular analysis of the isolates, the fungus was identified as N. phoenicum (Phillips et al. 2008). To confirm pathogenicity, five one-year-old potted plants were used for each isolate (RYCK-1 and RYCK-2) and the plants were inoculated by pricking the epidermis of the leaf with a needle. Five leaves of each plant were sprayed with 100 µl of a conidial suspension (1 × 106 conidia/ml) to the wounded surface for each plant. Sterilized distilled water was used as the control and the experiment was repeated. All the plants were incubated at 26 ± 2°C (12 h light/dark) and covered with plastic bags to maintain constant high humidity. After 14 days, all the inoculated leaves showed the same symptoms as those observed in the original diseased plants, but the control plants remained health. The reisolated fungus was identified as N. phoenicum by morphological and molecular characteristics. N. phoenicum is an important pathogen of Phoenix species plants worldwide, which have been reported to cause shoot blights and stalk rots on P. dactylifera and P. canariensis in Greece (Ligoxigakis et al. 2013) and root rot on P. dactylifera in Qatar (Nishad and Ahmed 2020). To our knowledge, this is first report N. phoenicum causing leaf spot on P. roebelenii in China.

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First Report of Golovinomyces cichoracearum Causing Powdery Mildew on Zinnia elegans in China

Plant Disease ◽

10.1094/pdis-11-20-2333-pdn ◽

2020 ◽

Author(s):

Mo Zhu ◽

Jie Ji ◽

Xiao Duan ◽

YongFang Li

Keyword(s):

Powdery Mildew ◽

Morphological Characteristics ◽

Signs And Symptoms ◽

Its Region ◽

Zinnia Elegans ◽

Width Ratio ◽

Post Inoculation ◽

First Report ◽

Golovinomyces Cichoracearum ◽

Length Width

Zinnia elegans, common zinnia, is an annual plant with highly ornamental values. It is widely planted in many nurseries, city parks, universities and home gardens in China. From August to October 2020, powdery mildew-like signs and symptoms were observed on leaves of Z. elegans growing on the campus of Henan Normal University, Henan Province, China. White powdery colonies in circular- or irregularly shaped-lesions were abundant on both surfaces of leaves and covered up to 95 % of the leaf area. Any infected leaves were chlorotic, deformed or senescence. More than 70 % of the monitored Z. elegans plants showed these signs and symptoms. Conidiophores (n = 20) were 100 to 200 × 9 to 13 μm and composed of foot cells, followed by straight cells and conidia. Mycelial appressoria were single and nipple-shaped. The oval-shaped conidia (n = 30) were 22 to 36 × 12 to 18 μm, with a length/width ratio of 1.4 to 2.7, and produced germ tubes from the polar ends of the spore. No chasmothecia were found. Based on these morphological characteristics, the pathogen was initially identified morphologically as Golovinomyces cichoracearum (Braun and Cook 2012). Structures of the pathogen were scraped from infected leaves and total genomic DNA was isolated using the method previously described by Zhu et al. (2019). The internal transcribed spacer (ITS) region of rDNA was amplified by PCR using the primers ITS1/ITS4 (White et al. 1990) and the amplicon was sequenced by Invitrogen (Shanghai, China). The sequence for the fungus was deposited into GenBank under Accession No. MW029904 and was 99.83 % identical (595/596 bp) to G. cichoracearum on Symphyotrichum novi-belgii (HM769725)(Mørk et al. 2011). To perform pathogenicity analysis, leaf surfaces of five healthy plants were fixed in a settling tower and then inoculated by blowing fungal conidia from mildew-infested leaves using pressurized air. Five non-inoculated plants served as a control. The inoculated and non-inoculated plants were separately maintained in two growth chambers (humidity, 60 %; light/dark, 16 h/8 h; temperature, 18 ℃). Eleven- to twelve-days post-inoculation, powdery mildew signs were conspicuous on inoculated plants, while control plants remained healthy. Similar results were obtained by conducting two repeated pathogenicity assays. Thus, based on the morphological characteristics and molecular analysis, the pathogen was identified and confirmed as G. cichoracearum. This pathogen has been reported on Z. elegans in India, Israel, Jordan, Korea, Nepal, Sri Lanka, Switzerland, and Turkey (Farr and Rossman 2020). To our best knowledge, this is the first report of G. cichoracearum on Z. elegans in China. The sudden outbreak of powdery mildew caused by G. cichoracearum on Z. elegans may adversely impact the plant health and ornamental value in China. Therefore, the confirmation of G. cichoracearum infecting Z. elegans expands the understanding of this pathogen and provides the fundamental knowledge for future powdery mildew control.

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First Report of Powdery Mildew Caused by Erysiphe heraclei on Peucedanum japonicum in Korea

Plant Disease ◽

10.1094/pdis-09-14-0895-pdn ◽

2015 ◽

Vol 99 (1) ◽

pp. 161-161 ◽

Cited By ~ 1

Author(s):

I. Y. Choi ◽

S. H. Hong ◽

S. E. Cho ◽

J. H. Park ◽

H. D. Shin

Keyword(s):

Powdery Mildew ◽

Organic Farming ◽

Disease Incidence ◽

Morphological Characteristics ◽

Its Region ◽

Petroselinum Crispum ◽

Width Ratio ◽

First Report ◽

Potted Plants ◽

Powdery Mildews

Peucedanum japonicum Thunb., belonging to the family Apiaceae, is distributed in many Asian countries, including Korea. This plant was recently developed as an edible green and is cultivated under organic farming in Korea. In June 2013, plants showing typical symptoms of powdery mildew were found with approximately 50% disease incidence in polyethylene-film-covered greenhouses in Iksan City, Korea. Symptoms first appeared as circular white colonies, which subsequently showed abundant mycelial growth on the leaves, often covering the whole surface. Infected plants were unmarketable mainly due to signs of white fungal growths and reddish discoloration on the leaves. The same symptoms were found on P. japonicum in poly-tunnels in Iksan City and Jinan County of Korea in 2014. Voucher specimens (n = 3) were deposited in the Korea University Herbarium (KUS). Appressoria were lobed, and solitary or in opposite pairs. Conidiophores were cylindrical, 80 to 145 × 8 to 10 μm, and composed of three to four cells. Foot-cells of conidiophores were straight to substraight, cylindrical, and 25 to 63 μm long. Singly produced conidia were oblong-elliptical to oblong, occasionally ovate, 35 to 50 × 13 to 16 μm with a length/width ratio of 2.3:3.1, with angular/rectangular wrinkling of outer walls, and lacked distinct fibrosin bodies. Germ tubes were produced on the perihilar position of conidia. Primary conidia were apically conical, basally truncate, and generally smaller than the secondary conidia. No chasmothecia were found. These structures are typical of the powdery mildew Pseudoidium anamorph of the genus Erysiphe. The specific measurements and morphological characteristics were consistent with those of E. heraclei DC. (2). To confirm the identification, the complete internal transcribed spacer (ITS) region of rDNA from KUS-F27872 was amplified with primers ITS1/ITS4 and sequenced. The resulting 560-bp sequence was deposited in GenBank (Accession No. KM491178). The obtained ITS sequence shared >99% similarity with those of E. heraclei from apiaceous hosts, e.g., Daucus carota (KC480605), Pimpinella affinis (AB104513), and Petroselinum crispum (KF931139). Pathogenicity was confirmed through inoculation by gently dusting conidia onto leaves of five healthy potted plants. Five non-inoculated plants served as controls. Inoculated plants developed symptoms after 6 days, whereas the control plants remained symptomless. The fungus present on the inoculated plants was identical in morphology to those observed in the field. Powdery mildew of P. japonicum caused by E. heraclei has been reported in Japan (4), and numerous reports of E. heraclei on various species of Peucedanum plants have been made in most part of Europe and East Asia (Japan and far eastern Russia) (1,3). However, this is the first report of powdery mildew caused by E. heraclei on P. japonicum in Korea. Occurrence of powdery mildews is a threat to the quality and marketability of this plant, especially in organic farming. References: (1) K. Amano. Host Range and Geographical Distribution of the Powdery Mildew Fungi. Japan Scientific Societies Press, Tokyo, 1986. (2) U. Braun and R. T. A. Cook. Taxonomic Manual of the Erysiphales (Powdery Mildews), CBS Biodiversity Series No.11. CBS, Utrecht, 2012. (3) D. F. Farr and A. Y. Rossman. Fungal Databases, Syst. Mycol. Microbiol. Lab., online publication. ARS, USDA. Retrieved August 18, 2014. (4) S. Tanda and C. Nakashima. J. Agric. Sci., Tokyo Univ. Agric. 47:54, 2002.

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