First Report of Foliar Blight Caused by Phytophthora capsici on Citrus reticulata Blanco cv. Nian Ju in Guangdong, China

Citrus reticulata Blanco cv. Nian Ju, an important ornamental plant, is traditionally displayed during the Chinese Spring Festival because its golden fruits are a symbol of auspiciousness. In the spring of 2012, foliar blight was observed on 10 to 30% of the Nian Ju plants at four nurseries in Yangjiang, Guangdong Province, China. Initial symptoms appeared as brown to black foliar lesions, followed by expansion of spots into blight. Some young branches also had necrosis. During frequent rainfall and prolonged wet periods at 22°C to 30°C, white and dense mycelia and sporangia were observed on the infected seedlings. To isolate the causal organism, leaves and stems were cut into sections. Each section included some partial lesion and adjacent asymptomatic tissues. They were surface-disinfested in 1% sodium hypochlorite for 60 s, rinsed three times with sterile water, and placed on V8 juice agar (V8A) at 25°C. After 3 days, 10 isolates were obtained and purified by single-zoospore method. These isolates were identified to species level by sequencing the rRNA internal transcribed spacer (ITS) region. Four representative isolates had an identical ITS sequence (GenBank Accession No. KF750568), which had 99% homology with Phytophthora capsici sequences in GenBank. In addition, all recovered isolates were identical in morphological characteristics. They produced caducous, papillate, and ovoid to ellipsoid sporangia (Length × width = 46.2 ± 7.7 × 23.6 ± 11.3 μm), often with a tapered base. The average length of pedicels was 33.3 ± 4.5 μm. All isolates are A2 mating type. They produced gametangia when paired with an A1 tester of P. capsici isolated from pepper on V8A. Plerotic oospores were 25.3 ± 2.1 μm in diameter. Amphigynous antheridia were 13.6 ± 2.8 μm long and 11.2 ± 0.9 μm wide. Oogonia were 27.4 ± 3.2 μm in diameter. To determine the pathogenicity, three 3-year-old potted C. reticulata cv. Nian Ju plants were sprayed with 20 ml of zoospore suspension from one representative isolate at 105 per ml. Two control plants were sprayed with 20 ml distilled water. All plants were then maintained at 90% relative humidity at 25°C with a 12-h photoperiod. Symptoms similar to those observed in the nurseries developed on all inoculated plants but not on any control plants after 10 days. The pathogenicity test was repeated once and similar results were obtained. P. capsici was recovered from all inoculated plants and resultant isolates had identical morphology to that of the isolates used for inoculation. P. capsici has a relatively broad host range including pumpkins, cucumbers, peppers, beans, squashes, and spinach (1,2). To our knowledge, this is the first report of foliar blight of C. reticulata cv. Nian Ju caused by P. capsici. This study indicates that P. capsici is potentially an important pathogen of C. reticulata cv. Nian Ju plants and further investigations into its epidemiology and development of site-specific integrated management programs for this new disease are warranted. References: (1) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. American Phytopathological Society, St. Paul, MN, 1996. (2) D. Tian and M. Babadoost. Plant Dis. 88:485. 2004.

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First Report of Stem and Foliage Blight of Chrysanthemum Caused by Phytophthora drechsleri in the United States

Plant Disease ◽

10.1094/pdis-03-21-0631-pdn ◽

2021 ◽

Author(s):

Charles Krasnow ◽

Nancy Rechcigl ◽

Jennifer Olson ◽

Linus Schmitz ◽

Steven N. Jeffers

Keyword(s):

United States ◽

South Carolina ◽

Sequence Similarity ◽

Morphological Characteristics ◽

The United States ◽

Universal Primers ◽

Broad Host Range ◽

Pathogenicity Test ◽

First Report ◽

Foliage Blight

Chrysanthemum (Chrysanthemum × morifolium) plants exhibiting stem and foliage blight were observed in a commercial nursery in eastern Oklahoma in June 2019. Disease symptoms were observed on ~10% of plants during a period of frequent rain and high temperatures (26-36°C). Dark brown lesions girdled the stems of symptomatic plants and leaves were wilted and necrotic. The crown and roots were asymptomatic and not discolored. A species of Phytophthora was consistently isolated from the stems of diseased plants on selective V8 agar (Lamour and Hausbeck 2000). The Phytophthora sp. produced ellipsoid to obpyriform sporangia that were non-papillate and persistent on V8 agar plugs submerged in distilled water for 8 h. Sporangia formed on long sporangiophores and measured 50.5 (45-60) × 29.8 (25-35) µm. Oospores and chlamydospores were not formed by individual isolates. Mycelium growth was present at 35°C. Isolates were tentatively identified as P. drechsleri using morphological characteristics and growth at 35°C (Erwin and Ribeiro 1996). DNA was extracted from mycelium of four isolates, and the internal transcribed spacer (ITS) region was amplified using universal primers ITS 4 and ITS 6. The PCR product was sequenced and a BLASTn search showed 100% sequence similarity to P. drechsleri (GenBank Accession Nos. KJ755118 and GU111625), a common species of Phytophthora that has been observed on ornamental and vegetable crops in the U.S. (Erwin and Ribeiro 1996). The gene sequences for each isolate were deposited in GenBank (accession Nos. MW315961, MW315962, MW315963, and MW315964). These four isolates were paired with known A1 and A2 isolates on super clarified V8 agar (Jeffers 2015), and all four were mating type A1. They also were sensitive to the fungicide mefenoxam at 100 ppm (Olson et al. 2013). To confirm pathogenicity, 4-week-old ‘Brandi Burgundy’ chrysanthemum plants were grown in 10-cm pots containing a peat potting medium. Plants (n = 7) were atomized with 1 ml of zoospore suspension containing 5 × 103 zoospores of each isolate. Control plants received sterile water. Plants were maintained at 100% RH for 24 h and then placed in a protected shade-structure where temperatures ranged from 19-32°C. All plants displayed symptoms of stem and foliage blight in 2-3 days. Symptoms that developed on infected plants were similar to those observed in the nursery. Several inoculated plants died, but stem blight, dieback, and foliar wilt were primarily observed. Disease severity averaged 50-60% on inoculated plants 15 days after inoculation. Control plants did not develop symptoms. The pathogen was consistently isolated from stems of symptomatic plants and verified as P. drechsleri based on morphology. The pathogenicity test was repeated with similar results. P. drechsleri has a broad host range (Erwin and Ribeiro 1996; Farr et al. 2021), including green beans (Phaseolus vulgaris), which are susceptible to seedling blight and pod rot in eastern Oklahoma. Previously, P. drechsleri has been reported on chrysanthemums in Argentina (Frezzi 1950), Pennsylvania (Molnar et al. 2020), and South Carolina (Camacho 2009). Chrysanthemums are widely grown in nurseries in the Midwest and other regions of the USA for local and national markets. This is the first report of P. drechsleri causing stem and foliage blight on chrysanthemum species in the United States. Identifying sources of primary inoculum may be necessary to limit economic loss from P. drechsleri.

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First Report of Phytophthora drechsleri Associated with Stem and Foliar Blight of Gynura bicolor in Taiwan

Plant Disease ◽

10.1094/pdis-12-10-0931 ◽

2011 ◽

Vol 95 (7) ◽

pp. 874-874 ◽

Cited By ~ 1

Author(s):

Y. M. Shen ◽

C. H. Chao ◽

H. L. Liu

Keyword(s):

Disease Incidence ◽

Morphological Characteristics ◽

Hyphal Growth ◽

Pathogenicity Test ◽

Distilled Water ◽

First Report ◽

Foliar Blight ◽

Dual Cultures ◽

Phytophthora Drechsleri ◽

Gynura Bicolor

Gynura bicolor (Roxb. ex Willd.) DC., known as Okinawa spinach or hong-feng-cai, is a commonly consumed vegetable in Asian countries. In May 2010, plants with blight and wilt symptoms were observed in commercial vegetable farms in Changhua, Taiwan. Light brown-to-black blight lesions developed from the top of the stems to the petioles and extended to the base of the leaves. Severely infected plants declined and eventually died. Disease incidence was approximately 20%. Samples of symptomatic tissues were surface sterilized in 0.6% NaOCl and plated on water agar. A Phytophthora sp. was consistently isolated and further plated on 10% unclarified V8 juice agar, with daily radial growths of 7.6, 8.6, 5.7, and 2.4 mm at 25, 30, 35, and 37°C, respectively. Four replicates were measured for each temperature. No hyphal growth was observed at 39°C. Intercalary hyphal swellings and proliferating sporangia were produced in culture plates flooded with sterile distilled water. Sporangia were nonpapillate, obpyriform to ellipsoid, base tapered or rounded, and 43.3 (27.5 to 59.3) × 27.6 (18.5 to 36.3) μm. Clamydospores and oospores were not observed. Oospores were present in dual cultures with an isolate of P. nicotianae (p731) (1) A2 mating type, indicating that the isolate was heterothallic. A portion of the internal transcribed spacer sequence was deposited in GenBank (Accession No. HQ717146). The sequence was 99% identical to that of P. drechsleri SCRP232 (ATCC46724) (3), a type isolate of the species. The pathogen was identified as P. drechsleri Tucker based on temperature growth, morphological characteristics, and ITS sequence homology (3). To evaluate pathogenicity, the isolated P. drechsleri was inoculated on greenhouse-potted G. bicolor plants. Inoculum was obtained by grinding two dishes of the pathogen cultured on potato dextrose agar (PDA) with sterile distilled water in a blender. After filtering through a gauze layer, the filtrate was aliquoted to 240 ml. The inoculum (approximately 180 sporangia/ml) was sprayed on 24 plants of G. bicolor. An equal number of plants treated with sterile PDA processed in the same way served as controls. After 1 week, incubation at an average temperature of 29°C, blight and wilt symptoms similar to those observed in the fields appeared on 12 inoculated plants. The pathogen was reisolated from the lesions of diseased stems and leaves, fulfilling Koch's postulates. The controls remained symptomless. The pathogenicity test was repeated once with similar results. G. bicolor in Taiwan has been recorded to be infected by P. cryptogea (1,2), a species that resembles P. drechsleri. The recorded isolates of P. cryptogea did not have a maximal growth temperature at or above 35°C (1,2), a distinctive characteristic to discriminate between the two species (3). To our knowledge, this is the first report of P. drechsleri being associated with stem and foliar blight of G. bicolor. References: (1) P. J. Ann. Plant Pathol. Bull. 5:146, 1996. (2) H. H. Ho et al. The Genus Phytophthora in Taiwan. Institute of Botany, Academia Sinica, Taipei, 1995. (3) R. Mostowfizadeh-Ghalamfarsa et al. Fungal Biol. 114:325, 2010.

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First Report of Podosphaera xanthii Causing Powdery Mildew on Mungbean (Vigna radiata) in Taiwan

Plant Disease ◽

10.1094/pdis-09-20-2092-pdn ◽

2021 ◽

Author(s):

Zong-ming Sheu ◽

Ming-hsueh Chiu ◽

Lawrence Kenyon

Keyword(s):

Powdery Mildew ◽

Vigna Radiata ◽

Morphological Characteristics ◽

Resistance Breeding ◽

Its Region ◽

Infected Leaf ◽

Pathogenicity Test ◽

Causal Organism ◽

First Report ◽

Powdery Mildew Pathogen

Mungbean (Vigna radiata L.) is routinely grown in the experimental fields at the headquarters of the World Vegetable Center (23°6'30.88"N, 120°17'51.31"E) for breeding, research and germplasm multiplication. In a spring 2016 mungbean trial, about 50% of the plants were affected with powdery mildew. The white, powdery-like patches first appeared on the upper leaf surfaces, and soon developed to grey patches on both sides of the leaves. Purple to brown discoloration appeared on the underside of the infected leaf. Microscopy examination revealed that the causal organism was not Erysiphe polygoni, which had previously been documented as the powdery mildew pathogen on mungbean in Taiwan (Hartman et al. 1993). The fungus produced typical structures of the powdery mildew Euoidium, anamorph of the genus Podosphaera. The mycelium consisted of septate, flexuous hyphae with indistinct appressoria. The erect conidiophores arising from superficial hyphae varied from straight or slightly curved to curled. Three to ten conidia were borne in long chains with crenate edges. Foot-cells were straight, cylindrical and measured 30 to 52 µm long. Conidia were hyaline, ellipsoid-ovoid to barrel-shaped, with fibrosin bodies, and measured 27 to 33 (mean = 30.4) × 15 to 20 (mean = 16.6) µm. Germ tubes were clavate and occasionally forked, and were produced from the lateral sites of the conidia. No chasmothecia were found in the samples. The morphological characteristics were consistent with P. xanthii (Castagne) U. Braun & Shishkoff (Braun & Cook 2012). To confirm the identity, the internal transcribed spacer (ITS) region of rDNA and partialβ-tubulin gene (TUB2) for the isolate MG3 were amplified with the primers ITS4/ITS5 (White et al. 1990) and BtuF5/BtuR7a (Ellingham et al. 2019), respectively. BLASTn analysis revealed the ITS sequence (MN833717) was 100% identical to many records of P. xanthii whereas the TUB2 sequence (MW363957) was 100% identical to a record of P. fusca (syn. P. xanthii; KC333362) in NCBI GenBank. A pathogenicity test was conducted by dusting conidia from an infected leaf onto six healthy four-week-old mungbean plants (cv ‘Tainan No. 3’). Another three plants were not inoculated and were used as control. All the plants were maintained in a greenhouse at 25 to 28°C. All inoculated plants developed powdery mildew symptoms after 10 days, whereas the control plants remained symptomless. To our knowledge, this is the first report of P. xanthii causing disease on mungbean in Taiwan. P. xanthii also has been reported on mungbean in Thailand (Meeboon et al. 2016), while other records referring to E. polygoni infecting Vigna spp. are from Brazil and Fiji (Farr & Rossman 2020). Although both P. xanthii and E. polygoni have now been reported as causing powdery mildew on mungbean in Taiwan, which species predominates or is more important remains unclear. A comprehensive survey with accurate species identification is required to develop effective management of the disease, particularly for resistance breeding.

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First report of Corynespora cassicola causing black spot on Sarcandra glabra in China

Plant Disease ◽

10.1094/pdis-02-21-0273-pdn ◽

2021 ◽

Author(s):

Jiang Ni ◽

B. R. Lin ◽

Lisha Song ◽

Guiyu Tan ◽

Jiang zhan Zhang ◽

...

Keyword(s):

Salvia Miltiorrhiza ◽

Pathogenic Fungus ◽

Morphological Characteristics ◽

Bootstrap Support ◽

Control Group ◽

Broad Host Range ◽

Pathogenicity Test ◽

Guangxi Province ◽

First Report ◽

Fungal Isolates

Sarcandra glabra is an important Chinese medicinal plant, which was widely cultivated under forest in south China. Guangxi province is the main producing areas of this herb. In June 2019, a serious leaf disease was found causing severe defoliation in the S. glabra plantation under bamboo forest in Rongan country, Guangxi province (109°13′N′′E). About 70% of the plants in the plantation (300 ha) showed the similar symptoms. Initially, circular lesions appeared on young leaves as black spots (about 1 to 2 mm). Then, the spots gradually enlarged usually with an obvious yellowish margin (6 to 8 mm). Finally, the lesions coalesced and formed irregular, black, and large necrotic areas, resulting in the leaf abscission. For pathogen isolation, small pieces of tissue (5×5 mm) taken from 25 diseased leaves were sterilized with 75% ethanol for 30 s, subsequently, soaked in 0.1% HgCl2 for 2 min, rinsed three times in sterile distilled water, dried, and then placed aseptically onto the potato dextrose agar (PDA) plates, and incubated at 28 °C (12 h/12 h light/dark). Three days later, the isolates were placed on a new PDA plate for subsequent purification and sporulation. 20 pure fungal isolates were obtained from single spores. Of which, 15 isolates showed similar morphological characteristics.The colonies on PDA were round, dense, gray edge and dark gray in center area. Conidia in culture were appeared light brown, cylindrical in shape, with 0 to 8 septa, and 55 to 165 μm × 5.2 to 13.5 μm in size (mean = 106.2 μm × 8.6 μm, n = 30). These morphological characteristics resemble those of Corynespora sp. (Berk. & M.A. Curtis) C.T. Wei (Ellis et al. 1971). A single-spore isolate (ZD5) was selected from the 15 fungal isolates for a subsequent molecular identification. The genes of internal transcribed spacer (ITS) of ribosomal DNA, β-tublin, and actin were amplified with the primer pairs ITS-1/ITS-4 (White et al. 1990), β-tubulin 2-Bt2a/Bt2b (Glass and Donaldson 1995), ACT-512F/ACT-783R (Carbone and Kohn 1999), respectively. And the ITS, β-tublin, and actin sequences were deposited in the GenBank database with the accession numbers MW362446, MW367029, and MW533122. Blast analysis and neighbor-joining analysis based on ITS, β-tublin, and actin sequences using MEGA 6 revealed that the isolate was placed in the same clade as C. cassicola with 100% bootstrap support. Pathogenicity test was performed on the two-year-old potted S. glabra. Six-mm-diameter mycelial plugs were attached to the healthy leaves of S. glabra for co-culture, while the control group was attached with PDA. All plants were covered with plastic bags for 2 days in order to maintain high humidity and cultured in a greenhouse at 28 °C with a 12-h/12-h light/dark cycle. The symptoms appeared 2 days after co-culture were identical to those observed in the field. The same fungus was re-isolated from the lesions, and further morphological characterization and molecular assays, as described above.The control leaves remained symptomless during the pathogenicity tests. According to the previous literatures, C. cassicola is a plant pathogenic fungus with a broad host range, which can damage diverse tropical plants including Salvia miltiorrhiza (Lu et al. 2019), Solanum americanum (Wagner and Louise 2019), Vitex rotundifolia (Yeh and Kirschner 2017), Cucumis sativus, Lycopersicon esculentum (Hsu et al. 2002), Carica papaya (Tsai et al. 2015),and so on. To our knowledge, this is the first report of C. cassicola causing leaf spot on S. glabra in China.

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First report of Cercospora cf. citrulina causing leaf spot of Ipomoea pes-caprae in China

Plant Disease ◽

10.1094/pdis-05-20-1081-pdn ◽

2020 ◽

Author(s):

Min Li ◽

Meijiao Hu ◽

Zhaoyin Gao ◽

Xiaoyu Hong ◽

Chao Zhao ◽

...

Keyword(s):

Molecular Identification ◽

Leaf Spot ◽

Plant Protection ◽

Morphological Characteristics ◽

Pathogenicity Test ◽

Conidial Suspension ◽

Xisha Islands ◽

First Report ◽

Initial Symptoms ◽

Leaf Surfaces

Ipomoea pes-caprae plays an important role in protecting the tropical and subtropical coastal beach of the world. In 2018, a leaf spot was observed on I. pes-caprae in Xisha islands of China, 13.2–25.8% of leaves were infected. The initial symptoms were small (1–3 mm diameter), single, circular, dark gray spots with a light-yellow center on the leaves. The lesions enlarged and were scattered or confluent, distinct and circular, subcircular or irregular, occasionally vein-limited, pale to dark gray-brown, with a narrow dark brown border surrounded by a diffuse yellow margin. Microscopic observations of the spots revealed that caespituli were dark brown and amphigenous, but abundant on the underside of the leaves. Mycelia were internal. Conidiophores were fasciculate, occasionally solitary, pale olivaceous-brown throughout, 0- to 3-septate, 27.9–115.8 (63.4±22.5) µm × 3.2–5.3 (4.3±0.87) µm (n=100). Conidial scars were conspicuously thickened. Conidia were solitary, hyaline, filiform, acicular to obclavate, straight to slightly curved, subacute to obtuse at the apex, truncate at the base, multi-septate, 21.0–125.5 (60.2±20.1) µm × 2.0–5.0 (3.8±0.83) µm (n=100). Single-conidium isolates were obtained from representative colonies grown on potato dextrose agar (PDA) incubated at 25℃ in the dark. The colonies grew slowly and were dense, white to gray and flat with aerial mycelium. Mycelia were initially white, and then became gray. Conidia were borne on the conidiophores directly. The pure isolate HTW-1 was selected for molecular identification and pathogenicity test, which were deposited in Microbiological Culture Collection Center of Environment and Plant Protection Institute, Chinese Academy of Tropical Agricultural Sciences. The internal transcribed spacer (ITS) region of rDNA, translation elongation factor 1-alpha (tef1) and histone H3 (his3) genes were amplified with ITS1/ITS4, EF-1 / EF-2, and CYLH3F / CYLH3R primers, respectively (Groenewald et al. 2013). The obtained sequences of HTW-1 were all deposited in GenBank with accession numbers MT410467 for ITS, MT418903 for tef1 and MT418904 for his3. The ITS, tef1 and his3 genes all showed 100% similarity for ITS (JX143582), tef1 (JX143340) and his3 (JX142602) with C. cf. citrulina (MUCC 588; MAFF 239409) from I. pes-caprae in Japan. Based on the morphological characteristics and molecular identification, the pathogen was identified as Cercospora cf. citrulina (Groenewald et al. 2013). The pathogenicity test was conducted by spraying conidial suspension (1×104 conidia/mL) on wounded and unwounded leaves for seedling of I. pes-caprae in greenhouse and in sterile vitro condition. The conidial suspension was prepared using conidia from 30-day-old culture grown on PDA at 25℃ in the dark. Leaf surfaces of seedling in greenhouse were wounded by lightly rubbing with a steel sponge and detached leaf surfaces were wounded by sterile needles. the treatments were sprayed with conidial suspensions on wounded and unwounded leaf surfaces. The control was sprayed with sterile water. After eight days, the typical symptoms of spots which were small, single, circular and dark gray appeared on the inoculated wounded leaves, while the inoculated unwounded leaves and the control leaves were symptomless. The pathogen was only re-isolated from the inoculated wounded leaves. The pathogen may be infected by wound. A total of 20 Cercospora and related species was found on Ipomoea spp. (García et al. 1996). Cercospora cf. citrulina has been reported on I. pes-caprae in Japan, although it was unclear if it was a pathogen or saprophyte (Groenewald et al. 2013). To our knowledge, this is the first report of C. cf. citrulina causing leaf spot of I. pes-caprae in China. This disease could threat the cultivation of I. pes-caprae in China.

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First Report of Powdery Mildew caused by Podosphaera xanthii on Euphorbia tithymaloides in Malaysia

Plant Disease ◽

10.1094/pdis-08-20-1700-pdn ◽

2020 ◽

Author(s):

Siti Izera Ismail ◽

Aziera Roslen

Keyword(s):

Powdery Mildew ◽

Sequence Data ◽

Morphological Characteristics ◽

Pathogenicity Test ◽

Podosphaera Xanthii ◽

Sequence Alignments ◽

Voucher Specimen ◽

First Report ◽

Initial Symptoms ◽

Young Leaves

Euphorbia tithymaloides L. (zig-zag plant) is a succulent, perennial shrub belonging to the Euphorbiaceae family and is widely cultivated in Malaysia for ornamental purposes and commercial values. In June 2019, typical symptoms of powdery mildew were observed on over 50% of the leaves of E. tithymaloides in a garden at Universiti Putra Malaysia, Serdang city of Selangor province, Malaysia. Initial symptoms included circular to irregular white powdery fungal colonies on both leaf surfaces and later covered the entire leaf surface. Severely infected leaves became necrotic, distorted and senesced. A voucher specimen Ma (PM001-Ma) was deposited in the Mycology laboratory, Faculty of Agriculture, UPM. Microscopic observation showed hyphae hyaline, branched, thin-walled, smooth, 3 to 6 µm wide with nipple-shaped appressoria. Conidiophores were straight, measured 30 to 90 μm long × 8 to 12 μm wide and composed of a cylindrical foot cell, 50 to 75 μm long. Conidia formed in chains were hyaline, ellipsoid to oval with fibrosin bodies, measured 25 to 36 × 16 to 20.1 μm in size and chasmothecia were not observed on the infected leaves. Genomic DNA was directly isolated from mycelia and conidia of isolate Ma using DNeasy Plant Mini Kit (Qiagen, USA). The universal primer pair ITS4/ITS5 of rDNA (White et al. 1990) was used for amplification and the resulting 569-bp sequence was deposited in GenBank (Accession no. MT704550). A BLAST nucleotide search revealed 100% similarity with that of Podosphaera xanthii on Momordica charantia wild from Taiwan (Accession no. KM505135) (Kirschner and Liu 2015). Both the morphological characteristics of the anamorph and ITS sequence data support the identification of this powdery mildew on E. tithymaloides as Podosphaera xanthii (Castagne) U. Braun & Shishkoff (Braun and Cook 2012). A pathogenicity test was conducted by gently pressing the infected leaves onto young leaves of five healthy potted plants. Five noninoculated plants were used as controls. The inoculated plants were maintained in a greenhouse at 25 ± 2°C and the test was repeated. Seven days after inoculation, white powdery symptoms were observed similar to those on the naturally infected leaves, while control plants remained asymptomatic. The fungus on the inoculated leaves was morphologically and molecularly identical to the fungus on the original specimens. Sequence alignments were made using MAFFT v.7.0 (Katoh et al. 2019) and a maximum likelihood phylogram was generated by MEGA v.7.0 (Kumar et al. 2016). Isolate Ma grouped in a strongly supported clade (100% bootstrap value) with the related species of P. xanthii available in GenBank based on the ITS region. Powdery mildew caused by P. xanthii has been reported as a damaging disease that can infect a broad range of plants worldwide (Farr and Rossman 2020). It also has been recently reported on Sonchus asper in China (Shi et al. 2020). According to our knowledge, this is the first report of powdery mildew caused by P. xanthii on E. tithymaloides worldwide. The occurrence of powdery mildew on E. tithymaloides could pose a serious threat to the health of this plant, resulting in death and premature senescence of young leaves.

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First Report of Vinca Blight Caused by Phytophthora nicotianae in Northwestern Mexico

Plant Disease ◽

10.1094/pdis-04-13-0400-pdn ◽

2013 ◽

Vol 97 (9) ◽

pp. 1257-1257 ◽

Cited By ~ 3

Author(s):

B. Alvarez-Rodriguez ◽

J. A. Ortiz-Meza ◽

I. Rojo-Baez ◽

I. Marquez-Zequera ◽

R. S. Garcia-Estrada ◽

...

Keyword(s):

Sequence Analysis ◽

Agar Medium ◽

Phytophthora Nicotianae ◽

Economic Losses ◽

Diseased Plant ◽

Pathogenicity Test ◽

First Report ◽

Foliar Blight ◽

Initial Symptoms ◽

Northwestern Mexico

Vinca (Catharantus roseus (L.) G. Don) is a common ornamental landscape plant. From July to September 2012, blighted and wilted vinca plants were found in retail stores, commercial nurseries, and urban landscape areas of Culiacan, Sinaloa, in northwestern Mexico. In several commercial nurseries and a retail store, incidence of the unknown disease on vinca plants ranged from 20 to 50%, resulting in significant economic losses. Symptoms of the disease started with a foliar blight, and if warm and wet conditions were present, the disease progressed, causing plant wilting and death. Surface-sterilized (0.5% NaOCl 1 min) diseased plant tissue was plated on V8 agar medium, and after 72 h of incubation at 25°C, white colonies of coenocytic mycelium were developed from the plated tissues. Isolates produced cottony colonies on V8 agar medium, grew well between 7 and 30°C (optimum of 25°C), and produced spherical, intercalary, and terminal chlamydospores (17 to 30 μm) and non caducous, papillate, spherical to ovoid sporangia of 30 to 39 × 21 to 31 μm. Based on these morphological characteristics, Phytophthora isolates were identified as Phytophthora nicotianae Breda de Haan (1,3). The identity of two representative isolates OV4 and OV11 was confirmed by sequence analysis of the rDNA internal transcribed spacers (ITS; GenBank Accession Nos. KC248202 and KC248201), and of the β-tubulin (β-tub; KC248404 and KC248403) and translation elongation factor 1-α (EF1-α; KC248206 and KC248205) genes. Comparative sequence analysis against the NCBI nucleotide database showed a high degree of identity with reference sequences of P. nicotianae (ITS, 99%; β-tub, 99%; EF1-α, 100%) (2). A pathogenicity test with a representative isolate of P. nicotianae was performed on 10-week-old healthy vinca seedlings (n = 10). An aliquot of 10 ml of a zoosporic suspension (104 zoospores/ml) was sprayed onto the seedlings' leaves. An equal number of non-inoculated control seedlings were sprayed with sterile distilled water. Seedlings were maintained in a moist chamber at 25°C with 80 to 90% relative humidity and watered as needed with sterile water. Inoculated plants showed initial symptoms of foliar blight after 4 days, whereas control plants remained healthy. Ten days after inoculation, inoculated plants showed severe wilting. P. nicotianae was reisolated only from inoculated plants, thus fulfilling Koch's postulates. To our knowledge, this is the first report of P. nicotianae attacking annual vinca in northwestern Mexico. References: (1) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phytopathological Society, St. Paul, MN, 1996. (2) L. P. Kroon et al. Fungal Genet Biol. 41:766, 2004. (3) F. N. Martin et al. Plant Dis. 96:1080, 2013.

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First Report of Hyphodermella rosae Causing Dry Fruit Rot Disease on Plum in Iran

Plant Disease ◽

10.1094/pdis-10-11-0825 ◽

2012 ◽

Vol 96 (8) ◽

pp. 1228-1228 ◽

Cited By ~ 1

Author(s):

M. Sayari ◽

V. Babaeizad ◽

M. A. T. Ghanbari ◽

H. Rahimian ◽

B. Borhani ◽

...

Keyword(s):

Sour Cherry ◽

Fruit Rot ◽

Pathogenicity Test ◽

Mazandaran Province ◽

Causal Organism ◽

Potato Dextrose Broth ◽

First Report ◽

Initial Symptoms ◽

Fruit Samples ◽

Dry Fruit

Plum (Prunus domestica) and peach (P. persica) are widely grown, often in alternate rows with citrus, in the Mazandaran Province of Iran. In June 2011, a dry fruit rot of plum was observed in several production regions in Mazandaran Province (35°47′N, 50°34′E). Initial symptoms at pit-hardening stage appeared as dark brown, circular, necrotic spots from 2 to 5 cm in diameter. They later developed into a dry fruit rot. Severe symptoms occurred during June and July when warm weather (temperature around 28°C) and high relative humidity (RH) (>85%) were present. Marketable yield losses reached 50% to almost 100% in many orchards. To isolate the causal organism, symptomatic fruits were surface disinfested for 1 min in 0.5% active chlorine, washed thoroughly with sterile distilled water, and segments were plated on potato dextrose agar (PDA) amended with 50 mg/liter of streptomycin sulfate and incubated at 25°C for 3 days. The fungus Hyphodermella rosae (Bresadola) Nakasone was consistently isolated (37 isolates from 79 samples) and identified on the basis of morphological characteristics on PDA. Basidiomata were effuse, resupinate, 15 × 10 mm, crustaceous, tubercules small with apical bristles, and light orange to grayish orange. Subhymenium was up to 30 μm thick, composed of vertically arranged, short-celled, nonagglutinated hyphae; subhymenial hyphae were 3 to 4 μm in diameter. Basidiospores were ellipsoid, 7.5 to 8.5 × 4.5 to 5.5 μm (100 determination), and their cell walls were thin, hyaline, and smooth (1). Genomic DNA was extracted from mycelium with a DNA extraction kit (Qiagen, Hilden, Germany) according to the manufacturer's directions and grown on potato dextrose broth for 4 days at 28°C. The rDNA region was amplified with the primers ITS4 (5′-TCCTCCGCTTATTGATATGC-3′) and ITS5 (5′- GGAAGTAAAAGTCGTAACAA-3′) (4) and the PCR product was sequenced. Nucleotide BLAST analysis of the amplified 627-bp fragment confirmed a 99% similarity with the sequence of H. rosae (GenBank Accession No. JN593086). A pathogenicity test was conducted with isolate MA4099 by placing 5-day-old mycelial plugs grown on PDA at the surface of healthy fruit (n = 6) incubated under >85% RH at 25°C for at least 4 days until the appearance of symptoms, which were similar to those displayed under orchard conditions. Control fruits, inoculated with blocks of PDA plugs, remained intact and symptomless. Reisolation from inoculated fruit samples consistently yielded the inoculated fungus, completing Koch's postulates. The genus Hyphodermella has been reported to be causing wood rot on apricot (2) and sweet and sour cherry (3). To our knowledge, this is the first report of H. rosae causing dry fruit rot on a stone fruit species in the world. References: (1) K. K. Nakasone. Mycologie, 29:231, 2008. (2) J. M. Ogawa et al. Diseases of Apricot (Prunus armeniaca L.). The American Phytopathological Society, St. Paul, MN, 2003. (3) J. K. Uyemoto et al. Diseases of Sweet Cherry (Prunus avium L.) and Sour Cherry (P. cerasus L.). IS-MPMInet, http://www.ismpminet.org/resources/common/comment/cherry.asp , accessed June 2012. (4) T. J. White et al. Page: 315 in: PCR Protocols: A Guide to Methods and Application. M.A. Innis et al., eds. Academic Press, San Diego, CA, 1990.

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First Report of Lasiodiplodia theobromae Causing Postharvest Fruit Rot on Guava (Psidium guajava) in Malaysia

Plant Disease ◽

10.1094/pdis-12-20-2732-pdn ◽

2021 ◽

Author(s):

Kar Yan Zee ◽

Norhayu Asib ◽

Siti Izera Ismail

Keyword(s):

Morphological Characteristics ◽

Its Region ◽

Psidium Guajava ◽

Fruit Rot ◽

Pathogenicity Test ◽

Tropical Fruit ◽

Lasiodiplodia Theobromae ◽

First Report ◽

Guava Fruit ◽

Initial Symptoms

Guava (Psidium guajava L.) is an economically important tropical fruit crop and is cultivated extensively in Malaysia. In September and October 2019, postharvest fruit rot symptoms were observed on 30% to 40% of guava fruit cv. Kampuchea in fruit markets of Puchong and Ipoh cities in the states of Selangor and Perak, Malaysia. Initial symptoms appeared as brown, irregular, water-soaked lesions on the upper portion of the fruit where it was attached to the peduncle. Subsequently, lesions then progressed to cover the whole fruit (Fig.1A). Lesions were covered with an abundance of black pycnidia and grayish mycelium. Ten symptomatic guava fruit were randomly collected from two local markets for our investigation. For fungal isolation, small fragments (5×5 mm) were excised from the lesion margin, surface sterilized with 0.5% NaOCl for 2 min, rinsed three times with sterile distilled water, placed on potato dextrose agar (PDA) and incubated at 25 °C with 12-h photoperiod for 2-3 days. Eight single-spore isolates with similar morphological characteristics were obtained and two representative isolates (P8 and S9) were characterized in depth. Colonies on PDA were initially composed of grayish-white aerial mycelium, but turned dark-gray after 7 days (Fig. 1B). Abundant black pycnidia were observed after incubation for 4 weeks. Immature conidia were hyaline, aseptate, ellipsoid, thick-walled, and mature conidia becoming dark brown and 1-septate with longitudinal striations, 25.0 − 27.0 ± 2.5 × 13.0 − 14.0 ± 1.0 μm (n = 30) (Fig.1C, D). On the basis of morphology, both representative isolates were identified as Lasiodiplodia theobromae (Pat.) Griffon & Maubl. (Alves et al. 2008). For molecular identification, genomic DNA of the two isolates was extracted using the DNeasy plant mini kit (Qiagen, USA). The internal transcribed spacer (ITS) region of rDNA and translation elongation factor 1-alpha (EF1-α) genes were amplified using ITS5/ITS4 and EF1-728F/EF1-986R primer set, respectively (White et al. 1990, Carbone and Kohn 1999). BLASTn analysis of the resulting ITS and EF1-α sequences indicated 100% identity to L. theobromae ex-type strain CBS 164.96 (GenBank accession nos: AY640255 and AY640258, respectively) (Phillips et al. 2013). The ITS (MW380428, MW380429) and EF1-α (MW387153, MW387154) sequences were deposited in GenBank. Phylogenetic analysis using the maximum likelihood based on the combined ITS-TEF sequences indicated that the isolates formed a strongly supported clade (100% bootstrap value) to the related L. theobromae (Kumar et al. 2016) (Fig.2). A pathogenicity test of two isolates was conducted on six healthy detached guava fruits per isolate. The fruit were surface sterilized using 70% ethanol and rinsed twice with sterile water prior inoculation. The fruit were wound-inoculated using a sterile needle according to the method of de Oliveira et al. (2014) and five-mm-diameter mycelial agar plugs from 7-days-old PDA culture of the isolates were placed onto the wounds. Six additional fruit were wound inoculated using sterile 5-mm-diameter PDA agar plugs to serve as controls. Inoculated fruit were placed in sterilized plastic container and incubated in a growth chamber at 25 ± 1 °C, 90% relative humidity with a photoperiod of 12-h. The experiment was conducted twice. Five days after inoculation, symptoms as described above developed on the inoculated sites and caused a fruit rot, while control treatment remained asymptomatic. L. theobromae was reisolated from all symptomatic tissues and confirmed by morphological characteristics and confirmed by PCR using ITS region. L. theobromae has recently been reported to cause fruit rot on rockmelon in Thailand (Suwannarach et al. 2020). To our knowledge, this is the first report of L. theobromae causing postharvest fruit rot on guava in Malaysia. The occurrence of this disease needs to be monitored as this disease can reduce the marketable yield of guava. Preventive strategies need to be developed in the field to reduce postharvest losses.

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First Report of Colletotrichum capsici Causing Anthracnose on Alocasia macrorrhizos in China

Plant Disease ◽

10.1094/pdis-06-20-1228-pdn ◽

2020 ◽

Author(s):

HaiYan Ben ◽

JianFei Huo ◽

YuRong Yao ◽

Wei Gao ◽

WanLi Wang ◽

...

Keyword(s):

Disease Incidence ◽

Morphological Characteristics ◽

Asia Pacific ◽

Stem Rot ◽

Perennial Herb ◽

Broad Host Range ◽

Distilled Water ◽

First Report ◽

Colletotrichum Capsici ◽

Initial Symptoms

Alocasia macrorrhizos (Linnaeus) G. Don is a perennial herb in the Araceae family. It is native to South Asia and the Asia-Pacific and has long been cultivated as it is an economically important medicinal and ornamental plant. During July 2012 and 2013, severe outbreaks of leaf spot and stem rot disease on this plant occurred in a greenhouse of Shunyi district, in Beijing, China (117°05’E, 40°13’N). The disease incidence was greater than 30%. The leaf spots first appeared as yellow dots. As lesions expanded, the symptoms were circular to subcircular, light brown lesions with darker brown edges, Around the lesions the leaf tissue was chlorotic causing the formation of a yellow halo (Suppl. Fig1). Initial symptoms on the stems were brown, round or fusiform spots . As the disease progressed, lesions enlarged and merged together. When humidity was high, black acervuli with grey brown cirrhus of conidia were rapidly produced in lesions. Infected plants eventually withered or collapsed from the stem rot (Suppl. Fig2). Infected tissues were surface-sterilized in 1% NaOCl for 1 min, washed three times with distilled water, and placed on potato dextrose agar (PDA). Colonies on PDA, growing at 25°C in darkness, showed grayish brown and grey brown conidial masses produced from acervuli with black seta (Suppl. Fig3). Acervuli (n=30) were dark brown to black and approximately round, 121 to 210 μm in diameter, averaging 166.5 μm (Suppl. Fig4). Setae (n=30) scattered in acervuli, black, septate, 94.4 to 128.4×3.4 to 4.7 μm, base inflated, and narrower toward the top (Suppl. Fig5). Conidiophores (n=50) were phialidic, hyaline, unicellular. Conidia (n=50) were hyaline, monospora, falcate, base obtuse, apices acute, and 20.5 to 24.7 ×2.8 to 3.4 μm (Suppl. Fig6). Six monoconidial isolates were made, and the morphological characteristics of the fungus were similar to those of Colletotrichum capsici (Syd.) Butler & Bisby (Mordue, 1971). In the greenhouse (25 to 30 °C, relative humidity 98%), pathogenicity tests were conducted by spraying a 106 spores /mL suspension on leaves and stems of 10 healthy potted A. macrorrhizos plants (3-year-old). A control was included that consisted of ten plants sprayed with sterile distilled water. All treated plants were covered with a plastic bag and removed 48 h later. After 12 days, all inoculated leaves and stems appeared with typical Anthracnose symptoms, whereas control plants remained healthy. The fungus was reisolated from diseased tissues, fulfilling Koch´s postulates. The ITS region of a representative isolate was amplified and sequenced using the primers ITS1/ITS4 (White et al. 1990).The obtained ITS sequence (GenBank Accession No. KJ018793.1) showed 100% similarity to Colletotrichum capsici (Accession No. HQ271469.1 and DQ454016.1). Colletotrichum capsici is synonymous to Colletotrichum truncatum. Colletotrichum capsici is a major phytopathogen with a broad host range which causes anthracnose disease. The first report of C. capsici as a pathogen of Alocasia macrorrhizos was reported in India in 1979 (Mathur, 1979). To our knowledge, this is the first record of C. capsici causing anthracnose on A. macrorrhizos in China.

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