First Report of Downy Mildew Caused by Peronospora belbahrii on Basil (Ocimum spp.) in Ontario

Basil (Ocimum spp.) is one of the most commercially significant fresh culinary herb crops worldwide. In Ontario, basil is grown both in the field and in the greenhouse. In the summer of 2011, basil plants grown in a research field at the Simcoe Research Station in Norfolk County, Ontario, Canada (44°15′N, 77°35′W), were infected with downy mildew. Infected leaves exhibited interveinal chlorotic lesions on the upper surface and clear to black sporulation on the abaxial leaf surfaces. Leaf senescence and defoliation occurred at high disease severity, which reduced marketable yield. Basil downy mildew symptoms were severe on leaves of cultivars Genovese and Sweet Basil, with 40 to 100% disease incidence. Based on morphological characteristics, the basil downy mildew causal agent was identified as Peronospora belbahrii Thines (4). Infected leaves were collected and microscopic observations of the sporulating lesions were carried out and the structures measured. Sporangiophores (n = 20) were hyaline with relatively long, straight trunks and were monopodially branched, with a length of 150 to 360 μm (average 285 μm). Sporangiophores ended with two slightly curved branchlets, the longer one measuring 15 to 27 μm (average 19 μm) and the shorter one 5 to 15 μm (average 9 μm). Sporangia (n = 50) were round, or slightly ovoid, olive to brown in color, and measured 29 × 25 μm (25 to 35 × 20 to 30 μm). Genomic DNA was extracted from 10 isolates and the nuclear ribosomal internal transcribed spacer (ITS) region was amplified with ITS4 and ITS5 primers and sequenced. The sequences of the 10 isolates were nearly identical. A BLAST search of the NCBI database with the ITS sequences (GenBank Accession No. KC756923) revealed a 98 to 100% similarity to the sequences of P. belbahrii (HQ730979, FJ436024, and HQ702191) isolated from sweet basil in Florida (3), California (1), and Hungary (2), respectively. To confirm pathogenicity, 5-week-old ‘Genovese’ seedlings were sprayed with a suspension of 1 × 105 sporangia/ml. Plants were kept in a growth chamber maintained at 23/18°C, 60 to 85% relative humidity, and 12/12 h light/dark. Non-inoculated plants served as controls. Basil downy mildew symptoms developed after 8 days on the inoculated plants and the pathogen was identified in association with symptoms consistent with downy mildew. The non-inoculated controls remained healthy. In North America, the occurrence of basil downy mildew has been reported since 2007 (3) and the disease has spread into several U.S. states. To our knowledge, this is the first report of downy mildew on sweet basil in Canada. References: (1) C. L. Blomquist et al. Plant Dis. 93:968, 2009. (2) G. Nagy and A. Horvath. Plant Dis. 95:1034, 2011. (3) P. D. Roberts et al. Plant Dis. 98:199, 2009. (4) M. Thines et al. Mycol. Res. 113:532, 2009.

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First Report of Downy Mildew Caused by Peronospora belbahrii on Sweet Basil (Ocimum basilicum) in Cyprus

Plant Disease ◽

10.1094/pdis-07-13-0759-pdn ◽

2014 ◽

Vol 98 (2) ◽

pp. 283-283 ◽

Cited By ~ 5

Author(s):

L. Kanetis ◽

A. Vasiliou ◽

G. Neophytou ◽

S. Samouel ◽

D. Tsaltas

Keyword(s):

Downy Mildew ◽

Disease Incidence ◽

Infected Plant ◽

Morphological Characteristics ◽

Ocimum Basilicum ◽

First Report ◽

Consensus Sequences ◽

Sweet Basil ◽

Plastic Bags ◽

Necrotic Spots

Sweet basil (Ocimum basilicum L.) is an economically important annual aromatic plant, grown mostly for culinary use for both fresh and dry consumption and as a source of essential oil. In Cyprus, approximately 4 ha are grown annually, either in greenhouses as a year-round crop or in open fields from April to November, and the majority of the production is exported to the European market. During May 2012, a sweet basil cv. Genovese Gigante greenhouse operation in the area of Limassol was severely affected by a foliar disease, causing almost 100% crop losses. Within a few days, a similar, heavy disease incidence was also reported from a nearby greenhouse facility on the Genovese-type cultivars Superbo, Aroma 2, and Bonazza, as well as on Thai basil (O. basilicum var. thyrsiflorum). Successively, destructive hits of similar symptomatology have been reported from other areas and since then the disease appears to have been well-established in the country, causing major economic damages. It is also noteworthy to mention that in greenhouse infections the disease remains active even during winter, considering the mild environmental conditions and the monoculture fashion followed. Symptoms appeared on the leaves initially as interveinal, zonal, chlorotic lesions, followed by the appearance of a fuzzy, purplish sporulation on the abaxial side. Progressively, infected leaves curled and sporadic necrotic spots were evident and finally abscised. Light microscopic examination of infected samples revealed the presence of straight, hyaline sporangiophores (n = 15) typical of downy mildew, 210 to 590 μm long (mean = 350.7 μm; SD ± 117.5 μm) × 12 to 15 μm wide (mean = 13.1 μm; SD ± 1.4 μm). Sporangiophores were monopodially branched three to five times, terminating with curved branchlets bearing single sporangia at their tips. The sporangia (n = 25) were purplish-grey, ovoid to subglobose, and measured 32 to 22 μm in length (mean = 27.2 μm; SD ± 2.8 μm) and 30 to 10 μm in breadth (mean = 21.7 μm; SD ± 4.8 μm). Based on these morphological characteristics, the causal agent was identified as Peronospora belbahrii Thines (1,4). Furthermore, genomic DNA was extracted from infected plant tissue from eight different samples according to Dellaporta et al. (2). The complete ITS rDNA region was amplified and sequenced using primers ITS5 and ITS4 (3). Two of the consensus sequences were deposited in GenBank (Accession Nos. KF419289 and KF419290) and a BLAST analysis in the NCBI database revealed 99% similarity to all of the P. belbahrii sequences and other Peronospora sp. previously reported on sweet basil (Accession Nos. AY831719, DQ479408, FJ394336, and FJ436024). In a pathogenicity trial, five 40-day-old potted sweet basil plants were spray-inoculated with a sporangial suspension (1 × 105 sporangia/ml) until runoff, bagged for 24 h, and placed in a growth chamber at 18°C. Subsequently, the plastic bags were removed and the plants were kept at 22°C with a 16-h photoperiod and 80% relative humidity. Additionally, five plants were water-sprayed and served as controls. Typical downy mildew symptoms appeared 6 to 8 days after inoculation, while the uninoculated plants remained disease-free. To our knowledge, this is first report of downy mildew on sweet basil in Cyprus. References: (1) L. Belbahri et al. Mycol. Res. 109:1276, 2005. (2) S. L. Dellaporta et al. Plant Mol. Biol. Rep., 1:19, 1983. (3) G. Nagy and A. Horvat, Plant Dis. 93:1999, 2009. (4) M. Thines et al. Mycol. Res. 113:532, 2009.

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First Report of Basil Downy Mildew Caused by Peronospora belbahrii in the Czech Republic

Plant Disease ◽

10.1094/pdis-06-14-0613-pdn ◽

2015 ◽

Vol 99 (3) ◽

pp. 418-418 ◽

Cited By ~ 3

Author(s):

I. Petrželová ◽

M. Kitner ◽

I. Doležalová ◽

V. Ondřej ◽

A. Lebeda

Keyword(s):

Czech Republic ◽

Downy Mildew ◽

Morphological Characteristics ◽

Its Region ◽

Width Ratio ◽

Natural Occurrence ◽

The Czech Republic ◽

First Report ◽

Sweet Basil ◽

The World

Sweet basil (Ocimum basilicum L.) is an annual aromatic and medicinal plant in the Lamiaceae that is originally native to India but is grown in warm regions all over the world. It is a popular culinary herb used fresh and dried, and is used in traditional folk medicine. In the Czech Republic, sweet basil is grown commercially in South Moravia or by home gardeners as a potted plant. In 2012, severe downy mildew was observed in a field of basil plants (cv. Dark Green) at the Crop Research Institute (CRI) in Olomouc, Czech Republic. Infected leaves each exhibited large, interveinal, chlorotic lesions, and violet-gray, fuzzy growth on the lower leaf surface. Within a few days, lesions turned necrotic and severely infected leaves dropped prematurely. Microscopic observations revealed hyaline conidiophores typical of Peronospora Corda, emerging from stomata. Conidiophores (n = 100) were usually 239.9 to 296.5 × 8.7 to 10.6 μm, straight, and were branched 4 or 5 times submonopodially at the upper ends. Ultimate branchlets (n = 100) were slightly curved and obtuse, with the longer branchlets usually 17.8 to 22.7 μm and the shorter branchlets 10.0 to 12.9 μm, and each bearing a single conidium. Conidia (n = 100) were olive-brown, mostly ellipsoidal to subglobose, and typically 29.0 to 31.0 × 23.2 to 25.4 μm, with a length/width ratio of 1.2 to 1.3. Oospores were not observed. Based on these morphological characteristics, the pathogen was identified as Peronospora belbahrii Thines (5). The specimen was deposited in a local herbarium at the CRI in Olomouc, as voucher PB-1. Genomic DNA was extracted from conidia, and the internal transcribed spacer (ITS) region of ribosomal DNA (rDNA) amplified with primers DC-6 (1) and LR-0 (4). A sequence was deposited in the NCBI database (GenBank Accession No. KJ960193). A BLAST search of the NCBI database revealed 99% identity to the deposited ITS sequences of P. belbahrii from basil and other host species (EU863410, FJ394334-7, GQ390794, GQ390795, HM462241, HM462242, HM486901, HQ702191, HQ730979, KC756923, KF419289, and KF419290). P. belbahrii was first described by Thines et al. (5) as a pathogen of sweet basil and coleus (Solenostemon scutellarioides), but can also infect Agastache spp. (2). There are many reports indicating the pathogen is spreading throughout the world (5). In Europe, chronologically, basil downy mildew has been reported from Italy, France, Switzerland, Germany, Hungary, and Cyprus (2,3,5). To our knowledge, this is the first report of natural occurrence of downy mildew on sweet basil in the Czech Republic. References: (1) D. E. L. Cooke et al. Fung. Genet. Biol. 30:17, 2000. (2) D. F. Farr and A. Y. Rossman. Fungal Databases. Systematic Mycology and Microbiology Laboratory, USDA ARS. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ , 16 June 2014. (3) A. Garibaldi et al. Plant Dis. 89:683, 2005. (4) O. Spring et al. Eur. J. Plant Pathol. 114:309, 2006. (5) M. Thines et al. Mycol. Res. 113:532, 2009.

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

Plant Disease ◽

10.1094/pdis-05-12-0436-pdn ◽

2012 ◽

Vol 96 (10) ◽

pp. 1580-1580

Author(s):

J. H. Park ◽

K. S. Han ◽

J. Y. Kim ◽

H. D. Shin

Keyword(s):

Leaf Spot ◽

Morphological Characteristics ◽

Its Region ◽

Culture Collection ◽

Distilled Water ◽

First Report ◽

Sweet Basil ◽

Organic Farm ◽

Leaf Spots ◽

Close Phylogenetic Relationship

Sweet basil, Ocimum basilicum L., is a fragrant herb belonging to the family Lamiaceae. Originated in India 5,000 years ago, sweet basil plays a significant role in diverse cuisines across the world, especially in Asian and Italian cooking. In October 2008, hundreds of plants showing symptoms of leaf spot with nearly 100% incidence were found in polyethylene tunnels at an organic farm in Icheon, Korea. Leaf spots were circular to subcircular, water-soaked, dark brown with grayish center, and reached 10 mm or more in diameter. Diseased leaves defoliated prematurely. The damage purportedly due to this disease has reappeared every year with confirmation of the causal agent made again in 2011. A cercosporoid fungus was consistently associated with disease symptoms. Stromata were brown, consisting of brown cells, and 10 to 40 μm in width. Conidiophores were fasciculate (n = 2 to 10), olivaceous brown, paler upwards, straight to mildly curved, not geniculate in shorter ones or one to two times geniculate in longer ones, 40 to 200 μm long, occasionally reaching up to 350 μm long, 3.5 to 6 μm wide, and two- to six-septate. Conidia were hyaline, acicular to cylindric, straight in shorter ones, flexuous to curved in longer ones, truncate to obconically truncate at the base, three- to 16-septate, and 50 to 300 × 3.5 to 4.5 μm. Morphological characteristics of the fungus were consistent with the previous reports of Cercospora guatemalensis A.S. Mull. & Chupp (1,3). Voucher specimens were housed at Korea University herbarium (KUS). An isolate from KUS-F23757 was deposited in the Korean Agricultural Culture Collection (Accession No. KACC43980). Fungal DNA was extracted with DNeasy Plant Mini DNA Extraction Kits (Qiagen Inc., Valencia, CA). The complete internal transcribed spacer (ITS) region of rDNA was amplified with the primers ITS1/ITS4 and sequenced. The resulting sequence of 548 bp was deposited in GenBank (Accession No. JQ995781). This showed >99% similarity with sequences of many Cercospora species, indicating their close phylogenetic relationship. Isolate of KACC43980 was used in the pathogenicity tests. Hyphal suspensions were prepared by grinding 3-week-old colonies grown on PDA with distilled water using a mortar and pestle. Five plants were inoculated with hyphal suspensions and five plants were sprayed with sterile distilled water. The plants were covered with plastic bags to maintain a relative humidity of 100% for 24 h and then transferred to a 25 ± 2°C greenhouse with a 12-h photoperiod. Typical symptoms of necrotic spots appeared on the inoculated leaves 6 days after inoculation, and were identical to the ones observed in the field. C. guatemalensis was reisolated from symptomatic leaf tissues, confirming Koch's postulates. No symptoms were observed on control plants. Previously, the disease was reported in Malawi, India, China, and Japan (2,3), but not in Korea. To our knowledge, this is the first report of C. guatemalensis on sweet basil in Korea. Since farming of sweet basil has recently started on a commercial scale in Korea, the disease poses a serious threat to safe production of this herb, especially in organic farming. References: (1) C. Chupp. A Monograph of the Fungus Genus Cercospora. Ithaca, NY, 1953. (2) D. F. Farr and A. Y. Rossman. Fungal Databases. Systematic Mycology & Microbiology Laboratory, ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ , May 5, 2012. (3) J. Nishikawa et al. J. Gen. Plant Pathol. 68:46, 2002.

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First Report of Fruit Rot of Melon Caused by Fusarium asiaticum in China

Plant Disease ◽

10.1094/pdis-08-20-1857-pdn ◽

2020 ◽

Author(s):

Fangmin Hao ◽

Quanyu Zang ◽

Weihong Ding ◽

Erlei Ma ◽

Yunping Huang ◽

...

Keyword(s):

Disease Incidence ◽

Elongation Factor ◽

Morphological Characteristics ◽

Its Region ◽

Fruit Rot ◽

Post Inoculation ◽

Basal Cells ◽

First Report ◽

Fusarium Asiaticum ◽

Sterile Needle

Melon (Cucumis melo L.) is a member of the Cucurbitaceae family, an important economical and horticultural crop, which is widely grown in China. In May 2020, fruit rot disease with water-soaked lesions and pink molds on cantaloupe melons was observed in several greenhouses with 50% disease incidence in Ningbo, Zhejiang Province in China. In order to know the causal agent, diseased fruits were cut into pieces, surface sterilized for 1 min with 1% sodium hypochlorite (NaClO), 2 min with 75% ethyl alcohol, rinsed in sterile distilled water three times (Zhou et al. 2018), and then placed on potato dextrose agar (PDA) medium amended with streptomycin sulfate (100 μg/ml) plates at 25°C for 4 days. The growing hyphae were transferred to new PDA plates using the hyphal tip method, putative Fusarium colonies were purified by single-sporing. Twenty-five fungal isolates were obtained and formed red colonies with white aerial mycelia at 25°C for 7 days, which were identified as Fusarium isolates based on the morphological characteristics and microscopic examination. The average radial mycelial growth rate of Fusarium isolate Fa-25 was 11.44 mm/day at 25°C in the dark on PDA. Macroconidia were stout with curved apical and basal cells, usually with 4 to 6 septa, and 29.5 to 44.2 × 3.7 to 5.2 μm on Spezieller Nährstoffarmer agar (SNA) medium at 25°C for 10 days (Leslie and Summerell 2006). To identify the species, the internal transcribed spacer (ITS) region and translational elongation factor 1-alpha (TEF1-α) gene of the isolates were amplified and cloned. ITS and TEF1-α was amplified using primers ITS1/ITS4 and EF1/EF2 (O’Donnell et al. 1998), respectively. Sequences of ITS (545 bp, GenBank Accession No. MT811812) and TEF1-α (707 bp, GenBank Acc. No. MT856659) for isolate Fa-25 were 100% and 99.72% identical to those of F. asiaticum strains MSBL-4 (ITS, GenBank Acc. MT322117.1) and Daya350-3 (TEF1-α, GenBank Acc. KT380124.1) in GenBank, respectively. A phylogenetic tree was established based on the TEF1-α sequences of Fa-25 and other Fusarium spp., and Fa-25 was clustered with F. asiaticum. Thus, both morphological and molecular characterizations supported the isolate as F. asiaticum. To confirm the pathogenicity, mycelium agar plugs (6 mm in diameter) removed from the colony margin of a 2-day-old culture of strain Fa-25 were used to inoculate melon fruits. Before inoculation, healthy melon fruits were selected, soaked in 2% NaClO solution for 2 min, and washed in sterile water. After wounding the melon fruits with a sterile needle, the fruits were inoculated by placing mycelium agar plugs on the wounds, and mock inoculation with mycelium-free PDA plugs was used as control. Five fruits were used in each treatment. The inoculated and mock-inoculated fruits were incubated at 25°C with high relative humidity. Symptoms were observed on all inoculated melon fruits 10 days post inoculation, which were similar to those naturally infected fruits, whereas the mock-inoculated fruits remained symptomless. The fungus re-isolated from the diseased fruits resembled colony morphology of the original isolate. The experiment was conducted three times and produced the same results. To our knowledge, this is the first report of fruit rot of melon caused by F. asiaticum in China.

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First Report of Leaf Blight Caused by Septoria allii on Garlic Chives in Korea

Plant Disease ◽

10.1094/pdis-08-12-0809-pdn ◽

2013 ◽

Vol 97 (1) ◽

pp. 147-147

Author(s):

J. H. Park ◽

S. E. Cho ◽

K. S. Han ◽

H. D. Shin

Keyword(s):

Disease Incidence ◽

Morphological Characteristics ◽

Its Region ◽

Leaf Blight ◽

Plant Diseases ◽

Cultivated Plants ◽

Sequence Information ◽

Conidial Suspension ◽

Korean Society ◽

First Report

Garlic chives, Allium tuberosum Roth., are widely cultivated in Asia and are the fourth most important Allium crop in Korea. In June 2011, a leaf blight of garlic chives associated with a Septoria spp. was observed on an organic farm in Hongcheon County, Korea. Similar symptoms were also found in fields within Samcheok City and Yangku County of Korea during the 2011 and 2012 seasons. Disease incidence (percentage of plants affected) was 5 to 10% in organic farms surveyed. Diseased voucher specimens (n = 5) were deposited at the Korea University Herbarium (KUS). The disease first appeared as yellowish specks on leaves, expanding to cause a leaf tip dieback. Half of the leaves may be diseased within a week, especially during wet weather. Pycnidia were directly observed in leaf lesions. Pycnidia were amphigenous, but mostly epigenous, scattered, dark brown to rusty brown, globose, embedded in host tissue or partly erumpent, separate, unilocular, 50 to 150 μm in diameter, with ostioles of 20 to 40 μm in diameter. Conidia were acicular, straight to sub-straight, truncate at the base, obtuse at the apex, hyaline, aguttulate, 22 to 44 × 1.8 to 3 μm, mostly 3-septate, occasionally 1- or 2-septate. These morphological characteristics matched those of Septoria allii Moesz, which is differentiated from S. alliacea on conidial dimensions (50 to 60 μm long) (1,2). A monoconidial isolate was cultured on potato dextrose agar (PDA). Two isolates have been deposited in the Korean Agricultural Culture Collection (Accession Nos. KACC46119 and 46688). Genomic DNA was extracted using the DNeasy Plant Mini DNA Extraction Kit (Qiagen Inc., Valencia, CA). The internal transcribed spacer (ITS) region of rDNA was amplified using the ITS1/ITS4 primers and sequenced. The resulting sequence of 482-bp was deposited in GenBank (JX531648 and JX531649). ITS sequence information was at least 99% similar to those of many Septoria species, however no information was available for S. allii. Pathogenicity was tested by spraying leaves of three potted young plants with a conidial suspension (2 × 105 conidia/ml), which was harvested from a 4-week-old culture on PDA. Control leaves were sprayed with sterile water. The plants were placed in humid chambers (relative humidity 100%) for the first 48 h. After 7 days, typical leaf blight symptoms started to develop on the leaves of inoculated plants. S. allii was reisolated from the lesions of inoculated plants, confirming Koch's postulates. No symptoms were observed on control plants. The host-parasite association of A. tuberosum and S. allii has been known only from China (1). S. alliacea has been recorded on several species of Allium, e.g. A. cepa, A. chinense, A. fistulosum, and A. tuberosum from Japan (4) and A. cepa from Korea (3). To the best of our knowledge, this is the first report of S. allii on garlic chives. No diseased plants were observed in commercial fields of garlic chives which involved regular application of fungicides. The disease therefore seems to be limited to organic garlic chive production. References: (1) P. K. Chi et al. Fungous Diseases on Cultivated Plants of Jilin Province, Science Press, Beijing, China, 1966. (2) P. A. Saccardo. Sylloge Fungorum Omnium Hucusque Congnitorum. XXV. Berlin, 1931. (3) The Korean Society of Plant Pathology. List of Plant Diseases in Korea, Suwon, Korea, 2009. (4) The Phytopathological Society of Japan. Common Names of Plant Diseases in Japan, Tokyo, Japan, 2000.

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First Report of Downy Mildew on Sweet Basil Caused By Peronospora belbahrii in India

Plant Disease ◽

10.1094/pdis-03-21-0621-pdn ◽

2021 ◽

Author(s):

Sujata Singh Yadav ◽

Priyanka Suryavanshi ◽

Indrajeet Nishad ◽

Soumya Sinha

Keyword(s):

Downy Mildew ◽

Disease Incidence ◽

Consensus Sequence ◽

Effective Control ◽

Broad Host Range ◽

Internal Transcribed Spacer Region ◽

Foliar Disease ◽

First Report ◽

Sweet Basil ◽

Initial Symptoms

Sweet basil (Ocimum basilicum L.; Family Lamiaceae) is an annual aromatic and medicinal plant grown in tropical and subtropical regions of the world. In India, it is cultivated as a commercial crop on ~8,000 ha. Aerial plant parts and essential oil of sweet basil are used in pharmaceutical, perfumery, food industries and in different formulations of traditional Ayurvedic and Unani medicines (Shahrajabian et al. 2020). The leaves have the highest concentrations of secondary metabolites such as terpenes and phenylpropanoids which provide the distinctive aroma (Viuda-Martos et al. 2011). During October 2020, severe foliar disease was observed in experimental fields of sweet basil at Council of Scientific and Industrial Research (CSIR)-Central Institute of Medicinal and Aromatic Plants (CIMAP) in Lucknow, India. Initial symptoms included large, interveinal chlorotic lesions on the adaxial surface of the leaves and black sporulation on the abaxial surface. Within a few days, the abaxial side of leaves turned necrotic, and leaf senescence and defoliation occurred on plants with severe symptoms. Disease incidence was 20 to 30% of plants. The pathogen was characterized morphologically using a light microscope. Sporangiophores were hyaline, dichotomously branched, 186.9 to 423.07 × 6.85 to 9.06 µm and, branched 3 to 5 times with each branch, terminating in two slightly curved branchlets, the longer one 7.05 to 25.31 µm and the shorter one 4.98 to 15.92 µm. Each branchlet had a single sporangium at the tip. Conidia were ellipsoidal to sub-globose, olive-brown in color, and typically measured 25.21 to 33.86 × 17.92 to 26.24 µm, each, without a pedicel. Based on these morphological characteristics, the foliar disease was identified as downy mildew was caused by Peronospora belbahrii (Thines et al. 2009). Eight symptomatic and two asymptomatic plant samples were collected from different locations in the field, and genomic DNA was extracted from the conidia of the eight naturally infected tissues of sweet basil samples as well as leaf tissues from two asymptomatic plants, using the CTAB method. The internal transcribed spacer region was amplified using ITS1 and ITS4 primers. Only eight infected samples amplified products of expected size (~ 700 bp) and two asymptomatic samples showed no amplification. Only five amplified PCR products were sequenced (White et al. 1990). All five sequences were identical and were a 98.1% match with five P. belbahrii isolates (MN450330.1, MN308051.1, MH620351.1, KJ960193, and MF693898). The consensus sequence was deposited into the NCBI database (GenBank Accession No. MW689257). Downy mildew caused by P. belbahrii previously has been reported on sweet basil from several countries (Wyenandt et al. 2015). To confirm the pathogenicity of these isolates on sweet basil (cv. CIM-Saumya), 25 - day-old sweet basil plants were sprayed with a suspension (1 × 105 sporangia/ml) of P. belbahrii. All plants were kept in a growth chamber with a 23/18°C diurnal cycle with 65 to 85% relative humidity for 24 h. Non-inoculated plants treated with sterile water served as a control treatment. After 8 days, typical symptoms of downy mildew appeared on all the inoculated plants while non-inoculated plants remained asymptomatic. Inoculated leaves with symptoms consistent of downy mildew were collected and the causal agent again identified as P. belbahrii on the basis of microscopic examination and ITS rDNA sequence data. To our knowledge, this is the first report of downy mildew caused by P. belbahrii on sweet basil in India. The pathogen has a broad host range and may pose a serious threat to the cultivation of this valuable crop in India. Thus, it is pertinent to develop effective control measures to avoid further spread and mitigate economic loss. References: Shahrajabian, M. H., et al. 2020. Int. J. Food Prop. 23:1961-1970. Wyenandt, C. A., et al. 2015. Phytopathology 105:885. Thines, M., et al. 2009. Mycol. Res. 113:532. White, T. J., et al. 1990. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Viuda-Martos, M., et al. 2011. Food Control. 22:1715.

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First Report of Downy Mildew Caused by Peronospora sp. on Basil (Ocimum basilicum) in France

Plant Disease ◽

10.1094/pd-89-0683c ◽

2005 ◽

Vol 89 (6) ◽

pp. 683-683 ◽

Cited By ~ 25

Author(s):

A. Garibaldi ◽

A. Minuto ◽

M. L. Gullino

Keyword(s):

Downy Mildew ◽

Leaf Surface ◽

Morphological Characteristics ◽

Sprinkler Irrigation ◽

Ocimum Basilicum ◽

Central Vein ◽

Pathogenicity Test ◽

First Report ◽

Sweet Basil ◽

Mediterranean Countries

Sweet basil (Ocimum basilicum) is an economically important herb in several Mediterranean countries. Approximately 30 ha are grown annually in France for fresh and processed consumption. During the spring and fall of 2004, a damaging foliar disease was observed in some crops near Saint Tropez in the French Riviera Region. More than 50% of plants were affected in an organically produced field-grown crop at an altitude of 250 m. Leaves of infected plants were initially slightly chlorotic, especially near the central vein. Within 2 to 3 days, a characteristic gray, furry growth was evident on the lower leaf surface and sometimes on the upper leaf surface. The appearance and severity of the disease was affected by overhead sprinkler irrigation. Basal leaves were severely affected. Microscopic observations revealed sporangiophores branching two to seven times. Sporangiophores, with a length of 250 to 500 μm (average 350 μm), ended with sterigmata bearing single sporangia. Sporangia measured 15 to 25 × 20 to 35 μm (average 22 × 28 μm), were elliptical and grayish in mass. The pathogen was identified as Peronospora sp. on the basis of its morphological characteristics (4). Pathogenicity was confirmed by inoculating leaves of 40-day-old healthy plants with a sporangial suspension (1 × 105 conidia/ml). Three containers with 150 plants each of O. basilicum cv Genovese gigante were used as replicates. Noninoculated plants served as controls. Plants were maintained in a growth chamber at 20°C (12 h of light per day) and 90 to 95% relative humidity. The pathogenicity test was carried out twice. After 6 days, typical symptoms of downy mildew developed on the inoculated plants, and Peronospora sp. was observed on the leaves. Noninoculated plants did not show symptoms. To our knowledge, this is the first report of Peronospora sp. on basil in France. Peronospora sp. was previously reported on sweet basil in Italy (1) and P. lamii on sweet basil in Uganda (3). Seed transmission (2) is suspected as the reason for recent outbreaks in Europe. References: (1) A. Garibaldi et al. Plant Dis. 88:312, 2004. (2) A. Garibaldi et al. Z. Pflanzenkr. Pflanzenschutz 111:465, 2004 (3) C. G. Hansford. Rev. Appl. Mycol. 12:421, 1933. (4) D. M. Spencer. The Downy Mildews. Academic Press, NY, 1981.

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First Report of Colletotrichum godetiae Causing Anthracnose and Twig Blight on Persian Walnut in Hungary

Plant Disease ◽

10.1094/pdis-03-20-0607-pdn ◽

2020 ◽

Author(s):

Virág Varjas ◽

Tamás Lakatos ◽

Tímea Tóth ◽

Csilla Kovács

Keyword(s):

Disease Incidence ◽

Juglans Regia ◽

Morphological Characteristics ◽

Unknown Origin ◽

Its Region ◽

Colletotrichum Acutatum ◽

Width Ratio ◽

Single Spore ◽

First Report ◽

Persian Walnut

Persian walnut (Juglans regia L.) fruit with preharvest anthracnose symptoms, necrotic fruit stalks, and twigs with necrotic buds, and peaks were collected in a Hungarian orchard next to Nágocs, in September 2018. Disease incidence was approximately 15% on a Hungarian bred walnut cultivar ‘Milotai 10’. Similar symptoms were found on Persian walnut in other locations (eg. Milota, Érd, Sarród, and Kocs). Acervuli were observed on necrotic lesions on fruit, and twigs with pale orange conidial masses. Conidia were hyaline, unicellular, and fusiform. Morphometric measurements of conidia showed mean length ± SD × width ± SD = 15.9 ± 1.7 × 4.5 ± 0.4 μm, length/width ratio 1:0.3 (n=100). The fungus was isolated from conidial masses on potato dextrose agar (PDA) medium amended with Chlorampenicol (25 mg/L). A total of 12 isolates were obtained as pure cultures by single-spore isolations and incubated at 23°C in dark for 10 days. The colonies were white to gray or grayish-orange on the upper side and with black spots on the reverse side. The isolates showed morphological characteristics of Colletotrichum acutatum in sensu lato (Jayawardena et al. 2016). Molecular analyses were conducted to identify the exact species. Internal transcribed spacer (ITS) region, actin (ACT), and calmodulin (CAL) partial genes were amplified by ITS1F/ITS4R, ACT512F/ACT783R and CAL1/CAL2 primers (White at al. 1990, Carbone and Kohn 1999, O’Donnell et al. 2000). The sequences of ITS region (GenBank Accession Nos: MK367398-99, MK367401-02) showed 100% identity with C. godetiae sequence. Based on ACT gene (GenBank Accession Nos: MK415991-92, MK415994-95) were 100% identity with the deposited C. godetiae type strains from walnut. The obtained sequences of CAL gene (GenBank Accession Nos: MK415998-99, MK416001-02) were same and showed 100% with other C. godetiae sequences from other host plants. The fungus was identified as Colletotrichum godetiae Neerg. Pathogenicity tests were accomplished in the field and under laboratory conditions (25°C on thermostat) on 10 green ‘Milotai 10’ walnut fruit, and 10 walnut twigs each. Tests were conducted on living trees, collected fruit, and two-year-old twigs by inserting mycelial agar plugs (5 mm in diameter) onto wounded pericarp tissues, which were then wrapped with wet cotton and parafilm. Wounded tissues on 5 fruit and 5 two-year-old twigs were treated with non-colonized PDA plugs as noninoculated controls. After 14 d necrotic lesions 9 to 17 mm in diameter developed on fruit on living trees. Lengths of 12 to 17 mm and width of 7 to 12 mm necrosis was measured on phloem of walnut twigs, and almost two times larger in cambium. No necrosis developed around control wounds. Koch's postulates were fulfilled with the reisolation of the pathogen from symptomatic tissues, isolates were identical morphologically and by sequence analysis of ITS region, ACT, and CAL partial genes to the original isolates. Damm et al. (2012) described two C. godetiae strains associated with walnut, one isolated in Austria and another one of unknown origin. An epidemic event of walnut anthracnose caused by Colletotrichum species mainly C. godetiae was reported in France (Da Lio et al. 2018). The pathogen was isolated from nuts, buds, insects, and stems. To our knowledge, this is the first report of anthracnose of walnut fruit caused by C. godetiae in Hungary. Anthracnose caused by C. godetiae, and previously reported C. fioriniae (Varjas et al. 2019) is becoming an increasing preharvest problem on Persian walnut in Hungary.

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First Report of Corynespora Leaf Spot Caused by Corynespora cassiicola on Gynura in China

Plant Disease ◽

10.1094/pdis-12-13-1284-pdn ◽

2014 ◽

Vol 98 (7) ◽

pp. 1007-1007 ◽

Cited By ~ 1

Author(s):

B. J. Li ◽

J. X. Chuan ◽

M. Yang ◽

G. F. Du

Keyword(s):

Leaf Spot ◽

Disease Incidence ◽

Morphological Characteristics ◽

Its Region ◽

Leaf Spot Disease ◽

Herbaceous Plant ◽

Distilled Water ◽

Corynespora Cassiicola ◽

Single Spore ◽

First Report

Gynura (Gynura bicolor DC.) is a perennial herbaceous plant in the family Compositae. It is an important Chinese vegetable, and is commonly used as a Chinese herbal medicine. In 2010, a severe leaf spot disease was observed on gynura grown in the main production areas in Tong Nan County, Chongqing City, China. Some farms experienced 60% disease incidence. Symptoms usually began on the lower leaves, as circular to elliptical or irregular spots with concentric rings. Individual spots were dark brown with grayish centers, sometimes coalescing and leading to extensive necrosis. The fungus associated with lesions was characterized as follows: Conidiophores were single or in clusters, straight or flexuous, unbranched, percurrent, cylindrical, pale to dark brown, 87.5 to 375.0 μm long and 5.0 to 10.5 μm wide. Conidia were solitary or catenate, straight to slightly curved, obclavate to cylindrical, 3 to 14 pseudoseptate, 82.8 to 237.5 μm long and 7.0 to 7.8 μm wide, and pale brown. The morphological characteristics of the conidia and conidiophores agreed with the descriptions for Corynespora cassiicola (1). To isolate the causal pathogen, surface-sterilized tissue at the margin of lesions was immersed in 75% ethanol for 30 s, rinsed in sterile water, dried in a laminar flow bench, transferred to PDA, and incubated at 28°C. Four single-spore cultures of the isolates were obtained and named from ZBTK10110637 to ZBTK10110640. All strains were identified as C. cassiicola. The isolate ZBTK10110637 was selected as representative for molecular identification. Genomic DNA was extracted by CTAB (2). The internal transcribed spacer (ITS) region of the rDNA was amplified using primers with ITS1 (5′-TCCGATGGTGAACCTGCGG-3′) and ITS4 (5′-TCCTCCGCTTATTGATATGC-3′). Amplicons were 433 bp (GenBank Accession No. JX867272) and shared 100% similarity with that of C. cassiicola (NRC2-1 No. AB539285.1). To confirm pathogenicity, four isolates were used to inoculate 12 gynura plants (6 weeks old) by mist spray-inoculation with 108 spores/ml suspension in sterile distilled water on the leaves. Control plants were misted with sterile distilled water. After inoculation, all plants were incubated in a greenhouse maintained at 20 to 28°C with relative humidity of 80 to 85%. Five days after inoculation, dark brown spots with a grayish center typical of field symptoms were observed on all inoculated plants. No symptoms were seen on water-treated control plants. The fungus was re-isolated from inoculated plants. The morphological characteristics of isolates were identical with the pathogen recovered originally. This is the first report of C. cassiicola on gynura. References: (1) M. B. Ellis. CMI Mycological Papers 65(9):1-15, 1957. (2) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.

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First Report of 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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