First Report of Rhizoctonia solani Subgroup AG 1-ID Causing Leaf Blight on Durian in Vietnam

During the rainy season in Vietnam, leaf blight disease caused by Rhizoctonia solani often occurs on 3- to 5-year-old durian (Durio zibethinus). Symptoms appear as large, pale brown, blighted lesions with an irregular border. In excessive moisture conditions, yellowish white hyphae appear on the lesions, and the affected leaves turn dark brown and wilt. There are no reports describing the anastomosis groups (AG) and subgroups of Rhizoctonia solani occurring in durian. In June of 2004, two isolates of R. solani were obtained from leaf blight lesions on durian growing in Binh Duong and Dong Nai provinces. The durian isolates were identified as AG 1 based on hyphal anastomosis. In pathogenicity tests, the durian isolates infected cucumber, mung bean, and leaf mustard seedlings grown on water agar in petri dishes. The rDNA-ITS sequence of the durian isolates was determined (GenBank Accession Nos. EF197797 and EF197798) and aligned with those of AG 1-IA, AG 1-IB, AG 1-IC, and AG 1-ID available in the GenBank database. The sequence similarity of the total rDNA-ITS region (including 5.8S) within the durian isolates was 99.9%. The sequence similarity of the durian isolates and AG 1-ID isolates was 99.1 to 100%, but similarity with other AG 1 subgroups was 89.1 to 94.0%. The results suggest that the two Vietnam durian isolates of R. solani are members of AG 1-ID. AG 1-ID has only been reported causing necrotic leaf spots on coffees in the Philippines (1). To our knowledge, this is the first report of R. solani AG 1-ID on durian and the first report of the presence of R. solani AG 1-ID in Vietnam. Reference: (1) A. Priyatmojo et al. Phytopathology 91:1054, 2001.

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First Report of Web Blight on Yellow-Sage (Lantana camara) Caused by Rhizoctonia solani in Europe

Plant Disease ◽

10.1094/pdis.2003.87.7.875a ◽

2003 ◽

Vol 87 (7) ◽

pp. 875-875 ◽

Cited By ~ 2

Author(s):

A. Garibaldi ◽

A. Minuto ◽

D. Bertetti ◽

R. Nicoletti ◽

M. L. Gullino

Keyword(s):

United States ◽

Rhizoctonia Solani ◽

The United States ◽

The Philippines ◽

Lantana Camara ◽

First Report ◽

Leaf Spots ◽

Entire Plant ◽

Pathogenicity Tests ◽

Web Blight

Lantana camara is increasingly grown in northern Italy as a potted plant and contributes to the diversification of offerings in the ornamental market. During the spring of 2001, selections of L. camara cuttings growing at a commercial farm located at Albenga (Riviera coast) exhibited tan leaf spots of irregular size and shape. Spots were at first isolated, 4 to 8 mm in diameter, and later coalesced and affected the entire plant. Heavily infected leaves, stems, and branches became blighted and were killed. Infected rooted cuttings also eventually died. Diseased cuttings showed a progressive reduction (to less than 20%) in rooting ability. Isolations from infected leaves and stems on potato dextrose agar (PDA), supplemented with 100 mg/liter of streptomycin sulphate, consistently yielded a fungus with mycelial and cultural characteristics resembling Rhizoctonia solani. The fungal isolates were further characterized as R. solani Kühn AG-4 based on hyphal anastomoses with several AG-4 tester isolates. Pathogenicity tests were performed by placing 5-day-old-fungal mycelial plugs, grown on PDA, at the base of five healthy yellow-sage stems and holding plants in a dew chamber at 18 to 22°C. After 2 days, foliage blight appeared on leaves of inoculated plants, and after 3 days, stems also became infected and entire plants wilted. Five noninoculated plants remained healthy. The fungal pathogen was reisolated from all inoculated plants. R. solani has been observed on L. camara in the United States (1) and the Philippines (2). To our knowledge, this is the first report of R. solani on L. camara in Europe. References: (1) D. F. Farr et al. Fungi on Plants and Plant Products in the United States. The American Phytopathological Society, St. Paul, MN, 1989. (2) F. T. Orillo and R. B. Valdez. Philipp. Agric. A. 42:292, 1958.

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First Report of Rhizoctonia solani AG4 HG-II Infecting Potato Stems in Idaho

Plant Disease ◽

10.1094/pdis-06-12-0568-pdn ◽

2012 ◽

Vol 96 (11) ◽

pp. 1701-1701 ◽

Cited By ~ 5

Author(s):

J. W. Woodhall ◽

P. S. Wharton ◽

J. C. Peters

Keyword(s):

Rhizoctonia Solani ◽

Seed Tuber ◽

Its Region ◽

Stem Canker ◽

Potato Production ◽

Koch’S Postulates ◽

First Report ◽

Rdna Its ◽

Plant Pathol ◽

Koch's Postulates

The fungus Rhizoctonia solani is the causal agent of stem canker and black scurf of potato (Solanum tuberosum). R. solani is a species complex consisting of 13 anastomosis groups (AGs) designated AG1 to 13 (2, 3). Stems of potato (cv. Russet Norkotah) with brown lesions were recovered from one field in Kimberley, Idaho, in August 2011. Using previously described methods (3), R. solani was recovered from the symptomatic stems and one representative isolate (J15) was selected for further characterization. Sequencing of the rDNA ITS region of isolate J15 was undertaken as previously described (3) and the resulting rDNA ITS sequence (HE667745) was 99% identical to sequences of other AG4 HG-II isolates in GenBank (AF354072 and AF354074). Pathogenicity of the isolate was determined by conducting the following experiment. Mini-tubers of cv. Santé were planted individually in 1-liter pots containing John Innes Number 3 compost (John Innes Manufacturers Association, Reading, UK). Pots were either inoculated with J15, an isolate of AG3-PT (Rs08), or were not inoculated. Each treatment was replicated four times. Inoculum consisted of five 10-mm-diameter potato dextrose agar plugs, fully colonized by the appropriate isolate, placed in the compost approximately 40 mm above each seed tuber. Pots were held in a controlled environment room at 21°C with 50% relative humidity and watered as required. After 21 days, plants were assessed for disease. No symptoms of the disease were present in non-inoculated plants. In the Rs08 (AG3-PT) inoculated plants, all stems displayed large brown lesions and 20% of the stems had been killed. No stem death was observed in J15 (AG4 HG-II) inoculated plants. However, brown lesions were observed in three of the four J15 (AG4 HG-II) inoculated plants. These lesions were less severe than in plants inoculated with the Rs08(AG3-PT) inoculated plants and were present in 40% of the main stems. In the J15 (AG4 HG-II) inoculated pots, R. solani AG4 HG-II was reisolated from the five symptomatic stems, thereby satisfying Koch's postulates. To our knowledge, this is the first report of AG4 HG-II causing disease on potatoes in Idaho. AG4 has been isolated from potato previously from North Dakota, although the subgroup was not identified (1). The only previous report where AG4 HG-II was specifically determined to cause disease on potato was in Finland, but the isolate could not be maintained and Koch's postulates were not completed (3). The present study shows that AG4 HG-II can cause stem disease in potatoes, although disease does not develop as severely or as consistently as for AG3-PT. However, as demonstrated with isolates of AG2-1 and AG5, even mild stem infection can reduce tuber yield by as much as 12% (4). AG4 HG-II is a pathogen of sugar beet in Idaho, which was grown previously in this field. This history may have contributed to high levels of soilborne inoculum required to produce disease on potato. References: (1) N. C. Gudmestad et al. Page 247 in: J. Vos et al. eds. Effects of Crop Rotation on Potato Production in the Temperate Zones. Kluwer, Dordrecht, Netherlands, 1989. (2) M. J. Lehtonen et al. Agric. Food Sci. 18:223, 2009. (3) J. W. Woodhall et al. Plant Pathol. 56:286, 2007. (4) J. W. Woodhall et al. Plant Pathol. 57:897, 2008.

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First Report of Aerial Blight of Peanut Caused by Rhizoctonia solani AG1-IA in Arkansas

Plant Disease ◽

10.1094/pdis-04-13-0414-pdn ◽

2013 ◽

Vol 97 (12) ◽

pp. 1658-1658 ◽

Cited By ~ 1

Author(s):

T. R. Faske ◽

T. N. Spurlock

Keyword(s):

Rhizoctonia Solani ◽

Fungal Pathogens ◽

Health Management ◽

Disease Incidence ◽

Its Region ◽

First Report ◽

Circular Pattern ◽

Rdna Its ◽

Randolph County ◽

Humidity Chamber

In early September 2012, symptoms similar to aerial blight were observed on runner peanut (cv. Georgia 09B) in a commercial field in Randolph County, Arkansas (3). Leaves within the canopy closest to the soil had water-soaked, gray to green lesions or tan to brown lesions. Localized areas of matted leaves with mycelium occurred on stems and hyphae extended along stems and newly affected leaves. Dark brown spherical sclerotia (1.5 to 4 mm diam.) were produced on the surface of symptomatic peanut tissue (3). Aerial blight symptoms were observed in two peanut fields (~4 to 6 ha) that were furrow irrigated. Symptomatic plants were localized in a single circular pattern (~20 × 25 m) near the lower end of each field with the final disease incidence of less than 5%. Isolations from surface-disinfected leaves on potato dextrose agar consistently yielded light brown to brown colonies with sclerotia typical of Rhizoctonia solani AG1-IA. The fungus was confirmed to be R. solani AG1 by anastomosis reaction (2) with known cultures of AG1-IA isolated from soybean and rice in Arkansas. Sequencing of the rDNA ITS region 5.8s with primers ITS1 and ITS4 (1) supported the identification of the R. solani isolates as AG1-IA. The BLAST search revealed that the sequence had a 96 to 97% maximum sequence identity to several R. solani AG1-IA isolates collected from rice sheaths in China and Arkansas. Eight-week-old peanut plants (cv. Georgia 09B) growing in pots were sprayed until runoff (2 ml/plant) with a solution containing approximately 1 × 105 hyphal fragments/ml. Five inoculated plants were placed in a humidity chamber within a greenhouse where temperatures ranged from 28 to 33°C. After 14 days, water soaked, gray to green or light brown lesions developed on all inoculated plants along with hyphal strands along inoculated sections of the peanut with dark brown sclerotia. None of the plants inoculated with sterile water expressed symptoms. Rhizoctonia solani was consistently reisolated from symptomatic tissue plated on PDA. Inoculations were repeated on peanut cv. Flavor Runner 458, Florida 07, FloRun 107, and Red River Runner with similar results. Although R. solani AG1-IA is a common pathogen on rice and soybean, causing sheath blight and aerial blight, respectively, to our knowledge this is the first report of aerial blight of peanut in the region. Currently, there is a renewed interest in peanut production in the state. Production practices include furrow irrigation, which can distribute floating sclerotia to peanut vines and the rotation practiced with soybean and, less frequently, rice, could potentially increase inoculum for the subsequent crop. Thus, this may be a significant disease problem in the region or Mid-South where peanut is planted after rice or soybean and furrow irrigated. References: (1) S. Kuninaga et al. Curr. Genet. 32:237, 1997. (2) G. C. MacNish et al. Phytopathology 83:922, 1993. (3) H. A. Melouk and P. A. Backman. Management of soilborne fungal pathogens. Pages 75-85 in: Peanut Health Management. H. A. Melouk and F. M. Stokes, eds. The American Phytopathological Society, St. Paul, MN, 1995.

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First Report of Leaf Blight on Woodland Sage Caused by Rhizoctonia solani AG 1 in Italy

Plant Disease ◽

10.1094/pdis-94-8-1071c ◽

2010 ◽

Vol 94 (8) ◽

pp. 1071-1071 ◽

Cited By ~ 2

Author(s):

A. Garibaldi ◽

G. Gilardi ◽

D. Bertetti ◽

M. L. Gullino

Keyword(s):

Rhizoctonia Solani ◽

Aqueous Suspension ◽

Its Region ◽

Morphological Characters ◽

Leaf Blight ◽

Pathogenicity Test ◽

First Report ◽

Leaf Petiole ◽

Perennial Plant ◽

Potted Plants

Woodland sage (Salvia nemorosa L.; Lamiaceae) is a hardy herbaceous perennial plant that is easy to grow and propagate and is used in parks and grown as potted plants. During the summer of 2009 in a nursery near Torino in northern Italy, a leaf blight was observed on 30-day-old plants of cv. Blau Koenigin grown in pots under shade. Semicircular, water-soaked lesions developed on leaves just above the soil line at the leaf-petiole junction and later along leaf margins. Lesions expanded along the midvein until the entire leaf was destroyed. Blighted leaves turned brown, withered, and clung to the shoots. No symptoms were observed on the roots. Severely infected plants died. Diseased tissue was disinfested for 10 s in 1% NaOCl, rinsed with sterile water, and plated on potato dextrose agar (PDA) amended with 25 mg/liter of streptomycin sulfate. A fungus with morphological characters of Rhizoctonia solani (3) was consistently recovered. Ten-day-old mycelium grown on PDA at 22 ± 1°C appeared light brown, rather compact, and with radial growth. Sclerotia were irregular and measured between 0.5 and 2 mm. Pairings were made with tester isolates of AG 1, 2, 3, 4, 5, 6, 7, 11, and AG B1. The only successful anastomosis was with tester isolate AG 1 (ATCC 58946). The hyphal diameter at the point of anastomosis was reduced, the anastomosis point was obvious, and cell death of adjacent cells was observed. Results were consistent with other reports on anastomosis reactions (2). The description of sclerotia of the isolate AG1 was typical for subgroup 1A Type 2 (3). The internal transcribed spacer (ITS) region of rDNA was amplified using primers ITS4/ITS6 and sequenced. BLASTn analysis (1) of the 688 bp showed a 100% homology with the sequence of R. solani AG-1A and the nucleotide sequence has been assigned (GenBank Accession No. HM044764). For pathogenicity tests, the inoculum of one isolate of R. solani from the nursery was prepared by growing the pathogen on PDA for 7 days. The foliage of 30-day-old potted plants of S. nemorosa cv. Blau Koenigin was artificially inoculated with an aqueous suspension of PDA and mycelium fragments (1 g per mycelium per plant) prepared from cultures with a blender. Plants were covered with plastic bags for 3 days. Plants inoculated with water and PDA fragments alone served as control treatments. Plants were maintained in a glasshouse at 20 to 25°C. The first symptoms, similar to those observed in the nursery, developed 7 days after foliar inoculation. R. solani was consistently reisolated from infected leaves. Control plants remained healthy. The pathogenicity test was carried out twice with similar results. To our knowledge, this is the first report of leaf blight of S. nemorosa caused by R. solani in Italy as well as worldwide. The importance of the disease is still unknown. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) D. E. Carling. Page 35 in: Rhizoctonia Species: Taxonomy, Molecular Biology, Ecology, Pathology and Disease Control. Kluwer Academic Publishers, the Netherlands, 1996. (3) B. Sneh et al. Identification of Rhizoctonia Species. The American Phytopathological Society, St Paul, MN, 1991.

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First report of Rhizoctonia solani Kühn AG 2-2 LP in Zoyzia japonica Steud in Brazil

Summa Phytopathologica ◽

10.1590/0100-5405/221916 ◽

2020 ◽

Vol 46 (4) ◽

pp. 289-298

Author(s):

Maria Aurea Saboya Chiaradia Picarelli ◽

Flavia Rodrigues Alves Patricio ◽

Ricardo Harakava ◽

Eliana Borges Rivas ◽

Addolorata Colariccio

Keyword(s):

Rhizoctonia Solani ◽

Mycelial Growth ◽

Its Region ◽

Anastomosis Group ◽

Large Patch ◽

First Report ◽

Rdna Its ◽

Rdna Its Region ◽

Important Disease

ABSTRACT The use of cultivated grasses in Brazil has grown by 40% between 2010 and 2015, and the species Zoysia japonica Steud, especially the cultivar ‘Esmeralda’, corresponds to 81% of the grass market in the country. The most important disease affecting zoysia grass, known as large patch, is caused by Rhizoctonia solani and occurs in the Brazilian lawns particularly during winter months. The aim of this study was to contribute to the identification and characterization of the anastomosis group of R. solani isolates from lesions typical of large patch collected from ‘Esmeralda’ grass at gardens and golf courses in the states of São Paulo and Bahia, Brazil. All 12 obtained isolates presented dark-brown colonies with aerial mycelial growth, multinucleated hyphae and absence of concentric zonation or sclerotia, and showed their greatest mycelial growth rate at 25°C. In pathogenicity experiments, except three out of R. solani isolates, reduced the growth of zoysia grass. Based on the analysis of sequences of the rDNA-ITS region, the isolates clustered with reference isolates of the anastomosis group AG 2-2 LP. Phylogenetic inference showed that the Brazilian isolates are grouped into two clades that shared the same common ancestral with 96% bootstrap. One of the clades includes only Brazilian isolates while the other one also includes American and Japanese R. solani isolates AG 2-2 LP. This is the first report and characterization of R. solani AG 2-2 LP in zoysiagrass in Brazil.

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First Report of Leaf Blight on Foxglove (Digitalis purpurea) Caused by Rhizoctonia solani AG-1-IA in Italy

Plant Disease ◽

10.1094/pdis-93-3-0318c ◽

2009 ◽

Vol 93 (3) ◽

pp. 318-318

Author(s):

A. Garibaldi ◽

G. Gilardi ◽

D. Bertetti ◽

M. L. Gullino

Keyword(s):

Rhizoctonia Solani ◽

Morphological Characteristics ◽

Its Region ◽

Leaf Blight ◽

Pathogenicity Test ◽

Uniform Size ◽

First Report ◽

Digitalis Purpurea ◽

Blastn Analysis ◽

Hyphal Diameter

Digitalis purpurea (Scrophulariaceae), foxglove, is used in flower gardens. In the spring of 2008, leaf blight was observed in a nursery near Biella (northern Italy) on 30% of potted 30-day-old plants grown in a peat substrate at temperatures from 20 to 25°C and relative humidity at 75 to 80%. Semicircular, water-soaked lesions developed on leaves just above the soil line at the blade-petiole junction and later along the leaf margins. Lesions expanded for several days along the midvein until the entire leaf was affected. Blighted leaves turned brown, withered, clung to the shoots, and matted on the surrounding foliage. Diseased tissue was disinfested for 10 s in 1% NaOCl, rinsed with sterile water, and plated on potato dextrose agar (PDA) amended with 100 mg/liter of streptomycin sulfate. A fungus with the morphological characteristics of Rhizoctonia solani was consistently and readily recovered, then transferred and maintained in pure culture (4). The isolates of R. solani obtained from affected plants successfully anastomosed with tester isolate AG 1 (ATCC 58946). The hyphal diameter at the point of anastomosis was reduced, the anastomosis point was obvious, and cell death of adjacent cells was observed. Results were consistent with other reports on anastomosis reactions (2). Pairings were also made with tester isolates AG 2, 3, 4, 6, 7, 11, and AG BI and anastomosis was not observed. Ten-day-old colonies grown on PDA appeared light brown, rather compact, and radial. Numerous sclerotia of uniform size (0.5 to 3 mm in diameter) and sometimes joined laterally were formed. Descriptions of mycelium and sclerotia were typical for subgroup IA Type 2 (4). The internal transcribed spacer (ITS) region of rDNA was amplified using primers ITS4/ITS6 and sequenced. BLASTn analysis (1) of the 724-bp fragment showed a 99% homology with the sequence of R. solani (GenBank Accession No. EU591800). The nucleotide sequence has been assigned GenBank Accession No. FJ467490. For pathogenicity tests, the inoculum of R. solani was prepared by growing the pathogen on PDA for 10 days. Plants of 30-day-old D. purpurea were grown in 10-liter containers (6 plants per container) in a steam disinfested peat/clay/perlite (70:20:10) substrate. Disks of PDA cultures were placed on leaves (1 cm2 of mycelium per plant). Plants inoculated with PDA alone served as control treatments. Three replicates were used. Plants were maintained in a growth chamber at 24 ± 1°C with 12 h light/dark. First symptoms developed 12 days after the artificial inoculation. R. solani was consistently reisolated from infected leaves and stems. Control plants remained healthy. The pathogenicity test was repeated twice. R. solani was isolated from a small percentage of infected seeds of D. purpurea in India (3). This is, to our knowledge, the first report of leaf blight of D. purpurea caused by R. solani in Italy as well as in Europe. The spread of R. solani in nurseries might cause a decrease in trade. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) D. E. Carling. Grouping in Rhizoctonia solani by hyphal anastomosis reactions in: Rhizoctonia Species: Taxonomy, Molecular Biology, Ecology, Pathology and Disease Control. Kluwer Academic Publishers, the Netherlands, 1996. (3) K. K. Janardhanan and D. Ganguly. Indian Phytopathol. 16:379, 1963. (4) B. Sneh et al. Identification of Rhizoctonia Species. The American Phytopathological Society, St Paul, MN, 1991.

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First Report of Leaf Blight on Hosta fortunei Caused by Rhizoctonia solani AG 4 in Italy

Plant Disease ◽

10.1094/pdis-93-4-0432c ◽

2009 ◽

Vol 93 (4) ◽

pp. 432-432 ◽

Cited By ~ 3

Author(s):

A. Garibaldi ◽

D. Bertetti ◽

M. L. Gullino

Keyword(s):

United States ◽

Rhizoctonia Solani ◽

Aqueous Suspension ◽

Its Region ◽

The United States ◽

Morphological Characters ◽

Leaf Blight ◽

Pathogenicity Test ◽

First Report ◽

Leaf Petiole

Hosta fortunei (Liliaceae) is used in semishaded areas of gardens for its lavender-colored flowers produced in midsummer. In April of 2008, in a greenhouse at the University of Torino, located in Grugliasco (northern Italy), a leaf blight was observed on 15% of potted 60-day-old plants growing at temperatures ranging between 20 and 25°C and relative humidity of 60 to 90%. Semicircular, water-soaked lesions developed on leaves just above the soil line at the leaf-petiole junction and later along leaf margins. Lesions expanded for several days along the midvein until the entire leaf was destroyed. Blighted leaves turned brown, withered, and clung to the shoots. Severely infected plants died. Diseased tissue was disinfested for 10 s in 1% NaOCl, rinsed with sterile water, and plated on potato dextrose agar (PDA) amended with 25 mg/liter streptomycin sulfate. A fungus with the morphological characters of Rhizoctonia solani (4) was consistently recovered, then transferred and maintained in pure culture. Ten-day-old mycelium grown on PDA at 22 ± 1°C appeared light brown, rather compact, and had radial growth. Sclerotia were not present. Isolates of R. solani obtained from affected plants were successfully anastomosed with tester isolate AG 4 (AG 4 RT 31 obtained from tobacco plants). Results were consistent with other reports on anastomosis reactions (2). Pairings were also made with tester isolates of AG 1, 2.1, 2.2, 3, 6, 7, 11, and BI, but no anastomosis was observed. The internal transcribed spacer (ITS) region of rDNA was amplified using primers ITS4/ITS6 and sequenced. BLASTn analysis (1) of the 646-bp fragment showed a 100% homology with the sequence of R. solani AG-4 AB000018. The nucleotide sequence has been assigned GenBank Accession No. FJ 534556. For pathogenicity tests, the inoculum of R. solani was prepared by growing the pathogen on PDA for 10 days. Six-month-old plants of H. fortunei were grown in 1-liter pots. Inoculum, which consisted of an aqueous suspension of PDA and mycelium disks (10 g of mycelium per pot), was placed at the collar of plants. Plants inoculated with water and PDA fragments alone served as control treatments. Five plants per treatment were used. Plants were maintained in a growth chamber at 20 ± 1°C. The first symptoms, similar to those observed in the nursery, developed 15 days after inoculation. R. solani was consistently reisolated from infected leaves and stems. Control plants remained healthy. The pathogenicity test was carried out twice with similar results. R. solani was reported on plants belonging to the genus Hosta in the United States (3). This is, to our knowledge, the first report of leaf blight of H. fortunei caused by R. solani in Italy as well as in Europe. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) D. E. Carling. Grouping in Rhizoctonia solani by hyphal anastomosis reactions. In: Rhizoctonia Species: Taxonomy, Molecular Biology, Ecology, Pathology and Disease Control. Kluwer Academic Publishers, The Netherlands, 1996. (3) D. F. Farr et al. Fungi on Plants and Products in the United States. The American Phytopathology Society, St Paul, MN, 1989. (4) B. Sneh et al. Identification of Rhizoctonia species. The American Phytopathological Society, St Paul, MN, 1991.

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First Report of Twig Blight Disease of Citrus Caused by Haematonectria haematococca in the Philippines

Plant Disease ◽

10.1094/pdis-12-10-0903 ◽

2011 ◽

Vol 95 (12) ◽

pp. 1590-1590 ◽

Cited By ~ 1

Author(s):

J. I. Yago ◽

K.-R. Chung

Keyword(s):

Sequence Similarity ◽

Its Region ◽

The Philippines ◽

Control Measures ◽

Its Sequence ◽

Post Inoculation ◽

Distilled Water ◽

First Report ◽

Fungal Propagules ◽

Twig Blight

Citrus has recently become one of the most important crops in the Philippines with approximately 151,000 t of production in 2005. A new disease affected citrus (Citrus reticulata Blanco cv. Satsuma) twigs and trunks in 2005. The problem has caused significant concerns to the industry because of its destruction and high severity of infection. Symptoms include twig drying, bark hardening, twig girdling, leaf chlorosis, and defoliation. Infection occurs at all stages of the plant where severe symptoms were observed from January to April. Average temperature of these months ranged from 20 to 28°C. Affected twigs were often covered with pink or salmon-colored fungal propagules (1). In April 2009, citrus twigs displaying the above-described symptoms and signs were collected from Malabing Valley, Kasibu, Nueva Vizcaya, the Philippines (16°20′4.6912″N, 121°16′41.0742″E). Fungal perithecia were surface sterilized and observed with a stereomicroscope. Single ascospore was picked and inoculated into the peptone-pentachloronitrobenzene medium (2) and the plates were incubated at 22 ± 2°C for 7 days with a 12-h light/dark regimen. Periodically, the plates were screened for the growth of mycelia or discrete colonies on the medium. The cultured fungus produced microconidia and multiple canoe-shaped macroconidia. With a hand-held sprayer, approximately 20 ml of distilled water containing fungal conidia (2 × 105 spores/ml) was inoculated each onto 1-year-old cv. Satsuma citrus (30 plants) bearing young twigs. Control plants were sprayed with sterile distilled water. After inoculation, plants were maintained at 20 to 28°C and 75 to 85% relative humidity, enclosed in clear plastic bags, placed under 70% woven shade cloth, and watered regularly. The inoculated plants started showing the initial twig blight symptoms in all inoculated branches at 37 days post inoculation (dpi). The occurrence of pink-to-orange perithecia of Haematonectria haematococca was observed at 45 dpi, which has similar morphological appearance to the perithecia collected from the field. Symptoms were not observed on branches treated with sterile water. H. haematococca was reisolated from the symptomatic twigs and the recovered isolates were morphologically similar to H. haematococca, thus completing Koch's postulates. Control plants remained healthy. DNA was extracted from all isolates, and the nuclear ribosomal internal transcribed spacer (ITS) region was amplified with primers ITS1 and ITS4 and sequenced. A portion of the ITS sequence has been deposited in the NCBI database (GenBank Accession No. HQ696788). A BLAST search of the NCBI database with the ITS sequence revealed H. haematococca (Berk. & Broome) Samuels & Nirenberg as the closest match with 100% sequence similarity. To our knowledge, this is the first report of H. haematococca causing citrus twig blight in the Philippines. To maintain the economic profitability of the citrus industry in the Philippines, control measures must be implemented to minimize tree loss. References: (1) P. J. Chester. Phil. Agri. Rev. 11:69, 1919. (2) S. M. Nash and W. C. Snyder. Phytopathology 52:567, 1962.

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First Report of Leaf Spot of Rocket (Eruca sativa) Caused by Fusarium equiseti in Italy

Plant Disease ◽

10.1094/pdis-03-11-0220 ◽

2011 ◽

Vol 95 (10) ◽

pp. 1315-1315 ◽

Cited By ~ 5

Author(s):

A. Garibaldi ◽

G. Gilardi ◽

C. Bertoldo ◽

M. L. Gullino

Keyword(s):

Elongation Factor ◽

Its Region ◽

Northern Italy ◽

Pathogenicity Test ◽

Eruca Sativa ◽

Single Spore ◽

Fusarium Equiseti ◽

First Report ◽

Rdna Its ◽

Leaf Spots

During the summer of 2010, rocket (Eruca sativa) plants grown in an open field and under a plastic tunnel in Piedmont (northern Italy) showed symptoms of a previously reported foliar disease. Symptoms were observed on 30-day-old plants and consisted of small, circular, brown leaf spots (1 to 3 to 10 to 12 mm in diameter), sometimes later becoming elliptical. Necrotic lesions were cracked in the center and showed a well-defined border, frequently surrounded by a violet-brown halo. Approximately 40% of the plants were affected with 30 to 40% of the leaves infected. An orange-brown colony with characteristics of Fusarium was isolated from leaf tissues of 30 infected plants on potato dextrose agar (PDA). Isolates were purified, subcultured on PDA, and single-spore cultures were obtained. On PDA, they produced orange-brown colonies and purple pigments. On Spezieller Nährstoffarmer agar (SNA) (1), the isolates produced hyaline macroconidia with dorsiventral curvature, five to seven septate, and measuring 36.2 to 49.3 × 3.4 to 5.3 (average 41.9 × 4.0) μm. Chlamydospores, solitary but also in short chains (two to three elements), measuring 7.2 to 15.3 (average 10.1) μm were produced on carnation leaf agar (CLA) after 10 days and became verrucose 20 days later. Macroconidia were produced on CLA in orange sporodochia from monophialides on branched conidiophores. Microconidia were not observed. Such characteristics are typical of the genus Fusarium (1). The rDNA ITS region (internal transcribed spacer) was amplified using the primers ITS1/ITS4 (2) and sequenced. BLASTn analysis of the 480-bp product obtained showed an E-value of 0.0 with Fusarium equiseti. The nucleotide sequence has been assigned the GenBank Accession No. JF460797. The translation elongation factor-1α (EF-1α) gene (GenBank Accession No. JN127347) was amplified using primers EF-1/EF-2 and sequenced. The 702-bp fragment showed 99% identity with F. equiseti (GenBank Accession No. FJ939673.1). To confirm pathogenicity, 20-day-old rocket plants were transplanted into 2-liter volume pots, filled with a steamed peat/perlite/sand (60:20:20 vol/vol) substrate and maintained in a growth chamber at 25 ± 1°C. Five pots per treatment were used, each pot containing two plants. The artificial inoculation was carried out either by spraying leaves with a spore suspension prepared from 15-day-old cultures of the pathogen on PDA or by applying CLA agar disks (6 mm in diameter) from 10-day-old cultures onto leaves. Control plants were inoculated with distilled water or with noninoculated agar disks. Plants were covered with plastic bags for 5 days. The first symptoms, consisting of chlorotic leaf halo and leaf spots surrounded by a violet-brown halo, developed 15 days after inoculation by foliar spraying and 5 days after inoculation by disks. Noninoculated plants remained healthy. F. equiseti was consistently isolated from symptomatic plants. The pathogenicity test was conducted twice. To our knowledge, this is the first report of F. equiseti on E. sativa in Italy. Currently, this disease is present in several farms in northern Italy. Its importance might increase because of the widespread cultivation of cultivated rocket in Italy. References: (1) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell, Ames, IA, 2006. (2) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, 1990.

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First Report of Leaf Blight and Stem Canker of Pachysandra terminalis Caused by Pseudonectria pachysandricola in Korea

Plant Disease ◽

10.1094/pdis-09-11-0813 ◽

2012 ◽

Vol 96 (2) ◽

pp. 287-287

Author(s):

K. S. Han ◽

J. H. Park ◽

S. E. Cho ◽

H. D. Shin

Keyword(s):

United States ◽

Its Region ◽

Stem Canker ◽

The United States ◽

Culture Collection ◽

Leaf Blight ◽

First Report ◽

Cornell University ◽

Plastic Bags ◽

Young Leaves

Pachysandra terminalis Siebold & Zucc., known as Japanese pachysandra, is a creeping evergreen perennial belonging to the family Buxaceae. In April 2011, hundreds of plants showing symptoms of leaf blight and stem canker with nearly 100% incidence were found in a private garden in Suwon, Korea. Plants with the same symptoms were found in Seoul in May and Hongcheon in August. Affected leaves contained tan-to-yellow brown blotches. Stem and stolon cankers first appeared as water soaked and developed into necrotic lesions. Sporodochia were solitary, erumpent, circular, 50 to 150 μm in diameter, salmon-colored, pink-orange when wet, and with or without setae. Setae were hyaline, acicular, 60 to 100 μm long, and had a base that was 4 to 6 μm wide. Conidiophores were in a dense fascicle, not branched, hyaline, aseptate or uniseptate, and 8 to 20 × 2 to 3.5 μm. Conidia were long, ellipsoid to cylindric, fusiform, rounded at the apex, subtruncate at the base, straight to slightly bent, guttulate, hyaline, aseptate, 11 to 26 × 2.5 to 4.0 μm. A single-conidial isolate formed cream-colored colonies that turned into salmon-colored colonies on potato dextrose agar (PDA). Morphological and cultural characteristics of the fungus were consistent with previous reports of Pseudonectria pachysandricola B.O. Dodge (1,3,4). Voucher specimens were housed at Korea University (KUS). Two isolates, KACC46110 (ex KUS-F25663) and KACC46111 (ex KUS-F25683), were accessioned in the Korean Agricultural Culture Collection. 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 using ABI Prism 337 automatic DNA sequencer (Applied Biosystems, Foster, CA). The resulting sequence of 487 bp was deposited in GenBank (Accession No. JN797821). This showed 100% similarity with a sequence of P. pachysandricola from the United States (HQ897807). Isolate KACC46110 was used in pathogenicity tests. Inoculum was prepared by harvesting conidia from 2-week-old cultures on PDA. Ten young leaves wounded with needles were sprayed with conidial suspensions (~1 × 106 conidia/ml). Ten young leaves that served as the control were treated with sterile distilled water. Plants were covered with plastic bags to maintain a relative humidity of 100% at 25 ± 2°C for 24 h. Typical symptoms of brown spots appeared on the inoculated leaves 4 days after inoculation and were identical to the ones observed in the field. P. pachysandricola was reisolated from 10 symptomatic leaf tissues, confirming Koch's postulates. No symptoms were observed on control plants. Previously, the disease was reported in the United States, Britain, Japan, and the Czech Republic (2,3), but not in Korea. To our knowledge, this is the first report of P. pachysandricola on Pachysandra terminalis in Korea. Since this plant is popular and widely planted in Korea, this disease could cause significant damage to nurseries and the landscape. References: (1) B. O. Dodge. Mycologia 36:532, 1944. (2) D. F. Farr and A. Y. Rossman. Fungal Databases. Systematic Mycology and Microbiology Laboratory, ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ , September 24, 2011. (3) I. Safrankova. Plant Prot. Sci. 43:10, 2007. (4) W. A. Sinclair and H. H. Lyon. Disease of Trees and Shrubs. 2nd ed. Cornell University Press, Ithaca, NY, 2005.

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