First Report of Damping-Off Caused by Rhizoctonia solani AG-4 on Mediterranean Fan Palm in Italy

Mediterranean fan palm (Chamaerops humilis L.), one of just two autochthonous European palms, is native to the western Mediterranean Region in southwestern Europe and northwestern Africa. It can be found growing wild in the Mediterranean area. In Europe, this species is very popular as an ornamental plant. In March 2009, a widespread damping-off was observed in a stock of approximately 30,000 potted 1-month-old plants of C. humilis cv. Vulcano in a nursery in eastern Sicily. Disease incidence was approximately 20%. Disease symptoms consisted of lesions at the seedling shoot (plumule). Stem lesions were initially orange, turned brown, and followed by death of the entire plumule or eophyll. A fungus with mycelial and morphological characteristics of Rhizoctonia solani Kühn was consistently isolated from lesions when plated on potato dextrose agar (PDA) amended with streptomycin sulfate at 100 μg/ml. Fungal colonies were initially white, turned brown with age, and produced irregularly shaped, brown sclerotia. Mycelium was branched at right angles with a septum near the branch and a slight constriction at the branch base. Hyphal cells removed from cultures grown at 25°C on 2% water agar were determined to be multinucleate when stained with 1% safranin O and 3% KOH solution (1) and examined at ×400. Anastomosis groups were determined by pairing isolates with tester strains AG-1 IA, AG-2-2-1, AG-2-2IIIB, AG-2-2IV, AG-3, AG-4, AG-5, AG-6, and AG-11 on 2% water agar in petri plates (3). Anastomosis was observed only with tester isolates of AG-4, giving both C2 and C3 reactions (2). One representative isolate obtained from symptomatic tissues was deposited at the Fungal Biodiversity Centre, Centraalbureau voor Schimmelcultures (CBS No. 125095). Pathogenicity tests were performed on container-grown, healthy, 1-month-old seedlings. Twenty plants of C. humilis cv. Vulcano were inoculated near the base of the stem with two 1-cm2 PDA plugs from 5-day-old mycelial cultures. The same number of plants served as uninoculated controls. Plants were incubated in a growth chamber and maintained at 25°C and 95% relative humidity on a 12-h fluorescent light/dark regimen. Symptoms identical to those observed in the nursery appeared 5 days after inoculation and all plants died within 20 days. No disease was observed on control plants. A fungus identical in culture morphology to R. solani AG-4 was consistently reisolated from symptomatic tissues, confirming its pathogenicity. To our knowledge, this is the first report in the world of R. solani causing damping-off on Mediterranean fan palm. References: (1) R. J. Bandoni. Mycologia 71:873, 1979. (2) D. E. Carling. Page 37 in: Grouping in Rhizoctonia solani by Hyphal Anastomosis Reactions. Kluwer Academic Publishers, the Netherlands, 1996. (3) C. C. Tu and J. W. Kimbrough. Mycologia 65:941, 1973.

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First Report of Crown and Root Rot Caused by Rhizoctonia solani AG-4 on Coprosma repens and C. lucida in Italy

Plant Disease ◽

10.1094/pdis-93-9-0972b ◽

2009 ◽

Vol 93 (9) ◽

pp. 972-972 ◽

Cited By ~ 2

Author(s):

G. Polizzi ◽

D. Aiello ◽

I. Castello ◽

A. Vitale

Keyword(s):

Rhizoctonia Solani ◽

Root Rot ◽

Disease Incidence ◽

Morphological Characteristics ◽

Fluorescent Light ◽

Centraalbureau Voor ◽

First Report ◽

Crown And Root Rot ◽

Stem Lesions ◽

Safranin O

Coprosma (J.R. Forster & G. Forster), a genus containing approximately 90 species, occurs principally in New Zealand, Hawaii, Australia, New Guinea, and islands of the Pacific. In Italy, some of these species, including many variegated varieties and hybrids, are grown as ornamental evergreen shrubs or small trees. In June 2008, a crown and root rot was observed in a stock of approximately 12,000 potted 3-year-old plants of Coprosma repens cv. Yvonne and C. lucida in a nursery in eastern Sicily. Disease incidence was approximately 30%. Disease symptoms consisted of water-soaked lesions at the crown of the trunk and a root rot. Successively, older stem lesions turned orange to brown. As a consequence, leaves initially became chlorotic, gradually became necrotic, and death of the plant followed. A fungus with mycelial and morphological characteristics of Rhizoctonia solani Kühn was consistently isolated from crown and root lesions when plated on potato dextrose agar (PDA) amended with streptomycin sulfate at 100 μg/ml. Fungal colonies were initially white, turned brown with age, and produced irregularly shaped, brown sclerotia. Mycelium was branched at right angles with a septum near the branch and a slight constriction at the branch base. Hyphal cells removed from cultures grown at 25°C on 2% water agar were determined to be multinucleate when stained with 1% safranin O and 3% KOH solution (1) and examined at ×400. Anastomosis groups were determined by pairing isolates on 2% water agar in petri plates (3). Pairings were made with tester strains of AG-1 IA, AG-2-2-1, AG-2-2IIIB, AG-2-2IV, AG-3, AG-4, AG-5, AG-6, and AG-11. Anastomosis was observed only with tester isolates of AG-4, giving C2 and C3 reactions (2). Two representative isolates obtained from symptomatic tissues of C. lucida and C. repens cv. Yvonne were deposited at the Fungal Biodiversity Centre, Centraalbureau voor Schimmelcultures (DISTEF CL1 = CBS-124593 and DISTEF CR1 = CBS-124594, respectively). Pathogenicity tests were performed on container-grown, healthy, 3-month-old cuttings. Ten plants of C. lucida and ten plants of C. repens cv. Yvonne were inoculated near the base of the stem with five 1-cm2 PDA plugs from 5-day-old mycelial cultures. The same number of plants served as uninoculated controls. Plants were maintained at 25°C and 95% relative humidity on a 12-h fluorescent light/dark regimen. Symptoms identical to ones observed in the nursery appeared 5 days after inoculation and all plants died within 15 days. No disease was observed on control plants. A fungus identical in culture morphology to R. solani AG-4 was consistently reisolated from symptomatic tissues, confirming its pathogenicity. To our knowledge, this is the first report of R. solani causing crown and root rot on the genus Coprosma. References: (1) R. J. Bandoni. Mycologia 71:873, 1979. (2) D. E. Carling. Page 37 in: Grouping in Rhizoctonia solani by Hyphal Anastomosis Reactions. Kluwer Academic Publishers, the Netherlands, 1996. (3) C. C. Tu and J. W. Kimbrough. Mycologia 65:941, 1973.

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First Report of Damping-Off Caused by Rhizoctonia solani AG-4 on Pink Ipê (Tabebuia impetiginosa) in Italy

Plant Disease ◽

10.1094/pdis-10-10-0748 ◽

2011 ◽

Vol 95 (1) ◽

pp. 78-78 ◽

Cited By ~ 2

Author(s):

G. Polizzi ◽

D. Aiello ◽

A. Vitale ◽

V. Guarnaccia ◽

A. Panebianco ◽

...

Keyword(s):

Rhizoctonia Solani ◽

Morphological Characteristics ◽

Fluorescent Light ◽

Damping Off ◽

First Report ◽

Initial Symptoms ◽

Public Areas ◽

Pathogenicity Tests ◽

Stem Lesions ◽

Safranin O

Pink ipê or pink lapacho (Tabebuia impetiginosa Martius ex DC., family Bignoniaceae) is one of the most attractive blooming trees in the world. In Europe, pink ipê is widely used as an ornamental tree in landscaped gardens and public areas. In August 2010, a widespread damping-off was observed in a stock of approximately 100,000 potted 2-month-old seedlings in a nursery in eastern Sicily (Italy). The seedlings were being watered with overhead irrigation. More than 5% of the seedlings showed disease symptoms. Initial symptoms were black lesions at the seedling crown that expanded rapidly to girdle the stem. On infected seedlings, leaves turned black and gradually died. Black extended stem lesions were followed by death of the entire seedling in a few days. A fungus with mycelial and morphological characteristics of Rhizoctonia solani Kühn was consistently isolated from crown and stem lesions when plated on potato dextrose agar (PDA) amended with streptomycin sulfate at 100 μg/ml. Fungal colonies were initially white, turned brown with age, and produced irregularly shaped, brown sclerotia. Mycelium was branched at right angles with a septum near the branch and a slight constriction at the branch base. Hyphal cells removed from cultures grown at 25°C on 2% water agar were determined to be multinucleate when stained with 1% safranin O and 3% KOH solution (1) and examined at ×400. Anastomosis groups were determined by pairing isolates with tester strains AG-1 IA, AG-2-2-1, AG-2-2IIIB, AG-2-2IV, AG-3, AG-4, AG-5, AG-6, and AG-11 on 2% water agar in petri plates (4). Anastomosis was observed only with tester isolates of AG-4, giving both C2 and C3 reactions (2). Pathogenicity tests were performed on container-grown, healthy, 3-month-old seedlings. Forty seedlings of T. impetiginosa were inoculated near the base of the stem with two 1-cm2 PDA plugs from 5-day-old mycelial cultures. The same number of plants only inoculated with PDA plugs served as controls. Plants were incubated in a growth chamber and maintained at 25°C and 95% relative humidity on a 12-h fluorescent light/dark regimen. Crown and stem lesions identical to those observed in the nursery appeared 5 days after inoculation and all plants died within 25 days. No disease was observed on control plants. R. solani AG-4 was reisolated from symptomatic tissues and identified as previously described. R. solani AG-4 was previously detected in the same nursery on Chamaerops humilis (3). To our knowledge, this is the first report of R. solani causing damping-off on T. impetiginosa. References: (1) R. J. Bandoni. Mycologia 71:873, 1979. (2) D. E. Carling. Page 37 in: Grouping in Rhizoctonia solani by Hyphal Anastomosis Reactions. Kluwer Academic Publishers, the Netherlands, 1996. (3) G. Polizzi et al. Plant Dis. 94:125, 2010. (4) C. C. Tu and J. W. Kimbrough. Mycologia 65:941, 1973.

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First Report of Damping-Off Caused by Rhizoctonia solani AG-4 on Lagunaria patersonii in Italy

Plant Disease ◽

10.1094/pdis-92-5-0836a ◽

2008 ◽

Vol 92 (5) ◽

pp. 836-836 ◽

Cited By ~ 3

Author(s):

D. Aiello ◽

G. Parlavecchio ◽

A. Vitale ◽

E. Lahoz ◽

R. Nicoletti ◽

...

Keyword(s):

Rhizoctonia Solani ◽

Morphological Characteristics ◽

Ruthenium Red ◽

Fluorescent Light ◽

Damping Off ◽

First Report ◽

Pectic Enzymes ◽

Electrophoretic Patterns ◽

Blue Solution ◽

Hyphal Diameter

Lagunaria patersonii (Adr.) G. Don (cow itch tree) is native to Australia and tolerates salted winds. During July 2007, damping-off of cow itch tree was observed on 4-month-old seedlings growing in a commercial nursery in eastern Sicily, Italy. More than 20% of the seedlings showed disease symptoms. First symptoms consisting of water-soaked lesions at the seedling base that expand rapidly girdle the stem and collapse the seedling in a few days. Diseased tissues were disinfested for 1 min in 1% NaOCl, rinsed in sterile water, plated on potato dextrose agar (PDA) amended with streptomycin sulphate at 100 mg/l, and then incubated at 25°C. A fungus with mycelial and morphological characteristics of Rhizoctonia solani Kühn was consistently yielded. Fungal colonies were initially white, turned brown with age, and produced irregularly shaped, brown sclerotia. Microscopic examination revealed that hyphae had a right-angle branching pattern, were constricted at the base of the branch near the union with main hyphae, and septate near the constriction. Basidia were not observed in the greenhouses or on the plates. Hyphal cells were determined to be multinucleate when stained with 0.5% aniline blue solution and examined at ×400 magnification with a microscope. Anastomosis groups were determined by pairing isolates on 2% water agar in petri plates (3). Pairings were made with tester strains of AG-1 IA, AG-2-2-1, AG-2-2IIIB, AG-2-2IV, AG-3, AG-4, AG-5, AG-6, AG-11. Anastomosis was observed only with tester isolates of AG-4 producing both C2 and C3 reactions. The hyphal diameter at the point of anastomosis was reduced, the anastomosis point was obvious, and cell death of adjacent cells was observed. These results were consistent with other reports on anastomosis reactions (1). The identification of group AG-4 within R. solani has been confirmed by electrophoretic patterns of pectic enzymes (polygalacturonases) in vertical pectin-acrylamide gel stained with ruthenium red (2). Pathogenicity tests were conducted on potted, healthy, 3-month-old seedlings of cow itch tree. Twenty plants were inoculated by placing plugs of PDA from 5-day-old mycelial cultures near the base of the stem. The same number of plants was treated with 1 cm2 PDA plugs as control. Plants were kept at 25°C and 95% relative humidity on a 12-h fluorescent light/dark regimen. Wilt symptoms due to basal stem rot, identical to ones observed in the nursery, appeared 10 days after inoculation and all inoculated plants showed symptoms within 1 month. Control plants remained healthy. The pathogen was reisolated from symptomatic tissues, completing Koch's postulates. To our knowledge, this is the first report in the world of R. solani causing disease on L. patersonii. References: (1) D. E. Carling. Page 37 in: Grouping in Rhizoctonia solani by Hyphal Anastomosis Reactions. Kluwer Academic Publishers, the Netherlands, 1996. (2) R. H. Cruickshank and G. C. Wade. Anal. Biochem. 107:177, 1980. (3) C. C. Tu and J. W. Kimbrough. Mycologia 65:941, 1973.

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First Report of Damping-Off on African Daisy Caused by Rhizoctonia solani AG-4 in Italy

Plant Disease ◽

10.1094/pdis-92-9-1367b ◽

2008 ◽

Vol 92 (9) ◽

pp. 1367-1367 ◽

Cited By ~ 2

Author(s):

D. Aiello ◽

I. Castello ◽

A. Vitale ◽

E. Lahoz ◽

R. Nicoletti ◽

...

Keyword(s):

Rhizoctonia Solani ◽

Morphological Characteristics ◽

Aerial Mycelium ◽

Ruthenium Red ◽

Fluorescent Light ◽

Damping Off ◽

First Report ◽

Perennial Plant ◽

Pectic Enzymes ◽

Electrophoretic Patterns

Osteospermum (African Daisy or Cape Daisy) is a genus belonging to the Calendulae with a large number of perennial plant species. In February of 2007, a severe damping-off occurred on 3- to 4-month-old potted cuttings of Osteospermum ‘Impassion Rose Purple’, ‘Impassion White’, ‘Impassion Purple’, and ‘Impassion White Rose’ cultivated in a nursery in eastern Sicily. More than 30% of the plants were infected. Disease symptoms consisted of extensive water-soaked lesions at the base of the stem followed by wilt and collapse of the plant. Isolations from diseased tissues on potato dextrose agar (PDA) amended with streptomycin sulfate at 100 mg/l consistently recovered a fungus with morphological characteristics of Rhizoctonia solani. Fungal colonies were initially white, turned brown after 2 to 3 days, and produced irregularly shaped, brown sclerotia after 1 week. Microscopic examination revealed that hyphae had right angle branching patterns, were constricted at the base of the branch near the union with main hyphae, and septate near the constriction. The number of nuclei per hyphal cell was determined on cultures grown at 25°C on 2% water agar in petri plates. Mycelium was stained with 0.5% aniline blue solution (4) and examined with a microscope at ×400. The hyphal cells were all multinucleate. Anastomosis groups were determined by pairing isolates (3) with tester isolates of AG-1 IA, AG-2-2-1, AG-2-2IIIB, AG-2-2IV, AG-3, AG-4, AG-5, AG-6, and AG-11. Anastomosis was observed only with tester isolates of AG-4, giving C2 reactions (1) at a high frequency. The identification of group AG-4 within R. solani had been obtained by electrophoretic patterns of pectic enzymes (polygalacturonases) in vertical pectin-acrylamide gel stained with ruthenium red (2). All isolates of R. solani collected from infected plants were paired in all combinations on PDA plus 1% activated charcoal and examined for somatic interaction. All paired colonies merged without producing visible tufts of aerial mycelium. Absence of tufts and the lack of formation of heterokaryon at the hyphal interaction zone indicated that all isolates belonged to the same mating type with the same mating alleles (3). Pathogenicity tests were performed by placing plugs of PDA from 5-day-old mycelial cultures in the soil near the base of the stem on 20 potted, healthy, 2-month-old cuttings of Osteospermum cv. Impassion Rose Purple. The same number of plants treated with 1/cm2 PDA plugs served as controls. Following inoculation, all plants were maintained in a growth chamber at 25°C and 95% relative humidity on a 12-h fluorescent light/dark regimen. Wilt symptoms and lesions at the base of stem identical to those observed in the nursery developed 7 days after inoculation, and all inoculated plants died within 20 days. Control plants remained symptomless. R. solani AG-4 was consistently reisolated from symptomatic tissues, completing Koch's postulates. To our knowledge, this is the first report of damping-off on the genus Osteospermum caused by R. solani. References: (1) D. E. Carling. Page 37 in: Grouping in Rhizoctonia solani by Hyphal Anastomosis Reactions. Kluwer Academic Publishers, the Netherlands, 1996. (2) R. H. Cruickshank and G. C. Wade. Anal. Biochem. 107:177, 1980. (3) M. C. Juliàn et al. Phytopathology 86:566, 1996. (4) C. C. Tu and J. W. Kimbrough. Mycologia 65:941, 1973.

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First Report of Crown and Root Rot Caused by Rhizoctonia solani AG-4 on Banana Passionflower (Passiflora mollissima) in Italy

Plant Disease ◽

10.1094/pdis-05-11-0411 ◽

2011 ◽

Vol 95 (9) ◽

pp. 1194-1194 ◽

Cited By ~ 2

Author(s):

G. Polizzi ◽

D. Aiello ◽

V. Guarnaccia ◽

A. Panebianco ◽

P. T. Formica

Keyword(s):

Rhizoctonia Solani ◽

Root Rot ◽

Disease Incidence ◽

Morphological Characteristics ◽

Fluorescent Light ◽

Disease Symptoms ◽

First Report ◽

Pathogenicity Tests ◽

Crown And Root Rot ◽

Safranin O

The genus Passiflora (Passifloraceae family) contains more than 500 species and several hybrids. In Italy, some of these species and hybrids are grown as ornamental evergreen vines or shrubs. During August and September 2010, a crown and root rot was observed in a stock of approximately 6,000 potted 2-year-old plants of Passiflora mollissima (Kunth) Bailey, commonly known as the banana passionflower, in a nursery located in eastern Sicily (southern Italy). Disease incidence was approximately 20%. Disease symptoms consisted of water-soaked lesions at the crown and a root rot. Successively, older crown lesions turned light brown to brown and expanded to girdle the stem. As crown and root rot progressed, basal leaves turned yellow and gradually became necrotic and infected plants wilted and died. A fungus with mycelial and morphological characteristics of Rhizoctonia solani Kühn was consistently isolated from crown lesions and brown decaying roots when plated on potato dextrose agar (PDA) amended with streptomycin sulfate at 100 μg/ml. Fungal colonies were initially white, turned brown with age, and produced irregularly shaped, brown sclerotia. Mycelium was branched at right angles with a septum near the branch with a slight constriction at the branch base. Hyphal cells removed from 10 representative cultures grown at 25°C on 2% water agar were determined to be multinucleate when stained with 1% safranin O and 3% KOH solution (1) and examined at ×400. Anastomosis groups were determined by pairing isolates on 2% water agar in petri plates (4). Pairings were made with tester strains of AG-1, AG-2, AG-3, AG-4, AG-5, AG-6, and AG-11. Anastomosis was observed only with tester isolates of AG-4 (3). Pathogenicity tests were performed on container-grown, healthy, 3-month-old cuttings. Twenty plants of P. mollissima were inoculated near the base of the stem with five 1-cm2 PDA plugs from 5-day-old mycelial plugs obtained from two representative cultures. The same number of plants served as uninoculated controls. Plants were maintained at 25°C and 95% relative humidity with a 12-h fluorescent light/dark regimen. Wilt symptoms due to crown and root rot, identical to ones observed in the nursery, appeared 7 to 8 days after inoculation with either of the two isolates and all plants died within 20 days. No disease was observed on control plants. R. solani AG-4 was reisolated from symptomatic tissues and identified as previously described, confirming its pathogenicity. Damping-off or crown and root rot due to R. solani were previously detected on P. edulis in Brazil, Africa, India, Oceania, and Australia (2). To our knowledge, this is the first report of R. solani causing crown and root rot on P. mollissima. References: (1) R. J. Bandoni. Mycologia 71:873, 1979. (2) J. L. Bezerra and M. L. Oliveira. Fitopathol. Brasil. 9:273, 1984. (3) D. E. Carling. Page 37 in: Grouping in Rhizoctonia solani by Hyphal Anastomosis Reactions. Kluwer Academic Publishers, the Netherlands, 1996. (4) C. C. Tu and J. W. Kimbrough. Mycologia 65:941, 1973.

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First Report on Ovate-leaf Atractylodes Damping-off Caused by Fusarium oxysporum species Complex in South Korea

Plant Disease ◽

10.1094/pdis-05-21-1107-pdn ◽

2021 ◽

Author(s):

Oliul Hassan ◽

Taehyun Chang

Keyword(s):

South Korea ◽

Fusarium Oxysporum ◽

Species Complex ◽

Disease Incidence ◽

Fusarium Species ◽

Morphological Characteristics ◽

Conidial Suspension ◽

Sterile Water ◽

Damping Off ◽

First Report

In South Korea, ovate-leaf atractylodes (OLA) (Atractylodes ovata) is cultivated for herbal medicine. During May to June 2019, a disease with damping off symptoms on OLA seedlings were observed at three farmer fields in Mungyeong, South Korea. Disease incidence was estimated as approximately 20% based on calculating the proportion of symptomatic seedlings in three randomly selected fields. Six randomly selected seedlings (two from each field) showing damping off symptoms were collected. Small pieces (1 cm2) were cut from infected roots, surface-sterilized (1 minute in 0.5% sodium hypochlorite), rinsed twice with sterile water, air-dried and then plated on potato dextrose agar (PDA, Difco, and Becton Dickinson). Hyphal tips were excised and transferred to fresh PDA. Six morphologically similar isolates were obtained from six samples. Seven-day-old colonies, incubated at 25 °C in the dark on PDA, were whitish with light purple mycelia on the upper side and white with light purple at the center on the reverse side. Macroconidia were 3–5 septate, curved, both ends were pointed, and were 19.8–36.62 × 3.3–4.7 µm (n= 30). Microconidia were cylindrical or ellipsoid and 5.5–11.6 × 2.5–3.8 µm (n=30). Chlamydospores were globose and 9.6 –16.3 × 9.4 – 15.0 µm (n=30). The morphological characteristics of present isolates were comparable with that of Fusarium species (Maryani et al. 2019). Genomic DNA was extracted from 4 days old cultures of each isolate of SRRM 4.2, SRRH3, and SRRH5, EF-1α and rpb2 region were amplified using EF792 + EF829, and RPB2-5f2 + RPB2-7cr primer sets, respectively (Carbone and Kohn, 1999; O'Donnell et al. 2010) and sequenced (GenBank accession number: LC569791- LC569793 and LC600806- LC600808). BLAST query against Fusarium loci sampled and multilocus sequence typing database revealed that 99–100% identity to corresponding sequences of the F. oxysporum species complex (strain NRRL 28395 and 26379). Maximum likelihood phylogenetic analysis with MEGA v. 6.0 using the concatenated sequencing data for EF-1α and rpb2 showed that the isolates belonged to F. oxysporum species complex. Each three healthy seedlings with similar sized (big flower sabju) were grown for 20 days in a plastic pot containing autoclaved peat soil was used for pathogenicity tests. Conidial suspensions (106 conidia mL−1) of 20 days old colonies per isolate (two isolates) were prepared in sterile water. Three pots per strain were inoculated either by pouring 50 ml of the conidial suspension or by the same quantity of sterile distilled water as control. After inoculation, all pots were incubated at 25 °C with a 16-hour light/8-hour dark cycle in a growth chamber. This experiment repeated twice. Inoculated seedlings were watered twice a week. Approximately 60% of the inoculated seedlings per strain wilted after 15 days of inoculation and control seedlings remained asymptomatic. Fusarium oxysporum was successfully isolated from infected seedling and identified based on morphology and EF-1α sequences data to confirm Koch’s postulates. Fusarium oxysporum is responsible for damping-off of many plant species, including larch, tomato, melon, bean, banana, cotton, chickpea, and Arabidopsis thaliana (Fourie et al. 2011; Hassan et al.2019). To the best of our knowledge, this is the first report on damping-off of ovate-leaf atractylodes caused by F. oxysporum in South Korea. This finding provides a basis for studying the epidemic and management of the disease.

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First Report of Stem and Foliar Blight of Sunflower Caused by Alternariaster helianthi in Louisiana

Plant Disease ◽

10.1094/pdis-01-12-0107-pdn ◽

2012 ◽

Vol 96 (5) ◽

pp. 761-761 ◽

Cited By ~ 1

Author(s):

R. Singh ◽

D. M. Ferrin

Keyword(s):

Disease Incidence ◽

Fluorescent Light ◽

Infected Leaf ◽

Stem Tissue ◽

First Report ◽

Foliar Blight ◽

Leaf Spots ◽

Plastic Bags ◽

Alternaria Helianthi ◽

Stem Lesions

During the fall of 2009, sunflower (Helianthus annuus L.) planted at the LSU AgCenter's Burden Center in Baton Rouge, LA exhibited severe stem and foliar blight symptoms. Symptoms on stems and petioles included elongated, slightly sunken lesions with dark brown margins. Leaf symptoms included irregular to circular, dark brown lesions with white centers and surrounded by a yellow halo. Several spots often coalesced to form large, blighted areas, and severely affected leaves turned yellow, followed by defoliation. The corolla and calyx exhibited similar lesions except for the yellow halo. Disease developed rapidly and the whole (100% disease incidence) field was blighted within a week following a rain (4 mm). Infected leaf and stem tissue was surface disinfested and plated on ¼-strength potato dextrose agar (PDA). Both leaf and stem tissue consistently produced dark olivaceous-to-black fungal colonies at room temperature under 12 h of fluorescent light per day. Conidia were 53 to 128 × 10 to 26 μm, borne singly on the conidiophores, hyaline to dark olivaceous, cylindrical, rounded at both ends, and with 6 to 10 transverse and 0 to 2 longitudinal septa. Conidiophores were single, unbranched, septate, hyaline to dark olivaceous, and measured 77 to 128 × 7 to 13 μm. Morphologically, the fungus was identified as Alternariaster helianthi (Hansf.) E.G. Simmons (= Alternaria helianthi [Hansf.] Tubaki & Nishih) (1). A single-spore isolate (PDC-4291) was obtained from the original culture and DNA from this isolate was extracted with a DNeasy Plant Mini Kit (Qiagen Inc., Valencia, CA). Primers ITS1 and ITS4 were used to amplify and sequence the internal transcribed spacer regions 1 and 2, and NCBI blast analysis of the 552-bp sequence (GenBank Accession No. JN208925) resulted in 100% homology with Alternaria helianthi isolated from sunflower infected with leaf spot and blight disease in India (GenBank Accession No. DQ156343). Pathogenicity was determined by inoculating 20 potted sunflower plants (Full Sun Improved TD, Fred C. Gloeckner and Company, Inc., Harrison, NY) with conidia from a 2-week-old culture of isolate PDC-4291. Each plant was sprayed with 25 ml of suspension containing 106 conidia/ml. Twenty control plants were sprayed with 25 ml of sterile distilled water. Inoculated and control plants were covered with plastic bags and maintained in a greenhouse at 28 ± 2°C. Plastic bags were removed 72 h after inoculation. Leaf spots similar to the original symptoms appeared on all 20 inoculated plants 5 days after inoculation. A few stem lesions were observed on 13 plants. Two weeks after inoculation, infected leaves turned yellow and blighted. Alternariaster helianthi (= Alternaria helianthi) was reisolated from the leaf spots and stem lesions. No symptoms developed on any of the 20 control plants. On the basis of morphology and sequence data, this pathogen was identified as A. helianthi, and to our knowledge, this is the first report of sunflower stem and foliar blight caused by A. helianthi in Louisiana. In Louisiana, sunflower is a popular ornamental that is grown in landscapes and gardens and by commercial flower growers who grow it for cut flower arrangements. Louisiana's hot, humid weather is ideal for disease development, which may discourage gardeners and commercial growers from planting sunflower. Reference: (1) E. G. Simmons. Alternaria: An Identification Manual. CBS Fungal Biodiversity Center, Utrecht, the Netherlands, 2007.

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First Report of Crown and Root Rot Caused by Rhizoctonia solani AG-4 on Orange Jessamine in Italy

Plant Disease ◽

10.1094/pdis-93-2-0204b ◽

2009 ◽

Vol 93 (2) ◽

pp. 204-204 ◽

Cited By ~ 1

Author(s):

D. Aiello ◽

A. Vitale ◽

E. Lahoz ◽

R. Nicoletti ◽

G. Polizzi

Keyword(s):

Rhizoctonia Solani ◽

Root Rot ◽

Morphological Characteristics ◽

Ruthenium Red ◽

Fluorescent Light ◽

First Report ◽

Pectic Enzymes ◽

Electrophoretic Patterns ◽

Crown And Root Rot ◽

Hyphal Diameter

Murraya paniculata (L.) Jack, commonly called orange jessamine or orange jasmine (Rutaceae), is a small tropical tree that is native to Asia. This species, closely related to Citrus, is grown as an ornamental tree or hedge. During October of 2007, crown and root rot was observed on approximately 12,000 pot-grown, 4-month-old plants in a nursery in eastern Sicily, Italy. Basal leaves turned yellow and gradually became necrotic, and infected plants often died. Disease symptoms were observed on 1,800 (15%) plants. Isolations from affected tissues on potato dextrose agar (PDA) amended with streptomycin sulfate at 100 mg/liter recovered a fungus with mycelial and morphological characteristics consistent with Rhizoctonia solani Kühn. Fungal colonies were initially white, turned brown with age, and produced irregularly shaped, brown sclerotia. Microscopic examination revealed that hyphae had a right-angle branching pattern, were constricted at the base of the branch near the union with main hyphae, and were septate near the constriction. The nuclear condition of hyphal cells was determined on cultures grown at 25°C on 2% water agar (WA) when stained with 3% safranin O solution and examined at ×400. Anastomosis groups were determined by pairing isolates on 2% WA in petri plates (4). Pairings were made with tester strains AG-1 IA, AG-2-2-1, AG-2-2IIIB, AG-2-2IV, AG-3, AG-4, AG-5, AG-6, and AG-11. Anastomosis was observed only with tester isolates of AG-4 producing both C2 and C3 reactions. The hyphal diameter at the point of anastomosis was reduced, the anastomosis point was obvious, and cell death of adjacent cells was observed. These results were consistent with other reports on anastomosis reactions (1). The identification of group AG-4 within R. solani has been confirmed by electrophoretic patterns of pectic enzymes (polygalacturonases) in vertical pectin-acrylamide gel stained with ruthenium red (2). Pathogenicity tests were conducted on potted, healthy, 6-month-old seedlings of orange jessamine. Twenty-five plants were inoculated by placing 1-cm2 PDA plugs from 5-day-old mycelial cultures near the base of the stem. The same number of plants inoculated with PDA plugs served as controls. Plants were maintained at 25°C and 95% relative humidity on a 12-h fluorescent light/dark regimen. Wilt symptoms, identical to ones observed in the nursery, developed 3 months after inoculation because of crown and root rot. Control plants remained disease free. The pathogen was reisolated from symptomatic tissues, completing Koch's postulates. Collar rot due to R. solani was previously detected on M. koenigii (3). To our knowledge, this is the first report of R. solani causing disease on M. paniculata. References: (1) D. E. Carling. Page 37 in: Grouping in Rhizoctonia solani by Hyphal Anastomosis Reactions. Kluwer Academic Publishers, the Netherlands, 1996. (2) R. H. Cruickshank and G. C. Wade. Anal. Biochem. 107:177, 1980. (3) A. C. Jain and K. A. Mahmud. Rev. Appl. Mycol. 32:460, 1953. (4) C. C. Tu and J. W. Kimbrough. Mycologia 65:941, 1973.

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First Report of Petunia Root Rot Caused by Rhizoctonia solani in Argentina

Plant Disease ◽

10.1094/pdis.2004.88.1.86b ◽

2004 ◽

Vol 88 (1) ◽

pp. 86-86

Author(s):

E. R. Wright ◽

M. C. Rivera ◽

K. Asciutto ◽

L. Gasoni ◽

V. Barrera ◽

...

Keyword(s):

Rhizoctonia Solani ◽

Root Rot ◽

Disease Incidence ◽

Common Garden ◽

Anastomosis Group ◽

Fluorescent Light ◽

First Report ◽

Blue Solution ◽

Crown And Root Rot ◽

Species Taxonomy

Common garden petunias (Petunia × hybrida Hort. Vilm.-Andr.) are herbaceous annual plants with brightly colored flowers up to 10 cm in diameter. During the winter of 2002, crown and root rot were observed on plants (cv. Ultra) growing in five greenhouses in Buenos Aires. Affected plants were randomly distributed in the greenhouses, and mean disease incidence in all the greenhouses was 26%. Basal leaves turned yellow and gradually became necrotic, and infected plants were often killed. Small pieces of affected tissues were disinfested in 2% sodium hypochlorite for 1 min and plated on 2% potato dextrose agar (PDA). Fifteen isolates identified to the genus Rhizoctonia were obtained. Fungal colonies were initially white, turned brown with age, and produced irregularly shaped, brown sclerotia. Hyphal branched at right angles, were constricted at the base of the branch near the union with main hyphae, and septate near the constriction. Basidia were not observed in the greenhouses or on the plates. Isolates were cultivated on water agar and incubated at 25°C for 3 days. Hyphal cells were determined to be multinucleate when stained with 1% aniline blue solution (2) and examined at ×400. Anastomosis group of one isolate was determined by using AG-4 HG II, AG-1 IA, AG-1 IB, AG-1 IC, AG-2 2-1, and AG-2 2IIIB tester strains of Rhizoctonia solani that includes isolates reported to be pathogenic on ornamentals (1). Anastomosis was observed only with strains of AG-4 HG II. Pathogenicity on this isolate was conducted on potted, healthy, adult plants that were 10 to 22 cm high and flowering. Thirty-five plants were inoculated by placing 1 cm2 pieces of PDA from 7-day-old mycelial cultures near the base of the stem. Twelve control plants were treated with 1 cm2 PDA plugs. Plants were kept at 22 to 24°C, >95% relative humidity, and 12 h of fluorescent light. Wilt symptoms due to basal stem rot appeared 7 days after inoculation, and all the inoculated plants died within 27 days. Control plants remained disease free. The pathogen was reisolated from symptomatic tissues, completing Koch's postulates. To our knowledge, this is the first report of R. solani causing disease on petunia in Argentina. References: (1) D. M. Benson and D. K. Cartwright. Ornamental diseases incited by Rhizoctonia spp. Pages 303–314 in: Rhizoctonia species: Taxonomy, Molecular Biology, Ecology, Pathology and Disease Control. B. Sneh et al., eds. Kluwer Academic Publishers, London, England, 1996. (2) C. C. Tu and J. W. Kimbrough. Mycologia 65:941, 1973.

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First Report of Damping-Off of Rhodiola sachalinensis Caused by Rhizoctonia solani AG-4 HG-II in China

Plant Disease ◽

10.1094/pdis-07-11-0559 ◽

2012 ◽

Vol 96 (1) ◽

pp. 142-142 ◽

Cited By ~ 1

Author(s):

Q. Bai ◽

Y. Xie ◽

X. Wang ◽

Y. Li ◽

J. Gao ◽

...

Keyword(s):

Rhizoctonia Solani ◽

Morphological Characteristics ◽

Its Region ◽

Herbaceous Plant ◽

Protective Effects ◽

Damping Off ◽

First Report ◽

Rhodiola Sachalinensis ◽

Perennial Herbaceous Plant ◽

Safranin O

Rhodiola sachalinensis A. Bor (family Crassulaceae), a perennial herbaceous plant, is distributed mainly in the mountainous areas of China, Japan, Korea, and Russia. It is widely used as a traditional Chinese medicine with adaptogenic properties, cardiopulmonary protective effects, and central nervous system activities (3). Currently, it is extensively cultivated in northeastern China. In August 2010, widespread (>60% of plants were symptomatic) damping-off was observed in a seedling field in Linjiang, China. Leaves and stems near the ground were affected first, with dark lesions forming on the stem and the lowest leaves exhibiting wilt. The wilt spread rapidly over the entire plant with leaves becoming grayish brown and water soaked and then turned black and died. Root rot, defoliation, and damping-off were also observed. Six isolates with morphological characteristics of Rhizoctonia solani Kühn were isolated from symptomatic stems when plated on potato dextrose agar (PDA). Mycelium was branched at right angles with a septum near the branch and a slight constriction at the branch base. Fungal colonies were initially white, turned brown with age, and produced irregularly shaped, brown sclerotia after 8 days on PDA. Hyphal cells removed from cultures grown at 25°C on 2% water agar were determined to be multinucleate when stained with 1% safranin O and 3% KOH solution (1) and examined at ×400 magnification. The internal transcribed spacer (ITS) region of the nuclear rDNA was amplified by using the primers ITS4/ITS5 (2). The ITS sequences (715 bp) were identical in these six isolates (GenBank Accession No. FR878087) and had 100% sequence identity with R. solani AG-4 HG-II (GenBank Accession No. HQ629873) along with numerous other accessions from this AG subgroup. Pathogenicity tests were performed on healthy, potted seedlings of R. sachalinensis. Twenty plants were inoculated near the base of the stem with a 0.6-cm-diameter mycelial plug from 3-day-old PDA cultures for each isolate. Twenty plants inoculated with only PDA plugs served as controls. The plants were covered with plastic bags and kept in a greenhouse at 20 to 25°C for 72 h. All inoculated plants showed characteristic symptoms as previously observed in the seedling field 13 days after inoculation, while control plants remained healthy. R. solani AG-4 HG-II was reisolated from symptomatic tissues on inoculated plants. To our knowledge, this is the first report of R. solani AG-4 HG-II causing damping-off on R. sachalinensis in China. References: (1) R. J. Bandoni. Mycologia 71:873, 1979. (2) D. E. L. Cooke et al. Mycol. Res. 101:667, 1997. (3) T. F. Yan et al. Conserv. Genet. 4:213, 2003.

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