First Report of Southern Blight of Manglietia decidua Caused by Sclerotium rolfsii in China

Manglietia decidua, named ‘Hua manglietia’, belonging to the Magnoliaceae family, is one of the most important ornamental plant in China. In 2019 and 2020, an unknown disease caused 3- to 12-month plants of M. decidua to wither and die in the field in Zhanjiang, Guangdong province（N21°9’3”;E110°17’47”）. Initially, the infected plants showed leaves dehydration, chlorosis and wilting with water-soaked lesions on stems at ground level. About 7 days later, the plants completely wilted, collapsed and died. Delayed and stunted growth with wilting of foliage continued through the whole year. Dense white mycelial mat and small white-to-brown spherical sclerotia were observed on the surface of the stalk lesion when weather conditions were warm and humid. Approximately 10% of plants were infected. Especially from July to October 2020, up to 30% of about 500 plants were infected and died. To identify the causal agents of the disease, infected tissue and sclerotia were collected, surface disinfected in 75% alcohol for 30s and 30% hydrogen peroxide solution for 5 min, and washed with sterile water for 1 min. The surface disinfected tissue and sclerotia were put on potato dextrose agar containing ampicillin (50mg/L) and kept in an incubator at 25°C in the dark. Fast growing fungus colonies with white mycelium and numerous sclerotia developed in the plates after 6 to 8 days of incubation. The hyphae were septate, hyaline and formed typically clamp connection after 10 days of growth. Sclerotia were initially white and became tan to dark brown over time and 1.0 to 3.0 mm (2.13 mm on average, n=124) in diameter at maturity. For molecular identification，the ITS region was amplified using primer pair ITS1/ITS4 (White et al. 1990). A 666 bp PCR product was sequenced (GenBank accession no. MW093622) and shared above 99% sequence identity with some Athelia rolfsii isolates (GenBank accession Nos. HQ895869, KX499470 and AB075290). Based on morphological and molecular characteristics, the fungus was identified as Sclerotium rolfsii (teleomorph A. rolfsii) （Paul et al. 2017，Xu et al. 2010. Pathogenicity tests were conducted by inoculating ten healthy 1-year-old M. decidua plants grown in pots. Five sclerotia and mycelial mat obtained from 15-day-old cultures were buried adjacent to the stem of each unwounded healthy plant. Non-inoculated plants served as controls. After inoculation, the plants were maintained in a 25-28 ℃ greenhouse and watered regularly to keep the soil moisture content at about 15%. Symptoms of southern blight were observed on all inoculated plants, which began to wilt 7 to 10 days after inoculation and died within 15 to 20 days. The control plants remained healthy. S. rolfsii was again isolated from the artificially inoculated plants, but not from non-inoculated plants. The pathogenicity test was repeated twice and the results were the same. S. rolfsii has an extensive host range worldwide and the common host ornamental plants are Iris, Chrysanthemum, CymbidiumTrifolium, Jasminum, Begonia, and Stevia etc. in China. To our knowledge, this is the first report of southern blight caused by S. rolfsiii on M. decidua in China. M. decidua is a horticultural plant which belongs to the protected and endangered tree species. This finding is important to alert growers to realize the proper management of this disease during species protection and cultivar extension.

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First Report of Southern Blight on Silverbush (Convolvulus cneorum) Caused by Sclerotium rolfsii in Italy

Plant Disease ◽

10.1094/pdis-94-1-0131a ◽

2010 ◽

Vol 94 (1) ◽

pp. 131-131 ◽

Cited By ~ 3

Author(s):

G. Polizzi ◽

D. Aiello ◽

V. Guarnaccia ◽

G. Parlavecchio ◽

A. Vitale

Keyword(s):

Sclerotium Rolfsii ◽

Ground Level ◽

Limiting Factor ◽

First Report ◽

Southern Blight ◽

The World ◽

Perennial Shrub ◽

Stem Lesions ◽

The Masses ◽

Tropical Climates

The genus Convolvulus has more than 200 species that are encountered in temperate to tropical climates all around the world. Convolvulus cneorum L., also known as silverbush, is a perennial shrub native to southern Europe (Sicily and Croatia) with dense, silver foliage and masses of large, circular, white flowers. During July of 2009, a widespread blight was observed on approximately 10% of 12,000 4-month-old potted silverbush plants. The plants were obtained from cuttings and produced by a commercial nursery in eastern Sicily, Italy. Symptomatic plants initially had sunken, tan lesions at the ground level that developed into typical southern blight. Circular and crescent-shaped patches were observed on the masses of weeds on the surface of the containers where silverbush were grown. At the soil line, white mycelia and small (1 to 2 mm in diameter), brown, spherical sclerotia with internally differentiated rind, cortex, and medulla characteristic of Sclerotium rolfsii Sacc. were observed. Crown and stem lesions were surface disinfested (1% NaOCl) for 1 min, rinsed in sterile water, and placed on potato dextrose agar. Isolation consistently yielded colonies of Sclerotium rolfsii (teleomorph Athelia rolfsii (Curzi) Tu & Kimbrough) with typical sclerotia produced within 6 to 7 days (2). Pathogenicity tests were performed on 20 plants by placing 10 sclerotia obtained from 10-days-old cultures in the soil below the crown portion on each of 5-month-old healthy cuttings of silverbush. The same number of plants served as noninoculated controls. All plants were maintained in a growth chamber at 25 ± 1°C and enclosed for 7 days in polyethylene bags. Plants were then moved to a greenhouse where temperatures ranged from 24 to 28°C. Symptoms of southern blight developed after 7 to 20 days on all inoculated plants. Control plants remained symptomless. S. rolfsii was reisolated from symptomatic plants. S. rolfsii was reported for the first time in Sicily in 2004 in an ornamental nursery (1). To our knowledge, this is the first report in the world of S. rolfsii on silverbush and it is the first outbreak of southern blight on Convolvulus species. The high susceptibility of silverbush to the pathogen could be a limiting factor for the cultivation in nursery of this indigenous wildflower plant. References: (1) G. Polizzi et al. Plant Dis. 88:310, 2004. (2) Z. K. Punja and A. Damiani. Mycologia 88:694, 1996.

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First Report of Southern Blight on Canadian Goldenrod (Solidago canadensis) Caused by Sclerotium rolfsii in China

Plant Disease ◽

10.1094/pdis-94-9-1172b ◽

2010 ◽

Vol 94 (9) ◽

pp. 1172-1172 ◽

Cited By ~ 7

Author(s):

W. Tang ◽

Y. Z. Zhu ◽

H. Q. He ◽

S. Qiang

Keyword(s):

Eastern China ◽

Sclerotium Rolfsii ◽

Soil Surface ◽

Pathogenicity Test ◽

Solidago Canadensis ◽

Uniform Size ◽

First Report ◽

Southern Blight ◽

Perennial Plant ◽

Nanjing City

Canadian goldenrod (Solidago canadensis L., Asteraceae) is a rhizomatous perennial plant native to North America that has invaded eastern China and continues to spread northward and westward. It is quite common on field borders, roadsides, and in undeveloped areas, posing a serious threat to native ecosystems and their biodiversity. During the late summers of 2007 and 2008, wilted Canadian goldenrod plants were occasionally found in invasive populations in the suburb of Nanjing city. Wilted plants were transplanted and maintained in a greenhouse at Nanjing Agricultural University. A white mass of fungal hyphae, which grew on the soil surface around the stem of the symptomatic S. canadesis plants and eventually covered the stem, was observed. Initially, the base of the stem became yellow, turned brown, and the light brown discoloration extended up the stem to a height of 3 to 7 cm. The leaves then collapsed, starting from the top until the entire plant wilted. The fungus produced numerous, small, roundish sclerotia of uniform size (0.7 to 2.0 mm in diameter), which were white at first and then became brown to dark brown. The fungus grew into the stems and downward into the rhizome area, but no sclerotia were detected inside the stem or root. Diseased tissue with sclerotia was disinfested for 1 min in 1% NaOCl and plated on potato dextrose agar amended with 100 mg/liter of streptomycin sulfate. On the basis of sclerotia morphology and the presence of clamp connections at hyphal septa, the fungus was identified as Sclerotium rolfsii. Pathogenicity of the isolate was confirmed by inoculating 1-year-old S. canadensis plants (average 1.5 m high) grown in pots. The inoculum consisted of cottonseed hulls infested with mycelium and sclerotia of the pathogen and was placed on the soil surface around the base of each unwounded plant. Noninoculated plants served as controls. The pathogenicity test was conducted twice. After inoculation, the plants were maintained at high humidity and 30°C for 3 days and then transferred to a greenhouse. All inoculated plants developed symptoms of southern blight. Inoculated plants developed symptoms of wilting 5 to 7 days after inoculation and were completely wilted within 15 to 20 days. Symptoms of wilting were soon followed by the appearance of white-to-light brown sclerotia on the collar region. Control plants remained symptomless and Sclerotium rolfsii was reisolated from inoculated plants. To our knowledge, this is the first report of southern blight of Canadian goldenrod caused by Sclerotium rolfsii in China.

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First Report of Chrysoporthe deuterocubensis Causing Canker on Syzygium samarangense in Taiwan

Plant Disease ◽

10.1094/pdis-03-13-0345-pdn ◽

2013 ◽

Vol 97 (11) ◽

pp. 1508-1508

Author(s):

M. C. Fan ◽

C. C. Huang ◽

J. S. Huang ◽

S. F. Tsai ◽

H. C. Yeh ◽

...

Keyword(s):

Apple Tree ◽

Average Length ◽

Its Region ◽

Fruit Production ◽

Molecular Characteristics ◽

Malt Extract ◽

Pathogenicity Test ◽

Total Production ◽

First Report ◽

Β Tubulin

Wax apple (Syzygium samarangense Merr. & Perry, syn. Eugenia javanica Lam.) belongs to the Myrtaceae family is an important economical tree fruit in Taiwan. The total production acreage of wax apple was 5,266 ha in which more than 77% were located in Pingtung County, southern Taiwan, in 2012. Since the winter of 2010, symptoms of withering leaves and cracking branches on wax apple trees were observed in some orchards in Nanjhou and Linbian Townships, Pingtung County. Diseased trees declined gradually and resulted in reduced fruit production. On the bark of diseased twigs and branches, black conidiamata with yellowish orange conidia were usually observed. For diagnosis, tissues from symptomatic branches were excised, surface sterilized with 0.5% sodium hypochlorite, and placed on 2% water agar in petri dishes. A total of four identical fungal isolates were obtained and maintained on potato dextrose agar (PDA). To fulfill Koch's postulates, three twigs of a wax apple tree were wounded with scalpel and inoculated with each of the four isolates, one tree per isolate. A 7-day-old hyphal mat (about 7 × 18 mm) of each fungal isolate was attached on the wound, wrapped with a wet absorbent cotton and Parafilm, and then covered with a layer of aluminum foil. For the control, the twigs of a wax apple tree were inoculated with PDA plugs. The pathogenicity test was repeated once. After 30 days, withering leaves and cracking twigs were observed on inoculated twigs and the same pathogen was reisolated. Conversely, all of the non-inoculated plants remained healthy. Identification of the pathogen was conducted using its morphological, physiological, and molecular characteristics. On malt extract agar, the colony was floccose and white with hazel hues. The optimal temperature for the mycelial growth was 30°C. Conidia were hyaline, and oblong, with the average size of 4.7 ± 0.6 × 2.7 ± 0.2 μm (100 conidia). Ascostromata were semi-immersed in the bark with fusoid asci, eight ascospores per ascus. Ascospores were hyaline, 2-celled, and tapered in both ends, with the average length of 6.8 ± 0.7 × 2.4 ± 0.3 μm (100 ascospores). For molecular identification, the internal transcribed spacer (ITS) of ribosomal DNA and β-tubulin genes was amplified using the ITS1/ITS4 (3), Bt1a/Bt1b, and Bt2a/Bt2b (1) primer pairs. The gene sequences were deposited in GenBank (Accessions KC792616, KC792617, KC792618, and KC792619 for the ITS region; KC792620, KC792621, KC792622, and KC792623 for Bt1 region, and KC812732, KC812733, KC812734, and KC812735 for Bt2 region) and showed 99 to 100% identity to the Chrysoporthe deuterocubensis isolate CMW12745 (DQ368764 for ITS region; GQ290183 for Bt1 region, and DQ368781 for Bt2 region). In addition, the Bt1 region of the β-tubulin gene consisted of two restriction sites for AvaI and one restriction site for HindIII. This is identical to the description of C. deuterocubensis, a cryptic species in C. cubensis, by Van Der Merwe et al. (2). According to these results, the pathogen was identified as C. deuterocubensis Gryzenh. & M. J. Wingf. To the best of our knowledge, this is the first report of canker disease caused by C. deuterocubensis on S. samarangense in Taiwan. References: (1) N. L. Glass and G. C. Donaldson. Appl. Environ. Microbiol. 61:1323, 1995. (2) N. A. Van Der Merwe et al. Fungal Biol. 114:966, 2010. (3) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.

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First Report of Southern Blight of Mexican Petunia (Ruellia brittoniana) Caused by Sclerotium rolfsii in Taiwan

Plant Disease ◽

10.1094/pdis-04-12-0412-pdn ◽

2012 ◽

Vol 96 (12) ◽

pp. 1822-1822

Author(s):

C. H. Fu ◽

Y. P. Huang ◽

F. Y. Lin

Keyword(s):

Sclerotium Rolfsii ◽

Stem Rot ◽

Pathogenicity Test ◽

Academic Press ◽

First Report ◽

Southern Blight ◽

Inoculation Techniques ◽

Pure Cultures ◽

Causal Fungus ◽

Garden Plant

Mexican petunia (Ruellia brittoniana) is an herbaceous flowering perennial with strikingly colored flowers, widely cultivated commercially as a potted plant and a popular garden plant. In July of 2010, root and stem rot that caused death was observed on Mexican petunia at the flower nursery of the Council of Agriculture & Chiayi County in Taiwan. Plants had rotted and girdled stem bases. Necrotic areas were covered with fans of white mycelium as well as abundant spherical sclerotia. A fungus was isolated from infected tissue and sclerotia, and maintained on potato dextrose agar (PDA) plates incubated at 25°C without light. Colonies were white, cottony, often forming fans; pure cultures were prepared by transferring hyphal tips to PDA. Sclerotia formed after 10 days, initially white becoming dark brown with age, and 0.5 to 0.6 mm in diameter. To confirm identity of the causal fungus, the complete internal transcribed spacer (ITS) rDNA region of the causal fungus was amplified using the primers ITS4 and ITS5 (2) and sequenced. The resulting sequence of 687 bp was uploaded in NCBI. The sequence was 98% similar to sequences of Athelia rolfsii (Sclerotium rolfsii) in NCBI (Accession No. JN543691.1). Koch's postulates were performed using two inoculation techniques. The soil near the base of healthy Mexican petunia plants (four plants per pot) were exposed to recently matured sclerotia (10 sclerotia per plant) developed from pure fungal cultures or 10-mm-diameter agar plugs of mycelium (one plug per plant). Noninoculated plants, in a separate pot, were used as a control. All plants were incubated in a growth chamber at 28 to 33°C. Disease symptoms occurred on all inoculated plants by 5 to 7 days and included yellowing of leaves, basal stem rot, and wilt. Ten days after inoculation, inoculated plants were dead whereas control plants remained healthy. The pathogenicity test was repeated twice with similar results and S. rolfsii was reisolated from infected plants in each test. The pathogen has been reported to cause substantial loss of Mexican petunia in Louisiana (1). The disease is becoming more common in Taiwan and could cause losses in Mexican petunia production. To our knowledge, this is the first report of disease on Mexican petunia caused by S. rolfsii in Taiwan. References: (1) G. E. Holcomb. Plant Dis. 88:770, 2004. (2) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, 1990.

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First Report of Web Blight on Rebutia perplexa Caused by Rhizoctonia solani AG 2-2-IIIB in Italy

Plant Disease ◽

10.1094/pdis-10-13-1063-pdn ◽

2014 ◽

Vol 98 (5) ◽

pp. 697-697 ◽

Cited By ~ 1

Author(s):

A. Garibaldi ◽

D. Bertetti ◽

P. Pensa ◽

G. Ortu ◽

M. L. Gullino

Keyword(s):

Rhizoctonia Solani ◽

Aerial Mycelium ◽

Its Region ◽

Molecular Characteristics ◽

Fluorescent Light ◽

Pathogenicity Test ◽

Mycelium Growth ◽

Daily Growth ◽

First Report ◽

Wheat Kernels

Rebutia perplexa, Cactaceae family, is a clumping fine thorny cactus, producing several flushes of pink flowers. In the spring of 2013, a blight was observed in a farm located near Imperia (northern Italy) on 2% of 2,000 3-year-old plants, grown in plastic pots. Affected plants showed pale brown discoloration of stems, starting from the base, and eventually collapsed. Flowers also rotted and wilted. In the presence of high relative humidity, a rare, whitish mycelium developed on the surface of the substrate. Eventually, infected plants died. Symptomatic tissues of the stem were taken from 10 plants and plated on potato dextrose agar (PDA). A fungus with the morphological characters of Rhizoctonia solani (3) was consistently recovered. Three representative isolates obtained from affected plants were successfully paired with tester strains of R. solani (AG 1, AG 2-2-IIIB, AG 2-2-IV, AG 4, AG 7, AG 11) (2) and examined microscopically. Three replicated pairings were made for each tester strain. The Rebutia isolates anastomosed only with AG 2-2-IIIB tester strain with high hyphal fusion frequency. The hyphal diameter at the point of anastomosis was reduced, the anastomosis point was obvious, and death of adjacent cells was observed, indicating anastomosis reactions (1). Tests were performed twice. Mycelium of 15-day-old isolates maintained at 27 to 30°C, appeared whitish or pale buff in color, coarse, with a concentric zonation, scarce aerial mycelium, and without sclerotia. The optimum temperature for mycelium growth was 30°C (daily growth rate: 24.6 mm) and isolates grew also at 35°C. The internal transcribed spacer (ITS) region of rDNA was amplified using the primers ITS1/ITS4 and sequenced. BLASTn analysis of the 523-bp amplicon (GenBank Accession No. KF719318) showed a 99% homology with the sequence of the R. solani AG 2-2-IIIB isolate GU811672. The nucleotide sequence has been assigned the GenBank Accession No. KF719318. Therefore, on the basis of molecular characteristics, anastomosis tests, temperature growth, and cultural characteristics, the isolates from R. perplexa were identified as R. solani AG 2-2-IIIB. For pathogenicity tests, 3 g of colonized wheat kernel from 10-day-old cultures of a representative isolate of the fungus was added per 1 l of substrate in 12 potted healthy plants of R. perplexa. The inoculum was prepared by inoculating wheat kernels with the mycelium of 10-day-old cultures of the fungus and incubating at 25 ± 1°C (12 h fluorescent light, 12 h dark). Twelve plants inoculated with non-infested wheat kernels served as controls. Plants were covered with plastic bags and maintained in a growth chamber at 25 ± 1°C. The first symptoms, similar to those observed in the farm, developed 5 days after inoculation. Fifteen days after the artificial inoculation, all inoculated plants were dead. R. solani was re-isolated only from the stems of symptomatic plants. Control plants remained healthy. The pathogenicity test was carried out twice with similar results. This is, to our knowledge, the first report of blight of R. perplexa caused by R. solani in Italy as well as worldwide. References: (1) 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, pp. 37-47, 1996. (2) A. Ogoshi. Ann. Rev. Phytopathol. 25:125, 1987. (3) B. Sneh et al. Identification of Rhizoctonia species. APS Press, St Paul, MN, 1991.

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First Report of Southern Blight Caused by Sclerotium rolfsii on St.-John's-Wort

Plant Disease ◽

10.1094/pdis.1999.83.7.696c ◽

1999 ◽

Vol 83 (7) ◽

pp. 696-696 ◽

Cited By ~ 2

Author(s):

A. P. Keinath ◽

J. W. Rushing ◽

R. J. Dufault

Keyword(s):

Sclerotium Rolfsii ◽

Soil Surface ◽

The United States ◽

Pathogenicity Test ◽

First Report ◽

Southern Blight ◽

St John’S Wort ◽

St John's Wort ◽

Pathogenicity Tests ◽

Hypericum Perforatum L

Interest in commercial production of common St.-John's-wort (Hypericum perforatum L.), an herb that is dried, processed, and used as an anti-depressant medication, is increasing. In August 1998, St.-John's-wort growing in the field at Charleston, SC, showed blight symptoms. Leaves on prostrate branches turned reddish-yellow, then brown, and then abscised. As the disease progressed, branches and approximately 10% of the plants were killed. Coarse, white mycelia were present on the bases of dead branches. Segments cut from symptomatic branches were disinfested in 0.5% sodium hypochlorite and placed on potato dextrose agar (PDA) at 25°C. Sclerotium rolfsii Sacc. was isolated from one of 12 branches with discolored leaves and six of six dead branches. For pathogenicity tests, sclerotia were harvested from 6-week-old cultures on PDA. Ten-week-old St.-John's-wort plants, growing in potting mix in 10-cm pots, were inoculated by placing four sclerotia on the soil surface 1 to 1.5 cm from the main stem of each plant. Plants were grown in a greenhouse at 90% relative humidity and 25 to 35°C. Single blighted branches were observed on three plants 12 days after inoculation and all plants were blighted 28 days after inoculation. S. rolfsii was recovered from 10 and 9 of 10 plants inoculated with isolates of S. rolfsii from St.-John's-wort and tomato, respectively. All 10 noninoculated plants remained symptomless. The pathogenicity test was repeated and the results were similar. This is the first report of S. rolfsii causing Southern blight on St.-John's-wort in the United States.

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First Report of Southern Blight on Firewheel Tree, Bay Laurel, Bird of Paradise, Mediterranean Fan Palm, and Liverwort Caused by Sclerotium rolfsii in Italy

Plant Disease ◽

10.1094/pdis-91-9-1199b ◽

2007 ◽

Vol 91 (9) ◽

pp. 1199-1199 ◽

Cited By ~ 2

Author(s):

G. Polizzi ◽

D. Aiello ◽

I. Castello ◽

A. Vitale ◽

G. Parlavecchio

Keyword(s):

Ornamental Plants ◽

Sclerotium Rolfsii ◽

Ground Level ◽

Marchantia Polymorpha ◽

First Report ◽

Southern Blight ◽

Potted Plants ◽

Inoculation Trial ◽

Athelia Rolfsii ◽

Strelitzia Reginae

During the summer of 2006, a widespread blight was observed on 6-month-old potted plants of firewheel tree (Stenocarpus sinuatus Endl.) and 3-month-old potted plants of bay laurel (Laurus nobilis L.) growing in a nursery in eastern Sicily, Italy. On both species, symptomatic plants initially had sunken, tan lesions at ground level where white mycelia and small (1 to 2 mm in diameter), brown, spherical sclerotia typical of Sclerotium rolfsii Sacc. were formed. As the disease progressed, the mycelia extended up the stem and entire plants collapsed. A sudden wilting affecting 4-month-old potted seedlings of bird of paradise (Strelitzia reginae Aiton) and 5-month-old potted seedlings of Mediterranean fan palm (Chamaerops humilis L.) was occasionally detected in other greenhouses of the same nursery. Liverwort (Marchantia polymorpha L.) was abundantly present on the surfaces of the containers where these plants were grown. Circular and crescent-shaped patches as much as 100 cm in diameter were observed on the massed liverwort plants. In these patches, the liverwort died and sclerotia typical of S. rolfsii were dispersed on white mycelial strands. Symptomatic tissues of the ornamental plants and liverwort were surface disinfested in 1% NaOCl for 1 min, rinsed in sterile water, and plated on potato dextrose agar. Tissues consistently yielded S. rolfsii (teleomorph Athelia rolfsii (Curzi) Tu & Kimbrough) and typical sclerotia with internally differentiated rind, cortex, and medulla were produced within 6 or 7 days (3). Pathogenicity tests were performed by placing 30 sclerotia obtained from 10-day-old cultures in the soil below the crown portion on each of 2-month-old healthy seedlings of Stenocarpus sinuatus, L. nobilis, Strelitzia reginae, and C. humilis (20 seedlings per host). In addition, liverwort growing in 10 pots (7 cm in diameter) was inoculated with 30 sclerotia per pot. For each species, the same number of plants or pots served as control. All ornamental plants and liverwort were maintained in a growth chamber at 25 ± 1°C and enclosed for 7 days in polyethylene bags and then moved to a greenhouse where temperatures ranged from 24 to 28°C. The inoculation trial was repeated once. Symptoms of southern blight developed after 5 to 20 days on all inoculated plants of Stenocarpus sinuatus and sporadically (two to five plants) after 20 days on L. nobilis, Strelitzia reginae, and C. humilis. After 5 days, liverwort in all inoculated pots was colonized and plants died within 12 days. Control plants of all species remained symptomless. S. rolfsii was reisolated from symptomatic plants. S. rolfsii was reported for the first time in Sicily in 2004 on ornamental plants (2). Strelitzia reginae was previously reported as a host of Corticium rolfsii (synonym S. rolfsii) in Portugal (1). To our knowledge, this is the first report of S. rolfsii on Stenocarpus sinuatus, L. nobilis, and C. humilis. In addition, this is the first report of the susceptibility of M. polimorpha to S. rolfsii. Liverwort could provide a food source for the fungus in container-grown nursery plants. References: (1) M. R. de Sousa Dias and M. T. Lusas. Bol. Soc. Brot. 53:469, 1980. (2) G. Polizzi et al. Plant Dis. 88:310, 2004. (3) Z. K. Punja and A. Damiani. Mycologia 88:694, 1996.

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First Report of Southern Blight Caused by Sclerotium rolfsii on Common Bean (Phaseolus vulgaris) in Italy

Plant Disease ◽

10.1094/pdis-12-12-1165-pdn ◽

2013 ◽

Vol 97 (10) ◽

pp. 1386-1386 ◽

Cited By ~ 2

Author(s):

A. Garibaldi ◽

G. Gilardi ◽

G. Ortu ◽

M. L. Gullino ◽

M. Testa

Keyword(s):

Phaseolus Vulgaris ◽

Common Bean ◽

Sclerotium Rolfsii ◽

Crown Rot ◽

Control Treatment ◽

Pathogenicity Test ◽

Green Beans ◽

First Report ◽

Southern Blight ◽

White Mycelium

Common bean (Phaseolus vulgaris L.) is grown worldwide for consumption of dry or green beans. During late spring of 2012, yellowing and wilting symptoms were observed in a commercial bean field cv. Lingua di fuoco in Cagliari Province (Sardinia, southern Italy) on 30% of plants 4 to 5 months after sowing. The first symptoms developed in May, when temperatures reached 18 to 30°C. Affected plants showed crown rot, necrosis of the cortex, and foliar chlorosis. As disease progressed, plants collapsed. In the presence of abundant moisture, white mycelium developed on the senescent tissue along with light to dark brown sclerotia (3.0 to 4.8 mm in diameter). Symptomatic tissue was disinfested for 1 min in 1% NaOCl and plated on potato dextrose agar (PDA) amended with 25 mg streptomycin sulfate/liter. The fungus that was isolated consistently from symptomatic plants onto PDA at 23°C grew rapidly in culture with silky-white, sterile mycelium, formed light to dark brown sclerotia (each 1.8 to 3.2 mm in diameter) after 7 days, and readily produced aerial hyphae. These morphological features are typical of Sclerotium rolfsii (2). The internal transcribed spacer (ITS) region of ribosomal DNA (rDNA) was amplified for one isolate using ITS1/ITS4 primers (4), and sequenced (GenBank Accession No. KF002510). BLASTn analysis (1) of the 656-bp segment showed 87% homology with the ITS sequence of an S. rolfsii isolate (JF819727). Pathogenicity of one isolate was confirmed by inoculating healthy P. vulgaris plants cv. Lingua di fuoco grown in 2-liter pots in a steamed potting mix containing 50% Tecno2 (70% white peat and 30% clay) and 50% Tiesse 3 (60% white peat, 20% clay, and 20% perlite) (Turco Silvestro terricci, Bastia d'Albenga, SV, Italy). Inoculum consisting of mycelium and sclerotia of the pathogen produced from 10-day-old cultures on PDA was mixed in the soil at 0.5 g/liter substrate. Four 7-day-old plants per pot, with three replicate pots, were used for inoculation. The same number of control plants grown in the same substrate were inoculated with non-colonized PDA as a negative control treatment. The pathogenicity test was repeated. Plants were kept in a growth chamber at 30°C and 85% RH. Inoculated plants developed symptoms of leaf yellowing within 10 days, followed by crown rot, appearance of white mycelium and sclerotia, and eventual wilting. Control plants remained asymptomatic. Isolations from inoculated plants demonstrated the absence of latent infections by the fungus S. rolfsii, but the fungus was not reisolated from non-inoculated control plants. To our knowledge, this is the first report of S. rolfsii infecting P. vulgaris in Italy. Southern blight has been reported on common bean in sub-tropical and tropical areas of the world (3), where it can cause severe crop losses. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) J. E. M. Mordue. CMI Descriptions of Pathogenic Fungi and Bacteria No. 410, 1974. (3) H. F. Schwartz et al. Page 20 in: Compendium of Bean Diseases. American Phytopathological Society Press, St. Paul, MN, 2005. (4) T. J. White et al. PCR Protocols. Page 315 in: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.

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First Report of Root and Stem Rot Caused by Phytophthora nicotianae on Peperomia tetraphylla in China

Plant Disease ◽

10.1094/pdis-94-9-1171c ◽

2010 ◽

Vol 94 (9) ◽

pp. 1171-1171 ◽

Cited By ~ 1

Author(s):

D. X. Zeng ◽

X. L. Wu ◽

Y. H. Li

Keyword(s):

Southern China ◽

Aerial Mycelium ◽

Its Region ◽

Soil Extract ◽

Phytophthora Nicotianae ◽

Stem Rot ◽

Molecular Characteristics ◽

Its Sequence ◽

First Report ◽

Pure Cultures

Peperomia tetraphylla, an evergreen herb, is becoming increasingly popular as a potted ornamental plant in southern China. In the summer of 2008, in some commercial flower nurseries in Shenzhen, Guangdong Province, P. tetraphylla showed extensive black stem and root rot, with leaves dropping from the rotten stem. Small pieces (approximately 3 mm2) of stems and leaves were excised from the margins of the black lesions, surface disinfected for 30 s to 1 min in 0.1% HgCl2, plated onto potato dextrose agar (PDA), and incubated at 25°C in the dark. All the plated samples yielded Phytophthora, and microscopic examination of pure cultures grown on PDA plates showed arachnoid colonies with abundant aerial mycelium, chlamydospores, and a few sporangia. Numerous sporangia were formed in sterile soil extract. Sporangia were ovoid or obpyriform, noncaducous, with prominent solitary papillae, and measured 31 to 52 μm (average 38 μm) × 21 to 34 μm (average 27 μm). Chlamydospores were spherical and 21 to 34 μm in diameter (average 28 μm). The internal transcribed spacer (ITS) region of rDNA of a single isolate was amplified using primers ITS4/ITS5 and sequenced (2). The ITS sequence, when submitted for a BLAST search in the NCBI database, showed 100% homology with the sequences of two reference isolates of Phytophthora nicotianae (Accession Nos. AY833526 and EU433396) and the consensus ITS sequence was deposited in the NCBI as Accession No. GQ499373. The isolate was identified as Phytophthora nicotianae on the basis of morphological and molecular characteristics (1). Pathogenicity of the isolate was confirmed by inoculating 1-year-old plants of P. tetraphylla growing in pots. The isolate was grown for 7 days on PDA plates and mycelial plugs, 5 mm in diameter and taken from the advancing margins of the colonies, were buried approximately 1 cm deep near the base of the stem in such a way that the mycelium on the plugs was in contact with the surface of the stem, which had been wiped earlier with 70% ethanol and gently wounded with a needle. Plants treated the same way but inoculated with sterile PDA plugs served as control plants. Three plants in each pot were inoculated and there were five replications each for the treatment and the control. All plants were kept in a greenhouse at 22 to 32°C. After 6 to 7 days, the inoculated plants showed black lesions around the mycelial plugs; symptoms of root and stem rot developed rapidly thereafter and the plants collapsed within 2 weeks. All symptoms on the inoculated plants were identical to those observed in naturally diseased plants, whereas the control plants remained healthy. The same fungus was consistently reisolated from the inoculated plants. To our knowledge, this is the first report of Phytophthora nicotianae on P. tetraphylla in China. References: (1) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phytopathological Society, St. Paul, MN, 1996. (2) J. B. Ristaino et al. Appl. Environ. Microbiol. 64:948, 1998.

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