scholarly journals First Report of Rhizoctonia solani Causing a Disease of Sunflower in India

Plant Disease ◽  
2010 ◽  
Vol 94 (4) ◽  
pp. 488-488 ◽  
Author(s):  
K. Srinivasan ◽  
S. Visalakchi

During the spring of 2009, symptoms including leaf yellowing and wilting, root rot, and death of plants were noted in sunflower (Helianthus annuus L.) crops in Dharmapuri District, Tamilnadu, India. In some fields, approximately 30% of the plants were affected. The disease began when plants were approximately 10 weeks old and occurred on scattered or adjacent plants. The presence of white mycelium was observed on necrotic crowns. Symptomatic tissue was surface disinfested in 70% alcohol for 30 s and 0.5% sodium hypochlorite for 1 min and plated onto potato dextrose agar (PDA) (1). One isolate (coded SV001) had near right-angle branching with basal constriction and adjacent septa and sclerotia typical of Rhizoctonia spp. (2). Cream-colored colonies produced irregular, light brown sclerotia that were 3.0 to 7.3 mm (average 3.8 mm) in diameter. Hyphae were 6.8 to 7.0 μm (average 6.9 μm) wide and multinucleate (8 to 15 nuclei per cell). On the basis of hyphal anastomosis with several known AG testers, the fungus was characterized as Rhizoctonia solani Kühn AG-IV (3). One culture was deposited at the Madras University Botany Laboratory, Center for Advanced Studies in Botany, University of Madras, Chennai, India. In a pathogenicity test, R. solani SV001 was grown on PDA for 5 days at 24°C in the dark. Five-millimeter-diameter disks were placed at the base of sunflower plants (cv. Mordan). Four sunflower plants in each of three pots were inoculated; noninoculated plants served as controls. Plants were placed in a glasshouse maintained at 25 to 27°C. Inoculated plants developed yellow foliage and crown rot and root rot symptoms after 7 to 12 days and died in 17 to 20 days. No symptoms were observed on noninoculated plants. The pathogen was reisolated from fragments of necrotic crown tissue of inoculated plants. To our knowledge, this is the first report of R. solani AG-IV causing a disease of sunflower plants in India. References: (1). R. C. Fenille et al. Plant Pathol. 54:325, 2005. (2). J. R. Parmeter et al. Phytopathology 59:1270, 1969. (3) B. Sneh et al. Identification of Rhizoctonia Species. The American Phytopathological Society, St Paul, MN, 1991.

Plant Disease ◽  
2003 ◽  
Vol 87 (4) ◽  
pp. 447-447 ◽  
Author(s):  
V. Vujanovic ◽  
C. Hamel ◽  
S. Jabaji-Hare ◽  
M. St-Arnaud

In August 2002, Phytophthora megasperma Drechs. was isolated from wilted plants of Asparagus officinalis L. cv. Guelph Millenium displaying spear and crown rot. Six affected plants were sampled in a commercial asparagus field located in the Saguenay-Lac-Saint-Jean Region (300 km northeast of Montreal, Quebec, Canada). The fungus was isolated from asparagus fern stalks, crown tissue, and spears after a rainy period and identified using morphological and cultural characteristics (2). In pinkish 4-week-old cultures, unbranched stalks bore abundant sporangia, which were ovoid to obpyriform in shape, 15 to 45 m long, and 10 to 30 m in diameter. Characteristic circular oospores >30 m in diameter were produced on V8 juice agar at 25°C in darkness after 1 month. Pathogenicity was tested on asparagus cvs. Guelph Millenium and Jersey Knight. A mycelium suspension (3 ml at 106 CFU/ml) prepared from 1-week-old shaken potato dextrose (PD) broth was sprayed on 30 1-week-old seedlings grown in petri plates filled with sterilized, moist, sandy soil and held at 20°C (day/night). Controls received sterile PD broth. Within 3 weeks of incubation in the dark, inoculated seedlings exhibited necrotic symptoms similar to those observed initially, while controls remained healthy. The pathogen was isolated from 75% of the ‘Guelph Millenium’ and 98% of the ‘Jersey Knight’ symptomatic seedlings, but not isolated from the control seedlings. In North America, disease caused by P. megasperma resulting in yield loss has been reported in California and New York (1,3). In Canada, the etiology of asparagus diseases is not well characterized. To our knowledge, this is the first report of P. megasperma on asparagus plants in Canada. References: (1) P. A. Ark and J. T. Barrett. Phytopathology 28:754, 1938. (2) D. C. Erwin et al. Phytophthora: Its Biology, Taxonomy, Ecology, and Pathology. The American Phytopathological Society, St Paul, MN, 1983. (3) T-L. Kuan and D. C. Erwin. Phytopathology 70:333, 1980.


Plant Disease ◽  
2006 ◽  
Vol 90 (8) ◽  
pp. 1109-1109 ◽  
Author(s):  
A. Garibaldi ◽  
G. Gilardi ◽  
M. L. Gullino

Lamb's lettuce or corn salad (Valerianella olitoria) is increasingly grown in Italy and used primarily in the preparation of mixed processed salad. In the fall of 2005, plants of lamb's lettuce, cv Trophy, exhibiting a basal rot were observed in some commercial greenhouses near Bergamo in northern Italy. The crown of diseased plants showed extensive necrosis, progressing to the basal leaves, with plants eventually dying. The first symptoms, consisting of water-soaked zonate lesions on basal leaves, were observed on 30-day-old plants during the month of October when temperatures ranged between 15 and 22°C. Disease was uniformly distributed in the greenhouses, progressed rapidly in circles, and 50% of the plants were affected. Diseased tissue was disinfested for 1 min in 1% NaOCl and plated on potato dextrose agar amended with 100 μg/liter of streptomycin sulfate. A fungus with the morphological characteristics of Rhizoctonia solani was consistently and readily isolated and maintained in pure culture after single-hyphal tipping (3). The five isolates of R. solani, obtained from affected plants successfully anastomosed with tester isolate AG 4, no. RT 31, received from R. Nicoletti of the Istituto Sperimentale per il Tabacco, Scafati, Italy (2). The hyphal diameter at the point of anastomosis was reduced, and cell death of adjacent cells occurred (1). Pairings were also made with AG 1, 2, 3, 5, 7, and 11 with no anastomoses observed between the five isolates and testers. For pathogenicity tests, the inoculum of R. solani (no. Rh. Vale 1) was grown on autoclaved wheat kernels at 25°C for 10 days. Plants of cv. Trophy were grown in 10-liter containers (20 × 50 cm, 15 plants per container) on a steam disinfested substrate (equal volume of peat and sand). Inoculations were made on 20-day-old plants by placing 2 g of infected wheat kernels at each corner of the container with 3 cm as the distance to the nearest plant. Plants inoculated with clean wheat kernels served as controls. Three replicates (containers) were used. Plants were maintained at 25°C in a growth chamber programmed for 12 h of irradiation at a relative humidity of 80%. The first symptoms, consisting of water-soaked lesions on the basal leaves, developed 5 days after inoculation with crown rot and plant kill in 2 weeks. Control plants remained healthy. R. solani was consistently reisolated from infected plants. The pathogenicity test was carried out twice with similar results. This is, to our knowledge, the first report of R. solani on lamb's lettuce in Italy as well as worldwide. The isolates were deposited at the AGROINNOVA fungal collection. The disease continues to spread in other greenhouses in northern Italy. References: (1) D. Carling. Rhizoctonia Species: Pages 37–47 in: Taxonomy, Molecular Biology, Ecology, Pathology and Disease Control. B. Sneh et al., eds. Kluwer Academic Publishers, the Netherlands, 1996. (2) J. Parmeter et al. Phytopathology, 59:1270, 1969. (3) B. Sneh et al. Identification of Rhizoctonia Species. The American Phytopathological Society, St. Paul, MN, 1996.


Plant Disease ◽  
2021 ◽  
Author(s):  
Jiahuai Hu

During August and September 2020, symptoms of leaf chlorosis, stunting, and wilting were observed on indoor hemp plants (Cannabis sativa L. cv. ‘Wedding Cake’) in a commercial indoor facility located in Coolidge, Arizona. Plants were grown in soilless coconut coir growing medium (Worm Factory COIR250G10), watered with 1.5 to 2.1 liters every 24 h through drip irrigation, and supplemented with 18 h of lighting. About 35% of plants displayed symptoms as described above and many symptomatic plants collapsed. To identify the causal agent, crown and root tissues from four symptomatic plants were harvested and rinsed with tap water. Tissue fragments (approx. 2 to 4 mm in size) were excised from the margins of the stem and root lesions, surface sterilized in 0.6% sodium hypochlorite for 1 min, rinsed well in sterile distilled water, blotted dry, and plated on potato dextrose agar (PDA) and on oomycete-selective clarified V8 media containing pimaricin, ampicillin, rifampicin, and pentachloronitrobenzene (PARP). Plates were incubated at room temperature (21-24 oC). Five isolates resembling Pythium were transferred after 3 days and maintained on clarified V8 media. Morphological characteristics were observed on grass blade cultures (Waterhouse 1967). Grass blades were placed on CV8 inoculated with the isolate. After a 1-day incubation at 25°C, the colonized blades were transferred to 8 ml of soil water extract in a Petri dish. Ten sporangia and oogonia were selected randomly and their diameters were measured under the microscope. Sporangia were mostly filamentous, undifferentiated or inflated lobulate, ranging from 7 to 17 µm in diameter. Knob-like appressoria were observed on branching clusters. Bulbous-like antheridia were formed on branched stalk with 1-8 antheridia per oogonium. Globose oogonia were terminal or intercalary and ranged from 21 to 33 µm in diameter. Globose oospores were mostly aplerotic and ranged from 15 to 21 μm in diameter. Based on these morphological characteristics, isolates were tentatively identified as Pythium myriotylum (Watanabe, 2002). Genomic DNA was extracted from mycelial mats of two isolates using DNeasy Plant Pro Kit (Qiagen Inc., Valencia, CA) according to the manufacturer’s instructions. The internal transcribed spacer (ITS) region of rDNA was amplified with primers ITS1/ITS4 and two identical nucleotide sequences were obtained and deposited under accession number MW380925. A BLASTn search revealed ≥ 98% query coverage and 100% match with sequences HQ237488.1, KY019264.1, and KM434129, which were isolates of P. myriotylum from palm, tobacco, and ginger, respectively. To fulfill Koch’s postulates, pathogenicity tests were conducted with 2 isolates using plants of ‘Wedding Cake’ grown in 12 1.9-liter pots filled with a steam-disinfested potting mix (Sungro Professional Growing Mix). Pots were placed in a plastic container and watered to flooding three times a week. Plants were maintained in a greenhouse with 18 h/10 h day/night supplemental light cycle (15-28 oC). Plants were fertilized weekly with Peters Professional fertilizer at 1mg/ml. Four plants were inoculated with each isolate at three weeks after seed sowing by placing two 5-mm mycelial plugs from active growing 4 days-old cultures on PDA media adjacent to the main root mass at an approximately 3 cm depth. Four plants were inoculated with blank PDA plugs as controls. Symptoms of leaf chlorosis, crown rot and wilting were observed after four weeks while control plants remained symptomless. P. myriotylum was re-isolated from necrotic roots of inoculated plants after surface-sterilization, but not from control plants. The pathogenicity test was repeated once. While P. myriotylum often occurs in warmer regions and has a wide host range of >100 host plant species including numerous economically important crops (Wang et al., 2003), there are only two reports of this pathogen on indoor hemp plants in a greenhouse in Connecticut (McGehee et al., 2019) and in Canada (Punja et al., 2019). This is the first report of P. myriotylum causing root and crown rot of indoor hemp in Arizona. A more careful water management in soilless growth medium to reduce periods of saturation would minimize the risk of Pythium root rot in indoor hemp production.


Plant Disease ◽  
2002 ◽  
Vol 86 (9) ◽  
pp. 1051-1051 ◽  
Author(s):  
B. de los Santos ◽  
M. Porras ◽  
C. Blanco ◽  
C. Barrau ◽  
F. Romero

Crown rot of strawberry (Fragaria × ananassa Duch. cv. Camarosa) was observed in three and two production fields in 2000 and 2001, respectively, in Huelva, southwestern Andalucia, Spain. Affected plants did not exhibit typical symptoms of red stele. Instead, there was an internal red-brown discoloration of the upper crown, a bluish discoloration of leaves, and the plants were wilted. Eventually, plants collapsed and died. Fungi were isolated from surface-disinfested necrotic crown tissue on P5ARPH medium (1). Plates were placed at 21°C for 5 to 10 days. One species was isolated consistently from symptomatic tissue. Microscopic observations revealed spherical oogonia with thin walls. Antheridia were paragynous and were attached to the oogonium near the oogonial stalk. Single oospores were spherical and had double-layered, yellow-brown walls (20 to 25 μm in diameter). Sporangia were usually borne terminally and were colorless and papillate (22 to 30 μm in diameter). Based on these characteristics, the causal agent was identified as Phytophthora cactorum (Lebert & Cohn) J. Schröt. (2). The fungus was transferred to V8 juice agar and maintained at 21°C in the dark. Disks (9-mm diameter) were removed from 7-day-old cultures of P. cactoru and used to inoculate five 2-month-old ‘Camarosa’ strawberry plants grown in sterilized peat in the greenhouse. Three disks were placed in the crown of each plant at soil level. Five noninoculated plants were similarly treated with sterile V8 juice agar disks only. After 2 weeks, the pathogen was reisolated from red-brown lesions visible on crowns of all inoculated plants. Noninoculated plants did not show any symptoms. To our knowledge, this is the first report of P. cactorum attacking strawberry plants in Spain. References: (1) S. N. Jeffers and S. B. Martin. Plant Dis. 70:1038, 1986. (2) G. M. Waterhouse and J. M. Waterston. No. 111 in: Descriptions of Pathogenic Fungi and Bacteria. CMI, Kew, UK, 1996.


Plant Disease ◽  
2006 ◽  
Vol 90 (8) ◽  
pp. 1114-1114
Author(s):  
G. E. Holcomb

Gazania rigens (L.) (treasure flower, Asteraceae) is grown as a winter and summer annual bedding plant in Louisiana and the lower southern United States. In March 2006, cv. Kiss Mix was observed in a wholesale nursery with symptoms of leaf yellowing, wilt, crown rot, and death. White mycelia and black sclerotia were present on some infected and dead plants. The plants had been grown outdoors and approximately 2% of 1,120 plants had been lost. Petiole and crown tissue from infected plants were surface disinfected in 70% ethyl alcohol, and sections were placed on acidified potato dextrose agar (PDA). A fungus that produced white mycelia and black sclerotia consistently grew from tissue pieces. Other characteristics included production of numerous sclerotia (32 to 71 per dish from 10 dishes) that were oval to oblong and formed in a ring at the periphery of culture dishes. Sclerotia measured 3 to 7 mm long (mean = 4.4, standard error = 0.15, N = 50) × 2 to 4 mm wide (mean = 3.0, standard error = 0.06, N = 50). Cells of sclerotial rinds were globose and lacked erect tomentum hyphae (1). Growth rate of the fungus at 26°C on PDA ranged from 1.3 to 3.1 cm/day (mean = 2.2, standard error = 0.05, N = 45) and mycelia covered the dishes after 3 days (2). On the basis of these characteristics, the fungus was identified as Sclerotinia sclerotiorum (Lib.) de Bary. Fungal inoculum for pathogenicity tests was grown on twice-sterilized wheat grains and 1 g of 10-day-old inoculum, consisting of fungus mycelia and sclerotia, was placed at the base of six G. rigens cv. Daybreak Mix plants. Inoculated and noninoculated control plants were placed in a dew chamber held at 22°C for 48 h and then moved to a greenhouse where temperatures ranged from 20 to 25°C. Leaf yellowing, wilt, and crown rot developed after 3 to 4 days on all inoculated plants followed by death after 6 days. S. sclerotiorum was reisolated from all inoculated plants. Noninoculated plants remained healthy. Sclerotinia crown rot of G. rigens was first reported in the United States from California (3) and has also been reported from Italy and Argentina (4). To our knowledge, this is the first report of Sclerotinia crown rot on G. rigens in Louisiana. References: (1) L. M. Kohn. Phytopathology 69:881, 1979. (2) G. Li et al. Mycol. Res. 104:232, 2000. (3) V. M. Muir and A. H. McCain. Calif. Plant Pathol. 16:1, 1973. (4) S. M. Wolcan. J. Plant Path. 86:263, 2004.


Plant Disease ◽  
2010 ◽  
Vol 94 (6) ◽  
pp. 788-788
Author(s):  
A. Garibaldi ◽  
D. Bertetti ◽  
M. T. Amatulli ◽  
M. L. Gullino

Rudbeckia fulgida (orange coneflower) is an herbaceous species (Asteraceae) grown in full sun in flower beds and borders in gardens. In the summer of 2009, a previously unknown leaf spot was observed on R. fulgida plants in three private gardens located near Biella (northern Italy). Leaves of infected plants showed extensive and irregular, dark brown, necrotic lesions that were slightly sunken with a well-defined border. Lesions initially ranged from 0.5 to 3 mm in diameter and eventually coalesced to cover the entire leaf, which curled without falling. At a later stage, stems were also affected, causing death of the plant. The disease affected 90% of plants. Dark brown pycnidia, 68 to 195 × 60 to 165 (average 135 × 117) μm in diameter, containing hyaline (light gray in mass), and ellipsoid, nonseptate conidia measuring 4.0 to 7.0 × 2.4 to 3.5 (average 5.4 × 3.0) μm were observed on symptomatic tissue. On the basis of these morphological characteristics, the fungus was related to the genus Phoma. Diseased tissue was excised from the margin of lesions, immersed in a solution containing 1% sodium hypochlorite for 2 to 3 s, rinsed in sterile distilled water, and then cultured on potato dextrose agar (PDA) medium. Fungal colonies initially produced a white mycelium that became greenish gray when incubated at temperatures ranging between 22 and 25°C under alternating daylight and darkness (13 h of light and 11 h of dark). After 14 days of incubation, unicellular, cylindrical or truncated cone-shaped, light brown chlamydospores measuring 6 to 12 μm in diameter developed in long chains. The internal transcribed spacer (ITS) region of rDNA was amplified using the primers ITS4/ITS6 and sequenced. BLAST analysis (1) of the 498-bp segment showed 100% homology with a sequence of a Phoma sp. (EF585395). The nucleotide sequence of our isolate was assigned GenBank Accession No. GU573979. Pathogenicity tests were performed by placing 100 ml of a water homogenate of mycelium (1 × 105 mycelial fragments/ml) obtained from 15-day-old PDA cultures of the fungus on leaves of three healthy 4-month-old potted R. fulgida plants. Three plants inoculated with a homogenate of PDA served as controls. Plants were maintained in a greenhouse, in a high humidity chamber for 7 days after inoculation, at temperatures ranging from 18 to 22°C and under high relative humidity conditions (70 to 90%). The first foliar lesions developed on leaves 7 days after inoculation, and after 10 to 12 days, 80% of leaves were severely infected. Control plants remained healthy. The organism reisolated on PDA from leaf lesions was identical in morphology to the isolate used for inoculation. The pathogenicity test was carried out twice. To our knowledge, this is the first report of the presence of a Phoma sp. on R. fulgida in Italy. Mycosphaerella ligulicola was reported on Rudbeckia sp. (2), while M. rudbeckiae and Phoma exigua have been reported on R. hirta (3). Currently, the economic importance of this disease is limited, but may become a more significant problem if the cultivation of this species increases. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) C. G. C. Chesters and J. P. Blakeman. Ann. Appl. Biol. 60:385, 1967. (3) D. F. Farr et al. Fungi on Plants and Plant Products in the United States. The American Phytopathological Society, St. Paul, MN, 1989.


Plant Disease ◽  
2013 ◽  
Vol 97 (10) ◽  
pp. 1386-1386 ◽  
Author(s):  
A. Garibaldi ◽  
G. Gilardi ◽  
G. Ortu ◽  
M. L. Gullino ◽  
M. Testa

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.


Plant Disease ◽  
2005 ◽  
Vol 89 (3) ◽  
pp. 340-340
Author(s):  
E. N. Rosskopf ◽  
C. B. Yandoc ◽  
B. Stange ◽  
E. M. Lamb ◽  
D. J. Mitchell

Polygonum odoratum (= Persicaria odorata), known as rau ram or sang hum, is native to southeastern Asia and is a common herb in Vietnamese cuisine (1). It has been studied most extensively for its aromatic compound content (2). In Florida, rau ram commonly is grown hydroponically in greenhouses using large, cement beds with recirculated water. The plants form dense mats from which new growth is trimmed for market. During January of 2002, a severe dieback was observed in one production house in Saint Lucie County, FL. Plants with less severe symptoms were yellowed and stunted. Roots of symptomatic plants were largely decayed with root symptoms beginning as a tip necrosis. The cortex of severely affected roots slipped off easily, leaving a stringy vascular system. Plating of symptomatic tissue from 20 randomly selected plant samples was performed with multiple general and selective media including potato dextrose agar, corn meal agar with pimaricin, ampicillin, rifampicin, and pentachloronitrobenzene (PARP) (3). All colonies produced were identified as Pythium helicoides Drechsler on the basis of sporangial, oogonial, and antheridial characteristics (4). Isolates had proliferous, obovoid, papillate sporangia, and were homothallic with smooth-walled oogonia and thick-walled, aplerotic oospores. Multiple antheridial attachments per oogonium were common with the antheridium attached along its entire length. Pathogenicity tests were conducted using P. odoratum plants grown from commercial transplants. Two tests were performed. Each test was conducted using eight inoculated and eight control plants. In the first test, plants were maintained in 10-cm pots immersed in sterilized pond water for the duration of the test. Plants were inoculated with five 7- × 70-mm sections of freshly growing mycelial culture per plant using 10-day-old cultures of Pythium helicoides grown on water agar. Chlorosis was observed at approximately 2 months after inoculation. Root necrosis was observed in inoculated plants approximately 5 months after inoculation. This test was performed in the greenhouse with temperatures ranging from 20 to 30°C. The second test was performed in growth chambers at 35 to 40°C. Plants were maintained in 10-cm pots immersed in Hoagland's solution and were inoculated with four 6-mm plugs per plant. Symptoms were observed on inoculated plants at this temperature within 1 week of inoculation. No chlorosis or root decay was observed in noninoculated, immersed plants. The pathogen was reisolated from inoculated, symptomatic tissue. To our knowledge, this is the first report of root rot of P. odoratum caused by Pythium helicoides. References: (1) R. E. Bond. Herbarist 55:34, 1989. (2) N. X. Dung et al. J. Essent. Oil Res. 7:339, 1995. (3) M. E. Kannwischer and D. J. Mitchell. Phytopathology 68:1760, 1978. (4) A. J. van der Plaats-Niterink. Monograph of the Genus Pythium. Vol. 21, Studies in Mycology. Centraalbureau voor Schimmelcutltures, Baarn, The Netherlands, 1981.


Plant Disease ◽  
2021 ◽  
Author(s):  
Yaxing Li ◽  
Yangfan Feng ◽  
Cuiping Wu ◽  
Junxin Xue ◽  
Binbin Jiao ◽  
...  

During a survey of pathogenic oomycetes in Nanjing, China from June 2019 to October 2020, at least ten adjacent Rhododendron pulchrum plants at a Jiangjun Mountain scenic spot showed symptoms of blight, and crown and root discoloration . Symptomatic root tissues collected from three 6-year-old plants were rinsed with water, cut into 10-mm pieces, surface sterilized with 70% ethanol for 1 min, and plated onto 10% clarified V8 PARP agar (cV8A-PARP) containing pimaricin (20 mg/liter), ampicillin (125 mg/liter), rifampicin (10 mg/liter), and pentachloronitrobenzene (20 mg/liter). Four Pythium-like isolates were recovered after three days of incubation at 26°C, and purified using hyphal-tipping. Ten agar plugs (2×2 mm2) of each isolate were grown in 10 mL of 10% clarified V8 juice (cV8) in a 10 cm plate at 26°C for 3 days to produce mycelial mats, and then the cV8 was replaced with sterile water. To stimulate sporangial production, three to five drops of soil extract solution were added to each plate. Sporangia were terminal, ovoid to globose, and the size is 24 to 45.6 (mean 34.7) (n=10.8) in length x 23.6 to 36.0 (mean 29.8) (n=6.2) in width. Gametangia were not observed in cV8A or liquid media after 30 days. For colony morphology, the isolates were sub-cultured onto three solid microbial media (cV8A-PARP, potato dextrose agar, corn meal agar) . All isolates had identical morphological features in the three media. Complete ITS and partial LSU and cox2 gene regions were amplified using primer pairs ITS1/ITS4, NL1/NL4, and FM58/FM66 , respectively. The ITS, LSU, and cox2 sequences of isolate PC-dj1 (GenBank Acc. No. MW205746, MW208002, MW208003) were 100.00% (936/936 nt), 100.00% (772/772 nt), and 99.64% (554/556 nt) identical to those of JX985743, MT042003, and GU133521, respectively. We built a maximum-likelihood tree of Phytopythium species using the concatenated dataset (ITS, LSU, cox2) to observe interspecific differences. Based on the morphological characters and sequences, isolate PC-djl was identified as Phytopythium litorale . As the four isolates (PC-dj1, PC-dj2, PC-dj3 and PC-dj4) tested had identical morphological characters and molecular marker sequences, the pathogenicity of the representative isolate, PC-dj1, was tested using two inoculation methods on ten one-year-old R. pulchrum plants. For the first inoculation method, plants were removed from the pot, and their roots were rinsed with tap water to remove the soil. Each of these plants was placed in a glass flask containing 250 mL of sterile water and 10 blocks (10 x 10 mm2) of mycelial mats harvested from a three-day-old culture of P. litorale, while the other plant was placed in sterile water as a control, and incubated at 26°C. After three days, symptoms including crown rot, root rot and blight was observed on the inoculated plants whereas the control remained asymptomatic. For the second inoculation method, ten plants were dug up to expose the root ball. Ten three-day-old cV8A plugs (5×5 mm2) from a PC-dj1 culture or sterile cV8A plugs were evenly insert into the root ball of a plant before it was planted back into the original pots. Both plants were maintained in a growth chamber set at 26°C with a 12/12 h light/dark cycle and irrigated as needed. After 14 to 21 days, the inoculated plant had symptoms resembling those in the field , while the control plant remained asymptomatic. Each inoculation method was repeated at triplicate and the outcomes were identical. Phytopythium isolates with morphological features and sequences identical to those of PC-dj1 were recovered from rotted crown and root tissues of all inoculated plants. Previously, P. litorale was found causing diseases of apple and Platanus orientalis in Turkey, fruit rot and seedling damping-off of yellow squash in southern Georgia, USA. This is the first report of this species causing crown and root rot on R. pulchrum, an important ornamental plant species in China. Additional surveys are ongoing to determine the distribution of P. litorale in the city of Nanjing.


Plant Disease ◽  
2004 ◽  
Vol 88 (6) ◽  
pp. 681-681
Author(s):  
A. Garibaldi ◽  
D. Bertetti ◽  
M. L. Gullino

Aquilegia flabellata Sieb. and Zucc. (columbine) is a perennial garden species belonging to the family Ranunculaceae. During the summer of 2003, a severe outbreak of a previously unknown powdery mildew was observed in several gardens near Biella (northern Italy). Upper surfaces of leaves were covered with a white mycelium and conidia, and as the disease progressed infected leaves turned yellow and died. Foot cell was cylindric and appressorium lobed. Conidia were hyaline, ellipsoid, and measured 31.2 to 47.5 × 14.4 to 33 μm (average 38.6 × 21.6 μm). Fibrosin bodies were not present. Cleistothecia were globose, brown, had simple appendages, ranged from 82 to 127 (average 105) μm in diameter, and contained one to two asci. Ascocarp appendages measured five to eight times the ascocarp diameter. Asci were cylindrical (ovoidal) and measured 45.3 to 58.2 × 30.4 to 40.2 μm. Ascospores (three to four per ascus) were ellipsoid or cylindrical and measured 28.3 to 31.0 × 14.0 to 15.0 μ;m. On the basis of its morphology, the pathogen was identified as Erysiphe aquilegiae var. aquilegiae (1). Pathogenicity was confirmed by gently pressing diseased leaves onto leaves of five, healthy A. flabellata plants. Five noninoculated plants served as controls. Inoculated and noninoculated plants were maintained in a garden where temperatures ranged between 20 and 30°C. After 10 days, typical powdery mildew symptoms developed on inoculated plants. Noninoculated plants did not show symptoms. To our knowledge, this is the first report of the presence of powdery mildew on Aquilegia flabellata in Italy. E. communis (Wallr.) Link and E. polygoni DC. were reported on several species of Aquilegia in the United States (2), while E. aquilegiae var. aquilegiae was previously observed on A. flabellata in Japan and the former Union of Soviet Socialist Republics (3). Specimens of this disease are available at the DIVAPRA Collection at the University of Torino. References: (1) U. Braun. Nova Hedwigia, 89:700, 1987. (2) D. F. Farr et al. Fungi on Plants and Plant Products in the United States. The American Phytopathological Society, St Paul, MN, 1989. (3) K. Hirata. Host Range and Geographical Distribution of the Powdery Mildews. Faculty of Agriculture, Niigata University, 1966.


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