scholarly journals First Report of Chaetomella raphigera Causing Leaf Spot on Rosa chinensis in China

Plant Disease ◽  
2014 ◽  
Vol 98 (4) ◽  
pp. 569-569 ◽  
Author(s):  
M. Zhang ◽  
J. J. Li ◽  
H. Y. Wu ◽  
Y. H. Geng ◽  
W. L. Han
Keyword(s):  
Leaf Spot ◽  
Botanical Garden ◽  
The United States ◽  
Henan Province ◽  
Its2 Region ◽  
First Report ◽  
Rosa Chinensis ◽  
China Rose ◽  
Chaetomella Raphigera ◽  
Over The Top

China rose, Rosa chinensis Jacq., is extensively cultivated as an ornamental plant in China (1). During the course of a disease survey of China rose in Henan Province, a leaf spot was observed on about 20 China roses, cultivated in a garden in Zhengzhou, Henan Province, in early October 2012. The early symptom appeared as small round, pale brown lesions on the leaves. Lesions expanded into 5 to 15-mm-diameter spots that were near round or irregular and brown. Both sporodochial and pycnidial conidiomata developed in necrotic areas of diseased leaves when placed in moist chambers. Pycnidia were elongated, reniform, with a single raphe over the top, pale to dark brown, and 260 to 350 × 150 to 210 μm. Sporodochia were pale luteous and 100 to 280 × 80 to 180 μm. Setae, conidiophores, conidiogenous cells, and conidia were the same between two types of conidioma. Setae were pale to dark brown, 0 to 2 septate, straight with rounded end, clavate to curved at apex, and 22 to 60 × 2 to 5 μm. Conidiophores were up to 120 × 1 to 2 μm, filiform, cylindric, and branched. Conidiogenous cells were enteroblastic, collar and channel minute. Conidia were nonseptate, hyaline, ellipsoid or cymbiform, smooth, guttulate, and 4 to 6.5 × 1.5 to 2.5 μm. Two pure cultures (zm12276-1 and zm12276-2) were obtained by picking spores from independent conidiomata on one leaf and then subsequently grown on potato dextrose agar (PDA), producing the same two kinds of conidiomata. The characteristics of conidial size and distinctly different conidiomata with setae are diagnostic of Chaetomella raphigera M.E. Swift (3,4). The identity of our fungus (zm12276-1) was confirmed to be C. raphigera by DNA sequencing of the ITS1-5.8S-ITS2 region. The DNA sequence was 99% identical to those of the other C. raphigera isolates (AY487076 and AY487085) (2). The ITS sequence from zm12276-1 was deposited in GenBank (KF483474). Pathogenicity was tested by inoculating 10 leaves of R. chinensis with mycelia plug from colony of zm12276-1 (0.5 cm in diameter). An equal number of fresh leaves inoculated with the plugs of non-colonized PDA medium served as the control. All leaves were incubated in clear plastic box with a dish of sterile distilled water at 25°C under ambient light. After 7 days, 90% of the inoculated leaves showed symptoms identical to those observed on R. chinensis leaves affected in the field. From each of the symptomatic leaves, C. raphigera was recovered, whereas controls remained symptom-free and no fungus was isolated from the control leaves. Koch's postulates were repeated three times with the same results using the pure culture of zm12276-1. C. raphigera has been previously reported on Rosa sp. in the United States (4). To our knowledge, this is the first report of C. raphigera infecting R. chinensis in China. The disease cycle and the control strategies in the regions are being further studied. References: (1) C. Z. Gu and K. R. Robertson. Pages 339-381 in: Flora of China, vol. 9. Science Press, Beijing and Missouri Botanical Garden, 2003. (2) A. Y. Rossman et al. Mycol. Progr. 3:275, 2004. (3) B. C. Sutton. The Coelomycetes. CAB International Publishing, New York, 1980. (4) M. E. Swift. Mycologia 22:165, 1930.

scholarly journals First Report of Botrytis cinerea Causing Leaf Spot of Aquilegia vulgaris in Japan

Plant Disease ◽  
2009 ◽  
Vol 93 (4) ◽  
pp. 425-425 ◽  
Author(s):  
M. Zhang ◽  
T. Tsukiboshi ◽  
I. Okabe
Keyword(s):  
United States ◽  
Botrytis Cinerea ◽  
Leaf Spot ◽  
Leaf Tissue ◽  
The United States ◽  
Plant Diseases ◽  
Its2 Region ◽  
Commonwealth Mycological Institute ◽  
First Report ◽  
Leaf Spots

European columbine, Aquilegia vulgaris L., Ranunculaceae, is an herbaceous flower widely used in gardens, parterres, and courtyards and is a traditional herbal plant. During the summer of 2008, leaf spots were observed on a plant cultivated along a roadside area in Nasushiobara, Tochigi, Japan. In some courtyards, the leaf spot affected more than 60% of the plants. Early symptoms appeared as small, round or elliptic, brown lesions on the leaves. Lesions expanded to 5 to 15 × 4 to 10 mm, irregular spots that were dark brown to black in the middle, with pale yellow-brown or purple-brown margins. In continuously wet or humid conditions, thick, gray mycelium and conidia appeared on the surface of leaf spots. Conidiophores were melanized at the base and hyaline near the apex, branched, and septated (approximately 3 mm × 16 to 18 μm). Conidia were hyaline, aseptate, ellipsoidal to obovoid, with a slightly protuberant hilum, and ranged from 9 to 14.5 × 5.5 to 6.5 μm. The pathogen was identified as Botrytis cinerea Pers.:Fr on the basis of morphology and sequence of ITS1-5.8s-ITS2 region of rDNA. The sequence (GenBank Accession No. FJ424510) exactly matched the sequences of two Botryotinia fuckeliana (anamorph Botrytis cinerea), (e.g., GenBank Accession Nos. AY686865 and FJ169666) (2). The fungus was isolated on potato dextrose agar (PDA) from a single conidium found on the symptomatic leaf tissue. Colonies of B. cinerea were first hyaline and later turned gray to black when the spores differentiated. Koch's postulates were performed with three whole plants of potted aquilegia. Leaves were inoculated with mycelia plugs harvested from the periphery of a 7-day-old colony; an equal number of plants were inoculated with the plugs of PDA medium only and served as controls. All plants were covered with plastic bags for 24 h to maintain high relative humidity and incubated at 25°C. After 8 days, all mycelium-inoculated plants showed symptoms identical to those observed on leaves from A. vulgaris infected in the field, whereas controls remained symptom free. Reisolation of the fungus from lesions on inoculated leaves confirmed that the causal agent was B. cinerea. B. cinerea has been previously reported on A. vulgaris in the United States and China (1,3). To our knowledge, this is the first report of leaf spots caused by B. cinerea on A. vulgaris in Japan. References: (1) Anonymous. Index of Plant Diseases in the United States. USDA Agric. Handb. No 165, 1960. (2) M. B. Ellis. Dematiaceous Hyphomycetes. Commonwealth Mycological Institute, Kew, England, 1971. (3) Z. Y. Zhang. Flora Fungorum Sinicorum. Vol. 26. Botrytis, Ramularia. Science Press, Beijing, 2006.

scholarly journals First Report of a Leaf Spot on Conyza sumatrensis Caused by Phoma macrostoma in China

Plant Disease ◽  
2012 ◽  
Vol 96 (1) ◽  
pp. 148-148 ◽  
Author(s):  
J. Liu ◽  
H. D. Luo ◽  
W. Z. Tan ◽  
L. Hu
Keyword(s):  
Leaf Spot ◽  
Fungal Pathogens ◽  
Biological Diversity ◽  
Biocontrol Agent ◽  
Continuous Light ◽  
The United States ◽  
Leaf Spot Disease ◽  
Pathogenicity Test ◽  
Dry Land ◽  
First Report

Conyza sumatrensis (Asteraceae), an annual or biennial plant, is native to North and South America. It is an invasive, noxious weed that is widespread in southern and southeastern China. It invades farm land and causes great losses to dry land crops, including wheat, corn, and beans. It also reduces biological diversity by crowding out native plants in the infested areas (3,4). During a search for fungal pathogens that could serve as potential biological control agents of C. sumatrensis, a leaf spot disease was observed in 2010 in Chongqing, China. An isolate (SMBC22) of a highly virulent fungus was obtained from diseased leaves. Pathogenicity tests were performed by placing 6-mm-diameter mycelial disks of 7-day-old potato dextrose agar (PDA) cultures of SMBC22 on leaves of 15 healthy greenhouse-grown plants of C. sumatrensis; the same number of control plants was treated with sterile PDA disks. Treated plants were covered with plastic bags for 24 h and maintained in a growth chamber with daily average temperatures of 24 to 26°C, continuous light (3,100 lux), and high relative humidity (>90%). Lesions similar to those observed in the field were first obvious on the SMBC22-inoculated leaves 3 days after inoculation. Symptoms became severe 7 to 9 days after inoculation. Control plants remained healthy. The fungus was reisolated from inoculated and diseased leaves and it was morphologically the same as SMBC22. The pathogenicity test was conducted three times. A survey of 10 southern and southeastern Chinese provinces revealed that the disease was widespread and it attacked leaves and stems of seedlings and mature plants of C. sumatrensis. Lesions on leaves were initially small, circular, and water soaked. The typical lesion was ovoid or fusiform, dark brown, and surrounded by a yellow halo. The spots coalesced to form large lesions and plants were often completely blighted. Fungal colonies of SMBC22 on PDA plates were initially white and turned dark gray. Colonies were circular with smooth edges with obvious rings of pycnidia on the surface. Aerial hyphae were short and dense. Pycnidia, black and immersed or semi-immersed in the medium, were visible after 12 days of incubation. Pycnidia were 72 to 140 μm in diameter. Conidia were produced in the pycnidia and were hyaline, unicellular, ellipsoidal, and 4.4 to 6.1 × 1.6 to 2.2 μm. To confirm identification of the fungus, genomic DNA was extracted from mycelia of a 7-day-old culture on PDA at 25°C (2). The internal transcribed spacer (ITS) gene of rDNA was amplified using primers ITS4/ITS5. The gene sequence was 524 bp long and registered in NCBI GenBank (No. HQ645974). BLAST analysis showed that the current sequence had 99% homology to an isolate of Phoma macrostoma (DQ 404792) from Cirsium arvense (Canada thistle) in Canada and reported to cause chlorotic symptoms on that host plant (1). To our knowledge, this is the first report of P. macrostoma causing disease on C. sumatrensis in China. P. macrostoma, thought of as a biocontrol agent of broadleaf weeds in Canada, has been patented in the United States. The current isolate of P. macrostoma is considered as a potential biocontrol agent of C. sumatrensis. References: (1) P. R. Graupner et al. J. Nat. Prod. 66:1558, 2004. (2) S. Takamatsu et al. Mycoscience 42:135, 2001. (3) W. Z. Tan et al. Page 177 in: Manual of Emergency Control Technology Invasive Pests in China. G. L. Zhang, ed. Science Press, Beijing, 2010. (4) C. Wang et al. J. Wuhan Bot. Res. 28:90, 2010.

scholarly journals First Report of Alternaria Leaf Spot of Banana Caused by Alternaria alternata in the United States

Plant Disease ◽  
2013 ◽  
Vol 97 (8) ◽  
pp. 1116-1116 ◽  
Author(s):  
V. Parkunan ◽  
S. Li ◽  
E. G. Fonsah ◽  
P. Ji
Keyword(s):  
United States ◽  
Alternaria Alternata ◽  
Leaf Spot ◽  
Morphological Characteristics ◽  
The United States ◽  
Its Sequencing ◽  
Single Spore ◽  
First Report ◽  
Alternaria Leaf Spot ◽  
Leaf Spots

Research efforts were initiated in 2003 to identify and introduce banana (Musa spp.) cultivars suitable for production in Georgia (1). Selected cultivars have been evaluated since 2009 in Tifton Banana Garden, Tifton, GA, comprising of cold hardy, short cycle, and ornamental types. In spring and summer of 2012, 7 out of 13 cultivars (African Red, Blue Torres Island, Cacambou, Chinese Cavendish, Novaria, Raja Puri, and Veinte Cohol) showed tiny, oval (0.5 to 1.0 mm long and 0.3 to 0.9 mm wide), light to dark brown spots on the adaxial surface of the leaves. Spots were more concentrated along the midrib than the rest of the leaf and occurred on all except the newly emerged leaves. Leaf spots did not expand much in size, but the numbers approximately doubled during the season. Disease incidences on the seven cultivars ranged from 10 to 63% (10% on Blue Torres Island and 63% on Novaria), with an average of 35% when a total of 52 plants were evaluated. Six cultivars including Belle, Ice Cream, Dwarf Namwah, Kandarian, Praying Hands, and Saba did not show any spots. Tissue from infected leaves of the seven cultivars were surface sterilized with 0.5% NaOCl, plated onto potato dextrose agar (PDA) media and incubated at 25°C in the dark for 5 days. The plates were then incubated at room temperature (23 ± 2°C) under a 12-hour photoperiod for 3 days. Grayish black colonies developed from all the samples, which were further identified as Alternaria spp. based on the dark, brown, obclavate to obpyriform catenulate conidia with longitudinal and transverse septa tapering to a prominent beak attached in chains on a simple and short conidiophore (2). Conidia were 23 to 73 μm long and 15 to 35 μm wide, with a beak length of 5 to 10 μm, and had 3 to 6 transverse and 0 to 5 longitudinal septa. Single spore cultures of four isolates from four different cultivars were obtained and genomic DNA was extracted and the internal transcribed spacer (ITS1-5.8S-ITS2) regions of rDNA (562 bp) were amplified and sequenced with primers ITS1 and ITS4. MegaBLAST analysis of the four sequences showed that they were 100% identical to two Alternaria alternata isolates (GQ916545 and GQ169766). ITS sequence of a representative isolate VCT1FT1 from cv. Veinte Cohol was submitted to GenBank (JX985742). Pathogenicity assay was conducted using 1-month-old banana plants (cv. Veinte Cohol) grown in pots under greenhouse conditions (25 to 27°C). Three plants were spray inoculated with the isolate VCT1FT1 (100 ml suspension per plant containing 105 spores per ml) and incubated under 100% humidity for 2 days and then kept in the greenhouse. Three plants sprayed with water were used as a control. Leaf spots identical to those observed in the field were developed in a week on the inoculated plants but not on the non-inoculated control. The fungus was reisolated from the inoculated plants and the identity was confirmed by morphological characteristics and ITS sequencing. To our knowledge, this is the first report of Alternaria leaf spot caused by A. alternata on banana in the United States. Occurrence of the disease on some banana cultivars in Georgia provides useful information to potential producers, and the cultivars that were observed to be resistant to the disease may be more suitable for production. References: (1) E. G. Fonsah et al. J. Food Distrib. Res. 37:2, 2006. (2) E. G. Simmons. Alternaria: An identification manual. CBS Fungal Biodiversity Center, Utrecht, Netherlands, 2007.

scholarly journals First Report of Leaf Spot of Rudbeckia hirta var. pulcherrima Caused by Septoria rudbeckiae in Korea

Plant Disease ◽  
2012 ◽  
Vol 96 (6) ◽  
pp. 911-911 ◽  
Author(s):  
J. H. Park ◽  
S. E. Cho ◽  
K. S. Han ◽  
H. D. Shin
Keyword(s):  
Leaf Spot ◽  
The United States ◽  
Flowering Plant ◽  
Leaf Spot Disease ◽  
Morphological Identification ◽  
Its Sequence ◽  
Conidial Suspension ◽  
First Report ◽  
Rudbeckia Hirta ◽  
Close Phylogenetic Relationship

Rudbeckia hirta L. var. pulcherrima Farw. (synonym R. bicolor Nutt.), known as the black-eyed Susan, is a flowering plant belonging to the family Asteraceae. The plant is native to North America and was introduced to Korea for ornamental purposes in the 1950s. In July 2011, a previously unknown leaf spot was first observed on the plants in a public garden in Namyangju, Korea. Leaf spot symptoms developed from lower leaves as small, blackish brown lesions, which enlarged to 6 mm in diameter. In the later stages of disease development, each lesion was usually surrounded with a yellow halo, detracting from the beauty of the green leaves of the plant. A number of black pycnidia were present in diseased leaf tissue. Later, the disease was observed in several locations in Korea, including Pyeongchang, Hoengseong, and Yangpyeong. Voucher specimens were deposited at the Korea University Herbarium (KUS-F25894 and KUS-F26180). An isolate was obtained from KUS-F26180 and deposited at the Korean Agricultural Culture Collection (Accession No. KACC46694). Pycnidia were amphigenous, but mostly hypogenous, scattered, dark brown-to-rusty brown, globose, embedded in host tissue or partly erumpent, 50 to 80 μm in diameter, with ostioles 15 to 25 μm in diameter. Conidia were substraight to mildly curved, guttulate, hyaline, 25 to 50 × 1.5 to 2.5 μm, and one- to three-septate. Based on the morphological characteristics, the fungus was consistent with Septoria rudbeckiae Ellis & Halst. (1,3,4). Morphological identification of the fungus was confirmed by molecular data. Genomic DNA was extracted using the DNeasy Plant Mini DNA Extraction Kit (Qiagen Inc., Valencia, CA.). The internal transcribed spacer (ITS) region of rDNA was amplified using the ITS1/ITS4 primers and sequenced. The resulting sequence of 528 bp was deposited in GenBank (Accession No. JQ677043). A BLAST search showed that there was no matching sequence of S. rudbeckiae; therefore, this is the first ITS sequence of the species submitted to GenBank. The ITS sequence showed >99% similarity with those of many Septoria species, indicating their close phylogenetic relationship. Pathogenicity was tested by spraying leaves of three potted young plants with a conidial suspension (2 × 105 conidia/ml), which was harvested from a 4-week-old culture on potato dextrose agar. Control leaves were sprayed with sterile water. The plants were covered with plastic bags to maintain 100% relative humidity (RH) for the first 24 h. Plants were then maintained in a greenhouse (22 to 28°C and 70 to 80% RH). After 5 days, leaf spot symptoms identical to those observed in the field started to develop on the leaves inoculated with the fungus. No symptoms were observed on control plants. S. rudbeckiae was reisolated from the lesions of inoculated plants, confirming Koch's postulates. A leaf spot disease associated with S. rudbeckiae has been reported on several species of Rudbeckia in the United States, Romania, and Bulgaria (1–4). To our knowledge, this is the first report of leaf spot on R. hirta var. pulcherrima caused by S. rudbeckiae in Korea. References: (1) J. B. Ellis and B. D. Halsted. J. Mycol. 6:33, 1890. (2) D. F. Farr and A. Y. Rossman. Fungal Databases. Systematic Mycology and Microbiology Laboratory, ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ February 2, 2012. (3) E. Radulescu et al. Septoriozele din Romania. Ed. Acad. Rep. Soc. Romania, Bucuresti, Romania, 1973. (4) S. G. Vanev et al. Fungi Bulgaricae 3:1, 1997.

scholarly journals First Report of a Leaf Spot Caused by Alternaria brassicae on the Invasive Weed Lepidium draba in North America

Plant Disease ◽  
2009 ◽  
Vol 93 (8) ◽  
pp. 846-846 ◽  
Author(s):  
A. J. Caesar ◽  
R. T. Lartey
Keyword(s):  
North America ◽  
Chinese Cabbage ◽  
Leaf Spot ◽  
The United States ◽  
Alternaria Brassicae ◽  
Voucher Specimen ◽  
First Report ◽  
Leaf Spots ◽  
Black Spots ◽  
Lepidium Draba

The exotic, rangeland weed Lepidium draba L., a brassicaceous perennial, is widely distributed in the United States. For example, Oregon contains 100,000 ha of land infested with L. draba (2). Because it is capable of aggressive spread and has the potential to reduce the value of wheat-growing land (4), it is the target of biological control research. The application of multiple pathogens has been advocated for control of other brassicaceous weeds, including the simultaneous application of biotrophic and necrotrophic pathogens (3). In pursuit of this approach, in 2007, we discovered the occurrence of leaf spots on approximately 90% of L. draba plants near Shepherd, MT, which were distinct from leaf lesions caused by Cercospora bizzozeriana (1). The lesions were initially tiny, black spots enlarging over time to become circular to irregular and cream-colored around the initial black spots and sometimes with dark brown borders or chlorotic halos. Conidia from the lesions were light brown, elongate and obclavate, produced singly from short conidia, with 8 to 12 transverse septa, and 2 to 6 longitudinal septa. The spore body measured 25 to 35 × 200 to 250 μm with a beak cell 42 to 100 μm long. On the basis of conidial and cultural characteristics, the fungus was identified as Alternaria brassicae (Berk.) Sacc. Leaf tissues bordering lesions were plated on acidified potato dextrose agar. Colonies on V8 and alfalfa seed agar were black with concentric rings, eventually appearing uniformly black after 10 to 14 days. The internal transcribed spacer region of rDNA was amplified using primers ITS1 and ITS4 and sequenced. BLAST analysis of the 575-bp fragment showed a 100% homology with a sequence of A. brassicae Strain B from mustard (GenBank Accession No. DQ156344). The nucleotide sequence has been assigned GenBank Accession No. FJ869872. For pathogenicity tests, aqueous spore suspensions approximately 105/ml were prepared from cultures grown at 20 to 25°C for 10 to 14 days on V8 agar and sprayed on leaves of three L. draba plants. Inoculated plants were enclosed in plastic bags and incubated at 20 to 22°C for 72 to 80 h. In addition, three plants of the following reported hosts of A. brassicae were inoculated: broccoli, canola, Chinese cabbage, collards, broccoli raab, kale, mustard greens, radish, rape kale, and turnip. Within 10 days, leaf spots similar to those described above developed on plants of radish, canola, Chinese cabbage, and turnip and A. brassicae was reisolated and identified. Control plants sprayed with distilled water remained symptomless. These inoculations were repeated and results were the same. To our knowledge, this is the first report of a leaf spot disease caused by A. brassicae on L. draba in North America. A voucher specimen has been deposited with the U.S. National Fungus Collections (BPI No. 878750A). References: (1) A. J. Caesar et al. Plant Dis. 93:108, 2009. (2) G. L. Kiemnec and M. L. McInnis. Weed Technol. 16:231, 2002. (3) A. Maxwell and J. K. Scott. Adv. Bot. Res. 43:143, 2005. (4) G. A. Mulligan and J. N. Findlay. Can. J. Plant Sci. 54:149, 1974.

scholarly journals First Report of Bacterial Spot of Peony Caused by a Xanthomonas sp. in the United States

Plant Disease ◽  
2012 ◽  
Vol 96 (4) ◽  
pp. 581-581 ◽  
Author(s):  
C. L. Oliver ◽  
R. Cai ◽  
B. A. Vinatzer ◽  
E. A. Bush ◽  
M. A. Hansen
Keyword(s):  
Leaf Spot ◽  
Consensus Sequence ◽  
Perfect Match ◽  
Leaf Tissue ◽  
The United States ◽  
Genetic Distances ◽  
Biochemical Tests ◽  
First Report ◽  
Herbaceous Perennials ◽  
Phosphate Buffered Saline

In early May 2008 and 2009, peony samples (Paeonia spp.) with symptoms of leaf spot and blight were submitted to the Virginia Tech Plant Disease Clinic. The 2008 peony was an unknown cultivar from a northern Virginia landscape. The three cultivars (Dr. Alexander Fleming, Felix Crousse, and Karl Rosenfield) submitted in 2009 were from a commercial nursery in southwestern Virginia that was reporting leaf spot progressing to severe blight, which rendered plants unsalable, on 75% of a 1,219 m2 block during a 10-day period of heavy rainfall. Bacterial streaming from spots was observed. On the basis of phenotypic and biochemical tests, the isolates were determined to be xanthomonads. Two isolates (one recovered from the 2008 sample and one from the 2009 sample) were used in the following work. Isolates were characterized by multilocus sequencing (MLST) (4). PCR reactions were prepared and cycled using 2X ImmoMix (Bioline, Tauton, MA) according to manufacturer's recommendations with an annealing temperature of 58°C. Template DNA was added by touching a single colony with a 20-μl pipette tip and placing the tip into the reaction mix for 1 min. Four bands of the expected size were visualized on an electrophoresis gel and cleaned products were sequenced in forward and reverse directions at the University of Chicago, Cancer Research Center DNA Sequencing Facility. Corresponding gene fragments of each isolate were identical. A consensus sequence (PAMDB Isolate ID No. 936) for each of the four gene fragments was constructed and compared with sequences in NCBI ( http://www.ncbi.nlm.nih.gov/nuccore/ ) and PAMDB ( http://genome.ppws.vt.edu/cgi-bin/MLST/home.pl ) (1) databases using Blastn (2). No perfect match was found. Genetic distances between the peony isolates and all strains in PAMDB were determined by MegAlign (Lasergene; DNAStar, Madison, WI). The Xanthomonas strain most similar to the isolates recovered from the peony samples was Xanthomonas hortorum pv. hederae ICMP 1661 with a genetic distance of 0.023; this strongly suggests that the peony isolates belong to X. hortorum. For Koch's postulates, six surface-disinfested young leaflets from Paeonia lactiflora ‘Karl Rosenfield’ were inoculated by forcefully spraying a phosphate-buffered saline suspension of each bacterial isolate (~4.3 × 109 CFU/ml) into the underside of the leaf until leaf tissue appeared water soaked. Controls were inoculated similarly with phosphate-buffered saline solution. Moist chambers with inoculated leaves were incubated at ambient temperature under two 48W fluorescent grow lights with 12 h of light and dark. Circular spots were observed on leaves inoculated with the 2009 and 2008 isolates in 18 and 20 days, respectively. No symptoms were observed on controls. Bacterial streaming from leaf spots was observed by phase-contrast microscopy; bacteria were isolated and confirmed to be identical to the original isolates by the methods described above. To our knowledge, this is the first report of a Xanthomonas sp. causing leaf spot and blight on peony. Although bacterial blight of peony has been attributed to a xanthomonad in recent years, the pathogen had not been further characterized (3). References: (1) N. F. Almeida et al. Phytopathology 100:208, 2010. (2) D. J. Altschul et al. J. Mol. Biol. 215:403, 1990. (3) M. L. Gleason et al. Diseases of Herbaceous Perennials. The American Phytopathological Society, St. Paul, MN. 2009. (4) J. M. Young et al. Syst. Appl. Microbiol. 31:366, 2008.

scholarly journals First Report of Powdery Mildew Caused by Blumeria graminis f. sp. bromi on Bromus catharticus in China

Plant Disease ◽  
2020 ◽  
Author(s):  
Mo Zhu ◽  
Jie Ji ◽  
Xiao Duan ◽  
Wenqi Shi ◽  
YongFang Li
Keyword(s):  
Powdery Mildew ◽  
Sequence Data ◽  
Morphological Characteristics ◽  
Signs And Symptoms ◽  
The United States ◽  
Blumeria Graminis ◽  
Henan Province ◽  
Width Ratio ◽  
Causal Organism ◽  
First Report

Bromus catharticus, rescuegrass, is a brome grass that has been cultivated for herbage production, and been widely naturalized in many provinces of China, including Henan province. During April and May 2020, powdery mildew was found on leaves of Br. catharticus on the campus of Henan Normal University, Xinxiang city (35.3°N; 113.9°E), Henan Province, China. Abundant white or grayish irregular or coalesced circular powdery colonies were scattered on the adaxial surface of leaves and 70% of the leaf areas were affected. Some of the infected leaves either were chlorotic or senescent. About 60% of the observed plants showed powdery mildew symptoms. Conidiophores (n = 25) were 32 to 45 μm × 7 to 15 μm and composed of foot cells and conidia (mostly 6 conidia) in chains. Conidia (n = 50) were 25 to 35 μm × 10 to 15 μm, on average 30 × 13 μm, with a length/width ratio of 2.3. Chasmothecia were not found. Based on these morphologic characteristics, the pathogen was initially identified as Blumeria graminis f. sp. bromi (Braun and Cook 2012; Troch et al. 2014). B. graminis mycelia and conidia were collected, and total genomic DNA was extracted (Zhu et al. 2019). The rDNA internal transcribed spacer (ITS) region was amplified with primer pairs ITS1/ITS4. The amplicon was cloned and sequenced. The sequence (574 bp) was deposited into GenBank under Accession No. MT892940. BLASTn analysis revealed that MT892940 was 100% identical to B. graminis f. sp. bromi on Br. catharticus (AB000935, 550 of 550 nucleotides) (Takamatsu et al. 1998). Phylogenetic analysis of MT892940 and ITS of other B. graminis ff. spp. clearly indicated least two phylogenetically distinct clades of B. graminis f. sp. bromi and that MT892940 clustered with the Takamatsu vouchers. Leaf surfaces of five healthy plants were fixed at the base of a settling tower and then inoculated by blowing conidia from diseased leaves using pressurized air. Five non-inoculated plants served as controls. The inoculated and non-inoculated plants were maintained separately in two growth chambers (humidity, 60%; light/dark, 16 h/8 h; temperature, 18℃). Thirteen- to fifteen-days after inoculation, B. graminis signs and symptoms were visible on inoculated leaves, whereas control plants remained asymptomatic. The pathogenicity assays were repeated twice with the same results. The observed signs and symptoms were morphologically identical to those of the originally infected leaves. Accordingly, the causal organism of the powdery mildew was confirmed as B. graminis f. sp. bromi by morphological characteristics and ITS sequence data. B. graminis has been reported on Br. catharticus in the United States (Klingeman et al. 2018), Japan (Inuma et al. 2007) and Argentina (Delhey et al. 2003). To our best knowledge, this is the first report of B. graminis on Br. catharticus in China. Since hybridization of B. graminis ff. spp. is a mechanism of adaptation to new hosts, Br. catharticus may serve as a primary inoculum reservoir of B. graminis to infect other species (Menardo et al. 2016). This report provides fundamental information for the powdery mildew that can be used to develop control management of the disease in Br. catharticus herbage production.

scholarly journals First Report of Leaf Spot Disease On Microstegium vimineum Caused by Bipolaris setariae in China

Plant Disease ◽  
2021 ◽  
Author(s):  
Min Tan ◽  
Qiong Huang ◽  
Hao Fan ◽  
Yun Wu ◽  
Richard C. Reardon ◽  
...  
Keyword(s):  
United States ◽  
Leaf Spot ◽  
The United States ◽  
Lethal Concentration ◽  
Reference Sequence ◽  
Microstegium Vimineum ◽  
Morphological Observation ◽  
Leaf Spot Disease ◽  
First Report ◽  
Spot Disease

Microstegium vimineum, a Poaceae annual C4 plant, occurred widely in crop fields, tea gardens, orchards, under forests and roadsides in most provinces and regions south of the Yellow River, China. It was introduced into the eastern USA causing ecological and environmental damage (Stricker, 2016). In October 2015, M. vimineum plants with leaf spots were observed on the roadside of Mingling Road (32.04521°E, 118.84323°N), Nanjing, China. In an early stage of disease development, light brown or brown, round or oval shaped lesions appeared on the upper surface of leaves. In a middle stage, the lesions gradually expanded and the edges of the diseased leaves were lightly curled. In a late stage, leaves were withered or curled and the entire plant died. Initial disease incidence was up to 85% among natural populations of the weed. Diseased leaves collected from field were surface disinfected (75% ethanol for 30s; 1% sodium hypochlorite solution for 30s; 75% ethanol for 30s; sterile deionized water for 1min) and placed on water agar (20g agar per liter) (Kleczewski et al., 2010). Plates were incubated in the dark at 28℃ for 3 days. Following incubation, leaves, spores and conidiophores were examined using light microscopy. Single spores were obtained by using the single-spore procedure, plating out a loopful of spores onto water agar, and then carving individual spores out with associated agar under a microscope. Single spores were isolated, plated onto MV-agar (30g M. vimineum leaves, 20g agar per liter), and placed under 365 nm wavelength black light. Fungal colonies were transferred onto PDA medium, after 4 days colonies measured between 83 to 86 mm in diameter, appeared flat and dark brown, with short, light gray aerial hyphae. Conidiophores were solitary or clustered, light brown to medium brown, with pale apical color and multiple septa. The upper part was usually geniculated, 5.5-9.5 μm wide. Conidia were light yellowish brown to medium yellowish brown, mostly fusiform, straight or curved, fusoid or navicular, often slightly curved, rarely straight, smooth, 5-9 (mostly 7) septa, 48-70×10-14.5 μm (average 57×12.5 μm); hilium slightly prominent, and truncated at the base. Through morphological observation, the fungus was preliminarily identified as Bipolaris sp.. Four to five seeds of M. vimineum were planted in pots (10 cm in diameter) filled with nutrient soil, placed in the greenhouse and watered regularly. Four pots were inoculated with a conidia suspension of 1×105 sp/mL, at 4-5 true stage. Inoculated seedlings were maintained under 80% humidity and 28℃ for 24h in the dark, and then transferred to a greenhouse. Three pots of uninoculated seedlings were used as controls. Two days after the inoculation, buff-colored, irregular-shaped spots appeared centered on leaf veins. Within a week, diseased leaves became crinkled and their edges were yellow to brown due to proliferation of the spots. By 15 days, large areas of brown spots appeared on the leaves, some leaves turned yellow-brown and severely curled, and 80% of the plants had died. The diseased symptoms were similar to that of the field sample. The fungus re-isolated resulted morphologically identical to the original isolate grown on PDA medium and used for inoculation, thus fulfilled Koch’s postulates. The CTAB method was used to extract DNA from isolates of diseased leaves taken directly from the field, and the internal transcribed spacer (ITS) and glyceraldehyde 3-phosphate dehydrogenase gene (GPDH) were amplified using primer pairs of ITS1/ITS4 and GPD/GPD2 (Manamgoda et al., 2014) respectively. The ITS amplified sequence (Genbank accession MW446193) shared 100% identity with the reference sequence of Bipolaris setariae (MN215638.1) and the GPDH amplified sequence (MW464364) shared 99.83% identity with the reference sequence of B. setariae (MK144540.1). Field experiments were conducted in Laboratory Base of Nanjing Agricultural University, where M. vimineum plants were planted. Spore suspensions with concentrations of 105, 104, 103, 102, and 101 sp/mL were prepared, distilled water was used for control, and there were four replicates of each treatment. Twenty four plots were randomly arranged, the experimental unit consisted of 50 to 60 plants in an area of 0.5m×0.6m. The interval distance between plots was about 20 cm so as to prevent the mutual influence among treatments. M. vimineum plants were inoculated at 3-4 true leaf stage. Inoculation was done at sunset, and 60 mL spore suspension was sprayed onto each plot. After spraying, the waterproof-breathable black cloth was used to cover the plots, and removed 36 hours later. The outdoor temperature was 20~28℃. After 10 days, the symptoms of M. vimineum were observed and the disease index was recorded. SPSS 20 software (SPSS Inc., Chicago, IL, USA) was used for variance analysis, and Origin 9.0 (OriginLab, Hampton, MA, USA) was used to calculate the half lethal concentration (ED50) and 90% lethal concentration (ED90) of the strain MLL-1-5 on M. vimineum. Symptoms appeared on inoculated M. vimineum seedlings immediately after dark treatment. Within a week, all seedlings inoculated with the highest spore concentration were dead. Plants sprayed with water remained healthy. ED50 and ED90 of the strain MLL-1-5 was 1.9×101 and 1.4×103 sp/mL respectively, which indicated aggressiveness of the strain MLL-1-5 B. setariae. After 28 days, infected M. vimineum plants did not recover. This is the first report of leaf spot disease on M. vimineum caused by B. setariae in China. M. vimineum is a widely distributed and extremely harmful weed in China and United States. No biocontrol agents against M. vimineum are currently available. B. setariae may have potential as a biocontrol agent against M. vimineum both in China and the United States.

scholarly journals First Report of Bacterial Leaf Spot of Spinach Caused by a Pseudomonas syringae Pathovar in California

Plant Disease ◽  
2002 ◽  
Vol 86 (8) ◽  
pp. 921-921 ◽  
Author(s):  
S. T. Koike ◽  
H. R. Azad ◽  
D. C. Cooksey
Keyword(s):  
Brassica Oleracea ◽  
Beta Vulgaris ◽  
Pseudomonas Syringae ◽  
Leaf Spot ◽  
The United States ◽  
Nutrient Broth ◽  
Bacterial Leaf Spot ◽  
First Report ◽  
Leaf Spots ◽  
Initial Symptoms

In 2000 and 2001, a new disease was observed on commercial spinach (Spinacia oleracea) in the Salinas Valley, Monterey County, CA. Initial symptoms were water-soaked, irregularly shaped leaf spots (2 to 3 mm diameter). As the disease developed, spots enlarged to as much as 1 to 2 cm, were vein-delimited, and turned dark brown. Faint chlorotic halos sometimes surrounded the spots. Death of large areas of the leaf occurred if spots coalesced. Spots were visible from the adaxial and abaxial sides of leaves, and no fungal structures were observed. The disease occurred on newly expanded and mature foliage. No fungi were isolated from the spots. However, cream-colored bacterial colonies were consistently isolated on sucrose peptone agar, and these strains were nonfluorescent on King's medium B. Strains were positive for levan and negative for oxidase, arginine dihydrolase, and nitrate reductase. Strains did not grow at 36°C, did not rot potato slices, but induced a hypersensitive reaction in tobacco (Nicotiana tabacum cv. Turk). These results suggested the bacterium was similar to Pseudomonas syringae. Fatty acid methyl ester (FAME) analysis (MIS-TSBA 4.10, MIDI Inc., Newark, DE) indicated the strains were highly similar (80.1 to 89.3%) to P. syringae pv. maculicola. However, in contrast to P. syringae pv. maculicola, the spinach strains did not utilize the carbon sources erythritol, L+tartrate, L lactate, and DL-homoserine. Pathogenicity of 10 strains was tested by growing inoculum in nutrient broth shake cultures for 48 h, diluting to 106 CFU/ml, and spraying 4-week-old plants of spinach cv. Bossanova. Control plants were sprayed with sterile nutrient broth. After 5 to 8 days in a greenhouse (24 to 26°C), leaf spots identical to those observed in the field developed on cotyledons and true leaves of inoculated plants. Strains were reisolated from the spots and identified as P. syringae. Control plants remained symptomless. The 10 strains were also inoculated on beet (Beta vulgaris), Swiss chard (Beta vulgaris subsp. cicla), cilantro (Coriandrum sativum), and spinach. Spinach showed leaf spots after 8 days; however, none of the other plants developed symptoms. Two strains were inoculated onto spinach cvs. Califlay, Lion, Nordic IV, Polka, Resistoflay, Rushmore, RZ 11, Spinnaker, Springfield, Viroflay, and Whitney. Leaf spot developed on all cultivars, and the pathogen was reisolated. Because the FAME data indicated a similarity between the spinach pathogen and P. syringae pv. maculicola, we inoculated sets of spinach cv. Bolero, cabbage (Brassica oleracea subsp. capitata cv. Grenedere), and cauliflower (Brassica oleracea subsp. botrytis cv. White Rock) with three P. syringae pv. maculicola and three spinach strains. Cabbage and cauliflower developed leaf spots only when inoculated with P. syringae pv. maculicola; spinach had leaf spots only when inoculated with the spinach strains. All inoculation experiments were done twice, and the results of the two tests were the same. To our knowledge, this is the first report of bacterial leaf spot of spinach in California caused by a nonfluorescent P. syringae, and the first record of this disease in the United States. Biochemical characteristics and limited host range of the pathogen indicate the California strains are likely the same as the P. syringae pv. spinaciae pathogen that was reported in Italy (1) and Japan (2). References: (1) C. Bazzi et al. Phytopathol. Mediterr. 27:103, 1988. (2) K. Ozaki et al. Ann. Phytopathol. Soc. Jpn. 64:264, 1998.

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