Macadamia Quick Decline Caused by Phytophthora tropicalis is Associated with Sap Bleeding, Frass, and Nectria in Hawaii

Macadamia quick decline (MQD) has been a persistent problem since 1986 when it started killing productive 14- to 36-year-old macadamia trees in the Hilo, HI area. Fungi including Nectria regulosa, Xylaria arbuscula, Phellinus gilvus, and Acremonium recifei have been attributed to MQD and could kill twigs on healthy macadamia trees after artificial inoculation (3). The oomycete originally called Phytophthora capsici and later reclassified as P. tropicalis was also considered to be involved in the MQD complex (3). However, the primary causal agent has never been determined and the issue continues to perplex the industry. Between 2005 and 2006, a mature macadamia field on the Waiakea Experiment Station planted with cv. HAES 333 began to experience a high frequency of MQD. Trees exhibiting dull green, yellow, or brown leaves within the tree canopy were observed. Sap bleeding from the trunk, Ambrosia beetles, and Nectria fruiting bodies were consistently associated with MQD. Disease incidence was 22%. Of 21 infected trees, 53% died within an average period of 6.8 months. Four branch samples were collected from four trees showing browning of leaves, sap bleeding, Ambrosia beetles, and Nectria, and seven P. tropicalis isolates were recovered from diseased tissue on water agar or V8 agar media. No other microorganisms were isolated from diseased branches. On the basis of the morphological characteristics described by Aragaki and Uchida (1), the isolates were identified as P. tropicalis. The morphological identification was confirmed by molecular analysis of the 5.8S subunit and flanking internal transcribed spacers (ITS1 and ITS2) of rDNA amplified from DNA extracted from single-zoospore cultures with the ITS1/ITS4 primers (2,4) and sequenced (GenBank No. FJ849839). Pathogenicity tests were conducted on four 12-year-old macadamia trees in the field. A 4 × 104 zoospore/ml suspension of P. tropicalis isolate L1 was injected into branches of cv. HAES 344 to incite MQD signs and symptoms. Branches inoculated with P. tropicalis started showing the initial sign of MQD, excessive sap bleeding, within 36 days postinoculation (dpi). The presence of Ambrosia beetle frass and the appearance of orange fruiting bodies of Nectria were visible within 110 dpi. No symptoms were noted on the four control tree branches inoculated by the same method but with sterilized distilled water. P. tropicalis was reisolated from the symptomatic macadamia branches, fulfilling Koch's postulates. To our knowledge, this is the first report of P. tropicalis as the primary causal agent of MQD and its association with sap bleeding, Ambrosia beetles, and a saprotrophic species of Nectria. After completion of our research, Ko (3) reported that the MQD P. capsici was P. tropicalis, supporting our finding in this study. Quick decline of macadamia trees continues to be a serious problem in Hawaii. Minimizing tree loss in mature orchards is critical for maintaining the economic viability of Hawaii's macadamia industry. Understanding the biology of this pathosystem will enable the development of control and prevention strategies. References: (1) M. Aragaki and J. Y. Uchida. Mycologia 93:137, 2001. (2) G. Caetano-Annolles et al. Curr. Genet. 39:346, 2001. (3) W.-H. Ko. Bot. Stud. 50:1, 2009. (4) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.

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First Report of Anthracnose Caused by Colletotrichum gloeosporioides on Pistachio (Pistacia vera) in China

Plant Disease ◽

10.1094/pdis-04-11-0269 ◽

2011 ◽

Vol 95 (10) ◽

pp. 1314-1314 ◽

Cited By ~ 6

Author(s):

S. Y. Yang ◽

S. C. Su ◽

T. Liu ◽

G. Fan ◽

J. Wang ◽

...

Keyword(s):

Dna Sequence ◽

Colletotrichum Gloeosporioides ◽

Disease Incidence ◽

Morphological Characteristics ◽

Northern China ◽

Gansu Province ◽

Internal Transcribed Spacers ◽

Pistacia Vera ◽

First Report ◽

Stem Cankers

In the 1990s, pistachio (Pistacia vera L. ‘Kerman’ and ‘Peters’) was introduced in China. They are found in many orchards in Beijing and Gansu and Hebei provinces, northern China. In 2009, a new disease was observed on leaves, stems, and fruits in pistachio orchards in Gansu Province. Disease incidence in 8- to 12-year-old orchards was 30%. Yield losses reached 25%. Symptoms began as discrete, sunken, black spots, approximately 10 mm in diameter, followed by circular lesions that eventually coalesced with tissue death recorded and orange fructifications developed on lesions. Pieces of diseased leaves, stems, and fruits were surfaced sterilized and placed on 2% potato dextrose agar (PDA) at 25°C. A fungus was consistently isolated. After 10 days, cultures on PDA showed aerial, white mycelium that turned gray to grayish black with a salmon-to-orange conidial mass at 25°C and a 12-h photoperiod. Brown, 80 to 120 μm long setae were observed in the acervulus. Conidia were hyaline, fusiform to nearly straight, and averaged 12 to 18 × 3 to 5 μm. On the basis of morphological characteristics, the fungus was identified as Colletotrichum gloeosporioides (Penz.) Sacc. (2). On PDA, 0.5 μg/ml of benomyl was applied for the sensitivity test (3). Benomyl completely inhibited the growth of the fungus. Mycelial DNA was extracted, PCR amplified using ITS1 and ITS4 primers for the ribosomal DNA internal transcribed spacers 1 and 2, and sequenced. The DNA sequence was recorded in GenBank as No. HQ631378. The DNA sequence was blasted showing 99% identity with Accession Nos. GQ144454 and GU004376, for C. gloeosporioides. Pathogenicity tests were conducted under greenhouse conditions at 25°C. Three replicates of 2-year-old ‘Kerman’ plants were inoculated with mycelial PDA plugs placed on 0.5-cm2 stem wounds and then wrapped with Parafilm. Controls were inoculated with PDA plugs without the fungus. After 3 weeks, stem cankers were observed on inoculated plants. Control plants remained healthy. Pathogenicity was also tested on injured leaves and fruits. A 10-μl drop of a spore suspension of 104 conidia/ml was applied on ‘Kerman’ and ‘Peters’ leaves and ‘Kerman’ fruits and placed on plates with a wet filter paper at 25°C. Small, black lesions were observed at 2 days after inoculation. At 7 days, necrotic lesions covered the entire surface. C. gloeosporioides was reisolated from necrotic lesions. Controls did not develop symptoms. C. acutatum has been reported on pistachio in Australia (1), but to our knowledge, this is the first report of anthracnose caused by C. gloeosporioides on pistachio. References: (1) G. J. Ash and V. M. Lanoiselet. Australas. Plant Pathol. 30:365, 2001. (2) J. Y. Lu. Plant Pathogenic Mycology. China Agricultural Press, Beijing, 2001. (3) N. A. R. Peres et al. Plant Dis. 86:620, 2002.

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First Report of Fruit Rot and Associated Branch Dieback of Almond in California Caused by a Phomopsis Species Tentatively Identified as P. amygdali

Plant Disease ◽

10.1094/pdis.1999.83.11.1073c ◽

1999 ◽

Vol 83 (11) ◽

pp. 1073-1073 ◽

Cited By ~ 7

Author(s):

J. E. Adaskaveg ◽

H. Förster ◽

J. H. Connell

Keyword(s):

Disease Incidence ◽

Morphological Characteristics ◽

The United States ◽

Tree Canopy ◽

Fruit Rot ◽

Control Treatment ◽

Field Inoculation ◽

First Report ◽

Branch Dieback ◽

Summer Fruit

A fruit rot of almond (Prunus dulcis (Mill.) D. Webb.) was observed in an orchard in Durham, CA (Butte County), in June of 1998 after an unusually wet spring with a total precipitation of 17.2 cm for April and May. Disease incidence on fully developed fruit of almond cv. Sonora was nearly 90% in the lower tree canopy by July. Almond cv. Nonpareil grown in alternate rows in the same orchard was much less affected. Fruit symptoms included extensive grayish brown discolored and shriveled hulls, often associated with a clear gum secretion and shriveled kernels. Affected fruit frequently abscised. Leaf symptoms and branch dieback were not associated with the disease in 1998. In May of 1999, however, extensive twig dieback was observed on almond cv. Sonora in the same orchard. Isolations from more than 100 symptomatic fruit were conducted from 9 sampling sites in the 9-ha orchard. Based on morphological characteristics, the same fungus was isolated from 93% of the fruit. The fungus also was isolated consistently from samples exhibiting twig dieback. During a major disease survey conducted in 1998, the fungus was only incidentally isolated from almond fruit from other California orchards. Ascomata were not observed in vivo or in vitro. The fungus produced alpha and beta spores in pycnidia when cultured on potato dextrose agar. Spore measurements were obtained from 10 spores for each of 3 isolates obtained from fruit or twig dieback of almond cv. Sonora. Conidial dimensions of fruit and twig isolates were very similar. Based on spore sizes, with alpha spores measuring 5.3 to 7.5 (to 8) × 1.7 to 2.5 μm and beta spores measuring12.8 to 29.8 × 0.6 to 0.7 μm, the fungus was tentatively identified as Phomopsis amygdali (Del.) Tuset & Portilla (2). Previous reports on this fungus (2), however, indicated that beta spores are not produced in culture, and disease symptoms have not been observed on fruit. The fungus was morphologically different from other species of Phomopsis reported from almond and other Prunus species, including P. mali Roberts, P. padina (Sacc. & Roum.) Died., P. parabolica Petrak, P. perniciosa Grove, P. pruni (Ellis & Dearn.) Wehm., P. prunorum (Cooke) Grove, P. ribetejana Camara, and P. stipata (Lib.) Sutton (3). Field inoculation studies were performed in May of 1999 on almond cvs. Carmel and Mission. Almond fruit were wounded (2 × 2 × 2 mm) or left unwounded and were sprayed with water (control) or a suspension of alpha spores (105 spores per ml). Branches were bagged for 4 days to maintain high humidity. Fruit symptoms on cv. Carmel were observed after 4 weeks on wounded and nonwounded inoculated fruit, and P. amygdali was successfully reisolated from diseased tissue. No symptoms were observed in the control treatment for almond cv. Carmel or in any treatment for cv. Mission. This is the first report of P. amygdali causing a late spring and summer fruit rot and associated branch dieback of almond in North America (1). References: (1) D. F. Farr et al. 1989. Fungi on Plants and Plant Products in the United States. The American Phytopathological Society, St. Paul, MN. (2) J. J. Tuset and M. T. Portilla. Taxonomic status of Fusicoccum amygdali and Phomopsis amygdalina. Can. J. Bot. 67:1275, 1989. (3) F. A. Uecker. 1988. A World List of Phomopsis Names with Notes on Nomenclature, Morphology, and Biology. Mycologia Memoir No. 13. J. Cramer, Berlin.

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First Report of Cercospora apiicola Causing Leaf Spots and Stem Lesions on Celery in China

Plant Disease ◽

10.1094/pdis-09-20-1879-pdn ◽

2020 ◽

Author(s):

Qian Zhao ◽

A LI CHAI ◽

Yanxia Shi ◽

Xuewen Xie ◽

Baoju Li

Keyword(s):

Disease Incidence ◽

Elongation Factor ◽

Morphological Characteristics ◽

Internal Transcribed Spacers ◽

Apium Graveolens ◽

Malt Extract ◽

Translation Elongation ◽

First Report ◽

Leaf Spots ◽

Stem Lesions

Celery (Apium graveolens L.) is a vegetable crop cultivated widely in the Mediterranean, Europe and parts of Asia. From March to May in 2014, leaf spots and stem lesions were observed on celery plants in Yanqing (116°03′E, 40°32′N), Beijing and Chengdu (104°06′E, 30°67′N), Sichuan Province. Plants developed 0.3-1.8 cm diameter subcircular leaf spots with brown centers surrounded by pale yellow halos. Spots on leaves were amphigenous. Necrotic areas on stems were subcircular to elongated, pale brown to brown. Plants in five greenhouses were surveyed with 30 to 60% disease incidence. Necrotic tissue from 8 stems and 12 leaves were cut from the margins of lesions and divided into two parts. One part was treated with lactophenol and used for microscopic examination. The other part was surface sterilized with 4% sodium hypochlorite for 2 min, rinsed three times in sterile water, placed onto 2% malt extract agar (MEA), and incubated at 26°C for seven days with natural daylight. Stromata on leaves and stems were not well developed. Four-to-ten conidiophores (15.3-56.5 × 2.8-5.5 μm) formed in fascicles, emerged through stomata or erupted through the cuticle. Conidia (n=50) were 60-135 × 2.5-4.5 μm, solitary, septate, cylindrical to obclavate-cylindrical, hila thickened and darkened. Colonies were white to smoke-gray, and aerial mycelia were sparse to moderate. Morphological characteristics of the pathogen were similar to Cercospora apiicola (Groenewald et al. 2006; Groenewald et al. 2013). The gDNA of 20 isolates was extracted from mycelium using the Plant Genomic DNA Kit (Tiangen, China). The internal transcribed spacers (ITS), actin (ACT), translation elongation factor 1-α (TEF1) and histone H3 (HIS3) regions were amplified with primer pairs ITS1/ITS4 (Groenewald et al. 2013), ACT-512F/ACT-783R (Carbone and Kohn 1999), EF1-728F/EF1-986R (Carbone and Kohn 1999), CYLH3F/CYLH3R (Crous et al. 2006). Phylogenetic analysis of multiple genes (Bakhshi et al. 2018) was conducted with the neighbor-joining method using MEGA7. The sequences of our isolate (QC14030702) and five published sequences of C. apiicola were clustered into one clade with a 99% confidence level. The sequences of QC14030702 have been deposited in GenBank with accessions KU870468 for ITS, KU870469 for ACT, KU870470 for TEF1, and KU870471 for HIS3. Pathogenicity of the isolates was tested on plants (cv. Jia Yuan Xi Yang Qin). Because the pathogen sporulated poorly on various media, mycelial fragments were sprayed on leaves in a suspension of 1×106 mL-1 in a greenhouse (temperature 26±0.5°C; RH 98%; photoperiod 12 h). Healthy plants were sprayed with sterilized water as controls. Three replicates of every isolate were conducted, and each replicate included 5 celery plants. After 7 days, leaf spots appeared on all inoculated plants, which were similar to those on celery in the field. All control plants remained asymptomatic. Re-isolation of the fungus from infected tissues showed same morphological and cultural characteristics of C. apiicola as the original isolates. C. apiicola has been reported in Greece, Korea, South Korea and Venezuela on celery, but never been reported in China (Farr and Rossman 2020). C. apiicola potential threatens celery production, and this the first report of the disease in China.

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First Report of Powdery Mildew on Carrot Caused by Erysiphe heraclei in Michoacan, Mexico

Plant Disease ◽

10.1094/pdis-94-4-0483b ◽

2010 ◽

Vol 94 (4) ◽

pp. 483-483 ◽

Cited By ~ 1

Author(s):

G. Rodríguez-Alvarado ◽

R. Rodríguez-Fernández ◽

A. Soto-Plancarte ◽

S. P. Fernández-Pavía

Keyword(s):

Powdery Mildew ◽

Morphological Characteristics ◽

Signs And Symptoms ◽

Internal Transcribed Spacers ◽

First Report ◽

Daucus Carota L ◽

5.8S Rdna ◽

Rdna Sequences ◽

Close Proximity ◽

Pathogenicity Tests

Carrot (Daucus carota L. subsp. sativus (Hoffm.) Arcang.) is planted as a home-grown vegetable in the central region of Michoacan, Mexico. Powdery mildew was observed on carrot plants cv. Nantesa at several locations near Morelia, Michoacan during March 2009. Affected plants had abundant, white, superficial conidia and mycelium on leaves and stems. All plants at each of five locations surveyed had powdery mildew symptoms with percent foliage coverage ranging from 50 to 80%. Mycelial growth was amphigenous, mainly on the upper leaf surface, covering the whole leaf and with irregular patches on inflorescences and stems. Hyphae were ectophytic with lobed appressoria. Conidiophores presented foot cells 22.5 to 35 (30) × 5.75 to 7 (6.3) μm followed by two cells, one shorter and one longer than the foot cell. Conidia were produced singly, most subcylindric to cylindric, lacked fibrosin bodies, and measured 31.2 to 42 (36.2) × 8.7 to 11.2 (10.5) μm. The teleomorph was not observed. Genomic DNA was extracted from infected leaves; sequences of the internal transcribed spacers (ITS) inclusive of 5.8S rDNA were amplified using previously described primers specific for Erysiphales (3). The ITS sequences shared 100% homology to Erysiphe heraclei specimen VPRI41227 from carrot in Australia (GenBank Accession No. EU371725). On the basis of the morphological characteristics observed and the ITS rDNA sequences, the pathogen was identified as E. heraclei DC. The ITS sequence was deposited in NCBI as Accession No. GU252368. Pathogenicity tests were conducted twice on a total of 10 healthy 8-week-old carrot plants cv. Nantesa. Infected plants were placed in close proximity to healthy plants and maintained in a greenhouse at 27 ± 5°C. Initial signs and symptoms were observed 3 weeks after inoculation and appeared as small, white colonies, which later coalesced and covered most of the foliage. Microscopic examination of the conidia and mycelial morphology matched the originally described pathogen, E. heraclei. Powdery mildew caused by this pathogen has been extensively reported on diverse species and genera of the Apiaceae in Europe and remains one of the most important diseases of carrot (2). The appearance of E. heraclei in diverse regions on a variety of umbelliferous crops indicates that formae speciales have spread, infecting different and specific hosts (1–3). Recently, E. heraclei has been reported on parsley in Puebla, Mexico (4). To our knowledge, this is the first report of E. heraclei causing powdery mildew on carrot in Michoacan, Mexico. This pathogen should be considered as a threat to commercial carrot crops in Mexico. Other crops in the Apiaceae may not be at risk in this area if this powdery mildew is specific for carrots. References: (1) B. J. Aegerter. Page 22 in: Compendium of Umbelliferous Crop Diseases. The American Phytopathological Society, St. Paul, MN, 2002. (2) U. Braun. The Powdery Mildew (Erysiphales) of Europe. Gustav Fischer-Verlag. Jena, Germany, 1995. (3) J. H. Cunnington et al. Australas. Plant Pathol. 32:421, 2003. (4) M. J. Yáñez-Morales et al. Schlechtendalia 19:47, 2009.

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First Report of Phomopsis Seed Decay of Soybean Caused by Phomopsis longicolla in South China

Plant Disease ◽

10.1094/pdis-04-12-0401-pdn ◽

2012 ◽

Vol 96 (11) ◽

pp. 1693-1693 ◽

Cited By ~ 3

Author(s):

Z. Shan ◽

S. Li ◽

Y. Liu ◽

Z. Yang ◽

C. Yang ◽

...

Keyword(s):

South China ◽

Seed Quality ◽

Morphological Characteristics ◽

Internal Transcribed Spacers ◽

Morphological Identification ◽

Pathogenicity Test ◽

Phomopsis Longicolla ◽

First Report ◽

Phomopsis Seed Decay ◽

Seed Decay

Phomopsis seed decay of soybean (Glycine max (L.) Merr.) causes poor seed quality and suppresses yield in most soybean-growing areas in the world. The disease is caused primarily by Phomopsis longicolla Hobbs. During the spring of 2010, soybean seeds without symptoms were planted in the fields but emergence was poor and the emergence rates ranged from 30 to 70% in south China. Approximately 3,000 symptomless seeds were randomly collected from seven fields at three locations in Guangzhou, Nanchang, and Wuhan. Seeds were surface disinfected with 1% sodium hypochlorite for 12 min, rinsed in sterile distilled water three to four times, and placed on 2% agar. Plates were then incubated at 26°C under 16/8-h photoperiod for 3 to 4 days. About 10 to 20% of the seeds produced white hyphae that spread rapidly and covered the whole seed. The hyphae from fungal isolates were transferred to potato dextrose agar (PDA) and incubated at 26°C in the dark. After 3 to 4 weeks, conidia were elliptical with two oil drops at both ends and hyaline (6.2 to 7.2 × 2.6 to 3.2 μm). The cultural and morphological characteristics of the isolates corresponded with the description of P. longicolla (2). Colonies on PDA were floccose, dense, and white. Stromata were large, black, and spreading. To confirm the morphological identification, the ribosomal internal transcribed spacers (ITS1-5.8S-ITS2) from three isolates were sequenced (GenBank Accession Nos. JQ899030, JQ899031, and JQ899032). BLAST analysis indicated that the isolates had 99% nucleotide sequence identity with P. longicolla (GenBank Accession Nos. AY857868.1, EF026104, and HQ130441.1). Pathogenicity tests were conducted on 2-week-old soybean seedlings (3). A mycelial plug (3 mm in diameter) from the margin of 1-week-old PDA culture of the Wuhan isolate was individually placed mycelial side down directly on the top of cut stem 1 to 2 cm above cotyledon node of the soybean seedling. PDA plugs without the fungus was used as the negative control. All seedlings were kept in a growth chamber at 26°C with 92 to 94% relative humidity. After 2 weeks, all inoculated seedlings showed browning, stem wilt, and the lesions were 0.3 to 2.0 cm long. No symptoms were observed in the control plants. P. longicolla was reisolated from the infected seedlings. The pathogenicity test was repeated three times. Soybean stem blight caused by P. longicolla has been reported in northeast China (1). To our knowledge, this is the first report of P. longicolla causing Phomopsis seed decay of soybean in south China. This report will establish a foundation for developing a program for screening germplasm for resistance to this disease in south China. References: (1) Y. L. Cui et al. Plant Pathol. 58:799, 2009. (2) T. W. Hobbs et al. Mycologia 77:535,1985. (3) S. Li et al. Plant Dis. 85:1031, 2001.

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Identification and control the causal agent of root rot disease of cucumber in Iraq: تشخيص ومكافحة مسبب مرض تعفن جذور محصول الخيار في العراق

Journal of agricultural, environmental and veterinary sciences - مجلة العلوم الزراعية والبيئية والبيطرية ◽

10.26389/ajsrp.n160319 ◽

2019 ◽

Vol 3 (3) ◽

Author(s):

Safaa Neamat Hussein

Keyword(s):

Root Rot ◽

Disease Incidence ◽

Morphological Characteristics ◽

Negative Control ◽

Significant Inhibition ◽

Causal Agent ◽

Root Rot Disease ◽

Rot Disease ◽

And Control

This study aimed to isolate the causal agent of the root rot disease of cucumber and control it biologically. Samples were collected in the cucumber fields in the Diyala and Saladin governorates of Iraq. Isolation test demonstrated associate fungi belong to the twelve geniuses. Fusarium solani exhibited highly percentage of appearance of 82.15% with frequency 54.00%. Seventy-seven isolates identified as F. solani according to their cultural and morphological characteristics while sixty-five isolates of them amplified successfully with specific primer of Fusarium spp using PCR technique. Isolate DF13 was most virulent isolated while exhibited 0% cucumber seed germination in vitro. The bio-agent Bacillus pumilus demonstrated significant inhibition ability against the fungal isolate DF13 in vitro of 100%. Under greenhouse condition B. pumilus decreased the disease incidence and severity to 30.55% and 20.75% respectively compared to the negative control which was 80.50%, 55.00% respectively.

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First Report of Colletotrichum aenigma Causing Anthracnose of Grape in Korea

Plant Disease ◽

10.1094/pdis-11-20-2458-pdn ◽

2021 ◽

Author(s):

Ju Sung Kim ◽

Oliul Hassan ◽

Taehyun Chang

Keyword(s):

South Korea ◽

Disease Incidence ◽

Juglans Regia ◽

Morphological Characteristics ◽

Causal Agent ◽

Effective Control ◽

Pathogenicity Test ◽

Conidial Suspension ◽

First Report ◽

Colletotrichum Species

Grape (cv. Kyoho) is one of the most popular dessert fruits in South Korea. Anthracnose caused by Colletotrichum species is a common and very destructive disease of grape in the country. In 2019, severe outbreaks of anthracnose was observed in different grape orchards in Gimcheon (36º09´N, 128º00´ E), South Korea. The disease incidence on fruit was up to 50% in the orchards with most severe outbreaks and infected fruit displayed typical anthracnose symptoms including sunken necrotic lesions with orange-like conidial mass. For isolation of putative causal agents, nine diseased fruits were collected from three commercial orchards. A total of nineisolates were made from nine of the infected fruit by spreading spore masses (1x106 conidia mL-1) from each fruit on water agar and collecting single germinated spores after incubation at 25 ºC overnigh. The single germinated spores were transferred on to fresh potato dextrose agar (PDA) (Difco, Becton Dickinson) and incubated at 25ºC in the dark. Seven day old colonies were cottony white on the upper side and gray at the center on the reverse side. Conidia were cylindrical with round ends and measured 13.9 – 20.1 × 5.4 – 8.1 μm (mean = 16.5 × 6.6 μm, n = 30). Appressoria were brownish, sub-cylindrical with a few lobes and 10.3 –16.7 × 6.6 – 10.9 μm (mean = 13.1 × 8.1 μm, n = 30). The morphological characteristics of the solates resembled those of Colletotrichum species within the C. gloeosporioides complex (Weir et al. 2012). DNA was amplified using the following primer pairs: ITS1/ITS4, GDF / GDR, ACT-512F / ACT-783R, Bt2a/ Bt2b, and CHS79-F/CHS-354R (Weir et al. 2012). Accession numbers, LC586811 to LC586825 were obtained after depositing all the resulting sequences in GenBank. A 50% majority rules phylogenetic tree (Bayesian phylogenic analysis) was constructed based on concatenated sequences of ITS, GAPDH, ACT, TUB, and CHS using MrBayes 3.2.10. The present isolates formed a single clade with the reference isolates of C. aenigma (isolate ICMP 18608 and ICMP 18686). For a pathogenicity test, healthy grapefruits were collected from an orchards, surface sterilized by dipping in 1% sodium hypochlorite, rinsed with sterilized water and dried by blotting. A conidial suspension (1×106 conidia mL-1) in sterilized water were prepared from one week old colonies of isolates GRAP10 and GRAP12. A small wound was made on sterilized detached fruit by punching with a sterile pin. A drop of the conidial suspension was placed on the wound, while the control fruit received a drop of sterile water. Similarly, unwounded fruit were also inoculated with a single droplet of conidial suspension. For each isolate and method (wounded and unwounded), ten fruit were inoculated, and ten non-inoculated fruit were used as control. All the treated fruit were kept in a plastic box containing moist tissue and incubated at 25º C in the dark. Typical anthracnose lesions appeared on all inoculated wounded fruit while non-inoculated and inoculated unwounded fruits remained asymptotic. Koch postulates were fulfilled by re-isolating and re-identifying the causal agent from inoculated fruit. Colletotrichum aenigma has been reported as the causal agent of anthracnose on Juglans regia, Camellia sinensis and Actinidia arguta in China (Weir et al. 2012; Wang et al. 2016; Wang et al. 2018). Previous studies reported four Colletotrichum species (C. acutatum, C. gloeosporioides, C. fructicola, and C. viniferum) to cause this disease on grapes in South Korea (Oo and Oh 2017; Lim et al. 2020). To the best of our knowledge, this is the first report on grape anthracnose caused by C. aenigma in South Korea. This finding may help to take effective control measures of this disease.

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Morphological and pathogenic characteristics of the fungus Cladobotryum dendroides, the causal agent of cobweb disease of the cultivated mushroom Agaricus bisporus in Serbia

Pesticidi i fitomedicina ◽

10.2298/pif0803175p ◽

2008 ◽

Vol 23 (3) ◽

pp. 175-181 ◽

Cited By ~ 7

Author(s):

Ivana Potocnik ◽

Emil Rekanovic ◽

Svetlana Milijasevic ◽

Biljana Todorovic ◽

Milos Stepanovic

Keyword(s):

Agaricus Bisporus ◽

Natural Infection ◽

Morphological Characteristics ◽

Causal Agent ◽

Fruiting Bodies ◽

Cultivated Mushroom ◽

Pathogenicity Tests ◽

Agar Media ◽

Cobweb Disease

Twenty isolates were isolated from diseased fruiting bodies of Agaricus bisporus collected from Serbian mushroom farms during 2003-2007. The isolates formed white, cottony, aerial colonies on agar media. With age, conidia and colonies turned yellow and redish. Pathogenicity of these isolates was confirmed by inoculation of harvested basidiomes of A. bisporus and by casing inoculation. Symptoms similar to natural infection were recorded. Based on pathogenicity tests and morphological characteristics, the isolates were identified as Cladobotryum dendroides (Bulliard : Fries) W. Gams & Hoozemans.

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First Report of Pestalotiopsis sydowiana Causing Leaf Necrosis of Myrica rubra in China

Plant Disease ◽

10.1094/pdis-01-12-0065-pdn ◽

2012 ◽

Vol 96 (5) ◽

pp. 764-764

Author(s):

X. R. Chen ◽

Y. P. Xing ◽

T. X. Zhang ◽

J. T. Zheng ◽

J. Y. Xu ◽

...

Keyword(s):

Southern China ◽

Morphological Characteristics ◽

Internal Transcribed Spacers ◽

Effective Control ◽

Morphological Identification ◽

Basal Cells ◽

Sterile Water ◽

First Report ◽

Myrica Rubra ◽

Leaf Necrosis

Red bayberry (Myrica rubra Seib. & Zucc.) has great economic importance in eastern and southern China. However, increasing cultivation of red bayberry has resulted in an increase in diseases such as leaf necrosis. In April 2011, a survey was conducted to identify the causal agents of leaf necrosis of red bayberry (cv. Biqi) in Cixi City, Zhejiang Province. Symptoms began with oval and pale brown lesions (2 mm in diameter) that developed into a round to irregular shape (4 to 12 mm in diameter) with pale brown centers and dark brown borders. After approximately 4 months, necrotic lesions expanded to the leaf tips or margins. Black acervuli developed on lesions at later stages. Leaf tissues were surface sterilized with 0.5% sodium hypochlorite for 3 min and rinsed in sterile water before plating onto potato dextrose agar (PDA). Seven isolates were obtained from four samples from four fields on PDA at 25°C. The colonies were cottony white with filiform edges and produced a honey yellow color into the agar at 7 days. Conidia were produced in ink-like fruiting bodies at 4 days at 25°C on PDA. Conidia were straight or slightly curved, fusiform, and five celled with constrictions at the septa. Conidia ranged from 18.7 to 25.8 × 6.2 to 7.7 μm with hyaline apical and basal cells. Thirteen percent of the apical cells had two and the rest had three hyaline appendages ranging from 11.2 to 26.0 μm long. Basal appendages were hyaline, straight, and varied from 3.6 to 5.8 μm long. The color of three median cells was light to dark brown and demonstrated versicolorous. These morphological characteristics matched those of Pestalotiopsis sydowiana (Bresadola) Sutton (1). The morphological identification of the fungus was confirmed by nucleotide blast analysis of the 5.8S subunit and flanking internal transcribed spacers (ITS1 and ITS2) of rDNA regions (GenBank Accession No. JQ322999), which revealed 100% with those of other P. sydowiana isolates (e.g., GenBank Accession No. FJ478105). Koch's postulates were confirmed with 20 healthy leaves of the same size on three branches of three plants in the field. Leaves were wounded by pressing slightly with sterile needles. Mycelial plugs (5 mm in diameter) obtained from the periphery of 7-day-old cultures were placed onto the wounds and covered with sterile-water-saturated cotton. Wounded leaves treated with sterile agar plugs served as controls. The inoculated leaves were sealed in moist plastic bags for 24 h to establish high humid conditions at 21 to 30°C. After 23 days, symptoms on all inoculated leaves were identical to those described above, whereas noninoculated control leaves did not show any symptoms. The fungus was consistently reisolated from the lesions. To our knowledge, this is the first report of P. sydowiana causing leaf necrosis of M. rubra in China. Results can help to better understand the diseases threatening red bayberry trees and develop effective control strategies for better fruit production. Reference: (1) E. F. Guba. Monograph of Monochaetia and Pestalotia. Harvard University Press, Cambridge, MA, 1961.

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First Report of Anthracnose on Camellia japonica Caused by Colletotrichum siamense in Zhejiang Province, China

Plant Disease ◽

10.1094/pdis-08-21-1600-pdn ◽

2021 ◽

Author(s):

Xiaojie Peng ◽

Yuxuan Yuan ◽

Shouke Zhang ◽

Xudong Zhou

Keyword(s):

Disease Incidence ◽

Morphological Characteristics ◽

Zhejiang Province ◽

Internal Transcribed Spacers ◽

Spore Suspension ◽

Likelihood Method ◽

Malt Extract ◽

Camellia Japonica ◽

First Report ◽

Initial Symptoms

Camellia japonica is an attractive flowering woody plant with great ornamental and medicinal value in China. However, typical anthracnose lesions on the leaves are usually observed in summer in Zhejiang province. A number of 100 trees have been investigated with over 70% of leaf disease incidence. The symptom initially develops from the tip or edge of the leaf and dark green infected spots appear. The diseased spots expand and become yellow brown. The lesions are covered with abundant, small and black acervuli at the center with yellow edges. The diseased leaves become brittle, cracked, and finally fall off. Sixty leaves with typical anthracnose symptoms were sampled from gardens in Lin’an, Zhejiang province. The diseased tissues were cut into pieces and incubated in moist chambers at 25°C. The spore mass was collected using a sterile needle under dissection microscope and put on 2% malt extract agar (MEA). The cultures were incubated at 25°C in the dark for one week. Thirty single spore cultures were obtained and grown on 2% MEA at 25°C for morphological characterization. White aerial mycelia and black conidiomata with orange masses of conidia developed seven days later. Conidia are cylindrical in shape, 12-19 μm, mean lengths ranging from 15.5 ± 1.0 to 16.0 ± 1.2 μm. The morphological characteristics are consistent with those of Colletotrichum species. DNA was extracted from three selected isolates (HT-71, J-5, J-20) for sequencing. The partial regions of ribosomal internal transcribed spacers (ITS), calmodulin (CAL), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), actin gene (ACT), beta-tubulin (TUB2), Apn2-Mat1-2 intergenic spacer and partial mating type gene (ApMat), and glutamine synthetase (GS) were amplified as described by Liu et al. (2015). Sequences of the above seven loci for the selected isolates were obtained, and deposited in the GenBank database (MZ014901 to MZ014905, MZ514915 to MZ514922, MZ514925 to MZ514930, MZ497332 and MZ497333). BLAST results indicate they represent Colletotricum siamense. Multi-locus phylogenetic analysis including ex-type of C. siamense (ICMP18578=CBS130417) and related species was conducted using Maximum Likelihood method, and C. acutatum (CBS 112996) served as the outgroup. The three obtained isolates clustered with the ex-type isolate of C. siamense. Eight leaves on two Camellia plants were inoculated to confirm the pathogenicity in the field. The leaves were surface sprayed with 75% ethanol and dried with sterilized filter paper. The leaves were inoculated using the wound/drop inoculation method: an aliquot of 10 μL of spore suspension (1.0 × 106 conidia per mL) was dropped on the left side of a leaf after wounding once by pin-pricking with a sterilized needle. The sterile water was dropped on the right side of the same leaf in parallel as control. The initial symptoms were observed seven days later, all inoculated leaves developed lesions similar to those observed in the field, and no symptoms observed in the control. The fungus was successfully re-isolated only from lesions inoculated with spore suspension exhibiting morphological characteristics resembling those in C. siamense, and further confirmed with sequence data. To our knowledge, this represents the first report of anthracnose on C. japonica caused by C. siamense worldwide. Confirmation of this pathogen in the region will be helpful for the disease management on C. japonica, considering previous report of C. camelliae-japonicae on the same host. References Fu, M., et al. 2019. Persoonia. 42: 1. https://doi.org/10.3767/persoonia.2019.42.01 Guarnaccia, V., et al. 2017. Persoonia. 39: 32. https://doi.org/10.3767/persoonia.2017.39.02 Hou, L. W., et al. 2016. Mycosphere. 7: 1111. Doi 10.5943/mycosphere/si/2c/4 Liu, F., et al. 2015. Persoonia. 35: 63. http://dx.doi.org/10.3767/003158515X687597 Vieira, A. D. S., et al. 2019. Mol. Phylogenet. Evol. https://doi.org/10.1016/j.ympev.2019.106694.

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