scholarly journals First Report of Alternaria dianthicola Causing Leaf Blight on Withania somnifera from India

Plant Disease ◽  
2007 ◽  
Vol 91 (4) ◽  
pp. 467-467 ◽  
Author(s):  
C. K. Maiti ◽  
S. Sen ◽  
A. K. Paul ◽  
K. Acharya

Withania somnifera (L.) Dunal, a potential medicinal plant used for the treatment of nervous disorders, intestinal infection, leprosy, and cancer, is a perennial herb belonging to Solanaceae and distributed throughout the drier parts of India. Leaf blight disease of this plant generally occurs during March in various districts of South Bengal, India. At the initial stage of infection, symptoms appear as small, light brown spots, gradually becoming irregular, dark brown, concentrically zonate with a diffuse margin, frequently surrounded by light yellow haloes, conspicuous brownish concentric rings in the advance stage of infection. A species of Alternaria was isolated from the lesions. The pathogen was isolated on potato dextrose agar (PDA) media. On PDA, the fungus grew slowly with colonies reaching approximately 35 to 40 mm in diameter in 7 days when incubated at 30°C. Conidiophores arose singly or in groups, straight or flexous, cylindrical, septate, pale to olivaceous brown, as much as 155 μm long, 4 to 5.5 μm thick; conidia were straight, obclavate, pale olivaceous brown, smooth, with up to 15 transverse and rarely 1 or 2 longitudinal or oblique septa and measured 50 to 115 × 5 to 10 μm. Pathogenicity tests were carried out three times on 6-month-old plants (n = 10). Plants were sprayed with a conidial suspension of 105 conidia/ml; control plants were sprayed with sterilized water. Plants were covered with polyethylene bags for 10 days. Disease symptoms appeared after 12 ± 1 day after inoculation. Symptoms on the leaves were similar to those of a naturally occurring diseased plant. The fungal pathogen was consistently reisolated from inoculated plants. The pathogen was identified as Alternaria dianthicola and further confirmed by the Agharkar Research Institute, Pune, India. A literature survey reports the occurrence of some fungal diseases (1), but to our knowledge, this is the first report of A. dianthicola on W. somnifera. References: (1) P. Sinha et al. Page 14 in: Recent Progress in Medicinal Plants. Vol. 6 Diseases and their Management. Sci Tech Publishing LLC, Houston, TX, 2000.

Plant Disease ◽  
2012 ◽  
Vol 96 (6) ◽  
pp. 907-907 ◽  
Author(s):  
D. D. M. Bassimba ◽  
J. L. Mira ◽  
C. Baixauli ◽  
A. Vicent

Fennel (Foeniculum vulgare Mill.) is an aromatic herb widely cultivated in Mediterranean areas for culinary and medicinal uses. In 2010, symptoms consisting of leaf blight and necrosis were observed in commercial organic fennel production areas in Valencia Province in east-central Spain. Disease incidence in affected fields was approximately 20%. Symptomatic leaves from four fields were surface disinfected with 0.5% NaOCl for 2 min, and small fragments from necrotic lesions were then plated on potato dextrose agar (PDA) amended with 0.5 g of streptomycin sulfate/liter. After 7 days at 25°C, isolates of the genus Alternaria were consistently isolated. Single conidium cultures were grown on PDA and V8 agar for morphological examination. On both agar media, colonies were dark olive brown without production of pigments. On V8 agar, conidia were solitary, darkly pigmented, and predominantly ovoid-subsphaeroid. Mature conidia were 25 to 59 × 12 to 23 μm with up to six to seven transepta and one to three longisepta. The 5.8S, ITS2, and 28S ribosomal RNA (rRNA) regions were amplified with the primers ITS3 and ITS4 (3) from DNA extracted from the isolate IVIA-A029, and sequenced (GenBank Accession No. JQ240204). The sequence had 100% identity (total score 399, 97% coverage) with that of Alternaria petroselini (Neergard) Simmons strain EGS 09-159 (GenBank Accession No. AF229454.1) (1). Pathogenicity tests were conducted on four 3-month-old fennel plants (cv. Giotto) by spraying a conidial suspension of the fungus (10 ml/plant, 103 conidia/ml of water). Four control plants were sprayed with sterile, distilled water. Plants were covered with plastic bags and incubated in a growth chamber for 72 h at 25°C. Leaf necrosis was visible on inoculated plants after 4 days, but symptoms were not observed on control plants. The fungus was reisolated from leaf lesions on inoculated plants, but not from leaves of control plants, confirming Koch's postulates. On the basis of the morphological (2), molecular, and pathogenicity data, the disease was identified as Alternaria leaf blight of fennel caused by A. petroselini. To our knowledge, this is the first report of A. petroselini in Spain. References: (1) B. M. Pryor and R. L. Gilbertson. Mycol. Res. 104:1312, 2000. (2) E. G. Simmons. Alternaria: An Identification Manual. CBS Fungal Biodiversity Centre, Utrecht, The Netherlands, 2007. (3) T. J. White et al. Pages 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.


Plant Disease ◽  
2014 ◽  
Vol 98 (9) ◽  
pp. 1270-1270 ◽  
Author(s):  
X. Wang ◽  
B. H. Lu ◽  
Y. Zhi ◽  
L. N. Yang ◽  
J. Gao

Eleutherococcus senticosus (Acanthopanax senticosus, manyprickle acathopanax) is a perennial herb belonging to the family Araliaceae and is mainly distributed in northeastern China, Siberia, Korea, and Japan. It is used for the treatment of rheumatism and neurasthenia. With the development of its cultivation, many diseases began to occur (2) and a previously unknown leaf blight on manyprickle was first observed in July of 2010 in Linjiang City, Jilin Province. The same symptoms were detected in other areas of Jilin Province, such as Baishan and Hunchun cities. The disease has resulted in serious loss of production of manyprickle acanthopanax, with 5 to 10% of leaves infected. The infection initially manifested as irregular lesions on the tips or margins of the leaves, which gradually developed into a V-shaped blight with concentric rings that was grayish brown in the center and dark brown at the margins. The blight eventually spread to cover one third of the entire leaf. Severely infected leaves were rolled or distorted, eventually desiccated and became brittle. Under continuously humid conditions, scattered gray mycelium and conidia appeared on the surface of affected leaf tissue. To isolate the causal agent, tissues were excised from diseased leaves, immersed in 0.1% mercuric chloride, suspended in sterile water, and plated on potato dextrose agar (PDA). Conidiophores arose singly or in groups, straight or flexuous, septate, with an inflated basal cell and dendriform near the apex, brown to light brown, and measured 5.0 to 10.0 × 100.0 to 150.0 μm (n = 50). Conidia were single-celled, globoid or oval-shaped, colorless, measuring 6.0 to 10.0 × 7.0 to 13.0 μm (n = 50). In culture, dark, irregular sclerotia were produced. The morphological descriptions and measurements of the fungi were similar to Botrytis cinerea (4). The ITS region of rDNA was amplified and sequenced. BLAST analysis of the 567-bp segment (JX840481) showed 100% identity with the sequence of Botryotinia fuckeliana (perfect stage of B. cinerea). To further identify the species of B. cinerea, three nuclear protein-coding genes (G3PDH, HSP60, and RPB2) (3) were sequenced and the sequences (KJ018759, KJ018757, and KJ018755) all showed 100% identity with those of B. fuckeliana. Pathogenicity tests were carried out on potted, healthy, 1-year-old plants (n = 10). A conidial suspension of 105 conidia/ml was sprayed with each strain (five strains total) on five leaves still on plants, and five plants were sprayed with water as controls. Plants were covered with polyethylene bags and incubated for 3 days at 25°C in a greenhouse. Symptoms appeared 7 days after inoculation, and were similar to those originally observed on plants under natural conditions, whereas control plants remained healthy. The pathogen was successfully re-isolated from inoculated leaves and was identified as B. cinerea on the basis of its morphological characteristics and related gene sequences. B. cinerea has been previously reported on E. senticosus in Korea (1). However, to our knowledge, this is the first report of Botrytis leaf blight of E. senticosus caused by B. cinerea in China. These results lay the foundation for the disease control. References: (1) K. J. Choi et al. Korean J. Med. Crop Sci. 15:199, 2007. (2) J. Gao et al. Plant Dis. 95:493, 2011. (3) M. Staats et al. Mol. Biol. Evol. 22:333, 2005. (4) Z. Y. Zhang. Flora Fungorum Sinicorum. 26. Botrytis, Ramularia. Science Press, Beijing, 2006.


Plant Disease ◽  
2014 ◽  
Vol 98 (5) ◽  
pp. 691-691 ◽  
Author(s):  
Y. H. Jeon ◽  
W. Cheon

Worldwide, Japanese yew (Taxus cuspidata Sieb. & Zucc.) is a popular garden tree, with large trees also being used for timber. In July 2012, leaf blight was observed on 10% of Japanese yew seedling leaves planted in a 500-m2 field in Andong, Gyeongsangbuk-do Province, South Korea. Typical symptoms included small, brown lesions that were first visible on the leaf margin, which enlarged and coalesced into the leaf becoming brown and blighted. To isolate potential pathogens from infected leaves, small sections of leaf tissue (5 to 10 mm2) were excised from lesion margins. Eight fungi were isolated from eight symptomatic trees, respectively. These fungi were hyphal tipped twice and transferred to potato dextrose agar (PDA) plates for incubation at 25°C. After 7 days, the fungi produced circular mats of white aerial mycelia. After 12 days, black acervuli containing slimy spore masses formed over the mycelial mats. Two representative isolates were further characterized. Their conidia were straight or slightly curved, fusiform to clavate, five-celled with constrictions at the septa, and 17.4 to 28.5 × 5.8 to 7.1 μm. Two to four 19.8- to 30.7-μm-long hyaline filamentous appendages (mostly three appendages) were attached to each apical cell, whereas one 3.7- to 7.1-μm-long hyaline appendage was attached to each basal cell, matching the description for Pestalotiopsis microspora (2). The pathogenicity of the two isolates was tested using 2-year-old plants (T. cuspidata var. nana Rehder; three plants per isolate) in 30-cm-diameter pots filled with soil under greenhouse conditions. The plants were inoculated by spraying the leaves with an atomizer with a conidial suspension (105 conidia/ml; ~50 ml on each plant) cultured for 10 days on PDA. As a control, three plants were inoculated with sterilized water. The plants were covered with plastic bags for 72 h to maintain high relative humidity (24 to 28°C). At 20 days after inoculation, small dark lesions enlarged into brown blight similar to that observed on naturally infected leaves. P. microspora was isolated from all inoculated plants, but not the controls. The fungus was confirmed by molecular analysis of the 5.8S subunit and flanking internal transcribed spaces (ITS1 and ITS2) of rDNA amplified from DNA extracted from single-spore cultures, and amplified with the ITS1/ITS4 primers and sequenced as previously described (4). Sequences were compared with other DNA sequences in GenBank using a BLASTN search. The P. microspora isolates were 99% homologous to other P. microspora (DQ456865, EU279435, FJ459951, and FJ459950). The morphological characteristics, pathogenicity, and molecular data assimilated in this study corresponded with the fungus P. microspora (2). This fungus has been previously reported as the causal agent of scab disease of Psidium guajava in Hawaii, the decline of Torreya taxifolia in Florida, and the leaf blight of Reineckea carnea in China (1,3). Therefore, this study presents the first report of P. microspora as a pathogen on T. cuspidata in Korea. The degree of pathogenicity of P. microspora to the Korean garden evergreen T. cuspidata requires quantification to determine its potential economic damage and to establish effective management practices. References: (1) D. F. Farr and A. Y. Rossman, Fungal Databases, Syst. Mycol. Microbiol. Lab. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ (2) L. M. Keith et al. Plant Dis. 90:16, 2006. (3) S. S. N. Maharachchikumbura. Fungal Diversity 50:167, 2011. (4) T. J. White et al. PCR Protocols. Academic Press, San Diego, CA, 1990.


Plant Disease ◽  
2021 ◽  
Author(s):  
Jiahao Lai ◽  
Guihong Xiong ◽  
Bing Liu ◽  
Weigang Kuang ◽  
Shuilin Song

Blueberry (Vaccinium virgatum), an economically important small fruit crop, is characterized by its highly nutritive compounds and high content and wide diversity of bioactive compounds (Miller et al. 2019). In September 2020, an unknown leaf blight disease was observed on Rabbiteye blueberry at the Agricultural Science and Technology Park of Jiangxi Agricultural University in Nanchang, China (28°45'51"N, 115°50'52"E). Disease surveys were conducted at that time, the results showed that disease incidence was 90% from a sampled population of 100 plants in the field, and this disease had not been found at other cultivation fields in Nanchang. Leaf blight disease on blueberry caused the leaves to shrivel and curl, or even fall off, which hindered floral bud development and subsequent yield potential. Symptoms of the disease initially appeared as irregular brown spots (1 to 7 mm in diameter) on the leaves, subsequently coalescing to form large irregular taupe lesions (4 to 15 mm in diameter) which became curly. As the disease progressed, irregular grey-brown and blighted lesion ran throughout the leaf lamina from leaf tip to entire leaf sheath and finally caused dieback and even shoot blight. To identify the causal agent, 15 small pieces (5 mm2) of symptomatic leaves were excised from the junction of diseased and healthy tissue, surface-sterilized in 75% ethanol solution for 30 sec and 0.1% mercuric chloride solution for 2 min, rinsed three times with sterile distilled water, and then incubated on potato dextrose agar (PDA) at 28°C for 5-7 days in darkness. Five fungal isolates showing similar morphological characteristics were obtained as pure cultures by single-spore isolation. All fungal colonies on PDA were white with sparse creeping hyphae. Pycnidia were spherical, light brown, and produced numerous conidia. Conidia were 10.60 to 20.12 × 1.98 to 3.11 µm (average 15.27 × 2.52 µm, n = 100), fusiform, sickle-shaped, light brown, without septa. Based on morphological characteristics, the fungal isolates were suspected to be Coniella castaneicola (Cui 2015). To further confirm the identity of this putative pathogen, two representative isolates LGZ2 and LGZ3 were selected for molecular identification. The internal transcribed spacer region (ITS) and large subunit (LSU) were amplified and sequenced using primers ITS1/ITS4 (Peever et al. 2004) and LROR/LR7 (Castlebury and Rossman 2002). The sequences of ITS region (GenBank accession nos. MW672530 and MW856809) showed 100% identity with accessions numbers KF564280 (576/576 bp), MW208111 (544/544 bp), MW208112 (544/544 bp) of C. castaneicola. LSU gene sequences (GenBank accession nos. MW856810 to 11) was 99.85% (1324/1326 bp, 1329/1331 bp) identical to the sequences of C. castaneicola (KY473971, KR232683 to 84). Pathogenicity was tested on three blueberry varieties (‘Rabbiteye’, ‘Double Peak’ and ‘Pink Lemonade’), and four healthy young leaves of a potted blueberry of each variety with and without injury were inoculated with 20 μl suspension of prepared spores (106 conidia/mL) derived from 7-day-old cultures of LGZ2, respectively. In addition, four leaves of each variety with and without injury were sprayed with sterile distilled water as a control, respectively. The experiment was repeated three times, and all plants were incubated in a growth chamber (a 12h light and 12h dark period, 25°C, RH greater than 80%). After 4 days, all the inoculated leaves started showing disease symptoms (large irregular grey-brown lesions) as those observed in the field and there was no difference in severity recorded between the blueberry varieties, whereas the control leaves showed no symptoms. The fungus was reisolated from the inoculated leaves and confirmed as C. castaneicola by morphological and molecular identification, fulfilling Koch’s postulates. To our knowledge, this is the first report of C. castaneicola causing leaf blight on blueberries in China. The discovery of this new disease and the identification of the pathogen will provide useful information for developing effective control strategies, reducing economic losses in blueberry production, and promoting the development of the blueberry industry.


Plant Disease ◽  
2020 ◽  
Vol 104 (1) ◽  
pp. 291
Author(s):  
X. Cao ◽  
Y. L. Du ◽  
X. S. Zhang ◽  
H. Y. Li ◽  
S. J. Guo ◽  
...  

Plant Disease ◽  
2020 ◽  
Author(s):  
Boda Praveen ◽  
A. Nagaraja ◽  
M. K. Prasanna Kumar ◽  
Devanna Pramesh ◽  
K. B. Palanna ◽  
...  

Little millet (LM) is a minor cereal crop grown in the Indian sub-continent. During October 2018, dark brown, circular to oval necrotic spots surrounded by concentric rings were observed on the upper leaf surface of the LM (cv. VS-13) grown in the fields of the University of Agricultural Sciences, Bengaluru, India (13.0784oN, 77.5793oE). As the disease progressed, infected leaves became blighted. Disease incidence up to 53% was recorded in 3 fields of 0.4-hectare area each. Thirty symptomatic leaves were collected to isolate the associated causal organism. The margins of diseased tissue were cut into 5 × 5-mm pieces, surface-sterilized in 75% ethanol for 45 seconds followed by 1% sodium hypochlorite for 1 min, finally rinsed in sterile distilled water five times and placed on PDA. After 7 days of incubation at 25°C, greyish fungal colonies appeared on PDA. Single-spore isolations were performed to obtain ten isolates. Pure cultures of the fungus initially produced light gray aerial mycelia that later turned to dark grey. All isolates formed obclavate to pyriform conidia measured 22.66-48.97μm long and 6.55-13.79µm wide with 1-3 longitudinal and 2-7 transverse septa with a short beak (2.55-13.26µm) (n=50). Based on the conidial morphology, the fungus was identified as Alternaria sp. Further, the taxonomic identity of all ten isolates was confirmed as A. alternata using species-specific primers (AAF2/AAR3, Konstantinova et al. 2002) in a PCR assay. Later, one of the isolate UASB1 was selected, and its internal transcribed spacer (ITS) region, glyceraldehyde-3-phosphate dehydrogenase (gapdh), major allergen Alt a 1 (Alt a 1), major endo-polygalacturonase (endoPG), OPA10-2, and KOG1058 genes were amplified in PCR (White et al. 1990; Berbee et al. 1999; Woudenberg et al. 2015), and the resultant products were sequenced and deposited in the NCBI GenBank (ITS, MN919390; gapdh, MT637185; Alt a 1, MT882339; endoPG, MT882340; OPA10-2, MT882341; KOG1058, MT882342). Blastn analysis of ITS, gapdh, Alt a 1, endoPG, OPA10-2, KOG1058 gene sequences showed 99.62% (with AF347031), 97.36% (with AY278808), 99.58% (with AY563301), 99.10% (with JQ811978), 99.05% (with KP124632) and 99.23% (with KP125233) respectively, identity with reference strain CBS916.96 of A. alternata, confirming UASB1 isolate to be A. alternata. For pathogenicity assay, conidial suspension of UASB1 isolate was spray inoculated to ten healthy LM (cv. VS-13) plants (45 days old) maintained under protected conditions. The spore suspension was sprayed until runoff on healthy leaves, and ten healthy plants sprayed with sterile water served as controls. Later, all inoculated and control plants were covered with transparent polyethylene bags and were maintained in a greenhouse at 28±2 ◦C and 90% RH. The pathogenicity test was repeated three times. After 8 days post-inoculation, inoculated plants showed leaf blight symptoms as observed in the field, whereas no disease symptoms were observed on non-inoculated plants. Re-isolations were performed from inoculated plants, and the re-isolated pathogen was confirmed as A. alternata based on morphological and PCR assay (Konstantinova et al. 2002). No pathogens were isolated from control plants. There is an increasing acreage of LM crop in India, and this first report indicates the need for further studies on leaf blight management and the disease impacts on crop yields.


Plant Disease ◽  
2013 ◽  
Vol 97 (1) ◽  
pp. 147-147
Author(s):  
J. H. Park ◽  
S. E. Cho ◽  
K. S. Han ◽  
H. D. Shin

Garlic chives, Allium tuberosum Roth., are widely cultivated in Asia and are the fourth most important Allium crop in Korea. In June 2011, a leaf blight of garlic chives associated with a Septoria spp. was observed on an organic farm in Hongcheon County, Korea. Similar symptoms were also found in fields within Samcheok City and Yangku County of Korea during the 2011 and 2012 seasons. Disease incidence (percentage of plants affected) was 5 to 10% in organic farms surveyed. Diseased voucher specimens (n = 5) were deposited at the Korea University Herbarium (KUS). The disease first appeared as yellowish specks on leaves, expanding to cause a leaf tip dieback. Half of the leaves may be diseased within a week, especially during wet weather. Pycnidia were directly observed in leaf lesions. Pycnidia were amphigenous, but mostly epigenous, scattered, dark brown to rusty brown, globose, embedded in host tissue or partly erumpent, separate, unilocular, 50 to 150 μm in diameter, with ostioles of 20 to 40 μm in diameter. Conidia were acicular, straight to sub-straight, truncate at the base, obtuse at the apex, hyaline, aguttulate, 22 to 44 × 1.8 to 3 μm, mostly 3-septate, occasionally 1- or 2-septate. These morphological characteristics matched those of Septoria allii Moesz, which is differentiated from S. alliacea on conidial dimensions (50 to 60 μm long) (1,2). A monoconidial isolate was cultured on potato dextrose agar (PDA). Two isolates have been deposited in the Korean Agricultural Culture Collection (Accession Nos. KACC46119 and 46688). 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 482-bp was deposited in GenBank (JX531648 and JX531649). ITS sequence information was at least 99% similar to those of many Septoria species, however no information was available for S. allii. 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 PDA. Control leaves were sprayed with sterile water. The plants were placed in humid chambers (relative humidity 100%) for the first 48 h. After 7 days, typical leaf blight symptoms started to develop on the leaves of inoculated plants. S. allii was reisolated from the lesions of inoculated plants, confirming Koch's postulates. No symptoms were observed on control plants. The host-parasite association of A. tuberosum and S. allii has been known only from China (1). S. alliacea has been recorded on several species of Allium, e.g. A. cepa, A. chinense, A. fistulosum, and A. tuberosum from Japan (4) and A. cepa from Korea (3). To the best of our knowledge, this is the first report of S. allii on garlic chives. No diseased plants were observed in commercial fields of garlic chives which involved regular application of fungicides. The disease therefore seems to be limited to organic garlic chive production. References: (1) P. K. Chi et al. Fungous Diseases on Cultivated Plants of Jilin Province, Science Press, Beijing, China, 1966. (2) P. A. Saccardo. Sylloge Fungorum Omnium Hucusque Congnitorum. XXV. Berlin, 1931. (3) The Korean Society of Plant Pathology. List of Plant Diseases in Korea, Suwon, Korea, 2009. (4) The Phytopathological Society of Japan. Common Names of Plant Diseases in Japan, Tokyo, Japan, 2000.


Plant Disease ◽  
2014 ◽  
Vol 98 (5) ◽  
pp. 686-686 ◽  
Author(s):  
L. M. Liu ◽  
S. W. Huang ◽  
L. Wang ◽  
E. Q. Hou ◽  
D. F. Xiao

Leaf-streak symptoms were observed on rice (Oryza sativa L.) starting at the booting stage through harvest in Zhejiang Province, China, in 2012. Based on Fuyang County, only 15% of the rice fields were estimated to show these symptoms. However, incidence could be 40 to 80% when the rice got infected. Typical symptoms started as green water-soaked streaks from the tip or edge of leaf blades, similar to bacterial leaf blight caused by Xanthomonas oryzae. Infected leaves turned yellow, then eventually became wilted and dry. No bacterial streaming was observed and no bacteria were isolated. Pieces of infected leaf tissue were surface sterilized using 0.1% (v/v) mercuric chloride, rinsed with sterilized water, then placed on water agar (WA). After 2 or 3 days on WA at 28°C, only fungal growth was observed from surface sterilized tissues. Fungi were isolated, purified by single spore separation process, and subcultured to potato dextrose agar (PDA) plates. Growing on PDA, the surface of the colony was circular, fluffy, and shiny velvety-black, whereas the under surface was dark Prussian blue. Conidiophores were single or fascicled, brown to dark brown, rarely branched, multiseptate, and straight or often geniculate near the apex. Conidia were brown, smooth, fusiform, geniculate or hook-shaped, 17.5 to 28.5 × 8.5 to 14.0 μm, and 3-septate, with the third cell from the base larger and darker than the others. Molecular identification was performed by analysis of the rDNA internal transcribed spacer region (ITS1-5.8S-ITS2). The rDNA-ITS region was amplified with primer pair ITS1 and ITS4 (5), sequenced, and deposited in GenBank (Accession No. KC462186). The sequence of rDNA-ITS (KC462186) showed 100% identity with Cochliobolus lunatus R.R. Nelson & Haasis (JN943422) after BLAST. Based on the results of morphological and molecular analyses, the fungus isolated from infected leaves was identified as C. lunatus (anamorph: Curvularia lunata (Wakk.) Boedijn) (3). Pathogenicity tests were conducted three times by spraying a conidial suspension (1 × 105 spores/ml) with 0.1% (v/v) Tween 20 on 12 healthy rice plants at late tillering stage. The same number of the healthy rice plants sprayed with sterilized water with 0.1% (v/v) Tween 20 were used as control. All plants were kept at 30°C and 75 to 85% relative humidity (RH) under a 12-h light/dark rotation. About 5 to 7 days after inoculation, green water-soaked streaks began to appear on inoculated plants. From 7 to 14 days after inoculation, the lesions developed quickly and the leaves began to wilt. After 14 days, inoculated plants showed symptoms similar to those originally observed in the field, while control plants (sprayed with sterilized water) remained healthy. C. lunatus was re-isolated from all inoculated plants, and re-identified by the same methods (morphological and molecular methods) as described above, thereby satisfying Koch's postulates, and confirming C. lunatus as the cause of the disease. C. lunatus is a pathogen of a wide range of plants and is common in paddy environments. It was reported as one of the causal agents of black kernel of rice (4) and rice spikelet rot disease (SRD) (1,2). The level of incidence observed in the affected fields suggest that this disease could potentially cause major losses under favorable weather conditions if susceptible cultivars are grown. To our knowledge, this is the first report of C. lunatus causing leaf blight of rice in China. References: (1) S. W. Huang et al. Crop Prot. 30:1, 2011. (2) S. W. Huang et al. Crop Prot. 30:10, 2011. (3) D. S. Manamgoda et al. Fungal Divers. 51:3. (4) S. H. Ou. Rice diseases [M]. CABI, 1985. (5) T. J. White et al. PCR Protocols: a Guide to Methods and Application. Academic Press, San Diego, CA, 1990.


Plant Disease ◽  
2013 ◽  
Vol 97 (1) ◽  
pp. 138-138 ◽  
Author(s):  
X. Y. Chen ◽  
J. D. Feng ◽  
Z. Su ◽  
C. Sui ◽  
X. Huang

Curcuma wenyujin Y.H. Chen & C. Ling is a traditional Chinese medicinal herb in the Zingiberaceae family. Commonly known as Wen yujin, the root is widely used for alleviating pain and protecting the liver. A severe leaf blight disease was observed in three C. wenyujin farms in Hainan Province of China in October 2010. The obvious symptoms of leaf blight, yellow to brown irregular lesions (1 to 20 cm) on C. wenyujin, usually began at the tips of leaves and the main veins. This disease, especially severe from August to October, caused heavy damage and 100% of mature plants (10 months old) in farms were infected. The disease was most severe when continuous cropping was performed and showed slight improvement when rotation was adopted. Farmers usually sprayed carbendazim (50% WP) and thiophanate-methyl (70% WP) to control this disease, but these treatments were not effective. To isolate the causal pathogen, diseased plants were collected in October 2010 from a field of the Hainan Branch Institute of Medicinal Plant Development in Hainan Province. Lesion tissue was removed from the border between symptomatic and healthy tissue, surface sterilized in 75% ethanol for 1 min, washed in three changes of sterile distilled water, transferred to potato dextrose agar (PDA) plates, and incubated at 28°C for 7 days. Single spore cultures of five isolates were obtained and identified as Curvularia clavata based on morphological characteristics (1). Conidia measured 20 to 29 × 7.5 to 10.5 μm (n = 100), were curved, 3-septate, and the third cell from the base was larger and darker than the others. Mycelia of single spore cultures growing on PDA for 5 days were used for DNA extraction using a plant genomic DNA kit (TIANGEN, Beijing). The internal transcribed spacer (ITS) region of the rDNA was amplified using primers ITS1 and ITS4. The amplicons were 562 bp in length (GenBank Accession No. JQ730852) and had 99% nucleotide identity with the GenBank Accession No. JN021115 and AF071336 of C. clavata. Pathogenicity tests were conducted using fresh and healthy detached Curcuma wenyujin leaves. Mycelial discs (10 mm) removed from a 5-day-old colony on PDA were used for inoculation. Each isolate was inoculated on three distinct leaves (two distinct inoculations per leaf). Three additional leaves inoculated with sterile PDA discs were used as control. Inoculated leaves were covered with a polythene film to maintain high humidity. Leaves in trays were kept in a growth chamber at 28°C and observed for symptom appearance every day. Five days after inoculation, inoculated leaves developed blight symptoms similar to those observed on naturally infected leaves. No symptoms were observed on non-inoculated leaves. C. clavata was reisolated from the inoculated leaves, thus fulfilling Koch's postulates. C. clavata has been previously reported to be economically important on a number of other hosts (2). To our knowledge, this is the first report of Curvularia leaf blight on Curcuma wenyujin caused by C. clavata in China. References: (1) A. M Mandokhot et al. Eur. J. Plant Pathol.78:65, 1972. (2) T. Y. Zhang et al. Flora fungorum sinicorum: Beijing, China, 2010.


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