First Report of Leaf Blight on Sweet Persimmon Tree by Pestalotiopsis theae in Spain

J. J. Tuset; C. Hinarejos; J. L. Mira

doi:10.1094/pdis.1999.83.11.1070c

First Report of Leaf Blight on Sweet Persimmon Tree by Pestalotiopsis theae in Spain

Plant Disease ◽

10.1094/pdis.1999.83.11.1070c ◽

1999 ◽

Vol 83 (11) ◽

pp. 1070-1070 ◽

Cited By ~ 10

Author(s):

J. J. Tuset ◽

C. Hinarejos ◽

J. L. Mira

Keyword(s):

Morphological Characteristics ◽

Climatic Conditions ◽

Leaf Blight ◽

Diospyros Kaki ◽

Conidial Suspension ◽

Basal Cells ◽

First Report ◽

Dry Conditions ◽

Agar Media ◽

Grey Blight

During July 1998, a leaf blight caused by Pestalotiopsis theae (Saw.) Stey. was observed at an incidence of 18 to 20% in sweet persimmon (Diospyros kaki L.fil.) orchards in Huelva Province (southwestern Spain). Symptoms appeared on leaves as large grayish brown circular ringspots. Usually, they were solitary, but occasionally, two to three spots occurred on an affected leaf. In severe cases, lesions developed on more than one-third of the leaf, resulting in defoliation. Small black acervular conidiomata were visible in the surface of spots. These conidiomata produced fusiform conidia that were straight or rarely curved, four five-celled euseptate, including three olivaceous or dark brown median cells, and hyaline apical and basal cells with appendages that were slightly constricted at septa. Conidiomata were up to 240 μm in diameter; conidiogenous cells were 6 to 13 × 1.2 to 2.8 μm; conidia were 24.7 × 7.8 μm; three median cells were 16.7 μm long; two to three apical appendages (rarely four) were 28.3 μm long; and straight basal appendage was 5.7 μm. P. theae was consistently isolated on potato dextrose agar from diseased leaves and conidiomata. To confirm pathogenicity, both mycelial plugs and a conidial suspension (1.5 × 106 conidia per ml) of the fungus were used as inocula. Young completely developed leaves from persimmon tree cvs. Sharon and Hanafuyu were inoculated in the laboratory and maintained in a moist chamber for 5 days. Lesions resembling symptoms that occurred in the field were observed on leaves after 5 days. Symptoms were not observed on control leaves inoculated with agar media or sprayed with water. The fungus reisolated from diseased leaves was identical to the original isolates. Based on the morphological characteristics of conidiomata and conidia as well as pathogenicity, the fungus was identified as P. theae (1). This is the first report of this fungus as a pathogen of D. kaki in Europe. Possibly the introduction of P. theae to Spain has been through young imported persimmon plants. Unusual climatic conditions (heavy rainfalls during 1997 in southwestern Spain) have been favorable for disease development. The hot and dry conditions that usually occur during flowering, growing, and maturation of persimmon fruits normally prevent dissemination of inoculum and infection of leaves. For these reasons, the wet areas of southwestern Spain could be more favorable for “grey blight” of persimmon trees. Reference: (1) T.-H. Chang et al. Korean J. Plant Pathol. 12:377, 1996.

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First Report of Leaf Blight of Japanese Yew Caused by Pestalotiopsis microspora in Korea

Plant Disease ◽

10.1094/pdis-08-13-0821-pdn ◽

2014 ◽

Vol 98 (5) ◽

pp. 691-691 ◽

Cited By ~ 2

Author(s):

Y. H. Jeon ◽

W. Cheon

Keyword(s):

Dna Sequences ◽

Apical Cell ◽

Morphological Characteristics ◽

Leaf Blight ◽

Economic Damage ◽

Leaf Margin ◽

Conidial Suspension ◽

First Report ◽

Pestalotiopsis Microspora ◽

Large Trees

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.

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First Report of Leaf Spot Caused by Botrytis cinerea on Cardamine hupingshanensis in China

Plant Disease ◽

10.1094/pdis-04-21-0698-pdn ◽

2021 ◽

Author(s):

Jun Guo ◽

Jin Chen ◽

Zhao Hu ◽

Jie Zhong ◽

Jun Zi Zhu

Keyword(s):

Botrytis Cinerea ◽

Morphological Characteristics ◽

Gray Mold ◽

Hunan Province ◽

Conidial Suspension ◽

Heat Shock Protein 60 ◽

Basal Cells ◽

Single Spore ◽

Internal Transcribed Spacer Region ◽

First Report

Cardamine hupingshanensis is a selenium (Se) and cadmium (Cd) hyperaccumulator plant distributed in wetlands along the Wuling Mountains of China (Zhou et al. 2018). In March of 2020, a disease with symptoms similar to gray mold was observed on leaves of C. hupingshanensis in a nursery located in Changsha, Hunan Province, China. Almost 40% of the C. hupingshanensis (200 plants) were infected. Initially, small spots were scattered across the leaf surface or margin. As disease progressed, small spots enlarged to dark brown lesions, with green-gray, conidia containing mold layer under humid conditions. Small leaf pieces were cut from the lesion margins and were sterilized with 70% ethanol for 10 s, 2% NaOCl for 2 min, rinsed with sterilized distilled water for three times, and then placed on potato dextrose agar (PDA) medium at 22°C in the dark. Seven similar colonies were consistently isolated from seven samples and further purified by single-spore isolation. Strains cultured on PDA were initially white, forming gray-white aerial mycelia, then turned gray and produced sclerotia after incubation for 2 weeks, which were brown to blackish, irregular, 0.8 to 3.0 × 1.2 to 3.5 mm (n=50). Conidia were unicellular, globose or oval, colourless, 7.5 to 12.0 × 5.5 to 8.3 μm (n=50). Conidiophores arose singly or in group, straight or flexuous, septate, brownish to light brown, with enlarged basal cells, 12.5 to 22.1 × 120.7 to 310.3 μm. Based on their morphological characteristics in culture, the isolates were putatively identified as Botrytis cinerea (Ellis 1971). Genomic DNA of four representative isolates, HNSMJ-1 to HNSMJ-4, were extracted by CTAB method. The internal transcribed spacer region (ITS), glyceraldehyde-3-phosphate dehydrogenase gene (G3PDH), heat-shock protein 60 gene (HSP60), ATP-dependent RNA helicaseDBP7 gene (MS547) and DNA-dependent RNA polymerase subunit II gene (RPB2) were amplified and sequenced using the primers described previously (Aktaruzzaman et al. 2018) (MW820311, MW831620, MW831628, MW831623 and MW831629 for HNSMJ-1; MW314722, MW316616, MW316617, MW316618 and MW316619 for HNSMJ-2; MW820519, MW831621, MW831627, MW831624 and MW831631 for HNSMJ-3; MW820601, MW831622, MW831626, MW831625 and MW831630 for HNSMJ-4). BLAST searches showed 99.43 to 99.90% identity to the corresponding sequences of B. cinerea strains, such as HJ-5 (MF426032.1, MN448500.1, MK791187.1, MH727700.1 and KX867998.1). A combined phylogenetic tree using the ITS, G3PDH, HSP60 and RPB2 sequences was constructed by neighbor-joining method in MEGA 6. It revealed that HNSMJ-1 to HNSMJ-4 clustered in the B. cinerea clade. Pathogenicity tests were performed on healthy pot-grown C. hupingshanensis plants. Leaves were surface-sterilized and sprayed with conidial suspension (106 conidia/ mL), with sterile water served as controls. All plants were kept in growth chamber with 85% humidity at 25℃ following a 16 h day-8 h night cycle. The experiment was repeated twice, with each three replications. After 4 to 7 days, symptoms similar to those observed in the field developed on the inoculated leaves, whereas controls remained healthy. The pathogen was reisolated from symptomatic tissues and identified using molecular methods, confirming Koch’s postulates. B. cinerea has already been reported from China on C. lyrate (Zhang 2006), a different species of C. hupingshanensis. To the best of our knowledge, this is the first report of B. cinerea causing gray mold on C. hupingshanensis in China and worldwide. Based on the widespread damage in the nursery, appropriate control strategies should be adopted. This study provides a basis for studying the epidemic and management of the disease.

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First Report of Leaf Blight Caused by Septoria allii on Garlic Chives in Korea

Plant Disease ◽

10.1094/pdis-08-12-0809-pdn ◽

2013 ◽

Vol 97 (1) ◽

pp. 147-147

Author(s):

J. H. Park ◽

S. E. Cho ◽

K. S. Han ◽

H. D. Shin

Keyword(s):

Disease Incidence ◽

Morphological Characteristics ◽

Its Region ◽

Leaf Blight ◽

Plant Diseases ◽

Cultivated Plants ◽

Sequence Information ◽

Conidial Suspension ◽

Korean Society ◽

First Report

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.

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First Report of Nigrospora Leaf Blight on Sesame Caused by Nigrospora sphaerica in China

Plant Disease ◽

10.1094/pdis-07-13-0684-pdn ◽

2014 ◽

Vol 98 (6) ◽

pp. 842-842 ◽

Cited By ~ 3

Author(s):

H. Zhao ◽

H. Y. Liu ◽

X. S. Yang ◽

Y. X. Liu ◽

Y. X. Ni ◽

...

Keyword(s):

Morphological Characteristics ◽

Leaf Blight ◽

Morphological Data ◽

Its2 Region ◽

Oilseed Crop ◽

Conidial Suspension ◽

First Report ◽

Initial Symptoms ◽

Central Regions ◽

Leaf Pathogen

Sesame (Sesamum indicum L.) is an important oilseed crop widely grown in the central regions of China. A new leaf blight has increasingly been observed in sesame fields in Anhui, Hubei, and Henan provinces since 2010. Approximately 30 to 40% of the plants were symptomatic in the affected fields. Initial symptoms were yellow to brown, irregularly shaped lesions. Lesions later expanded and the affected leaves tuned grayish to dark brown and wilted, with a layer of whitish mycelial growth on the underside. Severe blighting caused the center of lesions to fall out, leaving holes in the leaves. Sections of symptomatic leaf tissues were surface-sterilized in 75% ethanol for 30 s, then in 1% HgCl2 for 30 s, rinsed three times in sterile distilled water, and plated onto potato dextrose agar (PDA). The resulting fungal colonies were initially white, and then became grayish-brown with sporulation. Conidia were single-celled, black, smooth, spherical, 14.2 to 19.8 μm (average 17.1 μm) in diameter, and borne on a hyaline vesicle at the tip of each conidiophore. Morphological characteristics of the isolates were similar to those of Nigrospora sphaerica (1). To verify the identification based on morphological features, the ITS1-5.8S-ITS2 region of the ribosomal RNA was amplified using ITS1 (5′-TCCGTAGGTGAACCTGCGG-3′) and ITS4 (5′-TCCTCCGCTTATTGATATGC-3′) primers (3), and then sequenced and compared to the GenBank database through a BLAST search. Comparison of the sequence revealed 100% similarity to N. sphaerica (GenBank Accession No. JF817271.1). On the basis of morphological data and the ITS rDNA sequence, the isolate was determined to be N. sphaerica. Pathogenicity tests were conducted using fresh and healthy sesame leaves of 10 plants. A conidial suspension (106 conidia/ml) collected from a 7-day-old culture on PDA was used for inoculation. Leaves of 10 plants were spray-inoculated with the spore suspension at the 6-week-old growth stage, and an additional 10 plants were sprayed with sterile water. Inoculated plants were covered with polyethylene bags to maintain high humidity. Plants were kept at 28°C and observed for symptom every day. Ten to 15 days after inoculation, inoculated leaves developed blight symptoms similar to those observed on naturally infected leaves. No symptoms were observed on the control leaves. N. sphaerica was re-isolated from the inoculated leaves, thus fulfilling Koch's postulates. N. sphaerica has been reported as a leaf pathogen on several hosts worldwide (2). To our knowledge, this is the first report of Nigrospora leaf blight on sesame caused by N. sphaerica in China. References: (1) M. B. Ellis. Dematiaceous Hyphomycetes. CMI, Kew, Surrey, UK, 1971. (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/ . July 01, 2013. (3) M. A. Innis et al. PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.

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First Report of Volutella Blight on Pachysandra Caused by Volutella pachysandricola in China

Plant Disease ◽

10.1094/pdis-11-11-0997 ◽

2012 ◽

Vol 96 (4) ◽

pp. 584-584

Author(s):

Q. Bai ◽

Y. Xie ◽

R. Dong ◽

J. Gao ◽

Y. Li

Keyword(s):

Morphological Characteristics ◽

Stem Canker ◽

Botanical Garden ◽

The United States ◽

Culture Collection ◽

Leaf Blight ◽

Conidial Suspension ◽

First Report ◽

Leaf Spots ◽

Plastic Bags

Pachysandra (Pachysandra terminalis, Buxaceae) and Japanese Pachysandra, also called Japanese Spurge, is a woody ornamental groundcover plant distributed mostly in Zhejiang, Guizhou, Henan, Hubei, Sichuan, Shanxi, and Gansu provinces in China. In April 2010, P. terminalis asymptomatic plants were shipped from Beijing Botanical Garden Institute of Botany Chinese Academy of Science to the garden nursery of Jilin Agricultural University (43°48′N, 125°23′E), Jilin Province. In June 2011, Volutella blight (sometimes called leaf blight and stem canker) of P. terminalis was observed on these plants. Infected leaves showed circular or irregular, tan-to-brown spots often with concentric rings and dark margins. The spots eventually grew and coalesced until the entire leaf died. Cankers appeared as greenish brown and water-soaked diseased areas, subsequently turning brown or black, and shriveled and often girdled the stems and stolons. During wet, humid weather in autumn, reddish orange, cushion-like fruiting structures of the fungus appeared on the stem cankers and undersides of leaf spots. Symptoms of the disease were consistent with previous descriptions (2–4). Five isolates were obtained from necrotic tissue of leaf spots and cankers of stems and stolons and cultured on potato dextrose agar. The colony surface was salmon colored and slimy. Conidia were hyaline, one celled, spindle shaped, and 12.57 to 22.23 × 3.33 to 4.15 μm with rounded ends. Morphological characteristics of the fungus were consistent with the description by Dodge (2), and the fungus was identified as Volutella pachysandricola (telemorph Pseudonectria pachysandricola). The internal transcribed spacer (ITS) regions of the nuclear rDNA were amplified using primers ITS4/ITS5 (1). The ITS sequences were 553 bp long and identical among these five isolates (GenBank Accession No. HE612114). They were 100% identical to Pseudonectria pachysandricola voucher KUS-F25663 (Accession No. JN797821) and 99% identical to P. pachysandricola culture-collection DAOM (Accession No. HQ897807). Pathogenicity was confirmed by spraying leaves of clonally propagated cuttings of P. terminalis with a conidial suspension (1 × 106 conidia/ml) of the isolated V. pachysandricola. Control leaves were sprayed with sterile water. Plants were covered with plastic bags and kept in a greenhouse at 20 to 25°C for 72 h. After 5 to 8 days, typical disease symptoms appeared on leaves, while the control plants remained healthy. V. pachysandricola was reisolated from the leaf spots of inoculated plants. Pachysandra leaf blight and stem canker also called Volutella blight, is the most destructive disease of P. terminalis and previously reported in the northern humid areas of the United States (Illinois, Connecticut, Ohio, Indiana, Iowa, Massachusetts, Missouri, Kentucky, and Wisconsin), northern Europe (Britain, Germany, and Poland), and the Czech Republic. To our knowledge, this is the first report of the disease caused by V. pachysandricola in China. The disease may become a more significant problem in P. terminalis cultivation areas if the disease spreads on P. terminalis in nursery beds. References: (1) D. E. L. Cooke et al. Mycol. Res. 101:667, 1997. (2) B. O. Dodge. Mycologia 36:532, 1944. (3) S. M. Douglas. Online publication. Volutella Blight of Pachysandra. The Connecticut Agricultural Experiment Station, 2008. (4) I. Safrankova. Plant Protect. Sci.43:10, 2007.

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First Report of Leaf Blight on Rubus brasiliensis Caused by Colletotrichum acutatum in Brazil

Plant Disease ◽

10.1094/pdis-06-10-0441 ◽

2010 ◽

Vol 94 (11) ◽

pp. 1378-1378 ◽

Cited By ~ 1

Author(s):

U. P. Lopes ◽

L. Zambolim ◽

H. S. S. Duarte ◽

P. G. C. Cabral ◽

O. L. Pereira ◽

...

Keyword(s):

Native Species ◽

Morphological Characteristics ◽

Minas Gerais ◽

Leaf Blight ◽

Colletotrichum Acutatum ◽

Conidial Suspension ◽

Specific Primer ◽

First Report ◽

Young Leaves ◽

Necrotic Spots

There are more than 300 blackberry (Rubus) species worldwide. Rubus brasiliensis Mart. is a native Brazilian species found in tropical forests. In January 2009, samples of R. brasiliensis with severe leaf blight were collected from an area of rain forest in the city of São Miguel do Anta, State of Minas Gerais, Brazil. Dark spots began developing in the young leaves and progressed to necrotic spots with occasional twig dieback. From the spots, a fungus was isolated with the following morphology: acervuli that were 20 to 50.0 × 50 to 125.0 μm and hyaline amerospores that were ellipsoid and fusiform and 7.5 to 23.75 × 2.5 to 5.0 μm. On the basis of these morphological characteristics, the fungus was identified as Colletotrichum acutatum. In Brazil, C. acutatum is reported in apple, citrus, strawberry, peach, plum, nectarine, olive, medlar, and yerba-mate, but it was not reported as the causal agent of leaf blight in R. brasiliensis. A sample was deposited in the herbarium at the Universidade Federal de Viçosa, Minas Gerais, Brazil (VIC 31210). One representative isolate, OLP 571, was used for pathogenicity testing and molecular studies. Identity was confirmed by amplifying the internal transcribed spacer (ITS) regions of the ribosomal RNA with primers ITS4 (3), CaInt2 (a specific primer for C. acutatum [2]) and CgInt (a specific primer for C. gloeosporioides [1]). Isolates of C. acutatum (DAR78874 and DAR78876) and C. gloeosporioides (DAR78875) obtained from Australian olive trees were used as positive controls. The primers ITS4 and CaInt2 amplified a single DNA product of 500 bp expected for C. acutatum. OLP 571 was grown for 7 days on potato dextrose agar. Young leaves of R. brasiliensis were inoculated with a conidial suspension (106 conidia/ml) on young leaves. Inoculated plants were maintained in a moist chamber for 2 days and subsequently in a greenhouse at 25°C. Necrotic spots similar to those described were detected on young leaves 3 days after the inoculation. Control leaves, on which only water was sprayed, remained healthy. The same fungus was reisolated from the inoculated symptomatic tissues. To our knowledge, this is the first report of C. acutatum causing leaf blight in the native species of R. brasiliensis in Brazil. References: (1) P. R. Mills et al. FEMS Microbiol. Lett. 98:137, 1999. (2) S. Sreenivasaprasad et al. Plant Pathol. 45:650, 1996. (3) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.

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First Report of Lentil Ascochyta Blight Caused by Ascochyta lentis in Italy

Plant Disease ◽

10.1094/pdis.1999.83.1.77c ◽

1999 ◽

Vol 83 (1) ◽

pp. 77-77

Author(s):

C. Cappelli ◽

R. Buonaurio ◽

R. Torricelli

Keyword(s):

Lens Culinaris ◽

Ascochyta Blight ◽

Central Italy ◽

Morphological Characteristics ◽

Potato Dextrose Agar ◽

Conidial Suspension ◽

First Report ◽

Ascochyta Lentis ◽

Humidity Chamber ◽

Dry Conditions

In May 1997, ascochyta blight incited by Ascochyta lentis Vassiljevsky was observed at an incidence of less than 5% in lentil (Lens culinaris Medik.) fields in Umbria (Central Italy). Symptoms appeared on leaves and stems as tan spots surrounded by a dark margin. Small black pycnidia that produced a pink exudate containing hyaline, 1 septate, 14.2 to 15.8 × 3.5 μm conidia under high humidity were visible in the center of the spots. The fungus was consistently isolated on potato dextrose agar from diseased leaves or stems. To satisfy Koch's postulates, a conidial suspension (106 conidia per ml) of the fungus was sprayed on leaves of 20-day-old lentil plants (landrace Castelluccio) that were maintained in a humidity chamber for 96 h after inoculation. Lesions resembling symptoms that occurred in the field were observed on plants 3 weeks after inoculation. Symptoms were not observed on control plants sprayed with water. The fungus reisolated from the diseased plants was identical to the original isolates. Based on morphological characteristics of pycnidia and conidia as well as pathogenicity, the fungus was identified as A. lentis. A deep-freeze blotter method (2) was used to detect A. lentis in lentil seeds of 20 local landraces used by Umbrian farmers and two accessions from Canada and Turkey, as well as in seed collected from infected fields. The fungus was present only in the two lentil accessions with an incidence of about 5%. Although the fungus had been isolated from Italian seed germplasm in 1986 (1), this is the first report of ascochyta blight occurring in lentil crops in Italy. The heavy rainfalls that characterize the first stage of lentil cultivation in Umbria are favorable for disease development while hot and dry conditions that usually occur during flowering and maturation prevent the dissemination of inoculum and the infection of the seeds. For these reasons, some Umbrian areas could be more suitable for production of ascochyta-free lentil seeds. References: (1) W. J. Kaiser and R. M. Hannan. Phytopathology 76:355, 1986. (2) T. Limonard. Proc. Int. Seed Test. Assoc. 33:343, 1968.

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First Report of Botrytis Leaf Blight on Eleutherococcus senticosus Caused by Botrytis cinerea in China

Plant Disease ◽

10.1094/pdis-01-14-0031-pdn ◽

2014 ◽

Vol 98 (9) ◽

pp. 1270-1270 ◽

Cited By ~ 4

Author(s):

X. Wang ◽

B. H. Lu ◽

Y. Zhi ◽

L. N. Yang ◽

J. Gao

Keyword(s):

Botrytis Cinerea ◽

Morphological Characteristics ◽

Leaf Tissue ◽

Leaf Blight ◽

Jilin Province ◽

Perennial Herb ◽

Conidial Suspension ◽

Acanthopanax Senticosus ◽

Eleutherococcus Senticosus ◽

First Report

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.

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First Report of Botrytis Leaf Blight and Fruit Rot on Schisandra chinensis Caused by Botrytis cinerea in China

Plant Disease ◽

10.1094/pdis-02-11-0086 ◽

2011 ◽

Vol 95 (6) ◽

pp. 769-769

Author(s):

R. H. Yu ◽

J. Gao ◽

J. Wang ◽

X. Wang

Keyword(s):

Botrytis Cinerea ◽

Morphological Characteristics ◽

Gray Mold ◽

Leaf Blight ◽

Jilin Province ◽

Fruit Rot ◽

Conidial Suspension ◽

Schisandra Chinensis ◽

Mycelium Growth ◽

First Report

Schisandra (Schisandra chinensis (Turcz.) Baill) is a perennial plant belonging to Magnoliaceae. It is a very important medicinal herb in China and is mainly used for treatment of insomnia and memory decay. From July to September 2008, an unknown leaf blight and fruit rot on schisandra were first observed at Jingyu County, Jilin Province. The same symptoms were detected in other areas of Jilin Province, such as Ji'an City, Baishan City, and Hunchun City. Initially, some small, brown spots appeared on the tip or margin of the leaves. Light brown or brown necrotic lesions developed and eventually covered entire leaves. Seriously affected leaves were rolled or distorted and eventually became completely dry and brittle. Small spots appeared on the surface of mature fruits, coalesced, and the fruits finally dropped. Gray mycelia and conidiophores developed on the diseased leaves and fruits. To isolate the causal agent, conidia and conidiophores were scraped aseptically from the internal tissues, suspended in sterile water, and streaked onto the surface of potato dextrose agar (PDA). Single-hyphal tips were transferred on PDA and the isolated fungus was identified as Botrytis cinerea Pers.: Fr. on the basis of its morphological characteristics and internal transcribed spacer (ITS) sequence. Colonies of B. cinerea on PDA were colorless at first and became gray to brown 20 days later with the mycelium growth and conidia producing in cultures. Conidia are single celled, lemon shaped, colorless to a light color, and 4.4 to 15.0 × 7.0 to 10.0 μm. Sclerotia formed about 1 week later, were black-brown and varied in size (2.0 to 5.0 × 2.0 to 4.0 mm) and shape. The ITS region of rDNA was amplified from DNA extracted from single-spore isolate BC12 of B. cinerea using primers ITS4/ITS5 and sequenced (GenBank Accession No. GU724512), BLAST analysis (1) of the 535-bp segment showed 99% similarity with the sequence of Botryotinia fuckeliana (perfect stage of B. cinerea). Pathogenicity tests were carried out on healthy schisandra plants that were 4 years old. After the surface of the leaves and fruits was disinfected with 5% sodium hypochlorite, a conidial suspension of 105 conidia/ml was sprayed on 10 schisandra leaves, and plugs of the fungus obtained from the colony margins were transferred onto a 3- × 3-mm wound on the surface of disinfected fruit. Ten control schisandra leaves and 10 fruits were inoculated at the same time. Plants were covered with polyethylene bags and incubated at 25°C in a greenhouse with relative humidity of 85% for 3 days. Similar symptoms to those observed on diseased leaves and fruits in the field were observed on inoculated schisandra leaves and fruits 7 days after inoculation, whereas control leaves and fruits showed no symptoms. The pathogen was successfully reisolated. The gray mold disease caused by B. cinerea was reported in many plants, such as Lavandula stoechas and Chamelaucium uncinatum in Italy (2,3). However, to our knowledge, this is the first report of gray mold disease of schisandra caused by B. cinerea in China. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) A. Garibaldi et al. Plant Dis. 94:968, 2009. (3) A. Garibaldi et al. Plant Dis. 94:380, 2010.

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First Report of Paramyrothecium roridum causing leaf blight on water hyacinth (Eichhornia crassipes) in Malaysia

Plant Disease ◽

10.1094/pdis-04-21-0704-pdn ◽

2021 ◽

Author(s):

Iznee Rehanna Hassan ◽

Dzolkhifli Omar ◽

Samsudin Amit ◽

Siti Izera Ismail

Keyword(s):

Water Hyacinth ◽

Eichhornia Crassipes ◽

Morphological Characteristics ◽

Leaf Blight ◽

Post Inoculation ◽

Conidial Suspension ◽

Dark Cycle ◽

Leaf Quality ◽

First Report ◽

Wide Range

Water hyacinth (Eichhornia crassipes) is a free-floating aquatic plant and is also widely cultivated as an aquatic ornamental plant in Malaysia. In June 2018, a severe foliar disease with typical leaf blight symptoms were observed on leaves of water hyacinth plants (approximately 50%) in waterways adjacent to two rice fields located at Tanjung Karang and Sungai Besar, Selangor province, Malaysia. Symptoms appeared irregular necrotic lesions with concentric rings, later lesions expanded to entire leaves and became blighted. Twenty symptomatic leaves were collected from two sampling locations. Symptomatic leaf tissue was cut into small pieces (5 × 5 mm), surface sterilized with 0.5% sodium hypochlorite (NaOCl) for 2 min, rinsed three times with sterile distilled water, plated on potato dextrose agar (PDA), and incubated at 25 °C with a 12-h light/dark cycle for 7 days. Twenty single-spore isolates were recovered from sampled leaves, all isolates exhibited Paramyrothecium-like morphology and two representative isolates, PR1 and PR2 were used for further studies. Fungal colonies were initially white aerial mycelia with sporodochia bearing olivaceous green conidial masses formed on PDA after 5 days of incubation. Conidiogenous cells were phialidic, hyaline, smooth, straight to slightly curved, 13 to 20 × 1.0 to 1.8 μm and setae were absent. Conidia were aseptate, hyaline to pale green, smooth, cylindrical to ellipsoidal with rounded ends, and measured 5.8 to 8.0 μm × 1.8 to 2.2 μm (n=50). These morphological characteristics were consistent with the description of Paramyrothecium roridum (Tode) L. Lombard & Crous (Lombard et al. 2016). Total genomic DNA of the isolates was extracted from fresh mycelium using DNeasy Plant Mini kit (Qiagen, USA). The internal transcribed spacer (ITS) and calmodulin (cmdA) gene regions were amplified using the ITS5/ITS4 (White et al.1990) and CAL-228F/CAL2Rd primer sets (Carbone and Kohn 1999; Groenewald et al., 2013), respectively. BLASTn analysis showed that the ITS and cmdA sequences of the isolates were 100% identity with Paramyrothecium roridum ex-epitype strain CBS 357.89 (GenBank accession nos. KU846300 and KU846270), respectively. The resulting sequences were deposited in GenBank (ITS: Accession nos. MW850370, MW850371; cmdA Accession nos. MW854363, MW854364). Pathogenicity tests of the two isolates were performed by spray inoculation on healthy leaves of each five potted water hyacinth plants using a 3-ml conidial suspension (1 × 106 conidia/ml) produced on 7-day-old PDA cultures incubated at 25 °C with a 12-h light/dark cycle. Five potted water hyacinth plants inoculated with sterile water served as controls. Inoculated plants were covered with plastic bags for 48 h to maintain high humidity and kept in a growth chamber for 2 weeks at 25 ± 1°C, 95% relative humidity and a 12-h light/dark period. The experiment was repeated twice. Eight days post-inoculation, symptoms on inoculated leaves developed necrotic brown lesions similar to those observed in the field, while control leaves remained asymptomatic. After 2 weeks of inoculation, lesions enlarged into severe blighting until all leaves died. Paramyrothecium roridum was re-isolated from randomly selected symptomatic tissues and verified by morphology and sequencing of ITS (MZ675387, MZ706462) and cmdA (MZ686706, MZ712041) loci, confirming Koch’s postulates. The fungus was not re-isolated from non-inoculated control plants. Pa. roridum is distributed on a wide range of plants (Farr and Rossman 2021) and has been reported to cause leaf spot of water hyacinth in Nigeria (Okunowo et al. 2013) and Sri Lanka (Adikaram and Yakandawala 2020). To our knowledge, this is the first report of Pa. roridum causing leaf blight of water hyacinth in Malaysia. This disease is an emerging threat to water hyacinth and it reduces the leaf quality, therefore, appropriate management should be developed to control this disease.

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