First Report of Fusarium Wilt in Blueberry (Vaccinium corymbosum) Caused by Fusarium oxysporum in China

Blueberry (Vaccinium corymbosum) production is developing quickly in China with about 20,000 ha presently cultivated. In 2010 in Lin'an, Zhejiang Province, plants developed an apparently new disease of blueberry (cv. Duke) with symptoms consisting of wilting of foliage, stunting of plants, and reduced fruit yields. Internal vascular and cortical tissues of plant crowns showed a brown to orange discoloration. Approximately 3% of the plants in the commercial plantings were affected and eventually died after 50 to 60 days. Infected plant samples (stems and roots) collected from different fields were surface sterilized with 1.5% sodium hypochlorite for 2 min, rinsed in water, plated on 2% potato dextrose agar (PDA), and incubated at 25°C in the dark for 1 week. Single conidium cultures were consistently isolated and cultured on acidified PDA (APDA) for morphological characterization (1,2). Colonies were light with purple mycelia, and beige or orange reverse colony colors developed after 7 days incubation at 25°C. Colonies producing abundant microconidia and macroconidia. Microconidia were hyaline and oval-ellipsoid to cylindrical (3.9 to 9.6 × 1.1 to 3.4 μm). Macroconidia were 3 to 5 septate and fusoid-subulate with a pedicellate base (28.6 to 37.5 × 3.3 to 4.2 μm). Morphology and development of macroconidia and microconida were consistent with a description of Fusarium oxysporum Schltdl (1,2). The ribosomal internal transcribed spacers ITS1 and ITS2 of eight isolates were amplified using primers ITS1/ITS4 on DNA extracted from mycelium and nucleotide sequences showed 100% similarity to that of F. oxysporum. To confirm pathogenicity, 20 blueberry plants (cv. Duke) were inoculated by dipping the roots into a conidial suspension (107 conidia per ml) for 30 min. The inoculated plants were transplanted into pots containing sterilized peat and maintained at 25°C and 100% relative humidity in a growth chamber with a daily 12-h photoperiod of fluorescent light. The pathogenicity test was conducted twice. Within 40 days, all inoculated plants developed wilt symptoms similar to that observed in the field. No symptoms were observed on plants dipped into distilled water. The fungus was successfully re-isolated from crowns and roots cultured on APDA, exhibiting morphological characteristics identical to F. oxysporum (1,2), confirming Koch's postulates. To our knowledge, this is the first report of blueberry wilt caused by Fusarium. References: (1) P. M. Kirk et al. The Dictionary of the Fungi, 10th edition, page 159. CABI Bioscience, Wallingford, UK, 2008. (2) W. C. Snyder and H. N. Hansen. Am. J. Bot. 27:64, 1940.

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First Report of Fusarium Wilt Caused by Fusarium oxysporum on Strawberry in Spain

Plant Disease ◽

10.1094/pdis-93-3-0323b ◽

2009 ◽

Vol 93 (3) ◽

pp. 323-323 ◽

Cited By ~ 22

Author(s):

F. T. Arroyo ◽

Y. Llergo ◽

A. Aguado ◽

F. Romero

Keyword(s):

Fusarium Oxysporum ◽

Fusarium Wilt ◽

Potato Dextrose Agar ◽

Fluorescent Light ◽

Pathogenicity Test ◽

Conidial Suspension ◽

Soilless Culture ◽

Fragaria Ananassa ◽

Pcr Assay ◽

First Report

In the spring of 2007, wilted and dead strawberry plants (Fragaria × ananassa Duch. cvs. Camarosa and Ventana) were observed in a soilless culture system in Huelva, southwestern Spain. Approximately 8% of the plants in the field died. Isolations from necrotic crowns and roots and necrotic flowers were made on potato dextrose agar after disinfestation in 0.6% NaOCl for 30 s. Colonies with light purple mycelia and beige or orange reverse colony colors developed after 9 days of incubation at 25°C. Colonies produced abundant microconidia, macroconidia, and chlamydospores. Microconidia were hyaline and oval-ellipsoid to cylindrical (5.9 to 9.2 × 2.1 to 3.4 μm). Macroconidia were 3 to 5 septate and fusoid-subulate with a pedicellate base (28.8 to 37.3 × 3.2 to 4.3 μm). Morphology and growth matched descriptions of Fusarium oxysporum Schlechtend emend. Snyder & Hansen (2). A PCR assay for amplification of r-DNA using primers PFO2 and PFO3 established the identity of the isolate as F. oxysporum (1). To confirm the pathogenicity of the fungus, roots of 30-day-old strawberry cvs. Camarosa and Ventana (20 plants each) were inoculated by dipping the roots into a conidial suspension (107 conidia per ml) for 15 min. The inoculated plants were transplanted into plastic pots containing sterilized peat and maintained at 25°C and 100% relative humidity in a growth chamber with a daily 12-h photoperiod of fluorescent light. The pathogenicity test was conducted twice. Within 30 days, all inoculated plants developed wilt symptoms similar to that observed in the field and eventually 75% of the plants died. No symptoms were observed on plants dipped in distilled water. The fungus was successfully reisolated from crowns, roots, and necrotic flowers, fulfilling Koch's postulates. To our knowledge, this is the first report of the occurrence of Fusarium wilt caused by F. oxysporum on strawberry plants in Spain. References: (1) V. Edel et al. Mycol. Res. 104:518, 2000. (2) W. C. Snyder and H. N. Hansen. Am. J. Bot. 27:64, 1940.

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First Report of Basal Stem Rot of Apple Cactus (Cereus peruvianus monstruosus) Caused by Fusarium oxysporum in Italy

Plant Disease ◽

10.1094/pdis-03-11-0216 ◽

2011 ◽

Vol 95 (7) ◽

pp. 877-877

Author(s):

A. Garibaldi ◽

P. Pensa ◽

D. Bertetti ◽

A. Poli ◽

M. L. Gullino

Keyword(s):

United States ◽

Fusarium Oxysporum ◽

The United States ◽

Stem Rot ◽

Fluorescent Light ◽

Pathogenicity Test ◽

Conidial Suspension ◽

Single Spore ◽

Basal Stem Rot ◽

First Report

During the summer of 2010, 20% of 7,000 4-month-old plants of apple cactus (Cereus peruvianus monstruosus) showed symptoms of a basal stem rot in a commercial nursery located in Liguria (northern Italy). Affected plants showed yellow orange-to-pale brown color from the crown level to the stem apex and a water-soaked rot was observed on the stem starting from the base. Brown discoloration was observed in the vascular system. Eventually stems bent, plants collapsed and died, and affected tissues dried out. A Fusarium sp. was consistently and readily isolated from symptomatic tissue on Komada selective medium. Isolates were purified and subcultured on potato dextrose agar (PDA). Single-spore cultures on PDA, Spezieller Nährstoffarmer agar (SNA) (3), and carnation leaf-piece agar (CLA) (2) were incubated at 26 ± 1°C (12-h fluorescent light, 12-h dark). On PDA, cultures produced a thick growth of white-to-pink mycelium and pale pink pigments in the agar. On SNA, cultures produced short monophialides with unicellular, ovoid-elliptical microconidia measuring 4.3 to 8.2 × 2.3 to 3.8 (average 6.0 × 2.8) μm. Chlamydospores were abundant, single or paired, terminal and intercalary, rough walled, and 6 to 8 μm in diameter. On CLA, cultures produced orange sporodochia with macroconidia that were 3 to 4 septate, nearly straight with a foot-shaped basal cell and a short apical cell, and measured 31.1 to 51.5 × 4.4 to 3.5 (average 43.2 × 3.8) μm. Such characteristics are typical of Fusarium oxysporum (3). Amplification of the ITS (internal transcribed spacer) of the rDNA using primers ITS1/ITS4 (4) yielded a 498-bp band. Sequencing and BLASTn analysis of this band showed an E-value of 0.0 with F. oxysporum. The nucleotide sequence has been assigned GenBank Accession No. JF422071. To confirm pathogenicity, five 6-month-old healthy plants of C. peruvianus monstruosus were inoculated by dipping roots in a conidial suspension (2.4 × 106 CFU/ml) of F. oxysporum isolated from affected plants. Inoculum was obtained from pure cultures of three single-spore isolates grown for 10 days on casein hydrolysate liquid medium. Roots were not wounded before the inoculation. Plants were transplanted into pots filled with steam-sterilized substrate (sphagnum peat/perlite/pine bark/clay 50:20:20:10). Five noninoculated plants served as a control. Plants were placed in a climatic chamber at 25 ± 1°C (12-h fluorescent light, 12 h-dark). Basal stem rot and vascular discoloration in the crown and stem developed within 30 days on each inoculated plant. Noninoculated plants remained healthy. F. oxysporum was consistently isolated from symptomatic plants. The pathogenicity test was conducted twice. F. oxysporum has been reported on Cereus spp. in the United States (1). To our knowledge, this is the first report of F. oxysporum on C. peruvianus monstruosus in Italy as well as in Europe. Currently, this disease is present in a few nurseries in Liguria. References: (1) D. F. Farr et al. Fungi on Plants and Plant Products in the United States. The American Phytopathological Society, St Paul, MN, 1989. (2) N. L. Fisher et al. Phytopathology 72:151, 1982. (3) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell, Ames, IA, 2006. (4) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, 1990.

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First Report of Anthracnose Caused by Colletotrichum brasiliense on Violet Passion Fruit in China

Plant Disease ◽

10.1094/pdis-11-20-2485-pdn ◽

2021 ◽

Author(s):

G. Y. Shi ◽

Quan Zeng ◽

Y. W. Wei ◽

Chun Jin Hu ◽

X. L. Ye ◽

...

Keyword(s):

Large Scale ◽

Disease Incidence ◽

Morphological Characteristics ◽

Internal Transcribed Spacers ◽

Passion Fruit ◽

Fluorescent Light ◽

Distilled Water ◽

Conidial Suspension ◽

First Report ◽

Initial Symptoms

Violet passion fruit (Passiflora edulis Sims) is an important tropical and subtropical perennial evergreen vine with large-scale cultivation in Guangxi, China. Between May and September 2020, anthracnose symptoms occurred on passion fruit (cultivar Tainong No. 1) in Xingye county (22°77′13″N, 110°07′80″E) in Guangxi province, China. The disease incidence varied from 25 to 60% in different orchards. Initial symptoms on young fruits appeared as multiple tiny water-soaked, oval to irregular pale greenish spots. As the disease progressed, the lesions became medium brown, with sunken cavities. Under humid conditions, acervuli containing masses of conidia and dark setae were found on the lesions. The affected fruits became shriveled. Tissue pieces (5 × 5 mm) were cut out from infected fruits, surface sterilized in 75% ethanol for 15 s and 0.1% HgCl2 for 2 min, washed three times with sterile water, placed onto potato dextrose agar (PDA), and incubated at 28 °C for three days. Of the 29 Colletotrichum isolates obtained , the isolate B13 was selected for morphological characterization. B13 was purified by single spore isolation and incubated on PDA at 25°C under continuous fluorescent light irradiation, producing white to pale yellow colonies with dense aerial mycelia. The reverse side of the colony was pale yellowish to olive. Conidia were hyaline, unicellular, straight, cylindrical, with both ends slightly round or one end round and the other slightly pointed, measuring 10.5 to 18.8 (average 16.4) × 5.4 to 7.2 (average 6.3) µm (n = 50). Appressoria were light brown to dark black, smooth-walled, lobed, often with a roundish outline, sometimes also triangular, 7.2 to 10.9 (average 9.1) × 6.8 to 9.2 (average 8.2) µm (n = 50). Morphological characteristics of the isolate matched those of Colletotrichum brasiliense (Damm et al. 2012). The internal transcribed spacers (ITS), actin (ACT), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and beta-tubulin (TUB2) genes of strain B13 were sequenced using the method and primers of Damm et al. (2012). Sequences of the amplified DNA regions were submitted to GenBank (ITS: MW198820; ACT: MW266083; GAPDH: MW266084; and TUB2: MW266085). A concatenated maximum likelihood phylogenetic tree was built using MEGA 7.0.21 in which B13 clustered with C. brasiliense and clearly separated from other Colletotrichum spp. Pathogenicity of B13 was assayed using one-year-old plants of violet passion fruit cultivar ‘Tainong No. 1’. Conidial suspensions were prepared from 7-day-old cultures grown on PDA at 28°C Sterile distilled water was used to dislodge conidia from the culture dish and the conidial concentration was adjusted to 1 × 106 spores mL-1 using a haemocytometer. Fruits were rinsed with sterilized water and wounded with a sterile needle at three locations. Three fruits were inoculated by spraying with 20 mL of the conidial suspension. Control fruits were sprayed with distilled water. Fruits were then covered with plastic bags to maintain high relative humidity . After 9 days, all inoculated fruits developed brown spots with sunken cavities, resembling symptoms observed in the field, and controls remained symptomless. Fungal cultures with phenotypic features similar to C. brasiliense were re-isolated from the symptomatic fruits, verifying C. brasiliense as the causal agent of the disease based on Koch’s postulates. C. boninense, C. gloeosporioides, C.queenslandicum, C. brevisporum, and C. karstii were reported as causal agents of anthracnose on passion fruit (Júnior et al.2010; Power et al. 2010; James et al.2014; Du et al.2017; Ran et al.2020). To the best of our knowledge, this is the first report of C. brasiliense causing anthracnose on passion fruit in China.

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First Report of Fusarium oxysporum Causing Wilt on Jade Plant (Crassula ovata) in Italy

Plant Disease ◽

10.1094/pdis-12-10-0897 ◽

2011 ◽

Vol 95 (9) ◽

pp. 1191-1191 ◽

Cited By ~ 1

Author(s):

A. Garibaldi ◽

D. Bertetti ◽

P. Pensa ◽

A. Poli ◽

M. L. Gullino

Keyword(s):

Fusarium Oxysporum ◽

Vascular System ◽

Apical Cell ◽

Aerial Mycelium ◽

Selective Medium ◽

Fluorescent Light ◽

Pathogenicity Test ◽

Conidial Suspension ◽

Single Spore ◽

First Report

During summer 2010, symptoms of a wilt disease were observed in a commercial farm in northern Italy on Crassula ovata (jade plant). First symptoms consisted of chlorosis and premature drop of still turgid leaves. As the disease progressed, leaves turned yellow and wilted before dropping off and the stem wilted, bent, and eventually rotted starting from the base. In some cases, the stem broke or the basal portion of the leaf rotted. Brown discolorations were observed in the vascular system. Of 10,000 plants, 65% (cv. Mini) and 5% of 600 plants (cv. Magical Tree) were affected. Premature dropping of leaves was more frequent on cv. Magical Tree. Using the Komada's Fusarium-selective medium, a fungus was consistently and readily isolated from symptomatic vascular tissues of plants belonging to both cultivars. Isolates obtained from both cultivars were purified, subcultured on potato dextrose agar (PDA), and single-spore cultures were obtained. On PDA, both isolates produced pale violet, abundant, aerial mycelium, felted in old cultures, with purple pigments in the agar. The isolates were grown on Spezieller Nährstoffarmer agar for characterization of macroconidia and microconidia (1). Both isolates produced sparse, 3 to 5 septate, nearly straight macroconidia measuring 30 to 56 × 3 to 5 (average 40 × 4) μm with a short apical cell and a foot-shaped basal cell. Sporodochia were not observed. Unicellular, oval-elliptical microconidia measuring 5 to 13 × 3 to 4 (average 8 × 3) μm were produced on short monophialides. Chlamydospores were abundant, single and sometime in pairs, terminal and intercalary, rough walled, and measured 6 to 9 μm. Such characteristics are typical of Fusarium oxysporum (3). The ITS region (internal transcribed spacer) of rDNA was amplified with primers ITS1/ITS4 (4) and sequenced. BLASTn analysis of an isolate from C. ovata cv. Mini (515 bp, Accession No. HQ682196) and C. ovata cv. Magical Tree (509 bp, Accession No. HQ682197) showed an E-value of 0.0 with F. oxysporum. For these sequences, pairwise alignment of EMBOSS (E.B.I. - The European Bioinformatics Institute) revealed identity and similarity of 99.0%. To confirm pathogenicity, tests were conducted on 5-month-old plants of cvs. Mini and Magical Tree. Plants (three per treatment) were inoculated by dipping roots in a 1 × 106 CFU/ml conidial suspension of the two isolates of F. oxysporum prepared from 10-day-old cultures grown on casein liquid medium (2), shaken (90 rpm) for 10 days at 24°C ± 1 (12-h fluorescent light, 12-h dark). Inoculated plants were transplanted into pots filled with steamed mix (sphagnum peat/perlite/pine bark/clay; 50:20:20:10) and maintained in a plant growth chamber at 25 ± 1°C under a regimen of 12 h per day of fluorescent light. Inoculated plants belonging to both cultivars showed typical first symptoms of Fusarium wilt after 13 days. In the following days, leaves dropped, stems wilted, and plants died. Noninoculated plants remained healthy. F. oxysporum was reisolated from inoculated plants. The pathogenicity test was conducted twice. This is, to our knowledge, the first report of F. oxysporum on C. ovata in Italy or worldwide. References: (1) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Professional, Ames, IA, 2006. (2) A. Minuto et al. Phytoparasitica 36:294, 2008. (3) B. A. Summerell et al. Plant Dis. 87:117, 2003. (4) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, 1990.

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First Report of Ramularia coleosporii causing Leaf Spot on Perilla frutescens in Korea

Plant Disease ◽

10.1094/pdis-01-21-0223-pdn ◽

2021 ◽

Author(s):

Md Aktaruzzaman ◽

Tania Afroz ◽

Hyo-Won Choi ◽

Byung Sup Kim

Keyword(s):

Leaf Spot ◽

Ipomoea Batatas ◽

Rust Fungus ◽

Morphological Characteristics ◽

Perilla Frutescens ◽

Herbaceous Plant ◽

Pathogenicity Test ◽

Conidial Suspension ◽

Single Spore ◽

First Report

Perilla (Perilla frutescens var. japonica), a member of the family Labiatae, is an annual herbaceous plant native to Asia. Its fresh leaves are directly consumed and its seeds are used for cooking oil. In July 2018, leaf spots symptoms were observed in an experimental field at Gangneung-Wonju National University, Gangneung, Gangwon province, Korea. Approximately 30% of the perilla plants growing in an area of about 0.1 ha were affected. Small, circular to oval, necrotic spots with yellow borders were scattered across upper leaves. Masses of white spores were observed on the leaf underside. Ten small pieces of tissue were removed from the lesion margins of the lesions, surface disinfected with NaOCl (1% v/v) for 30 s, and then rinsed three times with distilled water for 60 s. The tissue pieces were then placed on potato dextrose agar (PDA) and incubated at 25°C for 7 days. Five single spore isolates were obtained and cultured on PDA. The fungus was slow-growing and produced 30-50 mm diameter, whitish colonies on PDA when incubated at 25ºC for 15 days. Conidia (n= 50) ranged from 5.5 to 21.3 × 3.5 to 5.8 μm, were catenate, in simple or branched chains, ellipsoid-ovoid, fusiform, and old conidia sometimes had 1 to 3 conspicuous hila. Conidiophores (n= 10) were 21.3 to 125.8 × 1.3 to 3.6 μm in size, unbranched, straight or flexuous, and hyaline. The morphological characteristics of five isolates were similar. Morphological characteristics were consistent with those described for Ramularia coleosporii (Braun, 1998). Two representative isolates (PLS 001 & PLS003) were deposited in the Korean Agricultural Culture Collection (KACC48670 & KACC 48671). For molecular identification, a multi-locus sequence analysis was conducted. The internal transcribed spacer (ITS) regions of the rDNA, partial actin (ACT) gene and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene were amplified using primer sets ITS1/4, ACT-512F/ACT-783R and gpd1/gpd2, respectively (Videira et al. 2016). Sequences obtained from each of the three loci for isolate PLS001 and PLS003 were deposited in GenBank with accession numbers MH974744, MW470869 (ITS); MW470867, MW470870 (ACT); and MW470868, MW470871 (GAPDH), respectively. Sequences for all three genes exhibited 100% identity with R. coleosporii, GenBank accession nos. GU214692 (ITS), KX287643 (ACT), and 288200 (GAPDH) for both isolates. A multi-locus phylogenetic tree, constructed by the neighbor-joining method with closely related reference sequences downloaded from the GenBank database and these two isolates demonstrated alignment with R. coleosporii. To confirm pathogenicity, 150 mL of a conidial suspension (2 × 105 spores per mL) was sprayed on five, 45 days old perilla plants. An additional five plants, to serve as controls, were sprayed with sterile water. All plants were placed in a humidity chamber (>90% relative humidity) at 25°C for 48 h after inoculation and then placed in a greenhouse at 22/28°C (night/day). After 15 days leaf spot symptoms, similar to the original symptoms, developed on the leaves of the inoculated plants, whereas the control plants remained symptomless. The pathogenicity test was repeated twice with similar results. A fungus was re-isolated from the leaf lesions on the inoculated plants which exhibited the same morphological characteristics as the original isolates, fulfilling Koch’s postulates. R. coleosporii has been reported as a hyperparasite on the rust fungus Coleosporium plumeriae in India & Thailand and also as a pathogen infecting leaves of Campanula rapunculoides in Armenia, Clematis gouriana in Taiwan, Ipomoea batatas in Puerto Rico, and Perilla frutescens var. acuta in China (Baiswar et al. 2015; Farr and Rossman 2021). To the best of our knowledge, this is the first report of R. coleosporii causing leaf spot on P. frutescens var. japonica in Korea. This disease poses a threat to production and management strategies to minimize leaf spot should be developed.

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First Report on Ovate-leaf Atractylodes Damping-off Caused by Fusarium oxysporum species Complex in South Korea

Plant Disease ◽

10.1094/pdis-05-21-1107-pdn ◽

2021 ◽

Author(s):

Oliul Hassan ◽

Taehyun Chang

Keyword(s):

South Korea ◽

Fusarium Oxysporum ◽

Species Complex ◽

Disease Incidence ◽

Fusarium Species ◽

Morphological Characteristics ◽

Conidial Suspension ◽

Sterile Water ◽

Damping Off ◽

First Report

In South Korea, ovate-leaf atractylodes (OLA) (Atractylodes ovata) is cultivated for herbal medicine. During May to June 2019, a disease with damping off symptoms on OLA seedlings were observed at three farmer fields in Mungyeong, South Korea. Disease incidence was estimated as approximately 20% based on calculating the proportion of symptomatic seedlings in three randomly selected fields. Six randomly selected seedlings (two from each field) showing damping off symptoms were collected. Small pieces (1 cm2) were cut from infected roots, surface-sterilized (1 minute in 0.5% sodium hypochlorite), rinsed twice with sterile water, air-dried and then plated on potato dextrose agar (PDA, Difco, and Becton Dickinson). Hyphal tips were excised and transferred to fresh PDA. Six morphologically similar isolates were obtained from six samples. Seven-day-old colonies, incubated at 25 °C in the dark on PDA, were whitish with light purple mycelia on the upper side and white with light purple at the center on the reverse side. Macroconidia were 3–5 septate, curved, both ends were pointed, and were 19.8–36.62 × 3.3–4.7 µm (n= 30). Microconidia were cylindrical or ellipsoid and 5.5–11.6 × 2.5–3.8 µm (n=30). Chlamydospores were globose and 9.6 –16.3 × 9.4 – 15.0 µm (n=30). The morphological characteristics of present isolates were comparable with that of Fusarium species (Maryani et al. 2019). Genomic DNA was extracted from 4 days old cultures of each isolate of SRRM 4.2, SRRH3, and SRRH5, EF-1α and rpb2 region were amplified using EF792 + EF829, and RPB2-5f2 + RPB2-7cr primer sets, respectively (Carbone and Kohn, 1999; O'Donnell et al. 2010) and sequenced (GenBank accession number: LC569791- LC569793 and LC600806- LC600808). BLAST query against Fusarium loci sampled and multilocus sequence typing database revealed that 99–100% identity to corresponding sequences of the F. oxysporum species complex (strain NRRL 28395 and 26379). Maximum likelihood phylogenetic analysis with MEGA v. 6.0 using the concatenated sequencing data for EF-1α and rpb2 showed that the isolates belonged to F. oxysporum species complex. Each three healthy seedlings with similar sized (big flower sabju) were grown for 20 days in a plastic pot containing autoclaved peat soil was used for pathogenicity tests. Conidial suspensions (106 conidia mL−1) of 20 days old colonies per isolate (two isolates) were prepared in sterile water. Three pots per strain were inoculated either by pouring 50 ml of the conidial suspension or by the same quantity of sterile distilled water as control. After inoculation, all pots were incubated at 25 °C with a 16-hour light/8-hour dark cycle in a growth chamber. This experiment repeated twice. Inoculated seedlings were watered twice a week. Approximately 60% of the inoculated seedlings per strain wilted after 15 days of inoculation and control seedlings remained asymptomatic. Fusarium oxysporum was successfully isolated from infected seedling and identified based on morphology and EF-1α sequences data to confirm Koch’s postulates. Fusarium oxysporum is responsible for damping-off of many plant species, including larch, tomato, melon, bean, banana, cotton, chickpea, and Arabidopsis thaliana (Fourie et al. 2011; Hassan et al.2019). To the best of our knowledge, this is the first report on damping-off of ovate-leaf atractylodes caused by F. oxysporum in South Korea. This finding provides a basis for studying the epidemic and management of the disease.

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First Report of Colletotrichum acutatum in Blueberry Plants in Spain

Plant Disease ◽

10.1094/pdis.2001.85.12.1285a ◽

2001 ◽

Vol 85 (12) ◽

pp. 1285-1285 ◽

Cited By ~ 3

Author(s):

C. Barrau ◽

B. de los Santos ◽

F. Romero

Keyword(s):

Pink Salmon ◽

Morphological Characteristics ◽

Vaccinium Corymbosum ◽

Fungal Species ◽

Colletotrichum Acutatum ◽

Enzyme Linked Immunosorbent Assay ◽

Conidial Suspension ◽

First Report ◽

Small Areas ◽

Leaf Tissues

An anthracnose disease was observed affecting leaves of high-bush blueberry plants (Vaccinium corymbosum L. ‘Sharpblue’) in small areas within two production fields in Huelva Province of Andalucía, in southwestern Spain. The first symptoms observed in late spring were circular, necrotic lesions, red to salmon in color, and ranging from 3 to 20 mm in diameter. Later, lesions became salmon colored in the center with a brilliant red halo. Fungal isolations were made from the lesions. Infected tissues were surface-disinfected in 1% sodium hypochlorite for 1 min, blotted dry on sterile filter paper, and plated on 2% water agar. The plates were incubated at 25°C for 5 to 10 days. Fungal colonies isolated from the tissues were transferred to potato dextrose yeast agar (PDYA). Only one fungal species was consistently isolated from affected leaf tissues and was identified as Colletotrichum acutatum J.H. Simmonds based on morphological characteristics (2) and enzyme-linked immunosorbent assay (1). Colonies of the fungus on PDYA showed a white-to-gray dense mycelium covered with salmon-colored spore masses. The reverse of the plates was a pink-salmon color. Colony diameter on PDYA averaged 50 mm after 7 days at 25°C. Conidia were hyaline, aseptate, fusiform to cylindrical, and 12.5 × 3.2 μm. Inoculation of leaves and fruits of blueberry cv. Misty with a conidial suspension (106 conidia per ml) of C. acutatum produced lesions on the leaves and fruits similar to those observed on diseased plants in the field. The pathogen was isolated from lesions on inoculated plants. To our knowledge, this is the first report of C. acutatum in high-bush blueberry plants in Spain. References: (1) T. A. Cooke et al. EPPO Bull. 25:57, 1995. (2) B. C. Sutton. The Coelomycetes. CMI, Kew, England, 1980.

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First Report of a Pestalotiopsis sp. Causing Leaf Spot of Blueberry in China

Plant Disease ◽

10.1094/pdis-92-1-0171a ◽

2008 ◽

Vol 92 (1) ◽

pp. 171-171 ◽

Cited By ~ 10

Author(s):

Y. S. Luan ◽

Z. T. Shang ◽

Q. Su ◽

L. Feng ◽

L. J. An

Keyword(s):

Leaf Spot ◽

Apical Cell ◽

Vaccinium Corymbosum ◽

Internal Transcribed Spacers ◽

Fluorescent Light ◽

Tubulin Gene ◽

First Report ◽

Plastic Bags ◽

Plant Nursery ◽

Β Tubulin

In August 2006, leaf spots were observed on half-high blueberry (Vaccinium corymbosum) in a plant nursery in Dalian, China. The symptomatic potted 1-year-old blueberry plants were located in parts of a plant nursery with poor ventilation. The primary symptom was a leaf spot, 0.4 to 0.8 cm in diameter, with brown margins that enlarged and coalesced. Mycelium grew from symptomatic and green leaf tissue removed from the margin of a necrotic leaf spot. Plant tissues were surface disinfested with 0.1% mercuric chloride for 3 min and 70% ethyl alcohol for 30 s before plating onto potato dextrose agar. The resulting colonies were white with a regular margin and a rough surface. The cultures were covered with black and globular acervuli with a diameter of 100 to 200 μm. The base of each conidiophore was swollen and globose with phialides growing from the apical end. Mature conidia were straight to fusiform, measuring 19.0 to 27.5 × 6.3 to 9.2 μm, and five-celled with the three middle cells brown and darker than the end cells. The apical cell was triangular and hyaline with three simple setulae that were 17.2 to 29.7 μm long. The base cell terminated in a point 4.0 to 8.6 μm long. Koch's postulates were fulfilled for the fungus by spray inoculating two healthy young plants with 2 × 105 conidia per ml of sterile distilled water. As a control, two similar plants were sprayed with sterile water. Plants were placed inside plastic bags to maintain humidity and incubated in a growth chamber at 26°C under fluorescent light for 14 h and at 20°C in darkness for 10 h. After 3 days, the plastic bags were removed and plants were maintained under the same conditions. More than 20 days after inoculation, symptoms on inoculated plants were similar to those previously described in the nursery. Control plants did not show any symptoms. Cultures isolated from the lesions were similar to those isolated previously from plants in the nursery. The morphological descriptions and measurements were similar to Pestalotiopsis clavispora (1). The 5.8S subunit and flanking internal transcribed spacers (ITS1 and ITS2) of rDNA and partial β-tubulin gene were amplified from DNA extracted from single-spore cultures using the ITS1/ITS4 and T1/Bt2b primers (2) respectively, and sequenced (GenBank Accession Nos. EF119336 and EF152585). The ITS sequences were most similar to the ITS regions of P. clavispora TA-8 (98%; GenBank Accession No. AY924264), P. clavispora TA-6 (98%; GenBank Accession No. AY924263), and P. clavispora PSHI 2002 Endo 389 (96%; GenBank Accession No. AY682929). The partial β-tubulin gene sequence was identical to Pestalotiopsis sp. isolate PSHI 2004 Endo 86 (100%; GenBank Accession No. DQ657901). The morphology and sequence data support the identity of the causal fungus as P. clavispora. To our knowledge, this is the first report on the presence of a Pestalotiopsis sp. causing a disease of blueberry in China. References: (1) E. F. Guba. Monograph of Monochaetia and Pestalotia. Harvard University Press, Cambridge, MA, 1961. (2) W. Tao et al. Mol. Cell Biol. 27:689, 2007.

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First Report of Fusarium Wilt of Lavandula pubescens Caused by Fusarium oxysporum in Saudi Arabia

Plant Disease ◽

10.1094/pdis-94-9-1163b ◽

2010 ◽

Vol 94 (9) ◽

pp. 1163-1163 ◽

Cited By ~ 2

Author(s):

K. Perveen ◽

N. Bokhari

Keyword(s):

Saudi Arabia ◽

Fusarium Oxysporum ◽

Vascular Tissue ◽

Agricultural Research ◽

Indian Agricultural Research Institute ◽

Pathogenicity Test ◽

Conidial Suspension ◽

Potential Threat ◽

King Saud University ◽

First Report

In November 2008, a wilt of lavender (Lavandula pubescens) seedlings was observed in the greenhouse at King Saud University, Riyadh, Saudi Arabia. Affected seedlings were wilted and the root system was poorly developed. Diseased stems developed a dark coloration that extended down to the roots. Vascular tissue of the affected seedlings appeared red or brown. Isolations consistently yielded a fungus growing from the discolored stem tissue when placed on potato dextrose agar. The macroscopic characteristics of the colony, as well as microscopic structures, were used to identify the fungus as Fusarium oxysporum (2). Oval to elliptical microconidia without septa and originating from short phialides were used to distinguish the species from F. solani (1). The fungus was authenticated by the ITCC (Indian Type Collection Centre), Indian Agricultural Research Institute, New Delhi, India, and given I.D. No. 7532.09. For conducting further experiments, healthy seedlings of L. pubescens were obtained from the botanical garden of the King Saud University and grown in steam-sterilized soil. Healthy seedlings of lavender were inoculated using a root-dip method with a conidial suspension (1 × 107 CFU/ml) of one strain of F. oxysporum obtained from infected plants. Inoculated seedlings were then transplanted into steam-sterilized soil. Plants inoculated with sterilized water (1 ml per plant) served as control treatments. Wilt symptoms and vascular discoloration in the roots and crown developed within 20 days on all plants inoculated with the pathogen, while control plants remained asymptomatic. F. oxysporum was consistently reisolated from symptomatic plants. The pathogenicity test was conducted twice. To our knowledge, this is the first report of F. oxysporum on L. pubescens in Saudi Arabia or elsewhere in the world, and this newly identified disease may be a potential threat to commercial production of lavender. References: (1) J. F. Leslie and B. A. Summerell. Page 212 in: The Fusarium Laboratory Manual. Blackwell Publishing Professional, Hoboken, NJ, 2006. (2) P. C. Nelson et al. Clin. Microbiol. Rev. 7:479, 1994.

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First Report of Passalora Leaf Spot Caused by Passalora bougainvilleae on Bougainvillea in Mexico

Plant Disease ◽

10.1094/pdis-11-10-0787 ◽

2011 ◽

Vol 95 (6) ◽

pp. 775-775 ◽

Cited By ~ 1

Author(s):

V. Ayala-Escobar ◽

V. Santiago-Santiago ◽

A. Madariaga-Navarrete ◽

A. Castañeda-Vildozola ◽

C. Nava-Diaz

Keyword(s):

Uv Light ◽

Disease Incidence ◽

Morphological Characteristics ◽

The United States ◽

Pathogenicity Test ◽

Commonwealth Mycological Institute ◽

Conidial Suspension ◽

Single Spore ◽

First Report ◽

Bougainvillea Spectabilis

Bougainvillea (Bougainvillea spectabilis Willd) growing in 28 gardens during 2009 showed 100% disease incidence and 3 to 7% disease severity. Bougainvilleas with white flowers were the most affected. Symptoms consisted of light brown spots with dark brown margins visible on adaxial and abaxial sides of the leaves. Spots were circular, 2 to 7 mm in diameter, often surrounded by a chlorotic halo, and delimited by major leaf veins. Single-spore cultures were incubated at 24°C under near UV light for 7 days to obtain conidia. Pathogenicity was confirmed by spraying a conidial suspension (1 × 104 spores/ml) on leaves of potted bougainvillea plants (white, red, yellow, and purple flowers), incubating the plants in a dew chamber for 48 h and maintaining them in a greenhouse (20 to 24°C). Identical symptoms to those observed at the residential gardens appeared on inoculated plants after 45 to 60 days. The fungus was reisolated from inoculated plants that showed typical symptoms. No symptoms developed on control plants treated with sterile distilled water. The fungus produced distinct stromata that were dark brown, spherical to irregular, and 20 to 24 μm in diameter. Conidiophores were simple, born from the stromata, loose to dense fascicles, brown, straight to curved, not branched, zero to two septate, 14 × 2 μm, with two to four conspicuous and darkened scars. The conidia formed singly, were brown, broad, ellipsoid, obclavate, straight to curved with three to four septa, 40 × 4 μm, and finely verrucous with thick hilum at the end. Fungal DNA from the single-spore cultures was obtained using a commercial DNA Extraction Kit (Qiagen, Valencia, CA); ribosomal DNA was amplified with ITS5 and ITS4 primers and sequenced. The sequence was deposited at the National Center for Biotechnology Information Database (GenBank Accession Nos. HQ231216 and HQ231217). The symptoms (4), morphological characteristics (1,2,4), and pathogenicity test confirm the identity of the fungus as Passalora bougainvilleae (Muntañola) Castañeda & Braun (= Cercosporidium bougainvilleae Muntañola). This pathogen has been reported from Argentina, Brazil, Brunei, China, Cuba, El Salvador, India, Indonesia, Jamaica, Japan, Thailand, the United States, and Venezuela (3). To our knowledge, this is the first report of this disease on B. spectabilis Willd in Mexico. P. bougainvilleae may become an important disease of bougainvillea plants in tropical and subtropical areas of Mexico. References: (1) U. Braun and R. R. Castañeda. Cryptogam. Bot. 2/3:289, 1991. (2) M. B. Ellis. More Dematiaceous Hypomycetes. Commonwealth Mycological Institute, Kew, Surrey, UK, 1976. (3) C. Nakashima et al. Fungal Divers. 26:257, 2007. (4) K. L. Nechet and B. A. Halfeld-Vieira. Acta Amazonica 38:585, 2008.

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