First Report of Meloidogyne javanica on Globe Artichoke in Florida, USA

Globe artichoke (Cynara cardunculus var. scolymus L.) is native to the Mediterranean region and cultivated worldwide for its edible flower buds and the medicinal value of its leaves (Pignone and Sonnante 2004). In 2019, artichokes were planted on 29 km2 predominantly in California, with a yield of over 100 million kg (USDA 2020). It has been grown as a specialty crop in Florida since 2017 (Agehara 2017a). Meloidogyne spp. (root-knot nematodes/RKNs) can lead to yield losses to artichoke (Greco et al. 2005). In June 2020, artichokes (cv. Imperial Star) with stunting, wilting, and galled-root symptoms were observed in a research field with sandy soil located at the University of Florida Gulf Coast Research and Education Center (UF/GCREC), Wimauma, Florida. The goal of this report was to identify the RKN species collected from two symptomatic artichoke roots. Morphological measurements (mean, standard deviation and range) of 15 second-stage juveniles (J2s) included body length = 409.1 ± 31.6 (360.3 - 471.3) µm, body width = 15.4 ± 1.6 (12.4 - 18.8) µm, and stylet length = 14.7 ± 0.7 (13.9 -16.1) µm. Perineal patterns of five matured females had a high dorsal arch and double lateral lines. Morphological characteristics of the RKN cultures were consistent with the description of M. javanica (Eisenback and Triantaphyllou 1991). DNA was extracted respectively from two RKN females isolated from the diseased artichoke roots. The nematode species was confirmed with primers Fjav/Rjav and resulted in ≈ 670 bp fragment (Zijlstra et al. 2000). The COXII region of mtDNA was amplified by C2F3/1108 (Powers and Harris 1993), and the sequencing results were submitted to the NCBI with GeneBank Accession No. MZ397905. The molecular sequences had 100% identity with M. javanica in COXII (MK033440 and MK033439). The pathogenicity test was conducted in the greenhouse at UF/GCREC from May to August 2021 (temperature = 26.7 ± 4.1°C, relative humidity = 83.9 ± 14.6 %). Each of the ten 6.5-in-diameter plastic pots containing 3.8-L pasteurized soil was seeded with one artichoke seed. Five pots were inoculated with 5000 eggs of the field RKN cultures 4-week after planting, and five pots served as the untreated control. Two months after inoculation, galled symptoms were only observed in inoculated plants with an average gall index (Bridge and Page 1980) of 6.2 ± 2.2; 99,240 ± 72,250 eggs were extracted from each root system, and the nematode reproduction factor was 19.9 ± 14.4. Meloidogyne spp. has been reported on artichoke in Europe, Asia, and South America (Greco et al. 2005). This is the first report of RKN on artichoke in the United States. Meloidogyne javanica caused severe root gall symptoms and visible aboveground damage in the form of chlorosis, stunting, and wilting of artichoke planted at the UF/GCREC research farm. Meloidogyne javanica is the predominant RKN species at the UF/GCREC research farm and one of the most common RKNs in Florida (Gu and Desaeger 2021). Artichoke is a new crop in Florida, and RKNs is likely to be one of the main soilborne problems for its production in the state. Its long growing season (October - May) (Agehara 2017b) allows for high nematode reproduction rates. Several new growers have already reported RKN as a problem in their fields. For artichoke to become a commodity in Florida, managing RKNs will be critical. This report provides new information on the risk that RKNs pose to artichoke, a newly established specialty crop in Florida.

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First Report of Stem and Foliage Blight of Chrysanthemum Caused by Phytophthora drechsleri in the United States

Plant Disease ◽

10.1094/pdis-03-21-0631-pdn ◽

2021 ◽

Author(s):

Charles Krasnow ◽

Nancy Rechcigl ◽

Jennifer Olson ◽

Linus Schmitz ◽

Steven N. Jeffers

Keyword(s):

United States ◽

South Carolina ◽

Sequence Similarity ◽

Morphological Characteristics ◽

The United States ◽

Universal Primers ◽

Broad Host Range ◽

Pathogenicity Test ◽

First Report ◽

Foliage Blight

Chrysanthemum (Chrysanthemum × morifolium) plants exhibiting stem and foliage blight were observed in a commercial nursery in eastern Oklahoma in June 2019. Disease symptoms were observed on ~10% of plants during a period of frequent rain and high temperatures (26-36°C). Dark brown lesions girdled the stems of symptomatic plants and leaves were wilted and necrotic. The crown and roots were asymptomatic and not discolored. A species of Phytophthora was consistently isolated from the stems of diseased plants on selective V8 agar (Lamour and Hausbeck 2000). The Phytophthora sp. produced ellipsoid to obpyriform sporangia that were non-papillate and persistent on V8 agar plugs submerged in distilled water for 8 h. Sporangia formed on long sporangiophores and measured 50.5 (45-60) × 29.8 (25-35) µm. Oospores and chlamydospores were not formed by individual isolates. Mycelium growth was present at 35°C. Isolates were tentatively identified as P. drechsleri using morphological characteristics and growth at 35°C (Erwin and Ribeiro 1996). DNA was extracted from mycelium of four isolates, and the internal transcribed spacer (ITS) region was amplified using universal primers ITS 4 and ITS 6. The PCR product was sequenced and a BLASTn search showed 100% sequence similarity to P. drechsleri (GenBank Accession Nos. KJ755118 and GU111625), a common species of Phytophthora that has been observed on ornamental and vegetable crops in the U.S. (Erwin and Ribeiro 1996). The gene sequences for each isolate were deposited in GenBank (accession Nos. MW315961, MW315962, MW315963, and MW315964). These four isolates were paired with known A1 and A2 isolates on super clarified V8 agar (Jeffers 2015), and all four were mating type A1. They also were sensitive to the fungicide mefenoxam at 100 ppm (Olson et al. 2013). To confirm pathogenicity, 4-week-old ‘Brandi Burgundy’ chrysanthemum plants were grown in 10-cm pots containing a peat potting medium. Plants (n = 7) were atomized with 1 ml of zoospore suspension containing 5 × 103 zoospores of each isolate. Control plants received sterile water. Plants were maintained at 100% RH for 24 h and then placed in a protected shade-structure where temperatures ranged from 19-32°C. All plants displayed symptoms of stem and foliage blight in 2-3 days. Symptoms that developed on infected plants were similar to those observed in the nursery. Several inoculated plants died, but stem blight, dieback, and foliar wilt were primarily observed. Disease severity averaged 50-60% on inoculated plants 15 days after inoculation. Control plants did not develop symptoms. The pathogen was consistently isolated from stems of symptomatic plants and verified as P. drechsleri based on morphology. The pathogenicity test was repeated with similar results. P. drechsleri has a broad host range (Erwin and Ribeiro 1996; Farr et al. 2021), including green beans (Phaseolus vulgaris), which are susceptible to seedling blight and pod rot in eastern Oklahoma. Previously, P. drechsleri has been reported on chrysanthemums in Argentina (Frezzi 1950), Pennsylvania (Molnar et al. 2020), and South Carolina (Camacho 2009). Chrysanthemums are widely grown in nurseries in the Midwest and other regions of the USA for local and national markets. This is the first report of P. drechsleri causing stem and foliage blight on chrysanthemum species in the United States. Identifying sources of primary inoculum may be necessary to limit economic loss from P. drechsleri.

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First Report of Leaf Blight on Fan Columbine (Aquilegia flabellata) Caused by Rhizoctonia solani AG 4 in Italy

Plant Disease ◽

10.1094/pdis-93-4-0433a ◽

2009 ◽

Vol 93 (4) ◽

pp. 433-433 ◽

Cited By ~ 2

Author(s):

A. Garibaldi ◽

G. Gilardi ◽

D. Bertetti ◽

M. L. Gullino

Keyword(s):

United States ◽

Rhizoctonia Solani ◽

Aqueous Suspension ◽

Morphological Characteristics ◽

The United States ◽

Leaf Blight ◽

Herbaceous Plant ◽

Pathogenicity Test ◽

First Report ◽

Leaf Petiole

Aquilegia flabellata (Ranunculaceae), fan columbine, is a perennial herbaceous plant with brilliant blue-purple flowers with white petal tips. It can also be grown for cut flower production. In April of 2008, in several nurseries located near Biella (northern Italy), a leaf blight was observed on 10 to 15% of potted 30-day-old plants grown on a sphagnum peat substrate at 15 to 20°C and relative humidity of 80 to 90%. Semicircular, water-soaked lesions developed on leaves just above the soil line at the leaf-petiole junction and later along the leaf margins. Lesions expanded over several days along the midvein until the entire leaf was destroyed. Blighted leaves turned brown, withered, and abscised. Severely infected plants died. Diseased tissue was disinfested for 10 s in 1% NaOCl, rinsed with sterile water, and plated on potato dextrose agar (PDA) amended with 25 mg/liter streptomycin sulfate. A fungus with the morphological characteristics of Rhizoctonia solani was consistently recovered, then transferred and maintained in pure culture. Ten-day-old mycelium grown on PDA at 22 ± 1°C appeared light brown, rather compact, and had radial growth. Sclerotia were not present. Isolates obtained from affected plants successfully anastomosed with tester isolate AG 4 (AG 4 RT 31, obtained from tobacco plants). Results were consistent with other reports on anastomosis reactions (2). Pairings were also made with tester isolates of AG 1, 2.1, 2.2, 3, 6, 7, 11, and BI with no anastomoses observed between the recovered and tester isolates. The internal transcribed spacer (ITS) region of rDNA was amplified using primers ITS4/ITS6 and sequenced. BLASTn analysis (1) of the 648-bp fragment showed a 100% homology with the sequence of R. solani AG-4 AB000018. The nucleotide sequence has been assigned GenBank Accession No. FJ 534555. For pathogenicity tests, the inoculum of R. solani was prepared by growing the pathogen on PDA for 10 days. Five plants of 30-day-old A. flabellata were grown in 3-liter pots. Inoculum consisting of an aqueous suspension of PDA and mycelium disks (5 g of mycelium + agar per plant) was placed at the collar of plants. Five plants inoculated with water and PDA fragments alone served as control treatments. Plants were maintained in a greenhouse at temperatures between 20 and 24°C. The first symptoms, similar to those observed in the nursery, developed 7 days after the artificial inoculation. R. solani was consistently reisolated from infected leaves and stems. Control plants remained healthy. The pathogenicity test was carried out twice with similar results. The presence of R. solani AG1-IB on A. flabellata has been reported in Japan (4), while in the United States, Rhizoctonia sp. is described on Aquilegia sp. (3). This is, to our knowledge, the first report of leaf blight of A. flabellata caused by R. solani in Italy as well as in Europe. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) D. E. Carling. Grouping in Rhizoctonia solani by hyphal anastomosis reactions. In: Rhizoctonia Species: Taxonomy, Molecular Biology, Ecology, Pathology and Disease Control. Kluwer Academic Publishers, The Netherlands, 1996. (3) D. F. Farr et al. Fungi on Plants and Products in the United States. The American Phytopathological Society, St Paul, MN, 1989. (4) E. Imaizumi et al. J. Gen. Plant Pathol. 66:210, 2000.

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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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First Report of Leaf Spot of Lettuce (Lactuca sativa L.) Caused by Phoma tropica in Italy

Plant Disease ◽

10.1094/pdis-04-12-0394-pdn ◽

2012 ◽

Vol 96 (9) ◽

pp. 1380-1380 ◽

Cited By ~ 4

Author(s):

A. Garibaldi ◽

G. Gilardi ◽

G. Ortu ◽

M. L. Gullino

Keyword(s):

Relative Humidity ◽

Lactuca Sativa ◽

Leaf Spot ◽

Morphological Characteristics ◽

The United States ◽

Fluorescent Light ◽

Malt Extract ◽

Pathogenicity Test ◽

First Report ◽

Lactuca Sativa L

Lettuce (Lactuca sativa L.) is widely grown in Italy, with the production for the preparation of ready-to-eat salads becoming increasingly important. During the spring of 2011, a previously unknown leaf spot was observed on L. sativa plants, cv Rubia, grown in several plastic tunnels in Lumbardy (northern Italy), 20 to 25 days after sowing. Thirty to forty per cent of leaves of the plants growing in the part of the tunnel with the highest relative humidity were affected. Leaves of infected plants showed extensive, irregular, dark brown, necrotic lesions with a chlorotic halo. Lesions initially ranged from 0.5 to 3 mm, then eventually coalesced, reaching 2 to 3 cm, showing a well-defined, dark brown border. Affected leaves senesced and withered. The crown was not affected by the disease. Diseased tissue was excised, immersed in a solution containing 1% sodium hypochlorite for 60 s, rinsed in water, then cultured on potato dextrose agar (PDA), amended with 25 mg/l of streptomycin sulphate. After 5 days, a fungus developed, producing a greenish grey mycelium with a white border when incubated under 12 h/day of fluorescent light at 21 to 23°C. In order to favor the production of conidia, the fungus was transferred on malt extract agar (MA) and maintained under 12 h/day of fluorescent light at 22°C. After 15 days, black pycnidia, 175 to 225 μm, developed, with hyaline, elliptical, unicellular conidia, measuring 3.21 to 6.7 × 1.08 to 3.2 (average 5.5 × 1.9) μm. On the basis of these morphological characteristics, the fungal causal agent of the disease could be related to the genus Phoma (2). The internal transcribed spacer (ITS) region of rDNA of the isolate PHT30 was amplified using the primers ITS1/ITS4 and sequenced. BLAST analysis (1) of the 466-bp segment showed a 99% similarity with the sequence of Phoma tropica (GenBank Accession No. JF923820.1). The nucleotide sequence has been assigned the GenBank Accession No. JQ954396. Pathogenicity tests were performed by spraying healthy 20-day-old lettuce plants, cv Rubia, with a spore suspension (1 × 105 conidia/ml) prepared from 14-day-old colonies of the strain PHT30 grown on MA cultures. Plants inoculated with water alone served as controls. Ten plants per isolate were used. Plants were covered with plastic bags for 5 days after inoculation and maintained in a growth chamber at 20°C and 80% relative humidity. The first foliar lesions, similar to those occurring on the naturally infected plants, developed on leaves 12 days after inoculation. Control plants remained healthy. The pathogen was consistently reisolated from leaf lesions. The pathogenicity test was completed twice. To our knowledge, this is the first report of the presence of P. tropica on lettuce in Italy as well as worldwide. In the United States, the presence of P. exigua was reported in 2006 (3). The economic importance of the disease at present is limited, probably also because symptoms can be confused with those caused by Botrytis cinerea. However, P. tropica could become a more significant problem because of the importance of the crop. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) G. H. Boerema. Trans. Br. Mycol. Soc. 67:289, 1976. (3) S. Y. Koike. Plant Dis. 90:1268, 2006.

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First Report of Leaf Spot of Fan Columbine (Aquilegia flabellata) Caused by Phoma aquilegiicola in Italy

Plant Disease ◽

10.1094/pdis-05-10-0391 ◽

2011 ◽

Vol 95 (7) ◽

pp. 880-880

Author(s):

A. Garibaldi ◽

D. Bertetti ◽

M. T. Amatulli ◽

M. L. Gullino

Keyword(s):

United States ◽

Morphological Characteristics ◽

Its Region ◽

Apical Part ◽

The United States ◽

Herbaceous Plant ◽

Pathogenicity Test ◽

First Report ◽

Perennial Herbaceous Plant ◽

Olive Green

Aquilegia flabellata (Ranunculaceae), fan columbine, is a perennial herbaceous plant with brilliant blue-purple flowers with white petal tips that is largely present in gardens. It can also be grown for cut flower production. In September of 2008 and 2009, in a private garden located near Biella (northern Italy), a leaf blight was observed. Leaves of infected plants showed extensive, irregular, brown, necrotic lesions, which were slightly sunken with a well-defined border and surrounded by a violet-brown halo. A hole frequently appeared in the center of dried tissues. Lesions, initially measuring 0.5 mm, later expanded up to 15 mm in diameter and eventually coalesced to cover the entire leaf, which curled without falling. At a later stage, stems were also affected, causing death of the apical part of the plant. The disease affected 90% of the plants in the garden. Dark brown, subglobose pycnidia, 116 to 145 μm, containing light gray, ellipsoid, nonseptate conidia measuring 9.0 to 16.2 × 2.6 to 4.2 (average 12.7 × 3.4) μm were observed on symptomatic tissue. On the basis of these morphological characteristics, the fungus was related to the genus Phoma (2). Diseased tissue was excised from the margin of lesions, rinsed in sterile distilled water, and then cultured on potato dextrose agar (PDA) medium at 23 ± 1°C under alternating daylight and darkness (12-h light and 12-h dark). Fungal colonies produced a pale olive green, lightly floccose mycelium, generating clusters of dark olive green swollen cells. The internal transcribed spacer (ITS) region of rDNA was amplified using the primers ITS4/ITS6 and sequenced. BLAST analysis (1) of the 504-bp segment showed 100% homology with a sequence of Phoma aquilegiicola (GenBank Accession No. GU237735). The nucleotide sequence of our isolate was assigned GenBank Accession No. HM222537. Pathogenicity tests were performed by spraying a mycelium suspension of a homogenate of mycelium (1 × 105 mycelial fragments per ml) obtained from 15-day-old PDA cultures of the fungus on leaves of six healthy 6-month-old potted A. flabellata plants. Six plants inoculated with a homogenate of PDA served as controls. Plants were maintained in a greenhouse in a high humidity chamber for 7 days after inoculation at 23 ± 1°C and under high relative humidity conditions (70 to 90%). The first foliar lesions developed on leaves 4 days after inoculation. After 15 days, 80% of the leaves were severely infected. Control plants remained healthy. The organism reisolated on PDA from leaf lesions was identical in morphology to the isolate used for inoculation. The pathogenicity test was carried out twice. To our knowledge, this is the first report of the presence of P. aquilegiicola on A. flabellata in Italy. Ascochyta aquilegiae (synonym P. aquilegiicola) has been reported on A. vulgaris in Germany (4) and Aquilegia spp. in the United States (3). Currently, the economic importance of this disease is limited, but may become a more significant problem if the use of A. flabellata in gardens increases. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) G. H. Boerema et al. Phoma Identification Manual. Differentiation of Specific and Infra-Specific Taxa in Culture. CABI Publishing, Wallingford, UK, 2004. (3) D. F. Farr et al. Fungi on Plants and Plant Products in the United States. The American Phytopathological Society, St. Paul, MN, 1989. (4) R. Laubert. Gartenwelt 34:621, 1930.

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The first report of Meloidogyne enterolobii on Thai basil in Florida, United States

Plant Disease ◽

10.1094/pdis-02-21-0293-pdn ◽

2021 ◽

Author(s):

Mengyi Gu ◽

Hung Xuan Bui ◽

Weimin Ye ◽

Johan Desaeger

Keyword(s):

Morphological Characteristics ◽

Outreach Program ◽

Vegetable Production ◽

Pathogenicity Test ◽

Limited Information ◽

Root Knot Nematode ◽

Nematode Reproduction ◽

Southeastern Us ◽

Florida Gulf Coast ◽

Vegetable Farm

Thai basil (Ocimum basilicum var. thyrsiflora) is an important ethnic aromatic herb native to Southeast Asia. According to the Vegetable Production Handbook of Florida 2020-2021, Asian vegetables are currently grown on more than 4,000 ha in Florida, and Thai basil is one of the most commonly grown among these. Meloidogyne spp. cause severe damage to different basil cultivars (Brito et al. 2007). During May-July 2020, plant stunting and galled root symptoms were observed on Thai basil plants sampled from a commercial Asian vegetable farm in Wimauma, Florida (27°44.951' N; 82°16.271’ E); 1,972 root-knot nematode second-stage juveniles (J2s) were extracted from 200 cm3 soil. A pathogenicity test was performed in September 2020 at the University of Florida Gulf Coast Research and Education Center, Wimauma, Florida. Ten of 20, three-week-old nematode-free Thai basil plants were inoculated with 5,000 eggs of field nematode cultures. Two months after inoculation (temperature = 22.8 ± 3.8 °C, relative humidity = 85.6 ± 14.0 %), average gall index (Bridge and Page 1980) = 5.4 ± 1.1 were only observed in inoculated plants, and 69,276 ± 18,904 eggs were extracted from roots using the NaClO method (Hussey and Barker 1973); 5 ± 7 J2s / 200 cc soil were recovered by the modified Baermann funnel technique (Forge and Kimpinski 2007). Nematode reproduction factor (RF) = 13.86 ± 3.78 (Nicol et al. 2010). Morphological measurements (mean, standard deviation and range) of J2s (n=20) included body length = 394.0 ± 22.3 (362.8 - 437.9) µm, body width = 15.7 ± 1.2 (13.6 - 18.3) µm, and stylet length = 12.8 ± 1.1 (10.4-14.5) µm. The perineal pattern of matured female (n=5) was oval-shaped with coarse and smooth striate; the dorsal arch was high and round; no lateral line presented. Morphological characteristics of females and J2s were consistent with those described for M. enterolobii (Yang and Eisenback 1983). DNA was extracted from a single female picked from infected Thai basil root using NaOH digestion method (Hübschen et al. 2004). The D2-D3 expansion segment of 28S rDNA and the COXII region on mitochondrial DNA were amplified by PCR using the primers 28S391a/28S501 and C2F3/1108 (Ye et al. 2020); the species was also confirmed with species-specific primers Me-F/Me-R (Ye et al. 2020). PCR products were sequenced by the Genomic Sciences Laboratory (North Carolina State University, Raleigh, NC, USA) and the results were recorded in the NCBI with GeneBank Accession Nos. MW488150 and MW507374. The sequences showed 100% identity with M. enterolobii in D2/D3 (KP901079, KP411230) and COXII (MN809527, KX214350). M. enterolobii (M. mayaguensis) has been reported on sweet basil in Florida (Brito et al. 2008). To our knowledge, this is the first detection of M. enterolobii on Thai basil in Hillsborough County, Florida. It is not clear to what extent M. enterolobii reduces the yield of Thai basil, but the RF value obtained in the pathogenicity test indicates the crop is certainly a very good host. Limited information is available on the distribution of M. enterolobii in Florida and the US. M. enterolobii is known to break down the root-knot resistance of crops including soybean, sweet potatoes, and tomatoes (Philbrick et al. 2020). This nematode is considered one of the major emerging threats to agriculture in the southeastern US. A multistate research and outreach program (FINDMe program) was initiated in 2019 to study the distribution and management of this nematode in the southeastern US.

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First Report of Downy Mildew Caused by Peronospora parasitica on Iberis sempervirens in Italy

Plant Disease ◽

10.1094/pd-90-1261c ◽

2006 ◽

Vol 90 (9) ◽

pp. 1261-1261

Author(s):

A. Garibaldi ◽

A. Minuto ◽

M. L. Gullino

Keyword(s):

New York ◽

Downy Mildew ◽

Morphological Characteristics ◽

Sprinkler Irrigation ◽

The United States ◽

Plant Diseases ◽

Pathogenicity Test ◽

Peronospora Parasitica ◽

First Report ◽

Downy Mildews

Iberis sempervirens (evergreen candytuft) is a garden species belonging to the Brassicaceae family. During June 2004, a damaging foliar disease was observed in several commercial farms near Albenga (northern Italy) on I. sempervirens plants grown outdoors in containers. More than 30% of the plants were affected. Symptoms appeared on both sides of leaves, buds, flowers, and fruits. Initially, leaves were slightly chlorotic, but within 5 to 7 days a characteristic whitish furry growth developed on the lower and upper leaf surfaces. The efflorescence was particularly evident on the lower surfaces of leaves and consisted of sporangiophores and sporangia. The appearance and severity of the disease increased because of overhead sprinkler irrigation. Microscopic observations revealed dichotomously branched sporangiophores with slender curved tips. Sporangiophores with a length of 115 to 410 μm (average 295 μm) ended with sterigmata bearing single sporangia. Sporangia were ovoid and measured 18 to 28 × 25 to 45 μm (average 22 × 35 μm). The pathogen was identified as Peronospora parasitica on the basis of its morphological characteristics (3). Pathogenicity was confirmed by inoculating leaves of 10 45-day-old healthy plants grown in 14-cm-diameter pots with a sporangial suspension (1 × 103 conidia/ml). Ten noninoculated plants served as controls. Plants were maintained outdoors at 50% light intensity with temperatures ranging between 16 and 25°C (average 18°C) and 85 to 100% relative humidity. The pathogenicity test was carried out twice. After 18 days, typical symptoms of downy mildew developed on the inoculated plants and P. parasitica was observed on the leaves. Noninoculated plants did not show symptoms. To our knowledge, this is the first report of P. parasitica on evergreen candytuft in Italy. P. parasitica was previously reported on I. sempervirens in the United Kingdom (1) and on I. amara in California (2). Voucher specimens are available at the AGROINNOVA Collection, University of Torino, Italy. References: (1) S. Francis and G. Waterhouse. Trans. Br. Mycol. Soc. 91:1, 1988. (2) P. R. Muller et al. Index of Plant Diseases in the United States. USDA Handbook No. 165, 1960. (3) D. M. Spencer. The Downy Mildews. Academic Press, New York, 1981.

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First Report of a Leaf Spot Caused by Alternaria compacta on Hydrangea anomala subsp. petiolaris in Italy

Plant Disease ◽

10.1094/pdis-92-1-0173c ◽

2008 ◽

Vol 92 (1) ◽

pp. 173-173 ◽

Cited By ~ 2

Author(s):

A. Garibaldi ◽

D. Bertetti ◽

M. L. Gullino

Keyword(s):

Morphological Characteristics ◽

The United States ◽

Plant Diseases ◽

Spore Suspension ◽

Pathogenicity Test ◽

Aesthetic Quality ◽

First Report ◽

Plastic Bags ◽

Alternaria Sp ◽

The Aesthetic

Hydrangea anomala subsp. petiolaris (synonym H. petiolaris and H. scandens), also known as climbing hydrangea, is cultivated as an ornamental for landscaping in parks and gardens. This species, belonging to the Hydrangeaceae and native to the woodlands of Japan and coastal China, is widely appreciated for its abundant, creamy white flowers with a sweet aroma, particularly in shade gardens. During the summer of 2006, extensive necroses were observed on leaves and young stems of 3-year-old plants grown outdoors in several gardens of Piedmont of northern Italy. In many cases, on the upper side of the leaves, necrotic spots (4 to 10 mm in diameter) turned progressively black. Lesions often coalesced, generating larger (2 to 6 cm in diameter) necrotic areas. Necroses initially developed mainly at leaf margins and near petioles, and severely affected plants were defoliated. Infected plants rarely died, but the presence of lesions reduced the aesthetic quality and subsequently the commercial value. The disease occurred on 50 of 100 plants. A fungus was consistently isolated from infected leaves on potato dextrose agar (PDA) and identified on the basis of its morphological characteristics as an Alternaria sp. Conidia were dark gray, multicellular, clavate to pear shaped, measuring 23 to 54 × 10 to 13 μm (average 38 × 12 μm), with five longitudinal crosswalls and a relatively short apical beak. DNA was extracted with a Nucleospin Plant Kit (Macherey Nagel, Brockville, ON, Canada) and PCR was carried out with ITS 6/ITS 4 primer (2). A 557-bp PCR product was sequenced, and a BLASTn search (1) confirmed that the sequence corresponded to Alternaria compacta (99% homology). The nucleotide sequence has been assigned GenBank Accession No. EU 128529. Pathogenicity tests were performed by spraying leaves of healthy 1-year-old potted H. anomala plants with an aqueous 105 CFU/ml spore suspension. The inoculum was obtained from cultures of the fungus grown on sterilized host leaves placed on PDA for 20 days in light/dark at 23 ± 1°C. Plants sprayed only with water served as controls. Five plants were used for each treatment. Plants were covered with plastic bags for 3 days after inoculation and maintained between 12 and 22°C. Lesions developed on leaves 8 days after inoculation with the spore suspension, whereas control plants remained healthy. A. compacta was consistently reisolated from these lesions. The pathogenicity test was repeated twice. The presence of an Alternaria sp. on H. macrophylla was reported in the United States (3), whereas A. hortensiae was observed in Spain on H. hortensis. Recently, A. alternata belonging to the alternata group was reported on H. macrophylla in Italy (4). This is, to our knowledge, the first report of A. compacta on H anomala subsp. petiolaris in Italy. References: (1) S. F. Altschud et al. Nucleic Acids Res. 25:3389, 1997. (2) D. E. L. Cooke and J. M. Duncan. Mycol. Res. 101:667, 1997. (3) M. L. Daughtrey et al. Page 9 in: Compendium of Flowering Potted Plant Diseases. American Phytopathological Society. St. Paul, MN, 1995. (4) A. Garibaldi et al. Plant Dis. 91:767, 2007.

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First Report of Leaf Spot of Milky Bellflower (Campanula lactiflora) Caused by a Phoma sp. in Italy

Plant Disease ◽

10.1094/pdis-94-5-0638a ◽

2010 ◽

Vol 94 (5) ◽

pp. 638-638

Author(s):

A. Garibaldi ◽

D. Bertetti ◽

C. Pellegrino ◽

M. L. Gullino

Keyword(s):

United States ◽

Leaf Spot ◽

Morphological Characteristics ◽

Its Region ◽

The United States ◽

Fluorescent Light ◽

Herbaceous Plant ◽

Pathogenicity Test ◽

First Report ◽

Perennial Herbaceous Plant

Campanula lactiflora (milky bellflower), a perennial herbaceous plant in the Campanulaceae, is used in park and gardens and sometimes cultivated for cut flower production. In June 2008, a previously unknown leaf spot was observed on C. lactiflora ‘New Hybrids’ plants from an experimental nursery located near Carmagnola (Torino, northern Italy). Leaves of infected plants showed extensive and irregular, dark brown, necrotic lesions that were slightly sunken with well-defined borders. Lesions initially ranged from 0.5 to 3 mm, eventually coalesced, and covered the entire leaf. Black pycnidia (107 to 116 μm in diameter) containing hyaline, ellipsoid, nonseptate conidia measuring 3.7 to 4.7 × 1.2 to 2.0 (average 4.3 × 1.6) μm were observed. On the basis of these morphological characteristics, the fungal causal agent of the disease could be related to the genus Phoma. In some cases, the basal leaves turned completely necrotic and the plant died. The disease affected 50% of plants. Diseased tissue was excised, immersed in a solution containing 1% sodium hypochlorite for 2 to 3 s, rinsed in water, and then cultured on potato dextrose agar (PDA) medium. A fungus developed that produced a greenish gray mycelium with a white border when incubated under 12 h/day of fluorescent light at 22 to 25°C. The internal transcribed spacer (ITS) region of rDNA was amplified using the primers ITS4/ITS6 and sequenced. BLAST analysis (1) of the 459-bp segment showed a 100% similarity with the sequence of a Didymella sp. (synonym Mycosphaerella), anamorphic stage of Phoma spp. The nucleotide sequence has been assigned GenBank Accession No. GU128503. Pathogenicity tests were performed by placing 8-mm-diameter mycelial disks removed from PDA cultures of the fungus isolated from infected plants on leaves of healthy potted 4-month-old C. lactiflora ‘New Hybrids’ plants. Eight disks were placed on each plant. Plants inoculated with PDA alone served as controls. Six plants per treatment were used. Plants were covered with plastic bags for 4 days after inoculation and maintained in a growth chamber with daily average temperatures ranging between 23 and 24°C. The first foliar lesions developed on leaves 5 days after inoculation, and after 8 days, 80% of leaves were severely infected. Control plants remained healthy. A Didymella sp. was consistently reisolated from leaf lesions. The pathogenicity test was completed twice. To our knowledge, this is the first report of the presence of a Didymella sp. on C. lactiflora in Italy. Mycosphaerella campanulae and M. minor were reported on C. americana and C. lasiocarpa in the United States (2). The economic importance of the disease currently is limited, but could become a more significant problem in the future if the cultivation of this species becomes more widespread. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) D. F. Farr et al. Fungi on Plants and Plant Products in the United States. The American Phytopathological Society, St. Paul, MN, 1989.

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First Report of Rust Caused by Pucciniastrum circaeae on Fuchsia × hybrida in Italy

Plant Disease ◽

10.1094/pdis-11-11-0976-pdn ◽

2012 ◽

Vol 96 (4) ◽

pp. 588-588 ◽

Cited By ~ 1

Author(s):

A. Garibaldi ◽

G. Gilardi ◽

G. Ortu ◽

M. L. Gullino

Keyword(s):

Phylogenetic Trees ◽

Aqueous Suspension ◽

Morphological Characteristics ◽

Pcr Amplification ◽

Signs And Symptoms ◽

The United States ◽

Pathogenicity Test ◽

First Report ◽

Potted Plants ◽

Plastic Bags

Fuchsia is a genus of flowering plants that is native to South America and New Zealand and belongs to the family Onagraceae. In September 2011, 2-year-old potted plants of Fuchsia × hybrida, cv. Citation, in a garden located near Biella (northern Italy) showed signs and symptoms of a previously unknown disease. Typically, infected plants showed leaf chlorosis followed by the appearance of necrosis on the adaxial leaf surfaces, while the abaxial surfaces showed orange uredinia irregularly distributed. As the disease progressed, infected leaves turned yellow and wilted. Affected plants showed a progressive phylloptosis and also flowering was negatively affected. Urediniospores were globose, yellow to orange, and measured 14.6 to 25.9 (average 19.6) μm. Teliospores were not observed. Morphological characteristics of the fungus corresponded to those of the genus Pucciniastrum. DNA extraction and PCR amplification were carried out with Terra PCR Direct Polymerase Mix (Clontech, Saint Germain-en-Laye, France) and primers ITS1/ITS4 (4). A 700-bp PCR product was sequenced and a BLASTn search (1) confirmed that the sequence corresponded with a 96% identity to Pucciniastrum circaeae. The nucleotide sequence has been assigned the GenBank Accession No. JQ029688. Pathogenicity tests were performed by spraying leaves of healthy 1-year-old potted Fuchsia × hybrida plants with an aqueous suspension of 1 × 103 urediniospores ml–1. The inoculum was obtained from infected leaves. Plants sprayed only with water served as controls. Three plants were used for each treatment. Plants were covered with plastic bags for 4 days after inoculation and maintained outdoors at temperatures ranging between 18 and 25°C. Lesions developed on leaves 20 days after inoculation with the urediniospore suspension, showing the same symptoms as the original plants, whereas control plants remained healthy. The organism that was recovered from the lesions after inoculation was the same as the one obtained from the diseased plants. The pathogenicity test was carried out twice with similar results. The presence of P. fuchsiae, later identified as P. epilobii, was repeatedly reported in the United States (3). P. epilobii and P. circaeae have closely related hosts and morphologically similar urediniospores. These species were reported to form a single group in molecular phylogenetic trees (2). This is, to our knowledge, the first report of P. circaeae on Fuchsia × hybrida in Italy. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997 (2) Y. M. Liang et al. Mycoscience 47:137, 2006. (3) L. B. Loring and L. F. Roth. Plant Dis. Rep. 48:99, 1964. (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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