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|a 10.1094/PDIS-06-23-1115-PDN
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|a eng
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|a Benitez, Brianna
|e verfasserin
|4 aut
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|a First report of a Herbaspirillum sp. causing leaf spots on Boston Fern (Nephrolepis exaltata) in Florida
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|c 2023
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|a Text
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|a ƒaComputermedien
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|a ƒa Online-Ressource
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|a Date Revised 04.08.2023
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|a published: Print-Electronic
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|a Citation Status Publisher
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|a Boston fern (Nephrolepis exaltata) samples were submitted by a nursery operation in Florida separately to the University of Florida Plant Diagnostic Center (UFPDC, Gainesville, FL) and to the North Carolina State University Plant and Pest Diagnostic Lab (NCSU PPDL, Raleigh, NC) in October 2021. Symptoms included tan spots on pinnules, some of which progressed into pinnule blight (Fig. S1). Bacterial streaming was noted from samples in both labs. Leaf spot margins were excised, macerated in sterile tap water, then streaked onto nutrient agar (NA) plates and incubated for 48 h at 27°C. Individual representative colonies that were opaque, creamy white, mucoid, and round with smooth margins were transferred and streaked onto additional NA plates. One strain from each lab (G21-1742, UFPDC and NC40101, NCSU PPDL) was selected for subsequent characterization. A suspension of each strain was adjusted to 108 CFU/mL and infiltrated into tobacco and tomato leaves, and confluent necrosis was observed 72 h after infiltration. The isolates were Gram-negative, oxidase-positive, HR-positive on tomato and tobacco, aerobic, not pectolytic, and nonfluorescent on King's Medium B. DNA was extracted from G21-1742 using Qiagen Stool kit (Qiagen cat#51604) and the 16S rRNA gene from strain G21-1742 was amplified using 16SrRNA universal primers UP1 (5'-TACGTGCCAGCAGCCGCGGTAATA-3') and UP2 (5'-AGTAAGGAGGGTATCCAACCGCA-3') (Kuppusamy et al. 2014). The amplicon was sequenced and submitted to NCBI (Genbank Accession No. OR004801). BLASTn analysis of 16S rRNA of G21-1742 resulted in 99.7% sequence identity to the type strain of Herbaspirillum huttiense subsp. huttiense ATCC 14670T (Genbank Accession NR_024698). The 16S rRNA sequence of NC40101 was identical to that of G21-1742. To determine if the G21-1742 strain was pathogenic, Boston fern plants were inoculated by suspending bacterial cells in tap water from a 24h culture grown on NA, adjusting the suspension to 108 CFU/mL and spraying the suspension on one three-week old frond from each of three healthy Boston fern plants. A second frond from each plant was sprayed with sterile tap water. Each treated frond was individually sealed in a clear plastic bag for 24h at approximately 25°C. Inoculated plants remained on the greenhouse bench after the plastic bags were removed. The inoculation experiment was repeated once. After 4 days, tan spots were observed on pinnules of inoculated plants that were identical to the original submitted samples, while no symptoms developed on water-inoculated plants. Bacterial strains were reisolated from symptomatic plants and were morphologically identical to G21-1742. The 16S rRNA sequence of the reisolated strain was identical to G21-1742. Additionally, we conducted MLSA analysis using 12 housekeeping genes (See Table S2 for housekeeping genes and accession numbers) from the fern strains and the corresponding housekeeping genes for the type strains of 13 Herbaspirillum species, which placed the fern strains most closely with H. huttiense (see Fig. S2). This is the first known report of a Herbaspirillum sp. on Boston fern, an important ornamental crop, that renders the plants aesthetically unsaleable. Previously, a Herbaspirillum sp. was reported in Florida to cause a leaf spot and blight on greenhouse grown tomato seedlings (Obradovic et al. 2007)
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|a Journal Article
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|a Causal Agent
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|a Crop Type
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|a Ornamentals
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|a Pathogen detection
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|a Prokaryotes
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|a Subject Areas
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|a herbaceous/flowering plants
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|a Poudel, Mousami
|e verfasserin
|4 aut
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|a Jones, Jeffrey B
|e verfasserin
|4 aut
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| 700 |
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|a Harmon, Carrie Lapaire
|e verfasserin
|4 aut
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