Soilborne pathogens are a significant economic problem for nursery production in the Southeastern United States. The goal of this study was to determine the impact of cover crops on soilborne disease suppressiveness in such systems. Soils from red maple (Acer rubrum L.) plantation fields grown with and without cover crops were sampled, either while the cover crops were growing (pre-disked) or post-season, following cover crop incorporation into the soil (post-disked). Greenhouse bioassays were conducted using red maple seeds on inoculated (with Rhizoctonia solani (J.G. Kühn) or Phytophthora nicotianae (Breda de Haan)) and non-inoculated field soils. The damping-off, root rot disease severity, percent recovery of Rhizoctonia and Phytophthora, and pseudomonad population were examined during the two years of the experiment. Results showed that cover crop incorporation was beneficial for inducing disease supressiveness characteristics of soil. Cover crop incorporation into the soil significantly or numerically reduced disease severity and pathogen recovery in infested soil compared to the bare soil treatment. Cover crop incorporation was found to be partially associated with the reduction of seedling damping-off. The pseudomonad microbial population was greater when cover crop was present, and is thought to be antagonist to soilborne pathogens. Therefore, cover crops can be integrated in field nursery production systems to suppress soilborne pathogens.
Diseases caused by soilborne pathogens are a major limitation to field grown nursery production. The application of cover crops for soilborne disease management has not been widely investigated in a woody ornamental nursery production system. The objective of this study was to explore the impact of winter cover crops usage on soilborne disease management in that system. Soils from established field plots of red maple (Acer rubrum L.) with and without winter cover crops (crimson clover (Trifolium incarnatum L.) or triticale (× Triticosecale W.)) were sampled following the senescence of the cover crops. Separate bioassays were performed using red maple cuttings on inoculated (with Phytopythium vexans, Phytophthora nicotianae or Rhizoctonia solani) and non-inoculated field soils. The results indicated that winter cover crop usage was helpful for inducing soil disease suppressiveness. There was lower disease severity and pathogen recovery when the cover crops were used compare to the non-cover cropped soil. However, there were no differences in maple plant fresh weight and root weight between the treatments. The rhizosphere pseudomonad microbial population was also greater when the cover crops were used. Similarly, the C:N ratio of the soil was improved with the cover crop usage. Thus, in addition to improving soil structure and reducing erosion, cover crops can provide improved management of soilborne diseases. Therefore, stakeholders can consider cover crop usage as an alternative sustainable management tool against soilborne diseases in field nursery production system.
Soilborne diseases caused by pathogens such as Phytophthora, Rhizoctonia, Fusarium, Verticillium, and Pythium species are the most important diseases of woody ornamentals. Ginkgo (Ginkgo biloba) and red maple (Acer rubrum) ‘October Glory’ plants grown in containers and fields in Tennessee have shown root and crown rot symptoms with dark brown to black lesions in 2017 and 2018. The objective of this research was to isolate and identify pathogens affecting ginkgo and red maple plants in nurseries of Tennessee and develop fungicide/biofungicide management recommendations for nursery producers. Isolations were made from the infected roots. Several Phytophthora-like colonies with spherical zoospores, filamentous to globose oogoni, and whitish mycelium, were isolated on V8-PARPH medium. For confirming identity, total genomic DNA was extracted followed by the sequence analysis of the internal transcribed spacer (ITS) regions, and large subunit (LSU) of the nuclear ribosomal RNA (rRNA) as well as cytochrome c oxidase subunit I (Cox I) and cytochrome c oxidase subunit II (Cox II) of mitochondrial DNA (mtDNA). Based on morphological and molecular analysis, Phytopythium vexans was described as a causal agent of crown and root rot from the infected ginkgo and red maple plants. To complete Koch’s postulates, a pathogenicity test was performed by drenching 100 ml V8 agar medium slurry of P. vexans inoculum on 1-year-old potted ginkgo plant root systems as well as red maple ‘October Glory’. Necrotic lesion development was observed in the root system 45 days after inoculation and P. vexans was re-isolated from the roots of both ginkgo and red maple. All control ginkgo and red maple plants remained disease-free and no pathogen was re-isolated. In addition, the efficacy of fungicides, biofungicides, fertilizer and host-plant defense inducers (traditionally recommended for management of oomycete diseases) for control of Phytopythium crown and root rot was evaluated on ginkgo and red maple ‘October Glory’ seedlings in greenhouse and field trials. The fungicides such as Empress Intrinsic, Pageant Intrinsic, Segovis and Subdue MAXX were effective in both greenhouse and field trials, and the biofungicide Stargus reduced the disease severity caused by pathogen P. vexans on ginkgo and red maple plants in greenhouse trials. These results will help nursery producers to make proper management decisions for newly reported Phytopythium crown and root rot disease of ginkgo and red maple plants.
Flowering cherry (Prunus serrulata Lindl. 'Kwanzan') rooted cuttings grown in propagation beds containing 40% coarse sand and 60% ground pine bark in a commercial propagation nursery in Warren County, Tennessee were exhibiting root and crown rot in December 2016. Dark brown to black soft lesions were observed in the roots as well as the crown region of flowering cherry rooted cuttings and those rooted cuttings were non-marketable due to lesions. Disease incidence was approximately 60% of 10,000 plants. Phytophthora ImmunoStrip test (Agdia Inc., Elkhart, IN, USA) was performed and the test result was positive. Diseased plant tissues were surface sterilized with 70% ethanol and washed twice with distilled water. Culturing the affected root and crown parts (1 cm pieces) on V8-PARPH, an oomycete-selective medium consistently yielded whitish radiate mycelial growth pattern with spherical zoospores, filamentous to globose oogoni, elongated, and cylindrical antheridia with constrictions (De Cock et al., 2015) after 7 days of incubation at 25°C in a 12-h fluorescent light and dark cycle, which is the typical morphology of Phytopythium vexans (de Bary) Abad, de Cock, Bala, Robideau, Lodhi & Lévesque. To confirm pathogen identity, total DNA was extracted using the UltraClean Microbial DNA Isolation Kit (MO BIO Laboratories, Inc., Carlsbad, CA, USA) directly from a 3-day old culture of isolate (FBG2017010) on V8 medium. The internal transcribed spacer (ITS) and 28S large subunit of ribosomal RNA, and cytochrome c oxidase subunit I (CoxI) of mitochondrial DNA (mtDNA) genes/ region were amplified by PCR using the primer pairs ITS1/ ITS4 (White et al., 1990), NL1/ NL4 (Baten et al., 2014), and Levup and Fm85mod (Robideau et al., 2011), respectively. The PCR products were sequenced and the sequences (GenBank accession nos. MT533275, MT533451, and MT547980) were compared to the voucher specimens. They were 99.23, 99.60, and 98.92% similar to those of P. vexans isolates in the NCBI database (HQ643400, KR092144, and HQ708996, respectively). To complete Koch’s postulates, 'Kwanzan' flowering cherry rooted cuttings grown on propagation substrate (10 cm pot containing 1 kg sterilized 40% coarse sand and 60% ground pine bark) were inoculated with identified pathogen and observations were taken on root rot disease symptoms. Five plants were inoculated with 100 ml of pathogen agar-slurry (1 plate of a 7-day old culture of isolate FBG2017010/1 L of sterilized water), and five control plants were drenched with agar slurry. The plants were maintained in the greenhouse condition (day/night temperature of 26/24°C), and irrigated twice a day for 2 min by overhead irrigation system. After 2 weeks, dark brown to black necrotic root lesions developed on all inoculated cuttings and P. vexans was consistently re-isolated from the inoculated plants. The morphology of the pathogen isolated on the V8-PARPH medium was identical to the original isolate. All control plants remained symptom-free and P. vexans was not isolated from the root tissue. To our knowledge, this is the first report of P. vexans causing root and crown rot in 'Kwanzan' flowering cherry in Tennessee, which can be a potential threat for the nursery crop production. The identification of P. vexans, the causal agent of Phytopythium root and crown rot is important in determination and implementation of effective management strategies.
Eastern ninebark (Physocarpus opulifolius (L.) Maxim.) is a popular native perennial plant used in landscapes because of its colorful foliage and spring flower display. Powdery mildew symptoms were observed on container-grown eastern ninebark ‘Mindia’ Coppertina® plants in a commercial nursery in DeKalb County, TN in May 2016. The disease severity was nearly 40% and the disease incidence was nearly 60% of 1,000 plants. Affected plants displayed witches’-brooms with cream to white colored, thickened shoots with stunted, curly leaves as well as patches of white powdery fungal growth on the surface of young and old leaves, inflorescences, infructescences and stems (Figures 1 and 2). Microscopic observation revealed masses of conidia and mycelium covering symptomatic tissues. Conidiophore foot cells measured 19.2 to 66.7 μm (mean = 38.3 μm) × 5.4 to 15.1 μm (mean = 9.7 μm) (n = 30). Conidia were ovoid and measured 11.4 to 28.5 μm (mean = 20.9 μm) (n = 30) in length and 8.2 to 14.8 μm (mean = 11.7 μm) (n = 30) in width. Conidiophores produced two to six conidia in chains. Fibrosin bodies were observed after treating conidia with a 3% KOH solution. Chasmothecia were numerous, 60.0 to 85.0 μm (mean = 74.2 μm) (n = 30) in size and contained one ascus [60.0 to 82.0 × 52.0 to 69.0 μm; mean = 73.4 × 59.4 μm (n = 30)] with 8 ascospores [25.2 to 28.0 × 14.8 to 16.0 μm; mean = 26.5 × 15.5 μm (n = 30)]. To confirm pathogen identity, total DNA was extracted directly from plant tissue with the UltraClean Microbial DNA Isolation Kit (MO BIO Laboratories, Inc., Carlsbad, CA) following the manufacturer's instructions. The ITS region of the ribosomal DNA was amplified by PCR using primer pair ITS1 and ITS4 (White et al. 1990). The sequence (GenBank acc. no. MT605142) of the amplicon had 100% coverage and 100% identity to that of Podosphaera physocarpi (U. Braun) U. Braun (= Podosphaera aphanis var. physocarpi (U. Braun) U. Braun & S. Takam.) (GenBank acc. no. MT106654). Pathogenicity was confirmed three times by inoculating leaf surfaces of five eastern ninebark ‘Mindia’ Coppertina® plants by tapping fungal spores from infected eastern ninebark leaves onto the surfaces of healthy leaves. Inoculated plants were maintained in a greenhouse (21 to 23°C) using drip irrigation system until symptoms developed. Five non-inoculated control plants were maintained in the same greenhouse. After two weeks, typical symptoms of powdery mildew developed on the inoculated plants and microscopic examination revealed the same pathogen morphology as the original isolate. All non-inoculated control plants remained disease-free. To our knowledge, this is the first report of powdery mildew caused by P. physocarpi on P. opulifolius in Tennessee. Powdery mildew is known to be a disease problem on eastern ninebark grown in its native range in landscape plantings. Lubell et al. (2011) reported varying levels of powdery mildew resistance among eastern ninebark cultivars. Timely application of fungicides with no phytotoxic effect will be necessary to manage this disease on susceptible eastern ninebark cultivars in affected nurseries.
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