Potential Biological Control Agents for Soilborne Fungal Pathogens in Tennessee Snap Bean Farms

Fungi isolated from snap bean roots and rhizosphere soil where fungicides are not used included Fusarium oxysporum, Fusarium equiseti, Fusarium subglutinans, Fusarium camptoceras, Fusarium chlamydosporum, Fusarium verticillioides, Fusarium proliferatum, Fusarium acuminatum, Fusarium solani, Peyronellaea pinodella, Macrophomina phaseolina, and Glomerella guttata. Only P. pinodella, M. phaseolina, and F. oxysporum were isolated on symptomatic plants. These soilborne fungi are common pathogens of diverse host plants. Pathogenicity tests under controlled environment demonstrated that these fungi were pathogenic on snap beans. Subsequently, bacterial endophytes isolated from snap bean roots, papaya roots and stems, and dogwood stems were evaluated as potential biological control agents against these diverse fungi. All bacteria isolated, including Bacillus vallismortis (PS), Bacillus amyloliquefaciens (Psl), Bacillus subtilis (Prt), Bacillus thuringiensis (Y and IMC8), Enterobacter sp. (E), Stenotrophomonas sp. (B17A), and Serratia sp. (B17B) suppressed growth of the fungal pathogens in vitro and formed clear inhibition zones in petri dish dual cultures. Growth media taken from the inhibition zones suppressed growth of the fungal pathogens in the absence of the bacterial cells, suggesting that the bacteria released unidentified antagonistic biochemical substances into the media. This study constitutes an initial screening of endophytes as biological control agents against diverse fungal pathogens and forms a basis for the discovery of novel strains that can be further developed and integrated into disease management systems for diverse fungal pathogens. Isolates B. vallismortis (PS), B. amyloliquefaciens (Psl), B. subtilis (Prt), and B. thuringiensis (Y IMC8) exhibited the best performance as potential biological control agents paving the way for larger-scale in vivo studies and characterization of their interactions with fungal pathogens

Data on plant defense enzyme activity associated with three endophytes against Cornus florida Erysiphe pulchra powdery mildew

Three bacteria endophytes that colonize flowering dogwood (Cornus florida) suppressed Erysiphe pulchra powdery mildew disease severity. The three bacteria identified as Stenotrophomonas sp. (B17A), Serratia marcescens (B17B), and Bacillus thuringiensis (IMC8) were assessed for plant defense enzymes associated with plant protection. Detached leaves inoculated with powdery mildew were spray treated with the selected bacterial isolates and incubated for 15 h, 26 h, 48 h and 72 h and then analyzed for activation of defense enzymes and Pathogenesis related (PR) proteins associated with induced systemic resistance (ISR) as a potential mode of action against powdery mildew. At each time point post treatment with the bacteria, leaf tissue was ground in liquid nitrogen and stored at -70°C for biochemical assay of enzyme activity. This data set presents the activation of enzyme activity for peroxidase (PO), polyphenol oxidase (PPO) and β -1,3-glucanase at 15 h, 26 h, 48 h and 72 h post treatment with bacteria as indicated by a change in absorbance min -1 mg-1 per gram fresh weight of leaves. The gene expression of the corresponding pathogenesis related (PR) protein for each bacterial treatment compared to the control was also analyzed using Real time PCR and five primers targeting PR1, PR2, and PR5. While changes for PO, PPO, and β -1,3-glucanase enzyme activities were observed at different time points post treatment with all three bacteria, expression of PR protein was detected for PR1, but it was negligible for PR2, and PR5

Analysis of Powdery Mildew-resistant Dogwood Accessions Using AFLP

Twenty-five dogwood accessions (one Cornus kousa, three C. kousa × C. florida hybrids, and 21 C. florida) were characterized using amplified fragment length polymorphism. Among the C. florida accessions, four were named cultivars and 17 were selections from Tennessee State University\u27s dogwood breeding program. Amplified fragment length polymorphism band profiles obtained from 13 EcoRI/MseI (+3/+3) primer pairs showed the presence of high genetic diversity between species and within the C. florida accessions. Each accession was distinctly different from each other, and the resistant clones clustered into distinct groups

Identification of Alternaria alternata as a Causal Agent for Leaf Blight in Syringa Species

While many isolates of Alternaria alternata are common saprophytes on trees and shrubs, this study clearly demonstrated that A. alternata is a primary pathogen in lilac (Syringa sp.), causing a leaf-blight that affects different Syringa species. Isolates of Alternaria sp. were collected from leaf blight samples of lilacs in the field. The internal transcribed spacer (ITS) region and morphological characterization were used to identify lilac blight pathogen. Based on 100% ITS nucleotide sequence identities to the Alternaria genus in the GenBank and morphological features, these isolates were identified as A. alternata. Disease symptoms were reproduced in lilac plants inoculated with A. alternata mycelial plugs and sprayed with a fungus-free culture filtrate, indicating that pathogenesis in lilac involves secondary metabolites or toxins. Diagnostic primers were developed to detect Alternaria sp. and A. alternata in lilac leaf blight based on ITS region and four known genes associated with pathogenesis in A. alternata: mixed-linked glucanase precursor, endopolygalacturonase, hsp70, and histone genes. The results from our study indicated A. alternata is a primary pathogen in lilac leaf blight, and these diagnostic primers can be used as a tool for the fast detection of A. alternata associated with lilac leaf blight

Evaluation of Biological Agents for Control of Macrophomina Root Rot and Powdery Mildew in Flowering Dogwood (Cornus florida L.)

Six biological control agents (BCAs) (two bacteria, two fungi, and two yeasts) that were previously shown to be effective against powdery mildew (Erysiphe pulchra) were tested for efficacy against Macrophomina phaseolina root rot on flowering dogwood (Cornus florida) in the greenhouse. Two of the bacterial isolates, Stenotrophomonas sp. (B17A) and Serratia sp. (B17B), were effective in controlling both macrophomina root rot and powdery mildew, similar to fungicide control thiophanate methyl, when roots were drenched with the six BCAs individually. In addition, the two bacterial BCAs improved plant growth with respect to stem diameter, stem length, dry weight, and green foliage compared with fungicide-treated plants or nontreated controls grown in sterile soil. These results confirm previous results in which B17A and B17B suppressed powdery mildew and also promoted plant growth in flowering dogwood. Although macrophomina root rot has been previously reported as a potential problem in flowering dogwood, especially in field conditions, simultaneous infection with macrophomina root rot and powdery mildew has not been previously reported. This study confirmed that M. phaseolina infection was characterized by stubby roots and black root lesions, and plants infected with both powdery mildew and macrophomina root rot had smaller root mass compared with fungicide-treated plants. Neither of the two pathogens killed their host plants, but compounded infections significantly reduced the plant root system and plant growth. The efficacy of the two bacterial isolates in controlling both powdery mildew and macrophomina root rot suggests their potential utilization in controlling both diseases in dogwood nursery production and in other plants that are hosts to both powdery mildew and macrophomina root rot. Plant growth promoted by the two BCAs may be attributed to powdery mildew and macrophomina root rot control, but comparisons between fungicide-treated plants and control plants not inoculated with BCAs or root rot pathogen suggested that the two BCAs may play a role as bio-stimulants in growth enhancement. These results also suggest that the two biocontrol agents are not phytotoxic to dogwood

Flowering Dogwood Infections with Macrophomina phaseolina

Macrophomina phaseolina was isolated from the crown region and roots of mature flowering dogwood (Cornus florida L.) trees in the landscape and nursery plantings. Although this pathogen has been reported in Cornus species, its occurrence and impact on C. florida has not been reported. Pathogenicity tests were conducted on dogwood seedlings, and all inoculated seedlings developed root necrotic lesions and no small lateral roots, whereas the non-inoculated control seedlings remained disease-free and developed numerous small roots. Seedlings inoculated with M. phaseolina exhibited numerous microsclerotia, but non-inoculated seedlings did not. In greenhouse experiments, plants inoculated on the stems near the soil line developed brown canker-like lesions and swellings around the inoculated area. These were not observed on non-inoculated plants

Sources of Resistance to Common Bacterial Blight and Charcoal Rot Disease for the Production of Mesoamerican Common Beans in the Southern United States

The gene pool of Mesoamerican common beans (Phaseolus vulgaris L.) includes genotypes in the small-to-medium-size seeded dry beans, as well as some snap beans from hotter environments adapted to the Southeastern United States. However, the warm and humid climate of the Southeastern United States is conducive to diseases such as Common Bacterial Blight (CBB) and Charcoal Rot (CR). The pathogens for these two diseases can survive long periods in infested soil or on seeds and are difficult to control through pesticides. Hence, field-level resistance would be the best management strategy for these diseases. The goals of this study were (1) to evaluate field-level resistance from the various commercial classes and subgroups represented in the Mesoamerican gene pool as sources for breeding beans for the region and (2) to evaluate genome-wide marker × trait associations (GWAS) using genetic markers for the genotypes. A total of 300 genotypes from the Mesoamerican Diversity Panel (MDP) were evaluated for CBB and CR in field experiments for three years. CBB resistance was also tested with a field isolate in controlled greenhouse conditions. The analysis of variance revealed the presence of variability in the MDP for the evaluated traits. We also identified adapted common bean genotypes that could be used directly in Southeastern production or that could be good parents in breeding programs for CBB and CR resistance. The GWAS detected 14 significant Single-Nucleotide Polymorphism (SNP) markers associated with CBB resistance distributed on five chromosomes, namely Pv02, Pv04, Pv08, Pv10, and Pv11, but no loci for resistance to CR. A total of 89 candidate genes were identified in close vicinity (±100 kb) to the significant CBB markers, some of which could be directly or indirectly involved in plant defense to diseases. These results provide a basis to further understand the complex inheritance of CBB resistance in Mesoamerican common beans and show that this biotic stress is unrelated to CR resistance, which was evident during a drought period. Genotypes with good yield potential for the Southeastern U.S. growing conditions were found with resistant to infection by the two diseases, as well as adaptation to the hot and humid conditions punctuated by droughts found in this region

Prevalence of Campylobacter and Salmonella in African food animals and meat: a systematic review and meta-analysis

Background: Campylobacter and Salmonella, particularly non-typhoidal Salmonella, are important bacterial enteric pathogens of humans which are often carried asymptomatically in animal reservoirs. Bacterial foodborne infections, including those derived from meat, are associated with illness and death globally but the burden is disproportionately high in Africa. Commercial meat production is increasing and intensifying in many African countries, creating opportunities and threats for food safety. Methods: Following Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) guidelines, we searched six databases for English language studies published through June 2016, that reported Campylobacter or Salmonella carriage or infection prevalence in food animals and contamination prevalence in food animal products from African countries. A random effects meta-analysis and multivariable logistic regression were used to estimate the species-specific prevalence of Salmonella and Campylobacter and assess relationships between sample type and region and the detection or isolation of either pathogen. Results: Seventy-three studies reporting Campylobacter and 187 studies reporting Salmonella across 27 African countries were represented. Adjusted prevalence calculations estimate Campylobacter detection in 37.7% (95% CI 31.6–44.3) of 11,828 poultry samples; 24.6% (95% CI 18.0–32.7) of 1975 pig samples; 17.8% (95% CI 12.6–24.5) of 2907 goat samples; 12.6% (95% CI 8.4–18.5) of 2382 sheep samples; and 12.3% (95% CI 9.5–15.8) of 6545 cattle samples. Salmonella were detected in 13.9% (95% CI 11.7–16.4) of 25,430 poultry samples; 13.1% (95% CI 9.3–18.3) of 5467 pig samples; 9.3% (95% CI 7.2–12.1) of 2988 camel samples; 5.3% (95% CI 4.0–6.8) of 72,292 cattle samples; 4.8% (95% CI 3.6–6.3) of 11,335 sheep samples; and 3.4% (95% CI 2.2–5.2) of 4904 goat samples. ‘External’ samples (e.g. hide, feathers) were significantly more likely to be contaminated by both pathogens than ‘gut’ (e.g. faeces, cloaca) while meat and organs were significantly less likely to be contaminated than gut samples. Conclusions: This study demonstrated widespread prevalence of Campylobacter species and Salmonella serovars in African food animals and meat, particularly in samples of poultry and pig origin. Source attribution studies could help ascertain which food animals are contributing to human campylobacteriosis and salmonellosis and direct potential food safety interventions

Spread of non-typhoidal Salmonella in the beef supply chain in northern Tanzania: Sensitivity in a probabilistic model integrating microbiological data and data from stakeholder interviews

East Africa is a hotspot for foodborne diseases, including infection by nontyphoidal Salmonella (NTS), a zoonotic pathogen that may originate from livestock. Urbanization and increased demand for animal protein drive intensification of livestock production and food processing, creating risks and opportunities for food safety. We built a probabilistic mathematical model, informed by prior beliefs and dedicated stakeholder interviews and microbiological research, to describe sources and prevalence of NTS along the beef supply chain in Moshi, Tanzania. The supply chain was conceptualized using a bow tie model, with terminal livestock markets as pinch point, and a forked pathway postmarket to compare traditional and emerging supply chains. NTS was detected in 36 (7.7%) of 467 samples throughout the supply chain. After combining prior belief and observational data, marginal estimates of true NTS prevalence were 4% in feces of cattle entering the beef supply and 20% in raw meat at butcheries. Based on our model and sensitivity analyses, true NTS prevalence was not significantly different between supply chains. Environmental contamination, associated with butchers and vendors, was estimated to be the most likely source of NTS in meat for human consumption. The model provides a framework for assessing the origin and propagation of NTS along meat supply chains. It can be used to inform decision making when economic factors cause changes in beef production and consumption, such as where to target interventions to reduce risks to consumers. Through sensitivity and value of information analyses, the model also helps to prioritize investment in additional research

Potential Biological Control Agents for Soilborne Fungal Pathogens in Tennessee Snap Bean Farms

Fungi isolated from snap bean roots and rhizosphere soil where fungicides are not used included Fusarium oxysporum, Fusarium equiseti, Fusarium subglutinans, Fusarium camptoceras, Fusarium chlamydosporum, Fusarium verticillioides, Fusarium proliferatum, Fusarium acuminatum, Fusarium solani, Peyronellaea pinodella, Macrophomina phaseolina, and Glomerella guttata. Only P. pinodella, M. phaseolina, and F. oxysporum were isolated on symptomatic plants. These soilborne fungi are common pathogens of diverse host plants. Pathogenicity tests under controlled environment demonstrated that these fungi were pathogenic on snap beans. Subsequently, bacterial endophytes isolated from snap bean roots, papaya roots and stems, and dogwood stems were evaluated as potential biological control agents against these diverse fungi. All bacteria isolated, including Bacillus vallismortis (PS), Bacillus amyloliquefaciens (Psl), Bacillus subtilis (Prt), Bacillus thuringiensis (Y and IMC8), Enterobacter sp. (E), Stenotrophomonas sp. (B17A), and Serratia sp. (B17B) suppressed growth of the fungal pathogens in vitro and formed clear inhibition zones in petri dish dual cultures. Growth media taken from the inhibition zones suppressed growth of the fungal pathogens in the absence of the bacterial cells, suggesting that the bacteria released unidentified antagonistic biochemical substances into the media. This study constitutes an initial screening of endophytes as biological control agents against diverse fungal pathogens and forms a basis for the discovery of novel strains that can be further developed and integrated into disease management systems for diverse fungal pathogens. Isolates B. vallismortis (PS), B. amyloliquefaciens (Psl), B. subtilis (Prt), and B. thuringiensis (Y IMC8) exhibited the best performance as potential biological control agents paving the way for larger-scale in vivo studies and characterization of their interactions with fungal pathogens