Carvacrol, a plant-derived volatile small molecule, is effective against various agents that can cause damage to humans, the food processing industry, and plants, and is considered a safe substance for human consumption. In this short communication, previous studies on the effectiveness of carvacrol against various agents, particularly plant pathogens and their associated mechanisms are described. In our study, carvacrol was found to be effective on media against several soilborne pathogens and in planta against three foliar pathogens (Xanthomonas perforans, Alternaria tomatophila, and Podosphaeraxanthii) of important vegetable crops in south Florida of the United States. Current research findings indicated that the effectiveness of carvacrol against various plant pathogens tested was associated with its direct bactericidal/fungicidal effect, which was affected greatly by its volatility. Development of new formulations to overcome the volatility and to prolong the effectiveness of carvacrol was also presented. Our studies on carvacrol suggested that, with advanced development of new formulations, carvacrol could be used as a promising tool in the integrated pest management for bacterial, fungal, and viral pathogens of important vegetable crops in Florida, the USA, and the world.

Bacterial spot caused by Xanthomonas spp. is one of the major diseases in tomato. Xanthomonas perforans is the main pathogen of bacterial spot on tomato in Florida. Currently, application of copper fungicides is the primary measure used to manage this disease. However, the development of copper resistance in X. perforans and accumulation of copper in the environment are major concerns for excessive use of copper-based products in agriculture. Due to its antibacterial properties and low environmental impact, N-acetylcysteine (NAC), a small molecule commonly used in medicine for human bacterial diseases, has been studied in agriculture for the control of plant bacterial pathogens, including X. citri and Xylella fastidiosa. This study evaluated the effect of NAC alone and in combination with copper on a copper-resistant X. perforans strain in vitro and its ability to control bacterial spot of tomato under greenhouse and field conditions. In vitro, the minimum inhibitory concentration of NAC against the X. perforans strain was 2,048 mg liter⁻¹. NAC increased sensitivity of the copper-resistant X. perforans to copper in vitro when application of NAC was followed by copper application after 6 h. In greenhouse assays, NAC applied alone or in combination with copper significantly (P < 0.05) reduced the disease severity of bacterial spot on tomato compared with the untreated control. NAC at 100 mg liter⁻¹ + copper at 300 mg liter⁻¹ consistently exhibited synergistic effects against bacterial spot. In the field trials, NAC at 1,000 mg liter⁻¹ + copper at 150 mg liter⁻¹ significantly reduced disease severity compared with the untreated control. Results from this study demonstrated that NAC significantly reduced the disease severity of bacterial spot of tomato and enhanced the efficacy of copper against copper-resistant X. perforans, indicating that NAC could be applied for the effective management of bacterial spot of tomato.

Bacterial spot of tomato continues to pose a significant problem to tomato production worldwide. In Florida, bacterial spot of tomato caused by Xanthomonas perforans is one of the most important diseases responsible for tomato yield loss. This disease is difficult to control, and new strategies are continually being investigated to combat the devastating effect of this disease. Recent efforts focusing on essential oils based on small molecules have spurred interests in the utilization of this class of chemicals for disease management. In this study, we evaluated the efficacy of eugenol for the management of bacterial spot of tomato caused by X. perforans. In the greenhouse experiments, eugenol applied as a foliar spray significantly (p < 0.5) reduced bacterial spot disease compared to the untreated control. In the field experiments, the area under the disease progress curve (AUDPC) was significantly (p < 0.5) lower in the plots treated with eugenol or eugenol combined with the surfactant Cohere than in the untreated control plots, and it was comparable to the copper-based treatments. To provide additional insights into the possible pathways of eugenol activities, we applied a liquid chromatography mass spectrometry (LC-MS)-based metabolomic study using a thermo Q-Exactive orbitrap mass spectrometer with Dionex ultra high-performance liquid chromatography (UHPLC) on X. perforans strain 91–118 treated with eugenol. Our results showed that eugenol affected metabolite production in multiple pathways critical to bacterial survival. For example, treatment of cells with eugenol resulted in the downregulation of the glutathione metabolism pathway and associated metabolites, except for 5-oxoproline, which accumulation is known to be toxic to living cells. While the peaks corresponding to the putatively identified sarmentosin showed the most significant impact and reduced in response to eugenol treatment, branched-chain amino acids, such as L-isoleucine, increased in production, suggesting that eugenol may not negatively affect the protein biosynthesis pathways. The results from our study demonstrated the efficacy of eugenol in the management of bacterial spot of tomato under greenhouse and field conditions and identified multiple pathways that are targeted.

Microbes are natural chemical factories and their metabolome comprise diverse arrays of chemicals. The genus Xanthomonas comprises some of the most important plant pathogens causing devastating yield losses globally and previous studies suggested that species in the genus are untapped chemical minefields. In this study, we applied an untargeted metabolomics approach to study the metabolome of a globally spread important xanthomonad, X. perforans. The pathogen is difficult to manage, but recent studies suggest that the small molecule carvacrol was efficient in disease control. Bacterial strains were treated with carvacrol, and samples were taken at time intervals (1 and 6 h). An untreated control was also included. There were five replicates for each sample and samples were prepared for metabolomics profiling using the standard procedure. Metabolomics profiling was carried out using a thermo Q-Exactive orbitrap mass spectrometer with Dionex ultra high-performance liquid chromatography (UHPLC) and an autosampler. Annotation of significant metabolites using the Metabolomics Standards Initiative level 2 identified an array of novel metabolites that were previously not reported in Xanthomonas perforans. These metabolites include methoxybrassinin and cyclobrassinone, which are known metabolites of brassicas; sarmentosin, a metabolite of the Passiflora-heliconiine butterfly system; and monatin, a naturally occurring sweetener found in Sclerochiton ilicifolius. To our knowledge, this is the first report of these metabolites in a microbial system. Other significant metabolites previously identified in non-Xanthomonas systems but reported in this study include maculosin; piperidine; β-carboline alkaloids, such as harman and derivatives; and several important medically relevant metabolites, such as valsartan, metharbital, pirbuterol, and ozagrel. This finding is consistent with convergent evolution found in reported biological systems. Analyses of the effect of carvacrol in time-series and associated pathways suggest that carvacrol has a global effect on the metabolome of X. perforans, showing marked changes in metabolites that are critical in energy biosynthesis and degradation pathways, amino acid pathways, nucleic acid pathways, as well as the newly identified metabolites whose pathways are unknown. This study provides the first insight into the X. perforans metabolome and additionally lays a metabolomics-guided foundation for characterization of novel metabolites and pathways in xanthomonad systems.

Halo blight, caused by Pseudomonas syringae pv. phaseolicola, is one of the major bacterial diseases on snap bean in Florida, and the outbreaks of this disease have occurred more often in recent years. Current management of this disease primarily depends on application of fixed copper-based bactericides but climate change and resistance development in the pathogen populations still cause hardship for management of this disease, especially in south Florida. In this study, nicotinamide adenine dinucleotide (NAD⁺) was evaluated in the greenhouse for its potential to reduce halo blight on snap bean. When NAD⁺ at 5 mM was applied by soil drench, foliar spray, or leaf infiltration, NAD⁺ significantly (P < 0.05) reduced disease severity of halo blight on snap bean compared with the untreated control. When NAD⁺ was applied by leaf infiltration, among the tested concentrations, NAD⁺ at 0.5 to 1.0 mM was most effective in decreasing halo blight disease. NAD⁺ at 2.5 mM applied as a foliar spray in rotation with Kocide 3000 (copper hydroxide) at 0.5 mg/ml further reduced disease severity compared with Kocide 3000 alone. In the in vitro study, no inhibitory effects of NAD⁺ were detected on the bacterial pathogen P. syringae pv. phaseolicola. Results of real-time PCR showed that the defense-related genes PR1, AZI1, EDS1, SARD1, PDF1.2, and PAL1 were upregulated in the NAD⁺ treatment. Taken together, these data indicated that NAD⁺ significantly suppressed halo blight on snap bean, and application of NAD⁺ has the potential in management of this important disease.

One negative consequence of international trade of agricultural commodities is the inadvertent global spread of crop diseases. Yam (Dioscorea spp.) is a staple food crop in many countries and is traded globally. Most of the commercially traded yams in the United States is imported. In late 2020, samples of yam tubers, from a commercial facility, were submitted to the plant diagnostic clinic at the UF/IFAS Tropical Research and Education Center in Homestead, Florida. Samples showed rotten symptoms and were drawn from lots that were marked to be destroyed because the source of the rotting symptoms was unknown. Preliminary isolation showed that a fungus was consistently associated with the symptoms and was confirmed in subsequent pathogenicity test as the causal agent. The fungus grew profusely on potato dextrose agar (PDA) with highly melanized hyphae. Matured conidia showed longitudinal striations. Based on its growth pattern and morphology, it was suspected that this fungus may be in the genus Lasiodiplodia. DNA-based identification using partial sequences of the internal transcribed spacer (ITS), β-tubulin (TUB2), 28S rDNA (LSU) and elongation factor alpha (EF1-α) genes confirmed the identity of the isolates as Lasiodiplodia iraniensis Abdollahz., Zare & A.J.L. Phillips (Synonym: L. iranensis). This is the first report of L. iraniensis affecting yam and has implications for international trade. This finding will provide important foundation for making quarantine decisions to prevent spread of this disease.