Pepper growth promotion and biocontrol against Xanthomonas euvesicatoria by Bacillus cereus and Bacillus thuringiensis formulations.

Background: Bacillus genus has been used in horticultural crops as a biocontrol agent against insect pests, microbial phytopathogens, and plant growth-promoting bacteria (PGPB), representing an alternative to agrochemicals. In particular, B. cereus (Bc) and B. thuringiensis (Bt) have been studied for their fungicidal and insecticidal activities. However, their use as biofertilizer formulations and biocontrol agents against phytopathogenic bacteria is limited. Objective: To evaluate Bc and Bt formulations as PGPB and biocontrol agents against the bacterial spot agent Xanthomonas euvesicatoria (Xe) in greenhouse-grown chili peppers. Methods: Bc and Bt isolates obtained from soil samples were identified and characterized using conventional biochemical and multiplex PCR identification methods. Bioassays to determine Bc and Bt isolates potential as PGPB were evaluated on chili pepper seedlings in seedbeds. In addition, formulations based on Bc (F-BC26 and F-BC08) and Bt (F-BT24) strains were assessed as biofertilizers on pepper, under controlled conditions. Furthermore, in vitro antagonism assays were performed by confronting Bc and Bt isolate formulations against Xe isolates in direct (foliage) and indirect (resistance induction) phytopathogen biocontrol assays on pepper plants, which were grown under controlled conditions for 15 d after formulations treatment. Results: Isolates were identified as Bc and Bt. Formulations significantly improved pepper growth in seedbeds and pots, whereas in vitro bioassays demonstrated the bactericidal effect of Bc and Bt strains against Xe isolates. Furthermore, assays showed significant plant protection by F-BC26, F-BC08, and F-BT24 formulated strains against Xe. Conclusion: Results indicated that F-BT24 and F-BC26 isolates formulations promoted pepper growth and protected it against Xanthomonas euvesicatoria.

Antagonist activities and phylogenetic relationships of actinomycetes isolated from an Artemisia habitat.

Diverse habitats have been screened for novel antimicrobial actinomycetes, while others remain unexplored. In this study, we analyzed the bioactivities of actinomycetes cultured from rhizosphere soils of the desert plant Artemisia tridentata and the nearby bulk soils. Actinomycetes were screened for antifungal and antibacterial activities toward a panel of plant pathogens; all comparisons were between activities of rhizosphere soil isolates toward those of its counterpart bulk soil. A selected group of the strongest antifungal isolates were also tested against two antifungal-drug resistant strains of Candida albicans. 16S rDNA partial sequences and phylogenetic analysis of isolates that showed broad-spectrum antifungal activities were performed. Forty-two out of 200 and two soil isolated actinomycetes were selected for their strong antifungal activities. The highest proportion of isolates (p<0.05) from rhizosphere soil of an old plant showed antagonism against gram-positive bacteria (0.483 and 0.224 proportions against Bacillus subtilis and Rathayibacter tritici, respectively), and phytopathogenic fungi (0.259, 0.431, and 0.345 proportions against Fusarium oxysporum, Rhizoctonia solani and Pythium ultimum, respectively), while the highest antagonism against the gram-negative bacteria predominated in isolates from the bulk soils. Isolates from a rhizosphere soil of a young plant were characterized for strong antagonist activities against Fusarium oxysporum (0.333 proportion, p<0.05). Phylogenetic analysis of 16S rDNA sequences showed that isolates that exhibited strong antifungal activity were genetically similar. We conclude that the rhizosphere soil of A. tridentata is an excellent source for discovery of actinomycetes with potentially novel antifungal compounds.

Purification and Characterization of Antibacterial Activity against Phytopathogenic Bacteria in Culture Fluids from Ganoderma lucidum.

Previous studies of Ganoderma lucidum have focused on its medicinal applications. Limited information is available about its antibacterial activity against plant pathogens. Thus, the goal of this study was to purify and characterize the antibacterial activity against plant pathogenic bacteria from culture fluids of G. lucidum. The nature of the bioactive components was determined using heat boiling, organic solvents, dialysis tubing, gel exclusion chromatography (GEC), proteinase sensitivity, HPLC, HPLC-APCI-MS, and GC-MS. The bioactive compounds were neither lipid, based on their solubility, nor proteic in nature, based on proteinase digestion and heat stability. The putative-bioactive polysaccharides have molecular weights that range from 3500 to 4500 Daltons as determined by dialysis tubing, GEC and APCI-MS analysis. The composition of the antibacterial compounds was determined by GC-MS. This is the first report of small polysaccharides produced by G. lucidum with activity against bacterial plant pathogens.

Prevalence of Xanthomonas euvesicatoria (formally X. perforans) associated with bacterial spot severity in Capsicum annuum crops in South Central Chihuahua, Mexico.

BACKGROUND: Xanthomonas spp. causes bacterial spot disease, which reduces quality and yield of pepper crops in Mexico. Identification of phytopathogen species is necessary to implement more effective control strategies. OBJECTIVE: The aim of this study was to isolate and identify infecting Xanthomonas species in South Central Chihuahua pepper-producing areas. METHODS: Diseased plants were collected from 30 cultivation lots and bacteria were isolated from damaged tissues. Potential causative agents were isolated, identified, and characterized by biochemical and molecular analysis. Pathogenicity tests from each isolate were then performed on 30-d-old pepper seedlings, exposing five leaves to 10 µL of 1 × 108 CFU/mL bacterial suspensions of each isolate, using sterile distilled water as a control. Disease severity was determined after 10 d by calculating leaf damage percentage. Furthermore, we evaluated the susceptibility of the highest bacterial spot severity-causing isolates (13 isolates) to copper sulphate (CuS), copper gluconate (CuG), copper oxychloride + oxytetracycline hydrochloride (Cu + Ox), gentamicin + oxytetracycline hydrochloride (Gen + Ox), and gentamicin sulphate (GenS). Copper-resistance genes (copLAB) were detected by PCR analysis among isolates. RESULTS: Thirty-seven foliage isolates were identified as Xanthomonas euvesicatoria (14%), which were associated with bacterial spot disease in jalapeño pepper. Tested Xanthomonas isolates were resistant to Cu-based compounds, but susceptible to Cu + Ox. All isolates were susceptible to Gen + Ox and GenS. CopLAB genes were detected in all but one strain. CONCLUSIONS: X. euvesicatoria (formally X. perforans) may be considered as an emerging pathogen of bacterial spot pepper in Mexico. Among disease management strategies, alternatives to copper should be taken into consideration.

Molecular and Biological Characterization of Recombinant Isolates of Potato virus Y Circulating in Potato Fields in Mexico.

Potato virus Y (PVY) is a significant threat to potato (Solanum tuberosum) production in Mexico. The presence of recombinant strains of PVY circulating in potato has been reported in the country, but no systematic study on the genetic diversity of PVY in potato and prevalence of PVY strains has been conducted yet. We report on a series of surveys in seed potato production areas in two states in Mexico, namely, Chihuahua and Jalisco, between 2011 and 2019. PVY was detected through the period of nine years in multiple potato cultivars in both states, often remaining asymptomatic in the most popular cultivars, such as 'Fianna' and 'Agata'. When typed to strain, all PVY samples studied were found to have N-serotype, and were all identified molecularly as isolates of the same recombinant strain, PVYNTN. Five of these PVY isolates were tested on tobacco (Nicotiana tabacum), where they induced vein necrosis supporting the molecular typing. This identification was also confirmed biologically on differential potato cultivars, where one PVYNTN isolate from the 2013 survey triggered the hypersensitive resistance conferred by the Nztbr gene in the cv. Maris Bard. Seven of these Mexican PVYNTN isolates, collected between 2013 and 2019, including two PVY isolates from potato tubers exhibiting potato tuber necrotic ringspot disease, were subjected to whole genome sequencing and found to show a typical PVYNTNa recombinant structure. When subjected to phylogenetic analysis, Mexican PVYNTN sequences clustered in more than three separate clades, suggesting multiple introductions of PVYNTN in the country. The wide circulation of the PVYNTN strain in Mexican potato should be considered by potato producers, to develop mitigation strategies for this PVY strain associated with tuber necrotic symptoms.

Molecular and biological characterization of a recombinant isolate of potato virus Y from Mexico.

An isolate of potato virus Y (PVY), PVY-M3, was subjected to biological characterization on potato indicators and to whole-genome sequencing. PVY-M3 induced a local and systemic hypersensitive resistance (HR) response in potato cultivar Maris Bard expressing the Nz gene while inducing no HR in potato cultivars Desiree and King Edward, carrying Ny and Nc genes, respectively. These HR responses, combined with a lack of vein necrosis in tobacco, clearly defined PVY-M3 as an isolate of the PVY(Z) strain. Recombination analysis demonstrated that PVY-M3 had a typical European PVY(NTN) genome with three recombinant junctions, and PVY(N) and PVY(O) were identified as parents.