A study was conducted to characterize the common Pythium spp. in greenhouses in Oman and their level of resistance to hymexazol, a widely used fungicide in the country. Pythium isolates were obtained from soil samples, cocopeat bags, and cucumber roots collected from seven regions in the country. Identification of 80 Pythium isolates to the species level using sequences of the internal transcribed spacer region of the ribosomal RNA showed that they belong to four species: Pythium aphanidermatum (77 isolates), P. spinosum (1 isolate), P. myriotylum (1 isolate), and P. catenulatum (1 isolate). Investigating the aggressiveness of three Pythium spp. on cucumber showed that P. aphanidermatum, P. myriotylum, and P. spinosum are pathogenic. Phylogenetic analysis of P. aphanidermatum isolates showed that most of the isolates obtained from cocopeat clustered separately from isolates obtained from soil and roots. This may indicate a difference in the origin of the cocopeat isolates. Evaluating the resistance of 27 P. aphanidermatum isolates to hymexazol showed that most isolates were sensitive (0.9 to 31.2 mg liter⁻¹) whereas one isolate was resistant (142.9 mg liter⁻¹). This study is the first to report P. myriotylum and P. catenulatum in Oman. It is also the first to report the development of resistance to hymexazol among P. aphanidermatum populations from greenhouses. Growers should use integrated disease management strategies to avoid further development of resistance to hymexazol.

Soil biofumigation is a widely used farming practice for soil-borne disease management in vegetable crops. Soil biofumigation is the practice of the incorporation of fresh plant materials from the members of the Brassicaceae family into the soil which release antimicrobial volatile organic compounds (VOCs), namely glucosinolates and their hydrolysis products. However, the VOCs may affect non-target beneficial soil biota including microbial biocontrol agents (MBAs) due to their broad-spectrum toxicity. In this study, native antagonistic rhizobacteria were isolated from cabbage plants grown in biofumigated fields and their potential in the management of Pythium aphanidermatum cucumber damping-off was evaluated with and also without biofumigation using cabbage leaf residue. A total of 20 morphologically distinct, culturable bacterial strains were recovered from the rhizosphere soil of cabbage cultivated in a biofumigated field, with the largest fraction of the isolates belonging to the genus Bacillus. The other bacterial genera isolated were Pseudomonas, Serratia, Stenotrophomonas, Microbacterium and Priestia. Of the 20 bacterial isolates, Pseudomonas aeruginosa B1-SQU, Pseudomonas indica B2-SQU, Serratia marcescens B3-SQU and Pseudomonas brenneri B4-SQU exhibited inhibitory activity against P. aphanidermatum in a co-culture assay. The deformation and shrinkage of P. aphanidermatum hyphae due to diffusible antimicrobial compounds from these bacterial strains were witnessed through scanning electron microscopy. A biofilm was formed by these bacterial strains in vitro and they exhibited compatibility with each other; however, they had no significant plant growth promotion effect on cucumber. These bacterial strains significantly reduced damping-off in cucumber under greenhouse conditions when applied to soil singly, but failed to show a significant effect when combined with biofumigation treatment, though the bacterial population in the soil was maintained at higher levels. Soil biofumigation alone was also not effective in suppressing the damping-off of cucumber. Gas chromatography–mass spectrometry analysis revealed that 1-butanol, n-butyl butyrate and butyl acetate were the predominant VOCs in the soil amended with cabbage leaf residue. The results suggest that soil biofumigation with cabbage leaf residue has no significant effect on P. aphanidermatum under high inoculum levels; furthermore, it affects the efficiency of the bacterial antagonists in controlling damping-off in cucumber.

In this study, endophytic and rhizospheric bacteria were isolated from Moringa olifera and M. perigreina from Oman, and their in vitro antagonistic activity against Pythium aphanidermatum was tested using a dual culture assay. The promising strains were tested further for their compatibility and potential for plant growth promotion, biofilm formation, antifungal volatile organic compound (VOC) production, and the biological control of P. aphanidermatum damping-off of cabbage (Brassica oleracea L.) under greenhouse conditions. A total of 12 endophytic and 27 rhizospheric bacteria were isolated from Moringa spp. Among them, Bacillus pumilus MPE1 showed the maximum antagonistic activity against P. aphanidermatum in the dual culture assay, followed by Paenibacillus glucanolyticus MPE3 and Pseudomonas indica MOR3 and MOR8. These bacterial isolates induced abundant morphological abnormalities in the hyphae of P. aphanidermatum, as observed via scanning electron microscopy. The in vitro cross-streak assay showed that these bacterial isolates were compatible among themselves, except for P. indica MOR8 × P. glucanolyticus MPE3. These antagonists released VOCs that restricted the growth of P. aphanidermatum in an in vitro assay. These antagonistic bacteria released 2,4-dimethylheptane and 4-methyloctane as the predominant volatile compounds. Of the four antagonistic bacterial strains, P. indica MOR8 was capable of forming biofilm, which is considered a trait that improves the efficacy of rhizosphere inoculants. The results of the greenhouse experiments showed that the soil treatment with B. pumilus MPE1 showed the highest reduction (59%) in the incidence of P. aphanidermatum damping-off in cabbage, evidencing its potential as a biological control agent for the management of this disease. Further research is needed to characterize the antifungal traits and activities of B. pumilus MPE1 and to assert its potential use against other soil-borne plant pathogens.

Potting media are commonly used by growers in different parts of the world for potted plants, raising seedlings and for improving soil characteristics. This study was conducted to characterize bacterial communities occurring in 13 commercial potting media products originating from seven countries. Bacteria were isolated using serial dilution. Identification to the species level was based on phylogenetic analysis of the 16S rRNA gene. The analysis showed the association of 13 bacterial species with the different potting media samples, namely Arthrobacter livingstonensis, Kocuria flava, Leifsonia lichenia, Bacillus vallismortis, Bacillus pumilus, Staphylococcus warneri, Burkholderia phenazinium, Burkholderia sp., Ralstonia pickettii, Rhodanobacter spathiphylli, Rhodanobacter sp., Pseudomonas thivervalensis and Chryseobacterium gallinarum. Bacterial densities in the samples ranged from 8 × 107 to 1.2 × 109 colony forming units per gram of substrate. The study shows the isolation of some potential plant and human bacterial pathogens. However, most of the isolated species were either biocontrol species or saprophytes. The study questions the ways by which these bacterial species were introduced into potting media. To the best of our knowledge, this appears to be the first report of most of the isolated bacteria from potting media, except B. pumilus.

Stemphylium lycopersici is an important pathogen causing leaf spot of tomatoes worldwide. Although much information is available about the pathogen, little is known about dynamics of S. lycopersici in tomato fields. Seventy-nine symptomatic leaf samples were collected from two tomato cultivars grown in a farm (Miral and Inbred line). Fungal species associated with the disease were isolated on potato dextrose agar. Seventy-nine isolates were obtained and identified as S. lycopersici based on sequence analysis of combined dataset of the internal transcribed spacer and glyceraldehyde-3-phosphate dehydrogenase regions. The 79 isolates were subjected to amplified fragment length polymorphism analysis using three primer combinations. The Stemphylium lycopersici population from the two cultivars was found to have a very low level of genetic diversity (H = 0.0948). Cluster analysis showed intermixing of isolates from the two cultivars. In addition, analysis of molecular variance showed the presence of a very low level of genetic differentiation between populations obtained from the two cultivars (Fst = 0.0206). These findings indicate the presence of a high rate of gene flow between the two populations and may suggest that the two populations originated from the same inoculum source. The implications of these findings on the management of Stemphylium-induced leaf spot of tomatoes are discussed.

In this study, bacteria residing in the gut of the rice weevils (Sitophilus oryzae L.) (Coleoptera: Curculionidae) feeding on aflatoxin-contaminated corn kernels were isolated and evaluated for their ability to suppress Aspergillus flavus and to remove/degrade aflatoxin B1 (AFB1). Four morphologically distinct S. oryzae gut-associated bacterial isolates were isolated and identified as Bacillus subtilis (RWGB1), Bacillus oceanisediminis (RWGB2), Bacillus firmus (RWGB3), and Pseudomonas aeruginosa (RWGB4) based on 16S rRNA gene sequence analysis. These bacterial isolates inhibited A. flavus growth in the dual culture assay and induced morphological deformities in the fungal hyphae, as confirmed by scanning electron microscopy. All four bacterial isolates were capable of removing AFB1 from the nutrient broth medium. In addition, culture supernatants of these bacterial isolates degraded AFB1, and the degradation of toxin molecules was confirmed by liquid chromatography-mass spectrometry. The bacterial isolates, B. subtilis RWGB1, B. oceanisediminis RWGB2, and P. aeruginosa RWGB4, were capable of producing antifungal volatile organic compounds that inhibited A. flavus growth. These results suggest that the bacterial isolates from S. oryzae gut have the potential to bind and/or degrade AFB1. Further research on their application in the food and feed industries could enhance the safety of food and feed production.