The efficacy of traditional control measures for the management of plant pathogens is decreasing, and the resistance of these pathogens to pesticides is increasing, which poses a serious threat to global food security. The exploration of novel and efficient management measures to combat plant disease is an urgent need at this time. In this study, fungal metabolites from three Trichoderma spp. (T. harzianum, T. virens and T. koningii) were prepared on three different growth media (STP, MOF and supermalt (SuM)). The fungal metabolites were tested in vitro and in vivo from March–April 2020 under greenhouse conditions in a pot experiment utilizing completely randomized design to test their management of the bacterial wilt disease caused by R. solanacearum in tomato plants. The effect of the fungal metabolites on bacterial cell morphology was also investigated through scanning electron microscopy (SEM) analysis. In vitro investigation showed that the fungal metabolites of T. harzianum obtained on the STP medium were the most effective in inhibiting in vitro bacterial growth and produced a 17.6 mm growth inhibition zone. SEM analysis confirms the rupture of the cell walls and cell membranes of the bacterium, along with the leakage of its cell contents. Generally, fungal metabolites obtained on an STP medium showed higher activity than those obtained on the other two media, and these metabolites were then evaluated in vivo according to three application times (0 days before transplantation (DBT), 4 DBT and 8 DBT) in a greenhouse trial to examine their ability to manage R. solanacearum in tomato plants. Consistent with in vitro results, the results from the greenhouse studies showed a level of higher anti-bacterial activity of T. harzianum metabolites than they did for the metabolites of other fungi, while among the three application times, the longest time (8 DBT) was more effective in controlling bacterial wilt disease in tomato plants. Metabolites of T. harzianum applied at 8 DBT caused the maximum decrease in soil bacterial population (1.526 log cfu/g), resulting in the lowest level of disease severity (area under disease progressive curve (AUDPC) value: 400), and maximum plant freshness (with a resulting biomass of 36.7 g, a root length of 18.3 cm and a plant height of 33.0 cm). It can be concluded that T. harzianum metabolites obtained on an STP medium, when applied after 8 DBT, can suppress soil bacterial population and enhance plant growth, and thus can be used as a safe, environmentally-conscious and consumer-friendly approach to managing bacterial wilt disease in tomato plants and possibly other crops.
Aims Because of severe economic losses and food security concerns caused by plant pathogenic bacteria, Ralstonia solanacearum, there is a need to develop novel control methods. This study was aimed to green synthesize the zinc oxide nanoparticles (ZnO NPs) through Withania coagulans leaf extracts and checked their antibacterial potential alone or in combination with W. coagulans leaf extract for the management of R. solanacearum causing bacterial wilt disease in tomato. Methods and Results ZnO NPs were synthesized through an eco‐friendly approach using leaves extract of W. coagulans and characterized through various spectroscopic approaches, that is Fourier transform infrared spectroscopic, UV–visible spectroscopy and energy dispersive spectroscopy. The antibacterial effect of W. coagulans leaf extract and ZnO NPs alone and in combination was tested in vitro and in vivo against bacterial wilt pathogen in tomato plants. The results showed that combine application of leaf extract and ZnO NPs inhibited in vitro growth of R. solanacearum more than applying alone. Three application times (0, 6 and 12 days before transplantation) of leaf extract, ZnONPs and their combine application were tested in vivo. The combine treatment and longest application time (12 days before transplantation) were more effective in suppressing soil population of R. solanacearum, reducing disease severity and enhancing plant growth than applying alone and smaller application time. Conclusion It is concluded that W. coagulans leaf extract and ZnO NPs have strong antibacterial potential against R. solanacearum in vitro and in vivo. Significance and Impact of Study The results of this study suggest the potential application of leaf extract and ZnO nanoparticles for controlling R. solanacearum as safe, eco‐friendly and less expensive integrated disease management strategy in tomato crop.
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