Characteristics of biological control and mechanisms of Pseudomonas chlororaphis zm-1 against peanut stem rot

Background Peanut stem rot is a serious plant disease that causes great economic losses. At present, there are no effective measures to prevent or control the occurrence of this plant disease. Biological control is one of the most promising plant disease control measures. In this study, Pseudomonas chlororaphis subsp. aurantiaca strain zm-1, a bacterial strain with potential biocontrol properties isolated by our team from the rhizosphere soil of Anemarrhena asphodeloides, was studied to control this plant disease. Methods We prepared extracts of Pseudomonas chloroaphis zm-1 extracellular antibacterial compounds (PECEs), determined their antifungal activities by confrontation assay, and identified their components by UPLC-MS/MS. The gene knockout strains were constructed by homologous recombination, and the biocontrol efficacy of P. chlororaphis zm-1 and its mutant strains were evaluated by pot experiments under greenhouse conditions and plot experiments, respectively. Results P. chlororaphis zm-1 could produce extracellular antifungal substances and inhibit the growth of Sclerotium rolfsii, the main pathogenic fungus causing peanut stem rot. The components of PECEs identified by UPLC-MS/MS showed that three kinds of phenazine compounds, i.e., 1-hydroxyphenazine, phenazine-1-carboxylic acid (PCA), and the core phenazine, were the principal components. In particular, 1-hydroxyphenazine produced by P. chlororaphis zm-1 showed antifungal activities against S. rolfsii, but 2-hydroxyphenazine did not. This is quite different with the previously reported. The extracellular compounds of two mutant strains, ΔphzH and ΔphzE, was analysed and showed that ΔphzE did not produce any phenazine compounds, and ΔphzH no longer produced 1-hydroxyphenazine but could still produce PCA and phenazine. Furthermore, the antagonistic ability of ΔphzH declined, and that of ΔphzE was almost completely abolished. According to the results of pot experiments under greenhouse conditions, the biocontrol efficacy of ΔphzH dramatically declined to 47.21% compared with that of wild-type P. chlororaphis zm-1 (75.63%). Moreover, ΔphzE almost completely lost its ability to inhibit S. rolfsii (its biocontrol efficacy was reduced to 6.19%). The results of the larger plot experiments were also consistent with these results. Conclusions P. chlororaphis zm-1 has the potential to prevent and control peanut stem rot disease. Phenazines produced and secreted by P. chlororaphis zm-1 play a key role in the control of peanut stem rot caused by S. rolfsii. These findings provide a new idea for the effective prevention and treatment of peanut stem rot.

Spent Substrate of Pleurotus ostreatus Has Potential for Managing Fusarium Wilt of Banana

A range of basidiomycetes including the edible mushroom Pleurotus ostreatus (Po) can suppress plant pathogens such as Fusarium spp. With the current increase in production and consumption of Po in Uganda, the spent Po substrate (SPoS) could be an alternative to manage Fusarium wilt of banana (FWB), caused by the soil borne pathogen Fusarium oxysporum f. sp. cubense, race 1 (Foc). This study determined the potential of SPoS to inhibit Foc in vitro and in potted plants. In vitro studies confirmed suppression of Foc in pure co-culture (Po vs. Foc) assays and media amended with different concentrations (0% to 50% w/v) of un-sterilized SPoS filtrates. Foc growth in the sterile SPoS filtrate was comparable to the water control, suggesting possible roles of biotic or thermolabile components of the SPoS. To further verify the suppressive effects of SPoS, pot experiments were carried out with a resistant (‘Mbwazirume’, AAA) and susceptible (‘Sukali Ndizi’, AAB) banana cultivar using both artificially and naturally infested soils. Independent of the inoculation method, SPoS significantly reduced the severity of FWB in pot experiments. Susceptible cultivar ‘Sukali Ndizi’ growing in substrates amended with SPoS showed lower (1.25) corm damage (Scale 0–5) than the un-amended control (3.75). No corm damage was observed in uninoculated controls. The resistant cultivar ‘Mbwazirume’, showed slight (0.25) corm damage only in the Foc-inoculated plants without SPoS. These findings suggest that SPoS could be used as part of the management practices to reduce the impact of FWB.

Evaluation of Preemergent Herbicides for Chloris virgata Control in Mungbean

Chloris virgata is a problematic weed in mungbean crops due to its high seed production, resistance to glyphosate and high dispersal ability. Pot and field experiments were conducted in 2020 and 2021 to evaluate a range of preemergent (PRE) herbicides for C. virgata control in mungbean. In the field and pot studies, isoxaflutole 75 g ai ha−1 caused crop injury, and in the field experiment, it reduced mungbean yield by 61% compared with the best treatment (pyroxasulfone 100 g ai ha−1). In the field and pot experiments, dimethenamid-P 720 g ai ha−1, pyroxasulfone 100 g ai ha−1 and S-metolachlor 1400 g ai ha−1 provided >88% control of C. virgata (for reduced biomass) and in the field experiment, these herbicides resulted in improved yield by 230%, 270% and 170%, respectively, compared with nontreated control (250 kg ha−1). Similarly, pendimethalin 1000 g ai ha−1 and trifluralin 600 g ai ha−1 provided >89% control (biomass) of C. virgata, and in the field experiment, these resulted in improved yields of 230% and 160%, respectively, compared with the nontreated control. PRE herbicides such as diuron 750 g ai ha−1, linuron 1100 g ai ha−1, metribuzin 360 g ha−1, terbuthylazine 750 g ai ha−1, imazapic 48 g ai ha−1 and imazethapyr 70 g ha−1 although did not cause crop injury; however, these herbicides did not control C. virgata. Flumioxazin 90 g ai ha−1 caused reduced biomass of C. virgata by 80% compared with the nontreated control, and in the field experiment, it resulted in improved yield by 140% compared with the nontreated control. This study suggests the potential use of herbicides, such as dimethenamid-P, pyroxasulfone and S-metolachlor in addition to pendimethalin and trifluralin, for C. virgata control in mungbean. Further studies are needed to determine the efficacy of dimethenamid-P, S-metolachlor and pyroxasulfone for controlling other troublesome weeds in mungbean.

Assessment of Ecotoxicity of Incinerated Sewage Sludge Ash (ISSA)

Combustion in fluidised bed boilers is one of the most commonly used methods of treatment of municipal sewage sludge. Fly ash (FA) and air pollution control (APC) residues are the solid by-products generated by flue gas treatment. There are significant differences in the chemical composition of these wastes. FA is composed of mainly SiO2, P2O5, CaO and metals such as Zn, Ti, Cu, Cr, Pb and Ni. APC residues mainly contain SO3 and Na2O. The leachability tests that were carried out indicate that these wastes display very low leachability of heavy metals (for example leachability of Pb was equal 0.0004 mg·dm−3 in both wastes, leachability of Cd was equal 0.0012 mg·dm−3 in FA an 0.00004 mg·dm−3 in APC). On the other hand, very high sulphate concentrations (49,375 mg·dm−3) were found in water extract for the APC residues. In order to determine the toxicity of these wastes for plants, pot experiments with different additions of waste to the soil were carried out (on Lepidium sativum and Sinapis alba). Tests based on seeds germinations (on Lepidium sativum) in water extracts from waste (in different concentrations) were also performed. The results obtained indicate the very high toxicity of APC residues. Complete inhibition of germination and growth of the test plants was found for all concentrations of the tested waste in water extract and for all additions of waste to the soil in pot experiments. Seed germination tests on water extracts from FA did not show any toxicity of this waste. Pot tests with FA showed their toxicity only with a high (30%) addition in soil.

Cadmium Uptake and Growth Responses of Potted Vegetables to the Cd-Contaminated Soil Inoculated with Cd-Tolerant Purpureocillium lilacinum N1

Bioremediation of Cd- (cadmium) contaminated soil using Cd-tolerant fungus is considered an eco-friendly and cost-effective technique. In this study, we isolated one fungal strain that was hyper-tolerant to Cd from a highly polluted river and conducted pot experiments to evaluate its effects on bioremediation. We found that the fungal strain belonging to the genus, Purpureocillium lilacinum, tolerated 12,000 mg/L Cd. SEM manifested that Cd can be bioaccumulated on the crumpled mycelial surface, generating plenty of metal precipitation particles. In addition, pot experiments showed that the inoculation of P. lilacinum N1 could reduce the total Cd content in soil (2.09% in low contaminated soil and 12.56% in high contaminated soil) and greatly promote plant growth (2.16~3.13 times). Although the Cd concentration of plants was increased by 112.8% in low contaminated soil and decreased by 9.5% in highly contaminated soil with the inoculation of P. lilacinum N1, the total uptake of Cd by plants was greatly improved—1.84~3.6 times higher than that in CK groups. All our results suggest that P. lilacinum N1 is a valuable candidate for the bioremediation of Cd-contaminated soils because of its dual effects on the total Cd content in soil and Cd uptake in plants.

Phytoremediation using Phyllostachys pubescens (Moso Bamboo) to reduce the risk of chromium exposure

In this study, a bamboo species, the Phyllostachys pubescens – Moso Bamboo (MB) -, was selected for its heavy metals accumulation and translocation potential to restore Cr-contaminated soil. In order to evaluate the potential for phytoremediation using MB to restore Cr-contaminated soil, pot experiments were carried out in simulated Mediterranean conditions, in laboratory, in a controlled environment, at a temperature of 20°C. Cr removal from soil was 43 % starting from a Cr content of approx. 200 mg/kg Dry Weight and the quantity of Cr per gram of root and rhizome was equal to 1.31 mg/g dw, while the quantity of Cr per gram of stem and leaves was equal to 0.86 mg/g dw, after 12 weeks. Pot experiments confirm that phytoremediation using plants such as MB provides an alternative approach for handling Cr-contaminated soil.

Inoculation of paddy soils with Rhodopseudomonas palustris enhanced heavy metal immobilisation

To investigate the effect of microbial inoculum on soil heavy metal immobilisation, pot experiments were conducted with paddy soils contaminated by cadmium (Cd), lead (Pb), arsenic (As), and mercury (Hg), respectively. The results showed that the inoculation of Rhodopseudomonas palustris was more effective in the immobilisation of Pb and Cd in soils than the composite of R. palustris and Bacillus subtilis. Interestingly, a lower dosage of inoculum immobilised significantly more heavy metals than the higher dosage, potentially due to the competition of bacteria with limited nutrients. The heavy metal contents in rice grains also supported this finding, as less Pb and Cd were accumulated under the lower dosage. However, there were limited effects of microbial inoculations on the immobilisation of Hg and As. In general, our study indicated the effectiveness of R. palustris in immobilising Pb and Cd in soils and highlighted the importance of determining the optimal dosage of inoculum in bioremediation.