Indole-3-acetic acid (IAA) is produced by microorganisms and plants via either tryptophan-dependent or tryptophan-independent pathways. Herein, we investigated the optimisation of IAA production by Streptomyces fradiae NKZ-259 and its formulation as a plant growth promoter to improve economic and agricultural development. The maximum IAA yield achieved using optimal conditions was 82.363 μg/mL in the presence of 2 g/L tryptophan after 6 days of incubation. Thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC) analysis of putative IAA revealed an RF value of 0.69 and a retention time of 11.842 min, comparable with the IAA standard. Regarding product formulation, kaolin-based powder achieved a suspension rate of 73.74% and a wetting time of 80 s. This carrier exhibited good shelf life stability for NKZ-259, and the cell population did not decrease obviously over 4 months of storage at 4 °C. In vivo analysis of plant growth promotion showed that tomato seedlings treated with kaolin powder containing NKZ-259 cells displayed a significant increase in root and shoot length of 7.97 cm and 32.77 cm, respectively, and an increase in fresh weight and dry weight of 6.72 g and 1.34 g. Compared to controls, plant growth parameters were increased almost it two-fold. Optimising the culture conditions resulted in an almost four-fold increase in IAA secretion by NKZ-259 cells. The results clearly demonstrate that S. fradiae NKZ-259 holds great potential for plant growth promotion and IAA production. Furthermore, kaolin-based powder is an effective carrier for NKZ-259 cells and may be useful for commercial applications.
Sclerotinia sclerotiorum (Lib.) de Bary, a destructive fungal pathogen with an extensive host range, causes various diseases with the potential to cause huge economic losses to crops worldwide. Streptomyces species produce secondary metabolites with variable structures and biological activities that offer possible control methods for crop diseases. Herein, we evaluated the inhibitory effects of wuyiencin, a secondary metabolite of Streptomyces albulus CK-15, against S. sclerotiorum. The results showed that wuyiencin markedly inhibited mycelial growth and germination and the formation of sclerotia. It also increased cell membrane permeability, resulting in leakage of intracellular substances in pathogen mycelia. Wuyiencin markedly decreased oxalic acid content and the activities of polygalacturonase and pectin methyl-galacturonic enzymes. Moreover, it downregulated Nox1, ITL, pph1, Caf1, and sca1, all genes related to growth and infection. Lesions were smaller and less pronounced on soybean (Glycine max [L.] Merr.) leaves pretreated with wuyiencin in vitro, and the inhibition rate reached 78.36%. The results suggest that wuyiencin holds promise for the management of diseases caused by S. sclerotiorum, and the findings provide clues on the mechanism of action.
Sclerotinia sclerotiorum (Lib) de Bary, a destructive fungal pathogen with an extensive host range, causes major economic losses to crop production activities globally. Streptomyces spp. produce secondary metabolites with diverse structures and biological activities with potential applications in the control of crop disease. This study explored the potential application of wuyiencin, a secondary metabolite of Streptomyces albulus CK-15, to induce defense responses in soybean against S. sclerotiorum. Lesion size was reduced by nearly 60% in wuyiencin-treated soybean plants compared with plants infected with S. sclerotiorum only in greenhouse experiments. Wuyiencin induced callose deposition at 6 h postinoculation and increased reactive-oxygen-scavenging enzyme activities, including superoxide dismutase, catalase, and peroxidase. Moreover, wuyiencin inoculated before S. sclerotiorum infection significantly increased polyphenol oxidase, phenylalanine ammonia lyase, chitinase, and β-1,3-glucanase activity, suggesting their involvement in soybean defense responses to S. sclerotiorum. Further, qRT-PCR results showed expression levels of the hormone signaling markers CO11, MYC2, PR4, PR1, NPR1, and ERF1 were upregulated in infected leaves treated with wuyiencin.
This study mainly evaluated the broad-spectrum fungicidal activity of ε-poly L lysine (ε-PL) against 12 pathogenic fungi. We further demonstrated synergistic antifungal activity of ε-PL combined with wuyiencin against Botryris cinerea. The combined bio-fungicide achieved an inhibition rate of 100% for mycelial growth using ε-PL at 500 μg/mL + wuyiencin at 50 μg/mL and for spore germination using ε-PL at 200 μg/mL + wuyiencin at 80 μg/mL in vitro. This synergistic spore and mycelia-damaging effect of the combination was confirmed using scanning electron microscopy. In vivo assays with combined bio-fungicide (1500 μg/mL ε-PL + 60 μg/mL wuyiencin) on detached leaves showed depressed growth and development of the spores of B. cinerea. The synergistic effect was further tested in combinations of ε-PL with wuyiencin by measuring the fractional inhibition concentration index (FICI) value below 0.5. Moreover, ε-PL and wuyiencin inoculation before B. cinerea infection significantly increased the superoxide dismutase, peroxidase, catalase, and phenylalanine ammonia-lyase activities, which suggested their involvement in tomato defense responses to disease to minimize damage to B. cinerea. These findings revealed that a combined bio-fungicide comprising ε-PL and wuyiencin had a good prospect for controlling plant fungal disease.