Lipopeptides from Bacillus species exhibit promising biological control activity against plant pathogens. This study aimed to explore the potential of purified fengycin to induce systemic resistance in tomato against Sclerotinia sclerotiorum. Bacillus amyloliquefaciens FZB42, its mutant AK1S, and their corresponding metabolites showed in vitro inhibition of S. sclerotiorum mycelium. Fengycin derived from an AK1S mutant was purified and identified through HPLC and MALDI-TOF-MS, respectively. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) revealed structural deformities in the fungal mycelium. Moreover, fengycin induced the accumulation of reactive oxygen species (ROS) in S. sclerotiorum mycelium and downregulated the expression of ROS-scavenging genes viz., superoxide dismutase (SsSOD1), peroxidase (SsPO), and catalase (SsCAT1) compared to the untreated control. Furthermore, the lesion size was dramatically reduced in fengycin-treated tomato plants compared to plants infected with S. sclerotiorum only in a greenhouse experiment. Additionally, the transcriptional regulation of defense-related genes GST, SOD, PAL, HMGR, and MPK3 showed the highest upsurge in expression at 48 h post-inoculation (hpi). However, their expression was subsequently decreased at 96 hpi in fengycin + S. sclerotiorum treatment compared to the plants treated with fengycin only. Conversely, the expression of PPO increased in a linear manner up to 96 hpi.
The ability of salicylic acid and jasmonic acid to suppress post-harvest infection with green mould Penicillium digitatum and blue mould P. italicum on three citrus species Citrus reticulata 'Kinnow', C. limon 'Meyer Lemon', and C. limetta 'Mosambi' was evaluated in a dose-response study. Salicylic acid (SA) and jasmonic acid (JA) were applied to the fruits as a post-harvest dip treatment followed by wound inoculation with the pathogens. Both resistance inducers caused a significantly lower disease severity compared with the infected but non-treated control, whereas disease incidence was not significantly lower than in the control. The efficacy of both SA and JA in reducing disease severity was concentration-dependent; the use of higher concentrations resulted in a greater degree of suppression. All the Citrus species tested in this study showed different responses in terms of disease development. C. limon 'Meyer Lemon' showed the highest disease development, and C. limetta 'Mosambi' the lowest. To get an insight into the mechanisms underlying the increase in resistance, the activity of defence-related enzymes -peroxidase (POD) and polyphenol oxidase (PPO) -was recorded in SA-and JA-treated fruit peelings. The activity of both enzymes was directly proportional to the concentration of the SA and JA applications. The highest activity of PPO and POD was observed in C. reticulata 'Kinnow' and the lowest in C. limon 'Meyer Lemon' fruits. This study is the first to document an increase in the activity of PPO and POD in SA-and JA-treated Citrus species in the presence of blue mould and green mould pathogens.Ke y wo r d s: enzymes, Penicillium digitatum, P. italicum, post-harvest, resistance inducers Abbreviations: BM − blue mould, GM − green mould, JA − jasmonic acid, POD − peroxidase, PPO − polyphenol oxidase, RH -relative humidity, SA − salicylic acid
Verticillium wilt caused by Verticillium dahliae is a threatening disease of cotton, causing economic loss worldwide. In this study, nine endophytic Bacillus strains isolated from cotton roots exhibited inhibitory activity against V. dahliae strain VD-080 in a dual culture assay. B. altitudinis HNH7 and B. velezensis HNH9 were chosen for further experiments based on their high antagonistic activity. The secondary metabolites of HNH7 and HNH9 also inhibited the growth of VD-080. Genetic marker-assisted detection revealed the presence of bacillibactin, surfactin, bacillomycin and fengycin encoding genes in the genome of HNH7 and HNH9 and their corresponding gene products were validated through LC-MS. Scanning electron microscopy revealed mycelial disintegration, curling and shrinkage of VD-080 hyphae after treatment with methanolic extracts of the isolated endophytes. Furthermore, a significant reduction in verticillium wilt severity was noticed in cotton plants treated with HNH7 and HNH9 as compared to control treatments. Moreover, the expression of defense-linked genes, viz., MPK3, GST, SOD, PAL, PPO and HMGR, was considerably higher in plants treated with endophytic Bacillus strains and inoculated with VD-080 as compared to control.
Sclerotinia sclerotiorum is a devastating necrotrophic pathogen that infects multiple crops, and its control is an unremitting challenge. In this work, we attempted to gain insights into the pivotal role of lipopeptides (LPs) in the antifungal activity of Bacillus amyloliquefaciens EZ1509. In a comparative study involving five Bacillus strains, B. amyloliquefaciens EZ1509 harboring four LPs biosynthetic genes (viz. surfactin, iturin, fengycin, and bacilysin) exhibited promising antifungal activity against S. sclerotiorum in a dual-culture assay. Our data demonstrated a remarkable upsurge in LPs biosynthetic gene expression through quantitative reverse transcription PCR during in vitro interaction assay with S. sclerotiorum. Maximum upregulation in LPs biosynthetic genes was observed on the second and third days of in vitro interaction, with iturin and fengycin being the highly expressed genes. Subsequently, Matrix-assisted laser desorption/ionization-time of flight-mass spectrometry analysis confirmed the presence of LPs in the inhibition zone. Scanning electron microscope analysis showed disintegration, shrinkage, plasmolysis, and breakdown of fungal hyphae. During in planta evaluation, S. sclerotiorum previously challenged with EZ1509 showed significant suppression in pathogenicity on detached leaves of tobacco and rapeseed. The oxalic acid synthesis was also significantly reduced in S. sclerotiorum previously confronted with antagonistic bacterium. The expression of major virulence genes of S. sclerotiorum, including endopolygalacturonase-3, oxalic acid hydrolase, and endopolygalacturonase-6, was significantly downregulated during in vitro confrontation with EZ1509.
Aims:The potential of endophytic Bacillus strains to improve plant growth and yield was evaluated.
Methods and results: Endophytic Bacillus altitudinis HNH7 and Bacillus velezensisHNH9 were evaluated for their growth-promoting traits. In an in vitro plate assay, HNH7 and HNH9 exhibited proteolytic, amylolytic, lipolytic and cellulolytic activity.HNH7 and HNH9 were able to solubilize iron by producing siderophores but were unable to solubilize insoluble phosphate. PCR confirmed the presence of four growthpromoting genes viz. pvd, budA, asbA and satA in the genome of HNH7, while HNH9 also possessed the same genes except for budA. In a greenhouse experiment, HNH7 and HNH9 promoted the growth of upland cotton plants by upregulating the expression of growth-linked genes, EXP6, ARF1, ARF18, IAA9, CKX6 and GID1b.However, the expression of genes involved in ethylene biosynthesis, that is ERF and ERF17 was downregulated after treating the plants with HNH7 and HNH9 compared to the control. Furthermore, cotton plants treated with HNH7 and HNH9 exhibited a significantly higher rate of photosynthesis and stomatal conductance.
Conclusion:HNH7 and HNH9 showed a promising potential to promote the growth of cotton plants.Significance and impact of study: Research on plant growth-promoting Bacillus strains can lead to the formation of biofertilizers.
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