Planta Experiments Research Articles

Antifungal efficacy and biofumigation potential of hydrophobic deep eutectic solvents: Postharvest treatment against Monilinia fructicola and Botrytis Cinerea.

Hydrophobic Deep Eutectic Solvents (HDES), as a subclass of Natural Deep Eutectic Solvents (NADES), present a green-chemistry alternative to toxic chemicals. As HDES are based on terpenoids, these solvents could potentially be effective antifungal agents against phytopathogens Monilinia fructicola and Botrytis cinerea that frequently cause diseases in sweet cherry fruit. To contribute to the disease prevention and management goals, as a part of this study, 30 different HDES were tested in the vapor phase, at identical concentrations of 25%, 50%, and 100%. In vitro experiments were conducted on Potato Dextrose Agar medium (PDA), while in planta experiments were carried out in hermetically sealed containers with inoculated sweet cherry fruits. All tested HDES demonstrated efficacy in suppressing the growth of M. fructicola colonies (66 - 100%) and B. cinerea colonies (37 - 100%). According to the Area Under the Disease Progress Curve (AUDPC), all HDES exhibited high efficacy in preventing disease occurrence in cherry fruits by the tested phytopathogens. This research provides the first insights into the antifungal potential of HDES in the vapor phase, with promising applications as biofumigants that minimize harmful impacts on the food - human - environment complex.

Enhancing agrochemical delivery in citrus leaves with geraniol: a promising strategy for controlling huanglongbing (HLB).

Citrus huanglongbing (HLB) is a devastating disease in citrus, caused by Candidatus Liberibacter asiaticus (CLas), which primarily resides in the phloem where chemicals cannot effectively reach, posing a significant challenge in controlling HLB. To address these challenges, plant essential oils (EOs), widely used as transdermal enhancers and known for their benefits for plant tissues, were investigated for their potential to enhance chemical permeation. In this study, seven EOs - eugenol, carvacrol, eucalyptol, geraniol, linalool, cinnamaldehyde, and d-limonene - were evaluated for their potential to enhance chemical penetration into citrus leaves. Preliminary screening with tracer methods revealed that geraniol provided the greatest enhancement of permeation, achieving the deepest and broadest distribution into the spongy tissue of the citrus leaf. The permeation levels of rhodamine B and crystal violet through the cuticular layer were elevated from 0.55 mg L-1 and 0.11 mg L-1 to 20.04 mg L-1 and 7.14 mg L-1, respectively, by the addition of geraniol. Further validation through high-performance liquid chromatography-mass spectrometry (HPLC-MS) confirmed that the penetration of fludioxonil and tetracycline into citrus leaf tissues was enhanced by 15.07-fold and 20.43-fold, respectively. In planta experiments confirmed that adding geraniol to tetracycline and oxytetracycline via foliar application eliminated CLas bacteria in HLB-affected citrus more efficiently than controls. Our study underscores the potential of geraniol as an effective EO-based permeation enhancer for combating HLB. By significantly improving the delivery and efficacy of bactericidal treatments, geraniol offers a promising strategy for managing this catastrophic citrus disease, potentially transforming the approach to controlling HLB. © 2024 Society of Chemical Industry.

Phage biocontrol success of bacterial wilt depends on synergistic interactions with resident rhizosphere microbiota.

Phages can successfully be used invitro and in planta to biocontrol the phytopathogenic Ralstonia solanacearum bacterium-the causal agent of bacterial wilt disease. However, phage biocontrol outcomes are still variable, and it is unclear what causes this. In this study, we assessed the efficiency of four phages in controlled invitro and in planta experiments in all one- and two-phage combinations. We found that using phages in combination did not improve the phage biocontrol efficiency relative to single phage treatments, while certain phages and their combinations were more effective than the others. High intra-treatment variability in phage efficiency was observed across all phage treatments, which was associated with clear shifts in microbiome composition, a reduction in R. solanacearum and an increase in phage densities. We further identified the bacterial taxa that were associated with these 'shifted' microbiomes and conducted additional plant growth experiments, demonstrating that some of the enriched bacterial species could protect plants from R. solanacearum infections-a pattern which was also observed using partial least squares path modelling (PLS-PM). Together, these results suggest that phages could open niche space for beneficial bacteria by reducing pathogen densities and that variability in phage biocontrol outcomes is rhizosphere microbiome-dependent, which can introduce between-replicate variation, even in controlled greenhouse conditions.

An implementation framework for evaluating the biocidal potential of essential oils in controlling Fusarium wilt in spinach: from in vitro to in planta.

Fusarium wilt, caused by Fusarium oxysporum f. sp. spinaciae, causes a significant challenge on vegetative spinach and seed production. Addressing this issue necessitates continuous research focused on innovative treatments and protocols through comprehensive bioassays. Recent studies have highlighted the potential of plant-based compounds in controlling fungal diseases. The present work aims to conduct a series of experiments, encompassing both in vitro and in planta assessments, to investigate the biocontrol capabilities of different essential oils (EOs) at various application rates, with the ultimate goal of reducing the incidence of Fusarium wilt in spinach. The inhibitory effect of four plant EOs (marjoram, thyme, oregano, and tea tree) was initially assessed on the spore germination of five unknown Fusarium strains. The outcomes revealed diverse sensitivities of Fusarium strains to EOs, with thyme exhibiting the broadest inhibition, followed by oregano at the highest concentration (6.66 μL/mL) in most strains. The tested compounds displayed a diverse range of median effective dose (ED50) values (0.69 to 7.53 µL/mL), with thyme and oregano consistently showing lower ED50 values. The direct and indirect inhibitory impact of these compounds on Fusarium mycelial growth ranged from ~14% to ~100%, wherein thyme and oregano consistently exhibiting the highest effectiveness. Following the results of five distinct inoculation approaches and molecular identification, the highly pathogenic strain F-17536 (F. oxysporum f.sp. spinaciae) was chosen for Fusarium wilt assessment in spinach seedlings, employing two promising EO candidates through seed and soil treatments. Our findings indicate that colonized grain (CG) proved to be a convenient and optimal inoculation method for consistent Fusarium wilt assessment under greenhouse conditions. Seed treatments with thyme and oregano EOs consistently resulted in significantly better disease reduction rates, approximately 54% and 36% respectively, compared to soil treatments (P > 0.05). Notably, thyme, applied at 6.66 µL/mL, exhibited a favorable emergence rate (ERI), exceeding seven, in both treatments, emphasizing its potential for effective disease control in spinach seedlings without inducing phytotoxic effects. This study successfully transitions from in vitro to in planta experiments, highlighting the potential incorporation of EOs into integrated disease management for Fusarium wilt in spinach production.

A Promising Biocontrol Agent of Bacillus velezensis VC3 against Magnaporthe oryzae and Colletotrichum gloeosporioides in Plants

Fungal diseases of plants are one of the key factors causing global crop losses. In this study, we isolated a Bacillus velezensis strain VC3, which was found to have a broad-spectrum inhibitory effect on a variety of phytopathogenic fungi through in vitro and in planta experiments, especially on Magnaporthe oryzae and Colletotrichum gloeosporioides. Further genomic and transcriptomic analyses revealed that the B. velezensis VC3 has multiple functional gene clusters encoding for the synthesis of a variety of antifungal secondary metabolites, including antimicrobial peptides (AMPs) and lipopeptides (LPs). In addition, AMPs and LPs were isolated and purified from B. velezensis VC3 fermentation broth and their antifungal activities were verified in this study. AMPs and LPs significantly inhibited spore germination, appressorium formation, and disease development, and AMPs have a better potential for controlling M. oryzae and C. gloeosporioides than LPs. These findings open new avenues for utilizing B. velezensis VC3 as biocontrol agents, providing potential sustainable solutions for agricultural production.

Biostimulant and antagonistic potential of endophytic fungi against fusarium wilt pathogen of tomato Fusarium oxysporum f. sp. lycopersici

Endophytic fungal-based biopesticides are sustainable and ecologically-friendly biocontrol agents of several pests and diseases. However, their potential in managing tomato fusarium wilt disease (FWD) remains unexploited. This study therefore evaluated effectiveness of nine fungal isolates against tomato fusarium wilt pathogen, Fusarium oxysporum f. sp. lycopersici (FOL) in vitro using dual culture and co-culture assays. The efficacy of three potent endophytes that inhibited the pathogen in vitro was assessed against FWD incidence, severity, and ability to enhance growth and yield of tomatoes in planta. The ability of endophytically-colonized tomato (Solanum lycopersicum L.) plants to systemically defend themselves upon exposure to FOL were also assessed through defence genes expression using qPCR. In vitro assays showed that endophytes inhibited and suppressed FOL mycelial growth better than entomopathogenic fungi (EPF). Endophytes Trichoderma asperellum M2RT4, Hypocrea lixii F3ST1, Trichoderma harzianum KF2R41, and Trichoderma atroviride ICIPE 710 had the highest (68.84–99.61%) suppression and FOL radial growth inhibition rates compared to EPF which exhibited lowest (27.05–40.63%) inhibition rates. Endophytes T. asperellum M2RT4, H. lixii F3ST1 and T. harzianum KF2R41 colonized all tomato plant parts. During the in planta experiment, endophytically-colonized and FOL-infected tomato plants showed significant reduction of FWD incidence and severity compared to non-inoculated plants. In addition, these endophytes contributed to improved growth promotion parameters and yield. Moreover, there was significantly higher expression of tomato defence genes in T. asperellum M2RT4 colonized than in un-inoculated tomato plants. These findings demonstrated that H. lixii F3ST1 and T. asperellum M2RT4 are effective biocontrol agents against FWD and could sustainably mitigate tomato yield losses associated with fusarium wilt.

Anti-oomycete activity of asparagus and olive by-products with potential to control Phytophthora cinnamomi root rot

The development of environmentally friendly control methods to mitigate the severe damages caused by Phytophthora cinnamomi in the Mediterranean climate-type ecosystems is essential. In this way, crop waste and by-products which represent between 13 and 65% of agriculture production, are a rich source of bioactive compounds with antifungal and biocide activity. The main objective of this work was to determine the biocide activity against P. cinnamomi of three organic extracts. These extracts enriched in bioactive compounds come from residues of asparagus (Asp) and olive crops (Oliv and OH, from fruits and leaves respectively). They were evaluated at two doses (0.15 and 0.10%) on the mycelial growth and sporangial production of P. cinnamomi by in vitro experiments. Mycelial growth and sporangial production were significant reduced from the three plant extracts at the two doses tested, reaching a total inhibition with Asp at both doses. In general, no phytotoxicity symptoms were observed on seed germination and plant development, except for a plant yield reduction in the substrate treated with Oliv and Asp at the highest dose. In experiments performed in artificially infested soil, Asp induced a reduction of chlamydospores viability greater than 75% compared to unamended soil. Additionally, in planta experiments showed a significant reduction in plant mortality in substrate amended with OH. These results suggest that soil application of Asp and OH can limit P. cinnamomi infectivity and survival, setting the first steps to develop a sustainable method to control the root disease based on agricultural waste circular economy.

Rhexocercosporidium matricariae sp. nov.: A New Pathogen on the Medicinal Plant Matricaria recutita.

A new disease was observed on chamomile plants (Matricaria recutita) in various commercially cultivated fields in 2016 and 2017 in Germany. From symptomatic stems and leaves, the fungal species described here was isolated repeatedly. Koch's postulates using in planta experiments were fulfilled by inoculation of chamomile plants proving the fungus to be the causal agent of the disease. Morphological studies and phylogenetic analyses using internal transcribed spacer, large subunit, and translation elongation factor-1α sequences suggested that the fungus represents a new species within the genus Rhexocercosporidium (Helotiales). The data are presented together with a description of the growth parameters and comprehensive illustrations of the new species, Rhexocercosporidium matricariae. All species so far assigned to Rhexocercosporidium are compared and discussed. The combination Rhexocercosporidium microsporum is validated.

Nisin-based therapy: a realistic and eco-friendly biocontrol strategy to contrast Xylella fastidiosa subsp. pauca infections in planta.

The lack of sustainable strategies for combating Xylella fastidiosa (Xf) highlights the pressing need for novel practical antibacterial tools. In this study, Lactococcus lactis subsp. lactis strain ATCC 11454 (L. lactis), known for its production of nisin A, was in vitro tested against Xf subsp. pauca. Preliminary investigations showed that nisin A was involved in a strong antagonistic activity exhibited by L. lactis against Xf. Thus, the efficacy of nisin A was comprehensively assessed through a combination of in vitro and in planta experiments. In vitro investigations employing viable-quantitative PCR, spot assay, turbidity reduction assay, fluorescence microscopy, and transmission electron microscopy demonstrated nisin's robust bactericidal effect on Xf at a minimal lethal concentration of 0.6 mg/mL. Moreover, results from fluorescence and transmission electron microscopies indicated that nisin directly and rapidly interacts with the membranes of Xf cells, leading to the destruction of bacterial cells in few minutes. In in planta tests, nisin also demonstrated the ability to tackle Xf infections within Nicotiana benthamiana plants that remained asymptomatic 74 days post inoculation. Furthermore, RPLC-ESI-MS/MS analyses showed that nisin translocated to all parts of the plants and remains intact for up to 9 days. For the first time, this study underscores the nisin-based strategy as a realistic and eco-friendly approach to be further investigated against Xf infections in the field.

Evaluation of fungicides for management of Botryosphaeriaceae associated with dieback in Australian walnut orchards

Dieback of fruiting spurs, stems and branches of walnut trees (Juglans regia L.), caused by Botryosphaeriaceae, is widespread in walnut orchards in Australia. Five species of Botryosphaeriaceae (Diplodia seriata, Dothiorella omnivora, Neofusicoccum macroclavatum, N. parvum, and Spencermartinsia viticola) were recovered from the Australian walnut orchards in a previous study, with D. seriata and N. parvum being the most prevalent. The present study evaluated inhibitory effects of ten fungicides on mycelium growth of those five species and on conidium germination of D. seriata and N. parvum. It investigated the preventative and curative efficacy of selected fungicides on disease incidence in glasshouse and field trials. In vitro experiments showed that nine of the fungicides reduced mycelium growth, and all ten inhibited conidium germination, but to varying extents. Tebuconazole, prochloraz manganese chloride, fluazinam, fludioxonil and pyraclostrobin were the most effective for inhibiting mycelium growth (EC50 < 0.14 µg a.i. mL-1), whereas pyraclostrobin, fluxapyroxad, fluopyram, penthiopyrad and tebuconazole were the most effective for inhibiting conidium germination (EC50 < 2.2 µg a.i. mL-1). In planta experiments with five fungicides confirmed that preventative treatments had greater efficacy than curative treatments. A field trial with four commercial fungicide formulations demonstrated that tebuconazole and tebuconazole + fluopyram provided protection of walnut trees for the longest period. The field trial also confirmed the efficacy of pyraclostrobin and the inhibitory effect of fluazinam. This study is the first in Australia to evaluate fungicides in different classes and with different modes of action for efficacy against Botryosphaeriaceae recovered from walnut orchards in Australia, and provides a wider selection of active ingredients for a fungicide rotation programme than that which is currently available to the Australian walnut industry.

Enhancing Botrytis disease management in tomato plants: insights from a Pseudomonas putida strain with biocontrol activity.

This study explores the biocontrol potential of Pseudomonas putida Z13 against Botrytis cinerea in tomato plants, addressing challenges posed by the pathogen's fungicide resistance. The aims of the study were to investigate the in vitro and in silico biocontrol traits of Z13, identify its plant-colonizing efficacy, evaluate the efficacy of different application strategies against B. cinerea in planta, and assess the capacity of Z13 to trigger induced systemic resistance (ISR) in plants. The in vitro experiments revealed that Z13 inhibits the growth of B. cinerea, produces siderophores, and exhibits swimming and swarming activity. Additionally, the Z13 genome harbors genes that encode compounds triggering ISR, such as pyoverdine and pyrroloquinoline quinone. The in planta experiments demonstrated Z13's efficacy in effectively colonizing the rhizosphere and leaves of tomato plants. Therefore, three application strategies of Z13 were evaluated against B. cinerea: root drenching, foliar spray, and the combination of root drenching and foliar spray. It was demonstrated that the most effective treatment of Z13 against B. cinerea was the combination of root drenching and foliar spray. Transcriptomic analysis showed that Z13 upregulates the expression of the plant defense-related genes PR1 and PIN2 upon B. cinerea inoculation. The results of the study demonstrated that Z13 possesses significant biocontrol traits, such as the production of siderophores, resulting in significant plant protection against B. cinerea when applied as a single treatment to the rhizosphere or in combination with leaf spraying. Additionally, it was shown that Z13 root colonization primes plant defenses against the pathogen.

Cytochrome P450 CitCYP97B modulates carotenoid accumulation diversity by hydroxylating β-cryptoxanthin in Citrus

Carotenoids in plant foods provide health benefits by functioning as provitamin A. One of the vital provitamin A carotenoids, β-cryptoxanthin, is typically plentiful in citrus fruit. However, little is known about the genetic basis of β-cryptoxanthin accumulation in citrus. Here, we performed a widely targeted metabolomic analysis of 65 major carotenoids and carotenoid derivatives to characterize carotenoid accumulation in Citrus and determine the taxonomic profile of β-cryptoxanthin. We used data from 81 newly sequenced representative accessions and 69 previously sequenced Citrus cultivars to reveal the genetic basis of β-cryptoxanthin accumulation through a genome-wide association study. We identified a causal gene, CitCYP97B, which encodes a cytochrome P450 protein whose substrate and metabolic pathways in land plants were undetermined. We subsequently demonstrated that CitCYP97B functions as a novel monooxygenase that specifically hydroxylates the β-ring of β-cryptoxanthin in a heterologous expression system. In planta experiments provided further evidence that CitCYP97B negatively regulates β-cryptoxanthin content. Using the sequenced Citrus accessions, we found that two critical structural cis-element variations contribute to increased expression of CitCYP97B, thereby altering β-cryptoxanthin accumulation in fruit. Hybridization/introgression appear to have contributed to the prevalence of two cis-element variations in different Citrus types during citrus evolution. Overall, these findings extend our understanding of the regulation and diversity of carotenoid metabolism in fruit crops and provide a genetic target for production of β-cryptoxanthin-biofortified products.

Evaluation of Bacillus amyloliquefaciens as a biocontrol agent against oak decline disease in Quercus trees

Aim of study: This study aimed to investigate the biocontrol potential of Bacillus amyloliquefaciens against Phytophthora cinnamomi infection in Quercus suber (cork oak). Both in vitro and in planta experiments were conducted to assess the effectiveness of B. amyloliquefaciens as a biocontrol agent. Area of study: The microorganism strains, B. amyloliquefaciens and P. cinnamomi, as well as the embryogenic lines of Q. suber used, have a Spanish origin. Material and methods: In vitro experiments involved evaluating the inhibitory effects of B. amyloliquefaciens on P. cinnamomi growth through dual-inoculated agar plates. In planta, dual inoculation tests were performed by co-inoculating plantlets with both P. cinnamomi and B. amyloliquefaciens. Physiological parameters, such as photosynthetic activity, chlorophyll content, and oxidative stress markers, were measured. All experiments were conducted under controlled conditions. Main results: In vitro experiments revealed the inhibitory effects of B. amyloliquefaciens on P. cinnamomi growth. Infected plantlets displayed symptoms of root infection. Dual inoculation tests resulted in plant survival against P. cinnamomi infection. Analysis of physiological parameters indicated variations among treatments and clones, highlighting the distinct response of Q. suber plantlets to the pathogen and underscoring the importance of genetic variability for disease management. Research highlights: This study provides insights into the potential of a strain of B. amyloliquefaciens as a biocontrol agent against P. cinnamomi infection in cork oak. Further investigations are warranted to elucidate the underlying mechanisms of susceptibility and resistance in different clones of Q. suber.

Stromule Geometry Allows Optimal Spatial Regulation ofOrganelle Interactions inthe Quasi-2D Cytoplasm.

In plant cells, plastids form elongated extensions called stromules, the regulation and purposes of which remain unclear. Here, we quantitatively explore how different stromule structures serve to enhance the ability of a plastid to interact with other organelles: increasing the effective space for interaction and biomolecular exchange between organelles. Interestingly, electron microscopy and confocal imaging showed that the cytoplasm in Arabidopsis thaliana and Nicotiana benthamiana epidermal cells is extremely thin (around 100 nm in regions without organelles), meaning that inter-organelle interactions effectively take place in 2D. We combine these imaging modalities with mathematical modeling and new in planta experiments to demonstrate how different stromule varieties (single or multiple, linear or branching) could be employed to optimize different aspects of inter-organelle interaction capacity in this 2D space. We found that stromule formation and branching provide a proportionally higher benefit to interaction capacity in 2D than in 3D. Additionally, this benefit depends on optimal plastid spacing. We hypothesize that cells can promote the formation of different stromule architectures in the quasi-2D cytoplasm to optimize their interaction interface to meet specific requirements. These results provide new insight into the mechanisms underlying the transition from low to high stromule numbers, the consequences for interaction with smaller organelles, how plastid access and plastid to nucleus signaling are balanced and the impact of plastid density on organelle interaction.

Plant commensal type VII secretion system causes iron leakage from roots to promote colonization.

Competition for iron is an important factor for microbial niche establishment in the rhizosphere. Pathogenic and beneficial symbiotic bacteria use various secretion systems to interact with their hosts and acquire limited resources from the environment. Bacillus spp. are important plant commensals that encode a type VII secretion system (T7SS). However, the function of this secretion system in rhizobacteria-plant interactions is unclear. Here we use the beneficial rhizobacterium Bacillus velezensis SQR9 to show that the T7SS and the major secreted protein YukE are critical for root colonization. In planta experiments and liposome-based experiments demonstrate that secreted YukE inserts into the plant plasma membrane and causes root iron leakage in the early stage of inoculation. The increased availability of iron promotes root colonization by SQR9. Overall, our work reveals a previously undescribed role of the T7SS in a beneficial rhizobacterium to promote colonization and thus plant-microbe interactions.

Colonization and endophytic effect of Beauveria bassiana (Bals.-Criv.) Vuill. UHSB-END1 against Myzus persicae (Sulzer) and Plutella xylostella (L.) in cabbage

BackgroundFungal microbial biocontrol agent playing an important role in sustainable pest management with low cost. Conidial contact with cuticle is sufficient to cause disease in insects by entomopathogenic fungi (EPF) compared to bacteria and viruses which required ingestion of contaminated food. The field level application of fungi resulted inconsistent in their efficacy against insects as they withstand in the agroecosystem with adverse environmental conditions. To overcome this bottleneck, endophytic EPF, Beauveria bassiana (Bals.-Criv.) Vuill. (Cordycipitaceae: Hypocreales), is well-studied for the management of insects of cultivated crops and got promising results. The colonization of endophytic isolates varied from one host plant to another and virulence against insects too. Hence, the study was undertaken to know the colonization of indigenous B. bassiana UHSB-END1 isolate in cabbage plant and its efficacy against major insects of cabbage.ResultsThe indigenous isolate of B. bassiana UHSB-END1 was able to colonize cabbage in all the methods of inoculations (seed treatment, seedling root dip, soil drenching, foliar spray and combination treatment) at 30, 45 and 60 days of post-inoculation (dpi). However, the colonization of the fungus inside the cabbage was restricted to tissues of inoculation, and movement from the site of inoculation to other parts of the plant was poor. In the present study, colonization was the highest at 30 dpi in all methods of colonization with restricted colonization to the site of exposure of fungus. Further, in vivo and in planta assay confirmed the effectiveness of B. bassiana UHSB-END1 colonized cabbage against Myzus persicae (Sulzer) and Plutella xylostella (Linnaeus). Under in vivo conditions the maximum mortality of M. persicae and P. xylostella was recorded at 30 dpi. It was decreased slightly at 45 dpi, and the lowest morality was recorded at 60 dpi. In planta experiment also proved better efficacy against both the test insects. Wherein mortality of M. persicae ranged from 22 to 36% five days after release (DAR), it was increased to 48–68% at 10 DAR and reached highest mortality rate at 15DAR (72–96%). Similarly, mortality of P. xylostella ranged from 14 to 24% after five DAR, and it was 27–44% at 10 DAR, and maximum mortality rate of larvae were recorded at 15 DAR (64–96%).ConclusionThe present study reported 100% colonization of cabbage plant by indigenous B. bassiana UHSB-END1 at 30 dpi in combination of treatment and showed a maximum efficacy in managing the population of major insects of cabbage. Introducing this indigenous isolate will pave a ray of hope in managing the both key insects (P. xylostella and M. persicae) in cabbage agroecosystem without disturbing the environment and further helps in production of cabbage with minimum pesticide residue for consumers.

Synergistic Action of Biosynthesized Silver Nanoparticles and Culture Supernatant of Bacillus amyloliquefacience against the Soft Rot Pathogen Dickeya dadantii.

Nanomaterials are increasingly being used for crop growth, especially as a new paradigm for plant disease management. Among the other nanomaterials, silver nanoparticles (AgNPs) draw a great deal of attention because of their unique features and multiple usages. Rapid expansion in nanotechnology and utilization of AgNPs in a large range of areas resulted in the substantial release of these nanoparticles into the soil and water environment, causing concern for the safety of ecosystems and phytosanitary. In an attempt to find an effective control measure for sweet potato soft rot disease, the pathogen Dickeya dadantii was exposed to AgNPs, the cell-free culture supernatant (CFCS) of Bacillus amyloliquefaciens alone, and both in combination. AgNPs were synthesized using CFCS of Bacillus amyloliquefaciens strain A3. The green synthesized AgNPs exhibited a characteristic surface plasmon resonance peak at 410-420 nm. Electron microscopy and X-ray diffraction spectroscopy determined the nanocrystalline nature and 20-100 nm diameters of AgNPs. Release of metal Ag+ ion from biosynthesized AgNPs increases with time. AgNPs and CFCS of B. amyloliquefaciens alone exhibited antibacterial activity against the growth, biofilm formation, swimming motility, and virulence of strain A3. The antibacterial activities elevated with the elevation in AgNPs and CFCS concentration. Similar antibacterial activities against D. dadantii were obtained with AgNPs at 50 µg·mL-1, 50% CFCS alone, and the combination of AgNPs at 12 µg·mL-1 and 12% CFCS of B. amyloliquefaciens. In planta experiments indicated that all the treatments reduced D. dadantii infection and increased plant growth. These findings suggest that AgNPs along with CFCS of B. amyloliquefaciens can be applied to minimize this bacterial disease by controlling pathogen-contaminated sweet potato tuber with minimum Ag nano-pollutant in the environment.

Aggressiveness and mycotoxin profile of Fusarium avenaceum isolates causing Fusarium seedling blight and Fusarium head blight in UK malting barley.

Fusarium avenaceum causing Fusarium seedling blight (FSB) and Fusarium head blight (FHB) on barley is associated with economic losses of crop yield and quality, and the accumulation of mycotoxins including the enniatins (ENNs) A, A1, B and B1. Although F. avenaceum is the main producer of ENNs, studies on the ability of isolates to cause severe Fusarium diseases or produce mycotoxins in barley are limited. In this work, we investigated the aggressiveness of nine isolates of F. avenaceum to two cultivars of malting barley, Moonshine and Quench, and defined their ENN mycotoxin profiles in in vitro and in planta experiments. We assessed and compared the severity of FSB and FHB caused by these isolates to disease severity by F. graminearum, F. tricinctum and F. poae. Quantitative real-time polymerase chain reaction and Liquid Chromatography Tandem Mass Spectrometry assays were used to quantify pathogen DNA and mycotoxin accumulation, respectively, in barley heads. Isolates of F. avenaceum were equally aggressive to barley stems and heads and caused the most severe FSB symptoms resulting in up to 55% reductions of stem and root length. Fusarium graminearum caused the most severe FHB disease, followed by the isolates of F. avenaceum with the most aggressive F. avenaceum isolates capable of causing similar bleaching of barley heads as F. avenaceum. Fusarium avenaceum isolates produced ENN B as the predominant mycotoxin, followed by ENN B1 and A1 in vitro. However, only the most aggressive isolates produced ENN A1 in planta and none produced ENN A or beauvericin (BEA) either in planta or in vitro. The capacity of F. avenaceum isolates to produce ENNs was related to the accumulation of pathogen DNA in barley heads, whilst FHB severity was related to the synthesis and accumulation of ENN A1 in planta. Cv. Moonshine was significantly more resistant than Quench to FSB or FHB, caused by any Fusarium isolate, and to the accumulation of pathogen DNA, ENNs or BEA. In conclusion, aggressive F. avenaceum isolates are potent ENN producers causing severe FSB and FHB with ENN A1 requiring further investigation as potential virulence factor for F. avenaceum in cereals.

Rhizosphere bacteria associated with Chenopodium quinoa promote resistance to Alternaria alternata in tomato

Microorganisms can interact with plants to promote plant growth and act as biocontrol agents. Associations with plant growth-promoting rhizobacteria (PGPR) enhance agricultural productivity by improving plant nutrition and enhancing protection from pathogens. Microbial applications can be an ideal substitute for pesticides or fungicides, which can pollute the environment and reduce biological diversity. In this study, we isolated 68 bacterial strains from the root-adhering soil of quinoa (Chenopodium quinoa) seedlings. Bacterial strains exhibited several PGPR activities in vitro, including nutrient solubilization, production of lytic enzymes (cellulase, pectinase and amylase) and siderophore synthesis. These bacteria were further found to suppress the mycelial growth of the fungal pathogen Alternaria alternata. Nine bacterial strains were selected with substantial antagonistic activity and plant growth-promotion potential. These strains were identified based on their 16S rRNA gene sequences and selected for in planta experiments with tomato (Solanum lycopersicum) to estimate their growth-promotion and disease-suppression activity. Among the selected strains, B. licheniformis and B. pumilus most effectively promoted tomato plant growth, decreased disease severity caused by A. alternata infection by enhancing the activities of antioxidant defense enzymes and contributed to induced systemic resistance. This investigation provides evidence for the effectiveness and viability of PGPR application, particularly of B. licheniformis and B. pumilus in tomato, to promote plant growth and induce systemic resistance, making these bacteria promising candidates for biofertilizers and biocontrol agents.

Isolation and evaluation of indigenous endophytic entomopathogenic fungus, Beauveria bassiana UHSB-END1 (Hypocreales: Cordycipitaceae), against Spodoptera litura Fabricius

BackgroundMicrobial biological control agents are gaining worldwide attention to manage insect pests as an alternative to synthetic insecticides. Entomopathogenic fungi (EPF) meet eco-friendly pest management’s demand since mere contact of infective propagules is sufficient to cause disease in insect pests, unlike other entomopathogens. However, epiphytic fungal isolates encounter multiple challenges including direct exposure of conidia to sunlight and UV light, high temperature, and low moisture content that reduce their efficacy at the field level. Therefore, utilization of endophytic EPF is becoming more popular because they get protection from adverse conditions compared to the epiphytic EPF as they reside inside the host tissue. In addition, the endophytic EPF also give protection against crop diseases and promote plant growth, degradation of heavy metals, and tolerance to abiotic stress.ResultsThe increased mortality of Spodoptera litura (Fab.) (Noctuidae: Lepidoptera) was achieved through endophytic colonization of indigenous Beauveria bassiana UHSB-END-1 (OM131742). The bioassay proved the highest mortality 2nd instar larvae of S. litura at 40 dpi, both in vivo and in planta experiment. Further, larvae fed with fungal colonized leaves of tomato plant ended with abnormal growth and developmental process. The recovery of B. bassiana from different plant parts (stem, leaves, and roots) was the highest (100%) in all the methods of colonization at 14, 40, 60, 80 dpi, and it was decreased at 120 dpi (80%). The colonization rate was again increased in the next-generation seeds and seedlings (25 days old). This isolate gets vertically transmitted to their progenies via seeds, and it is the first report in tomato crop.ConclusionAfter ensuring the safeness of this isolate against non-target organisms, it can be one of the constitutes in sustainable cost-effective strategy for management of pests affecting tomato as one of the components in integrated pests management. Inoculation of endophytic EPF into seed/seedling reduces environmental impacts and also easy, economical, and sustainable approach for pest management in horticulture crops which are often consumed as raw. Although field studies are required to support the present finding, this appears to be an interesting tool that should be considered for pest biocontrol.