Global citrus root microbiota unravels assembly cues and core members.

Introduction: Citrus is one of the most important fruit crops worldwide, and the root-associated microbiota can have a profound impact on tree health and growth. Methods: In a collaborative effort, the International Citrus Microbiome Consortium investigated the global citrus root microbiota with samples collected from nine citrus-producing countries across six continents. We analyzed 16S rDNA and ITS2 amplicon sequencing data to identify predominant prokaryotic and fungal taxa in citrus root samples. Comparative analyses were conducted between root-associated microbial communities and those from the corresponding rhizosphere and bulk soil samples. Additionally, genotype-based group-wise comparisons were performed to assess the impact of citrus genotype on root microbiota composition. Results: Ten predominant prokaryotic phyla, containing nine bacterial phyla including Proteobacteria, Actinobacteria, Acidobacteria, and Bacteroidetes and one archaeal phylum (Thaumarchaeota), and multiple fungal phyla including Ascomycota and Basidiomycota were identified in the citrus root samples. Compared with the microbial communities from the corresponding rhizosphere and bulk soil samples from the same trees, the prokaryotic and fungal communities in the roots exhibited lower diversity and complexity but greater modularity compared to those in the rhizosphere. In total, 30 root-enriched and 150 root-depleted genera in bacterial community were identified, whereas 21 fungal genera were enriched, and 147 fungal genera were depleted in the root niche compared with the rhizosphere. The citrus genotype significantly affected the root prokaryotic and fungal communities. In addition, we have identified the core root prokaryotic genera comprising Acidibacter, Allorhizobium, Bradyrhizobium, Chitinophaga, Cupriavidus, Devosia, Dongia, Niastella, Pseudomonas, Sphingobium, Steroidobacter and Streptomyces, and the core fungal genera including Acrocalymma, Cladosporium, Fusarium, Gibberella, Mortierella, Neocosmospora and Volutella. The potential functions of these core genera of root microbiota were predicted. Conclusion: Overall, this study provides new insights into the assembly of microbial communities and identifies core members of citrus root microbiota across a wide geographic range. The findings offer valuable information for manipulating root microbiota to enhance plant growth and health.

Systematics and phylogeny of the entomopathogenic nematobacterial complexes Steinernema-Xenorhabdus and Heterorhabditis-Photorhabdus.

Entomopathogenic nematodes of the genera Steinernema and Heterorhabditis, along with their bacterial symbionts from the genera Xenorhabdus and Photorhabdus, respectively, are important biological control agents against agricultural pests. Rapid progress in the development of genomic tools has catalyzed a transformation of the systematics of these organisms, reshaping our understanding of their phylogenetic and cophlylogenetic relationships. In this review, we discuss the major historical events in the taxonomy and systematics of this group of organisms, highlighting the latest advancements in these fields. Additionally, we synthesize information on nematode-bacteria associations and assess the existing evidence regarding their cophylogenetic relationships.

Search for biocontrol agents among endophytic lipopeptide-synthesizing bacteria Bacillus spp. to protect wheat plants against Greenbug aphid (Schizaphis graminum).

Beneficial endophytic bacteria can suppress the development of insect pests through direct antagonism, with the help of metabolites, or indirectly by the induction of systemic resistance through the regulation of hormonal signaling pathways. Lipopeptides are bacterial metabolites that exhibit direct antagonistic activity against many organisms, including insects. Also, lipopeptides are able to trigger induced systemic resistance (ISR) in plants against harmful organisms, but the physiological mechanisms of their action are just beginning to be studied. In this work, we studied ten strains of bacteria isolated from the tissues of wheat and potatoes. Sequencing of the 16S rRNA gene showed that all isolates belong to the genus Bacillus and to two species, B. subtilis and B. velezensis. The genes for lipopeptide synthetase - surfactin synthetase (Bs_srf ), iturin synthetase (Bs_ituA, Bs_ituB) and fengycin synthetase (Bs_fenD) - were identified in all bacterial isolates using PCR. All strains had high aphicidal activity against the Greenbug aphid (Schizaphis graminum Rond.) due to the synthesis of lipopeptides, which was proven using lipopeptide-rich fractions (LRFs) isolated from the strains. Endophytic lipopeptide-synthesizing strains of Bacillus spp. indirectly affected the viability of aphids, the endurance of plants against aphids and triggered ISR in plants, which manifested itself in the regulation of oxidative metabolism and the accumulation of transcripts of the Pr1, Pr2, Pr3, Pr6 and Pr9 genes due to the synthesis of lipopeptides, which was proven using LRF isolated from three strains: B. subtilis 26D, B. subtilis 11VM, and B. thuringiensis B-6066. We have for the first time demonstrated the aphicidal effect of fengycin and the ability of the fengycin-synthesizing strains and isolates, B. subtilis Ttl2, Bacillus sp. Stl7 and B. thuringiensis B-6066, to regulate components of the pro-/antioxidant system of aphid-infested plants. In addition, this work is the first to demonstrate an elicitor role of fengycin in triggering a systemic resistance to S. graminum in wheat plants. We have discovered new promising strains and isolates of endophytes of the genus Bacillus, which may be included in the composition of new biocontrol agents against aphids. One of the criteria for searching for new bacteria active against phloem-feeding insects can be the presence of lipopeptide synthetase genes in the bacterial genome.

Integration of mathematical modeling and target-based application of biocontrol agents for the control of Botrytis cinerea in vineyards.

BACKGROUND: Biocontrol agents (BCAs) are alternatives to synthetic fungicides with low risk to the environment and human health. Although several studies on the biocontrol of gray mold in vineyards have been performed, it is necessary to improve the usage of BCAs in fields conditions. Therefore, in the present study, BCAs were used both in calendar-based [based on four growth stages (GSs), i.e., flowering, pre-bunch closure, veraison, and before harvest] and predictive model-based strategies (only when Botrytis cinerea infection risk was predicted by the model). The BCAs applied during the seasons were selected considering the grapevine GSs. Treatments performed with BCAs were compared with synthetic fungicide treatments and an untreated control. The trials were conducted in three experimental vineyards with four epidemics. To evaluate the level of gray mold control of each treatment, disease severity was assessed at harvest and the presence of latent infection was evaluated. RESULTS: The integrative use of the predictive model and BCAs provided satisfactory levels of gray mold control, with gray mold severity levels significantly lower (P < 0.001) than those of the untreated control, which had severity values (< 7%) similar to those observed with synthetic fungicides following both calendar and model-based strategies. CONCLUSIONS: The integrative use of the predictive model and BCAs represents a valid alternative to conventional methods of gray mold control in vineyards, with more than 75% reduction in fungicide usage. © 2024 Society of Chemical Industry.

An Integrated Pest Management Strategy Approach for the Management of the Stable Fly Stomoxys calcitrans (Diptera: Muscidae).

Stable flies, Stomoxys calcitrans, stand as formidable pests with a global impact, inflicting significant economic losses on the livestock sector. Larval development occurs in diverse substrates, including decomposing plant material and manure, while emerged adults pose a threat through blood-feeding on both animals and humans. Conventional chemical control methods, predominantly reliant on insecticides, not only pose environmental risks but also face challenges of resistance among stable fly populations. To address this pressing issue, we propose an integrated pest management (IPM) strategy for stable fly control. This approach involved a combination of sanitary-cultural practices, animal protection, the release of natural enemies targeting immature stages, and a specialized trapping system for adults. The Stomoxycc® trap, designed for mass trapping of adult Stomoxys, was employed alongside the release of the predatory mite Macrocheles robustulus and two wasp parasitoids, Spalangia cameroni and Muscidifurax raptor (under the commercial brands Biomite® and Biowasp®) on animal bedding as a key component of this IPM strategy. The implementation of this initiative has been undertaken at a significant sanctuary for donkeys and mules in western Spain. In this publication, we present the application and results of the IPM strategy utilized and provide insights into its use as a sustainable and environmentally friendly option for controlling stable fly populations.

Enhancing tomato disease resistance through endogenous antifungal proteins and introduced nematode-targeting dsRNA of biocontrol agent Bacillus velezensis HS-3.

BACKGROUND: As a type of biological control agent (BCA), Bacillus velezensis possesses the efficacy of inhibiting pathogenic microorganisms, promoting plant growth, and overcoming continuous cropping obstacles (CCOs). However, there is limited reporting on the optimization of the cultivation conditions for such biocontrol agents and their role as double-stranded RNA (dsRNA) delivery vectors. RESULTS: In this study, a Bacillus velezensis strain HS-3 was isolated from the root zone of tomato plants with in vitro anti-Botrytis cinerea activity. The investigation into active compounds revealed that HS-3 predominantly employs proteins with molecular weights greater than 3 kDa for its antifungal activity. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis identified various proteases and chitosanase, further suggesting that HS-3 most likely employs these enzymes to degrade fungal cell walls for its antifungal effect. To optimize the production of extracellular proteins, fermentation parameters for HS-3 were systematically optimized, leading to an optimized medium (OP-M). HS-3 cultured in OP-M demonstrated enhanced capacity to assist tomato plants in withstanding CCOs. However, the presence of excessive nematodes in diseased soil resulted in the disease severity index (DSI) remaining high. An RNA interference mechanism was further introduced to HS-3, targeting the nematode tyrosine phosphatase (TP) gene. Ultimately, HS-3 expressing dsRNA of TP in OP-M effectively assisted tomatoes in mitigating CCOs, reducing DSI to 2.2% and 17.8% of the control after 45 and 90 days of growth, respectively. CONCLUSION: The advantages of Bacillus velezensis in crop disease management and the mitigation of CCOs become even more pronounced when utilizing both optimized levels of endogenous enzymes and introduced nematode-targeting dsRNA. © 2024 Society of Chemical Industry.

Rhizosphere Microorganisms Supply Availability of Soil Nutrients and Induce Plant Defense.

Plant health is necessary for food security, which is a key determinant of secure and sustainable food production systems. Deficiency of soil nutrients and invasion of plant pathogens or insects are the main destroyers of the world's food production. Synthetic fertilizers and chemical-based pesticides are frequently employed to combat the problems. However, these have negative impacts on microbial ecosystems and ecosystem functioning. Rhizosphere microorganisms have demonstrated their potency to improve or manage plant nutrients to encourage plant growth, resulting in increased yield and quality by converting organic and inorganic substances around the rhizosphere zone into available plant nutrients. Besides regulating nutrient availability and plant growth enhancement, rhizobacteria or fungi can restrict plant pathogens that cause disease by secreting inhibitory chemicals and boosting plant immunity to combat pests or pathogens. Thus, rhizosphere microorganisms are viewed as viable, alluring economic approaches for sustainable agriculture as biofertilizers and biopesticides. This review provides an overview of the role of rhizosphere microorganisms in soil nutrients and inducing of plant defenses. Moreover, a discussion is presented surrounding the recent consequences of employing these microorganisms and a sustainable strategy towards improving fertilization effectiveness, and encouraging stronger, more pest-resistant plants.

On the Way to the Technological Development of Newly Selected Non-Saccharomyces Yeasts Selected as Innovative Biocontrol Agents in Table Grapes.

Post-harvest decay of fresh table grapes causes considerable annual production losses. The main fungal agents of decay both in pre- and post-harvest are B. cinerea, Penicillium spp., Aspergillus spp., Alternaria spp., and Cladosporium spp. To date, the use of agrochemicals and SO2 are the main methods to control grape molds in pre- and postharvest, respectively. Significant improvements, however, have already been made in to apply innovative and more environmentally sustainable control strategies, such as Biological Control Agents (BCAs), which can reduce disease severity in both pre- and post-harvest. In this study, 31 new non-Saccharomyces yeast strains, isolated from berries of native Apulian table grape genotypes, were tested for their in vivo effectiveness against grey mold of table grapes, resulting in two St. bacillaris ('N22_I1' and 'S13_I3'), one S. diversa ('N22_I3'), one A. pullulans ('OLB_9.1_VL') and one H. uvarum ('OLB_9.1_BR') yeast strains that were marked as efficient and good BCAs. Their mechanisms of action were characterized through in vitro assays, and additional characteristics were evaluated to assess the economic feasibility and viability for future technological employment. Their effectiveness was tested by reducing the working concentration, their antagonistic effect on a wide range of fungal pathogens, their ability to survive in formulations with long shelf life, and their safety to human health.

Biocontrol of almond canker diseases caused by Botryosphaeriaceae fungi.

BACKGROUND: Botryosphaeria dieback is a canker disease caused by fungal species of the Botryosphaeriaceae family that threatens almond productivity. The most common control measure to prevent canker development is the application of fungicides which are being phased out by European Union regulations. In the present study, two sets of bacterial strains were evaluated for their antifungal activity against pathogenic Botryosphaeriaceae species through in vitro and in vivo antagonism assays. RESULTS: The rhizospheric bacteria Pseudomonas aeruginosa AC17 and Bacillus velezensis ACH16, as well as the endophytic bacteria Bacillus mobilis Sol 1-2, respectively inhibited 87, 95, and 63% of the mycelial growth of Neofusicoccum parvum, Botryosphaeria dothidea, Diplodia seriata, and Macrophomina phaseolina. Additionally, they significantly reduced the length of lesions caused by N. parvum and B. dothidea in artificially inoculated detached almond twigs. All these bacterial strains produce hydrolytic enzymes that are able to degrade the fungal cell wall. P. aeruginosa AC17 also produces toxic volatile compounds, such as hydrogen cyanide. This strain was the most effective in controlling Botryosphaeria dieback in planta under controlled conditions at a level similar to the biocontrol agent Trichoderma atroviride and standard chemical fungicide treatments. CONCLUSION: Pseudomonas aeruginosa AC17 is the best candidate to be considered as a potential biocontrol agent against Botryosphaeriaceae fungi affecting almond. © 2023 Society of Chemical Industry.

The application of entomopathogenic nematode modified microbial communities within nesting mounds of the red imported fire ants, Solenopsis invicta.

Entomopathogenic microorganisms (e.g., fungi, bacteria, nematodes) have been widely used in biological control of soil-dwelling pests, including the red imported fire ant (RIFA), Solenopsis invicta, a notorious invasive pest worldwide. The application of large amounts of entomopathogenic microorganisms to soil may affect the indigenous soil microbial communities. However, reports about the effect of entomopathogenic nematodes (EPN) on soil microbial communities are very few. In this study, the effects of EPN on RIFA populations and microbial communities in mounds were investigated. Our results showed that the application of the EPN Steinernema carpocapsae. All strain on mounds efficaciously suppressed RIFA worker populations, without forming significantly more satellite mounds compared with the control treatment. The application of EPN did not impact the bacterial and fungal diversity in soils derived from the RIFA mounds. However, it slightly altered the taxonomic make-up of the bacterial communities, but significantly altered the taxonomic composition of fungal communities at the phylum, family, and genus levels. The abundances of some beneficial bacteria and fungi, such as Streptomyces, decreased, while those of plant and animal pathogenic bacteria and fungi, dramatically increased, after EPN treatment. On the other hand, the abundances of some entomopathogenic fungi, such as Fusicolla, Clonostachys, and Mortierella, increased. Redundancy analysis or canonical correspondence analysis revealed a positive correlation between the efficacious EPN control and the presence of the insect-resistant bacteria, Sinomonas, as well as entomopathogenic fungi Fusicolla and Mortierella. This suggests that the interactions between EPN and entomopathogenic fungi may play a role in the biological control of RIFA. Our discoveries shed light on the interactions among EPN, RIFA, and soil microbial communities, and emphasize a possible mutualistic relationship between EPN and entomopathogenic fungi in the biological control of RIFA.

Diversity of parasitoid wasps (Insecta, Hymenoptera) in oilseed rape fields in Serbia.

Background: Oilseed rape is an important crop grown worldwide and used for various purposes, including oil extraction and animal feed. In Europe, there are six major pest species and several other minor pests that can significantly affect oilseed rape production, requiring growers to effectively control them in order to ensure crop yield. The host-parasitoid complexes of these pests have been studied in detail and recorded mainly in western, central and northern Europe. As an abundant source of pollen and nectar, oilseed rape may also be attractive to other parasitoids that do not have direct trophic interactions with oilseed rape pest species. The aim of this study is to fill the knowledge gap regarding the wider parasitoid community in oilseed rape fields, particularly in southern Europe. New information: During the two-year study, a total of 3135 specimens of primary and secondary parasitoids were sampled, of which 2855 were found in oilseed rape fields and 280 in semi-natural habitats. We found 153 taxa, of which 119 were found in oilseed rape fields and 87 in semi-natural habitats. We identified 31 genera (33 species) as parasitoids of oilseed rape pests, 54 genera (97 species) parasitising non-pest species and 10 genera (23 species) as possible parasitoids of oilseed rape pests. This study shows that the parasitoid community in oilseed rape fields is very diverse and that includes parasitoids of both oilseed rape pest and non-pest species.

Microbiome-Mediated Protection against Pathogens in Woody Plants.

Pathogens, especially invasive species, have caused significant global ecological, economic, and social losses in forests. Plant disease research has traditionally focused on direct interactions between plants and pathogens in an appropriate environment. However, recent research indicates that the microbiome can interact with the plant host and pathogens to modulate plant resistance or pathogen pathogenicity, thereby altering the outcome of plant-pathogen interactions. Thus, this presents new opportunities for studying the microbial management of forest diseases. Compared to parallel studies on human and crop microbiomes, research into the forest tree microbiome and its critical role in forest disease progression has lagged. The rapid development of microbiome sequencing and analysis technologies has resulted in the rapid accumulation of a large body of evidence regarding the association between forest microbiomes and diseases. These data will aid the development of innovative, effective, and environmentally sustainable methods for the microbial management of forest diseases. Herein, we summarize the most recent findings on the dynamic structure and composition of forest tree microbiomes in belowground and aboveground plant tissues (i.e., rhizosphere, endosphere, and phyllosphere), as well as their pleiotropic impact on plant immunity and pathogen pathogenicity, highlighting representative examples of biological control agents used to modulate relevant tree microbiomes. Lastly, we discuss the potential application of forest tree microbiomes in disease control as well as their future prospects and challenges.

Biocontrol of Botrytis cinerea as Influenced by Grapevine Growth Stages and Environmental Conditions.

The growth of four commercial biocontrol agents (BCAs: Bacillus amyloliquefaciens (BAD), Aureobasidium pullulans (APD), Metschnikowia fructicola (MFN), and Trichoderma atroviride (TAS)) was evaluated using turbidimetric assays on artificial substrates mimicking the chemical berry composition at four stages: pea-sized berries, veraison, softening, and ripe berries. The response of BCA growth differed among BCAs. Subsequently, the BCAs' population size was assessed after 1 to 13 days of incubation on the substrate mimicking ripe berries at 15 to 35 °C. The population size of BAD increased with temperatures, while that of MFN decreased; the population sizes of APD and TAS showed bell-shaped patterns with lower growth at 15 or 35 °C. Finally, the BCAs were applied to ripe berries and then incubated at 15 to 30 °C. After 1 to 13 days, the berries were inoculated with B. cinerea and incubated for 7 days, after which the BCA control efficacy was assessed. The highest control was observed at 25 °C for BAD and APD, at 15 to 20 °C for MFN, and at 25 to 30 °C for TAS. The results confirm that the plant substrates and temperature affect the population size of the BCA following application; temperature also affects the preventative efficacy of BCA against B. cinerea.

Predators as biocontrol agents of mosquito larvae in small and large habitats in Chiang Mai, Thailand.

Controlling mosquito-borne disease is a major global challenge due to the rise of insecticide-resistant mosquitoes. In response, we conducted a study in Chiang Mai Province, Thailand, which is one of the largest and the most popular cities for tourists in Southeast Asia, to explore the potential of local species as biological control agents for mosquito larvae. Mosquito larvae and aquatic predators were sampled from large and small habitats, while relevant physico-chemical parameters were measured. The study identified 560 predators and 1,572 mosquitoes, with most mosquito species belonging to the genus Culex. Additionally, the study identified 16 predator taxa, including four fish taxa and 12 taxa of predatory aquatic insects belonging to four orders: Coleoptera, Hemiptera, Odonata, and Diptera. The study found that several locally occurring predator species, namely Poecillia, Laccophilus, Lutzia, Toxorhynchites splendens, Agrionoptera, and Pseudarion, shared habitats with mosquitoes, indicating their potential as effective biological control agents for mosquito control. Conductivity, dissolved oxygen, and pH were the important physico-chemical parameters that affect both predators and mosquito larvae. Consequently, promoting native predators and reducing mosquito larvae through habitat management would be a sustainable and ecologically friendly approach in large habitats where it is not possible to remove mosquito oviposition sites. In smaller habitats, releasing local aquatic predators and removing oviposition sites may be a suitable strategy.

Stenotrophomonas rhizophila Ep2.2 inhibits growth of Botrytis cinerea through the emission of volatile organic compounds, restricts leaf infection and primes defense genes.

The bacterium Stenotrophomonas rhizophila is known to be beneficial for plants and has been frequently isolated from the rhizosphere of crops. In the present work, we isolated from the phyllosphere of an ornamental plant an epiphytic strain of S. rhizophila that we named Ep2.2 and investigated its possible application in crop protection. Compared to S. maltophilia LMG 958, a well-known plant beneficial species which behaves as opportunistic human pathogen, S. rhizophila Ep2.2 showed distinctive features, such as different motility, a generally reduced capacity to use carbon sources, a greater sensitivity to fusidic acid and potassium tellurite, and the inability to grow at the human body temperature. S. rhizophila Ep2.2 was able to inhibit in vitro growth of the plant pathogenic fungi Alternaria alternata and Botrytis cinerea through the emission of volatile compounds. Simultaneous PTR-MS and GC-MS analyses revealed the emission, by S. rhizophila Ep2.2, of volatile organic compounds (VOCs) with well-documented antifungal activity, such as furans, sulphur-containing compounds and terpenes. When sprayed on tomato leaves and plants, S. rhizophila Ep2.2 was able to restrict B. cinerea infection and to prime the expression of Pti5, GluA and PR1 plant defense genes.

Taxonomic and molecular characterization of a new entomopathogenic nematode species, Heterorhabditis casmirica n. sp., and whole genome sequencing of its associated bacterial symbiont.

BACKGROUND: Nematodes of the genus Heterorhabditis are important biocontrol agents as they form a lethal combination with their symbiotic Photorhabdus bacteria against agricultural insect pests. This study describes a new species of Heterorhabditis. METHODS: Six Heterorhabditis nematode populations were recovered from agricultural soils in Jammu and Kashmir, India. An initial examination using mitochondrial and nuclear genes showed that they belong to a new species. To describe this new species, a variety of analyses were conducted, including reconstructing phylogenetic relationships based on multiple genes, characterizing the nematodes at the morphological and morphometric levels, performing self-crossing and cross-hybridization experiments, and isolating and characterizing their symbiotic bacteria. RESULTS: The newly discovered species, Heterorhabditis casmirica n. sp., shares 94% mitochondrial cytochrome C oxidase subunit I gene (COI) sequence identity with Heterorhabditis bacteriophora and Heterorhabditis ruandica, and 93% with Heterorhabditis zacatecana. Morphologically, it differs from H. bacteriophora in its infective juvenile phasmids (present vs. inconspicuous) and bacterial pouch visibility in the ventricular portion of the intestine (invisible vs. visible); genital papilla 1 (GP1) position (at manubrium level vs. more anterior), and in its b ratio (body length/neck length), c ratio (tail length/bulb width), and D% [(excretory pore/neck length) × 100]. Other morphological differences include anterior end to the nerve ring distance (77-100 vs. 121-130 µm), V% [(anterior end of vulva/body length) × 100] (46-57 vs. 41-47) in hermaphroditic females; rectum size (slightly longer than the anal body diameter vs. about three times longer), phasmids (smaller vs. inconspicuous), body length (0.13-2.0 vs. 0.32-0.39 mm), body diameter (73-150 vs. 160-220 µm), anterior end to the excretory pore distance (135-157 vs. 174-214 µm), and demanian ratios in amphimictic females. Morphological differences with H. ruandica and H. zacatecana were also observed. Furthermore, H. casmirica n. sp. did not mate or produce fertile progeny with other Heterorhabditis nematodes reported from India. It was also discovered that H. casmirica n. sp. is associated with Photorhabdus luminescence subsp. clarkei symbiotic bacteria. CONCLUSIONS: The discovery of H. casmirica n. sp. provides novel insights into the diversity and evolution of Heterorhabditis nematodes and their symbiotic bacteria. This new species adds to the catalog of entomopathogenic nematodes in India.

Hyperspectral Imaging and Selected Biological Control Agents for the Management of Fusarium Head Blight in Spring Wheat.

Fusarium spp. are important pathogens on cereals, capable of causing considerable yield losses and significantly reducing the quality of harvested grains due to contamination with mycotoxins. The European Union intends to reduce the use of chemical-synthetic plant protection products (csPPP) by up to 50% by the year 2030. To realize this endeavor without significant economic losses for farmers, it is crucial to have both precise early detection of pathogens and effective alternatives for csPPP. To investigate both the early detection of Fusarium head blight (FHB) and the efficacy of selected biological control agents (BCAs), a pot experiment with spring wheat (cv. 'Servus') was conducted under semi-field conditions. Spikes were sprayed with different BCAs prior to inoculation with a mixture of F. graminearum and F. culmorum conidia. While early detection of FHB was investigated by hyperspectral imaging (HSI), the efficiency of the fungal (Trichoderma sp. T10, T. harzianum T16, T. asperellum T23 and Clonostachys rosea CRP1104) and bacterial (Bacillus subtilis HG77 and Pseudomonas fluorescens G308) BCAs was assessed by visual monitoring. Evaluation of the hyperspectral images using linear discriminant analysis (LDA) resulted in a pathogen detection nine days post inoculation (dpi) with the pathogen, and thus four days before the first symptoms could be visually detected. Furthermore, support vector machines (SVM) and a combination of LDA and distance classifier (DC) were also able to detect FHB symptoms earlier than manual rating. Scoring the spikes at 13 and 17 dpi with the pathogen showed no significant differences in the FHB incidence among the treatments. Nevertheless, there is a trend suggesting that all BCAs exhibit a diminishing effect against FHB, with fungal isolates demonstrating greater efficacy compared to bacterial ones.

Evaluation of Trichoderma spp. on Fusarium oxysporum f. sp. asparagi and Fusarium wilt Control in Asparagus Crop.

Among the key diseases affecting the asparagus crop (Asparagus officinalis L.), vascular wilting of asparagus caused by Fusarium oxysporum f. sp. asparagi stands out worldwide. This disease significantly shortens the longevity of the crop and limits economic production. Traditional control measures have been largely ineffective, and chemical control methods are difficult to apply, making biological control approaches, specifically the use of Trichoderma, an economical, effective, and risk-free alternative. This study aimed to identify the main factors that affect the efficacy of biopesticides studied as Biological Control Agents (BCAs) against Fusarium wilt in asparagus and to assess the efficacy of Trichoderma-based biopesticides under greenhouse and semi-field conditions. We evaluated the response of three Trichoderma spp. (T. atroviride, T. asperellum, and T. saturnisporum) to environmental variables, such as temperature and water activity, and their antagonistic capacity against Fusarium oxysporum f. sp. asparagi. All three Trichoderma species inhibited the growth of the pathogen in vitro. A decrease in water activity led to a greater reduction in the growth rate. The efficacy of the three biological control agents decreased with higher temperatures, resulting in minimal inhibition, particularly under conditions of restricted available water in the environment. The effect of the fungal inoculum density was also analyzed at two different temperatures. A direct correlation between the amount of inoculum and the score on the Disease Severity Index (DSI) was observed. A notable reduction in DSI was evident in treatments with high inoculum density (106 conidium/mL) for all three species of Trichoderma tested at both temperatures. In greenhouse and semi-field tests, we observed less disease control than expected, although T. asperellum and T. atroviride showed lower disease severity indices and increased the dry weight of seedlings and crowns, whereas T. saturnisporum resulted in the highest disease rate and lowest dry weight. This work highlights that the efficacy of Trichoderma as BCAs is influenced by various factors, including the quantity of soil inocula, and environmental conditions. The study findings have strong implications for selecting appropriate Trichoderma species for controlling specific pathogens under specific environmental conditions.

Toxicity of fungal-derived volatile organic compounds against root-knot nematodes.

BACKGROUND: Root-knot nematodes (RKNs), including Meloidogyne species, are among the most destructive plant-parasites worldwide. Recent evidence suggests that entomopathogenic fungi (EPF) can antagonize RKNs. Such antagonistic effects are likely mediated by toxic metabolites, including volatile organic compounds (VOCs), produced by the fungi. However, how widespread these effects are across EPF species, and which VOCs mediate negative interactions between EPF and RKNs needs to be further elucidated. RESULTS: First, we evaluated the nematicidal effect of VOCs emitted by 46 EPF isolates against Meloidogyne incognita and found variable toxicity depending on the isolate. Second, we measured the nematicidal effect of highly toxic isolates, including species in the genus Talaromyces, Aspergillus, Clonostachys, and Purpureocillium and, third, we analyzed the nematicidal effect of major VOCs, including 2-methyl-1-propanol, 3-methyl-1-butanol, isopropyl alcohol and 2-methyl-3-pentanone. The mortality of M. incognita juveniles (J2s) was generally high (50%) either via airborne or in-solution contact with VOCs. Moreover, the tested VOCs significantly inhibited egg hatching, and repelled J2s away from the VOCs. CONCLUSION: This study not only provides insights into the ecological function of VOCs in the rhizosphere, but also provides new approaches for developing environmentally friendly control methods of RKNs in agroecosystems. © 2023 Society of Chemical Industry.

Establishment of a rapid detection method for plutella xylostella granulovirus based on qPCR.

Plutella xylostella granulovirus (PlxyGV) biopesticide is an effective tool to control the long-term damage of Plutella xylostella (Linnaeus) to cruciferous vegetables. In China, PlxyGV can be produced on a large scale using host insects, and its products have been registered in 2008. In experiments and biopesticide production, the routine enumeration method of PlxyGV virus particles is to use the Petroff-Hausser counting chamber in dark field microscope. However, the accuracy and repeatability of granulovirus (GV) counting are affected due to the small particle size of GV occlusion bodies (OBs), the limitations of optical microscope, the judgment of different operators, host impurities, the addition of biological products. This limits the convenience of its production, product quality, trading and field application. Here we use PlxyGV as an example, the method based on Real-time fluorescence quantitative PCR (qPCR) was optimized from two aspects of sample treatment and specific primers design, which improved the repeatability and accuracy of absolute quantitative OBs of GV. This study provides basic information for accurate quantitative PlxyGV by qPCR method.