Biocontrol Research Articles

Impact of Zinnia elegans Cultivation on the Control Efficacy and Distribution of Aphidius colemani Viereck (Hymenoptera: Braconidae) against Aphis gossypii Glover (Hemiptera: Aphididae) in Cucumber Greenhouses.

This study aimed to evaluate the enhancement of A. gossypii control by A. colemani when Z. elegans was planted as a companion crop in cucumber greenhouses. The density and spatial distribution of A. gossypii and parasitized mummies were investigated across three treatment plots: (1) the simultaneous application of A. colemani and cultivation of Z. elegans (parasitoid-zinnia plot); (2) the application of A. colemani alone (parasitoid plot); and (3) a control plot (no application of both). A. gossypii maintained low densities in the parasitoid-zinnia plots, while its densities in the parasitoid plots initially decreased but rapidly increased thereafter. The spatial distribution patterns of A. gossypii and parasitized mummies showed similar trends across treatments. However, the parasitism rate of A. gossypii exhibited random distribution in parasitoid and control plots, while showing uniform distribution in the parasitoid-zinnia treatment. These results supported the idea that cultivating Z. elegans alongside cucumber could enhance the effectiveness of A. colemani as a biocontrol agent against A. gossypii, highlighting the importance of such companion planting in pest management strategies.

Evaluation of Bioagents against Stemphylium vesicarium Inducing Stemphylium Blight in Onions (Allium cepa L.)

Stemphylium blight of onion incited by S. vesicarium is one of the economically important diseases, causing severe yield losses in onion crop. The present investigation was carried out to evaluate efficacy of different bioagents against S. vesicarium under in vitro conditions. The experiment was laid out in a Completely Randomized Design (CRD) with three replications per treatment. Five fungal biocontrol agents viz., Trichoderma harzianum, T. viride, T. hamatum, T. virens and a fungal consortium from MPKV, Rahuri along with two bacterial biocontrol agents namely Pseudomonas fluorescens and Bacillus subtilis were evaluated for their effectiveness against S. vesicarium. The results showed significant variation in the inhibitory effects of bioagents on the mycelial growth of the test pathogen with percent growth inhibition ranging between 53.70 to 85.19 per cent. T. harzianum was reported to be most effective, showing a minimum colony diameter of 13.33 mm with the highest growth inhibition (85.19%) thus reported as superior to all other treatments whereas, Pseudomonas fluorescens was reported with a least percent growth inhibition (53.70%).

Assessment of Trichoderma Species Isolated From Volcanic Soil of a Durian Field in Sisaket Province, Thailand for Plant Growth Promotion and Biocontrol Potential

Trichoderma species are ubiquitous saprophytic fungi commonly found in soil, with potential as fungal biocontrol agents for plant disease control and growth promotion. This research aimed to assess the attributes of Trichoderma sp. as plant growth promoters, focusing on nitrogen fixation, siderophore production, plant nutrient solubilization and indole-3-acetic acid production. Out of 19 isolates tested, 6 (T05, T19, T25, T27, T28 and T33) showed superior plant growth promotion abilities. These isolates were further evaluated for their capacity to enhance the germination of Khao Dawk Mali 105 rice seeds. Seeds were soaked in a suspension of conidia at a concentration of 1×1012 conidia/mL, and germination rates were measured. The germination rate ranged between 77.67 - 86.67 %, statistically significantly higher than the control rate of 33.33 %. Among the isolates, T05 exhibited the highest promotion of plant growth, evidenced by root length (7.27 cm), shoot length (8.50 cm), fresh weight (0.54 g) and dry weight (0.27 g), all significantly different from the control. Additionally, the potential of isolates to inhibit the fungal pathogen Colletotrichum sp. MN-3 was assessed using a dual culture method, with T35 showing the highest inhibition percentage (60.90 %). Morphological classification and nucleotide sequencing of the ITS1-5.8S-ITS2 region of rDNA gene identified the isolates as T. harzianum, T. reesei, T. asperellum and T. longibrachiatum. These findings underscore the effectiveness of Trichoderma sp. in promoting plant growth and suppressing plant pathogenic fungi. HIGHLIGHTSSix Trichoderma isolates (T05, T19, T25, T27, T28 and T33) exhibited superior plant growth promotion abilities. These isolates significantly enhanced germination rates of Khao Dawk Mali 105 rice seeds compared to the method. Among the isolates, T05 showed the greatest promotion of plant growth, as evidenced by root length, shoot length, fresh weight and dry weight. Trichoderma isolates demonstrated potential for inhibiting the fungal pathogen Colletotrichum sp. MN-3. Morphological classification divided the Trichoderma sp. into 4 groups, and species identification through nucleotide sequencing revealed T. harzianum, T. reesei, T. asperellum and T. longibrachiatum. GRAPHICAL ABSTRACT

Occurrence, distribution and description of Harmonia dimidiata (Fabricius), a predacious Coccinellid from West Bengal, India

Aim: To provide information about the occurrence and distribution of Harmonia dimidiata from West Bengal and facilitate the identification of species based on morpho-taxonomy. Methodology: Survey was done in different agro-climatic zones of West Bengal. Specimens were procured from their associated host plants, brought to the laboratory and investigated for taxonomic characters. Results: The morpho-taxonomy of H. dimidiata, as well as its host record, and distribution, were fully explained in the present investigation. It is most common in hill, terai, and old alluvial zone of West Bengal. They have been observed preying on Rhopalosiphum maidis (Fitch) in corn and Aphis gossypii (Glover). Interpretation: H. dimidiata population were recorded during survey from four different agro-climatic zones of West Bengal. They were observed to be preyed upon A. gossypii and R. maidis. The information about the biodiversity of H. dimidiata and their role as predator of numerous agriculturally important crop pests are extremely valuable that enrich our knowledge and immensely help in devising a pest management programme considering them as an important biocontrol agent. Furthermore, research on their biodiversity aids in learning about the predaceous behaviour in connection with the pest and crop populations, their reproduction and occurrence. This information is indeed praiseworthy while exploiting the ladybird species, H. dimidiata for a commercial biocontrol operation. Key words: Coleoptera, Coccinellidae, Harmonia dimidiata, Predaceous, West Bengal

Endophyte Bacillus vallismortis BL01 to Control Fungal and Bacterial Phytopathogens of Tomato (Solanum lycopersicum L.) Plants

Some strains of Bacillus vallismortis have been reported to be efficient biocontrol agents against tomato pathogens. The aim of our study was to assess the biocontrol ability of the endophytic strain BL01 Bacillus vallismortis through in vitro and field trials, as well as to verify its plant colonization ability and analyze the bacterial genome in order to find genes responsible for the biocontrol activity. We demonstrated in a gnotobiotic system and by confocal laser microscopy that the endophytic strain BL01 was able to colonize the endosphere and rhizosphere of tomato, winter wheat and oilseed rape. In vitro experiments demonstrated the inhibition activity of BL01 against a wide range of phytopathogenic fungi and bacteria. BL01 showed biological efficacy in two-year field experiments with tomato plants against black bacterial spotting by 40–70.8% and against late blight by 47.1% and increased tomato harvest by 24.9% or 10.9 tons per hectare compared to the control. Genome analysis revealed the presence of genes that are responsible for the synthesis of biologically active secondary metabolites, which could be responsible for the biocontrol action. Strain BL01 B. vallismortis can be considered an effective biocontrol agent to control both fungal and bacterial diseases in tomato plants.

Bacillus amyloliquefaciens strain BaNCT02: An antagonist with multiple mechanisms of action against Meloidogyne incognita

AbstractBacillus species are among the most studied and commercially exploited biocontrol agents of plant‐parasitic nematodes. These antagonists may control nematodes by producing toxic or repellent substances, inducing systemic resistance in plants and disrupting nematode chemotaxis. Understanding the mechanism of action of antagonists increases the likelihood of success for microbiological nematicides under field conditions. Therefore, the objective of this work was to elucidate the mechanisms of action of Bacillus amyloliquefaciens strain BaNCT02 against Meloidogyne incognita. We assessed the potential of this strain for forming biofilm in roots and producing volatile and non‐volatile toxic substances. BaNCT02 did not induce systemic resistance in tomato plants against M. incognita. Scanning electron microscopy and chemotaxis analyses revealed that the bacteria form a biofilm on soybean roots and 67% of second‐stage juveniles of M. incognita are repelled by common bean roots treated with the antagonist. Additionally, BaNCT02 produces substances with toxic effects against eggs and juveniles of M. incognita, including volatile organic compounds such as 2‐undecanone and 2‐heptanone, as well as non‐volatile metabolites like proteases and chitinases. The suppressive effect of these substances on hatching and mortality of juveniles varied from approximately 70% to more than 90%. We concluded that B. amyloliquefaciens BaNCT02 forms a biofilm on root surfaces and produces volatile and non‐volatile metabolites with nematicidal and repellent effects against M. incognita.

A potential biocontrol and growth-promoting agent Streptomyces luteoverticillatus B4 for managing cabbage Fusarium wilt and cucumber Alternaria leaf blight

Cabbage Fusarium wilt and cucumber Alternaria leaf blight are two widespread diseases caused by Fusarium oxysporum and Alternaria cucumerina that severely affect cabbage and cucumber output across the world. However, very few biocontrol actinomycete resources have been identified for these two diseases. An antagonistic actinomycete B4 was isolated and identified as Streptomyces luteoverticillatus. It is effective against a wide range of plant pathogens. The purpose of this study was to look into the inhibitory impact of S. luteoverticillatus B4 on F. oxysporum and A. cucumerina, as well as to create the theoretical groundwork and research technologies for the use of B4 as a biopesticide. It was found that B4 has the ability to detoxify organic phosphorus and produce siderophores, IAA, and glucoamylase. In the greenhouse pot test, B4 had a control efficiency of 56 % against F. oxysporum and 76 % against A. cucumerina. Further investigation into the inhibitory mechanism indicated that once the B4 fermentation broth operated on the two pathogens, their mycelial morphology was altered, and mycelial growth was suppressed, particularly for F. oxysporum. B4 could create volatile organic compounds that may have an inhibiting impact. In addition, B4 fermentation broth dramatically increased seed germination and seedling development in both cabbage and cucumber. S. luteoverticillatus B4 is a possible biocontrol agent that inhibits F. oxysporum and A. cucumerina while also encouraging cabbage and cucumber development.

Mitigating root rot in Panax notoginseng: The synergistic effects of biochar and Chaetomium globosum YIM PH30719

Root rot is the main soilborne disease that causes the replant failure of Panax notoginseng, a famous medicinal plant in Asia. Biochar and biocontrol agents have been shown to reduce replant failure separately; however, a knowledge gap still exists regarding the effectiveness and mechanisms of their combined application on the root rot incidence of P. notoginseng. In this study, we conducted a pot experiment using a randomized block design, biochar derived from tobacco stems was applied at 0, 1, 2, and 3 % (T0, T1, T2, T3) to the soil that cultivating P. notoginseng for 10 years continuously; moreover, Chaetomium globosum YIM PH30719, a newly screened biocontrol strain, was applied at 0, 1.0×107, and 1.0×1010 cfu·mL−1 (C0, C1, C2) to the soil. Six months post-transplantation of P. notoginseng seedlings, root rot incidence, soil physicochemical properties, fungal communities, and abundance of the primary pathogen Fusarium oxysporum were investigated. The results showed that, comparing with the single application of YIM PH30719, the combined application with biochar was more efficient in reducing the root rot incidence (P<0.05). P. notoginseng experienced a root rot incidence of 65.4 % under the T0+C0 treatment, however, under T3+C2 treatment, the root rot incidence was only 5.6 % (P<0.05). The reduction in root rot incidence might be linked to changes in soil pH, organic matter, and essential nutrient contents, as well as the increased soil fungal diversity and the relative abundance (RA) of beneficial Chaetomium and Trichoderma, and the decreased RA of pathogenic Fusarium and Alternaria. Consequently, the joint use of biochar and biocontrol agents (YIM PH30719) synergistically decreased the occurrence of root rot by modifying soil physicochemical characteristics and fungal communities. Since root rot is commonly found in agriculture, the results of this study may help in controlling root rot disease in other crops.

Synergistic interactions of biocontrol agents and chemical fungicides enhance the disease resistance in Vigna radiata (L.) against Macrophomina phaseolina-associated with dry root rot

Macrophomina phaseolina, a necrotrophic fungus responsible for dry root rot disease in Vigna radiata L. (greengram) causes extensive yield losses. Biocontrol agents are reported to suppress phytopathogens, mitigate biotic stress, and improve growth and yield of plants. Here, Bacillus subtilis PSB-5 and Trichoderma sp. TD-6 along with chemical fungicides; mancozeb (MZB) and carbendazim (CBZM) were evaluated alone/or in combination for their effectiveness against M. phaseolina-induced root rot disease in greengram. B. subtilis and Trichoderma sp. suppressed phyto-fungal growth by 72 and 78 %, respectively. Both bioagents synthesized indole-3-acetic acid, ammonia, and solubilized insoluble phosphate, released antimicrobial substances including siderophore, hydrogen cyanide, and various extracellular lytic enzymes. Further, efficacy of bioagents and fungicides (at 50 and 100 ppm) were tested (single and combined) against M. phaseolina-infected greengram cultivated in pot soils. While evaluating, bioagents and fungicides increased plant height, biomass, and vigor indices of infected greengram in order: PSB-5 + TD-6 > PSB-5 > TD-6 > MZB + CBZM (100 ppm) > CBZM (100 ppm) > MZB (100 ppm). Further, among treatment, dual inoculation of bioagents (B. subtilis + Trichoderma), maximally increased total chlorophyll (59.4 %), carotenoid (68 %), root phosphorous (69.6 %), root nitrogen (65.7 %), total soluble protein (80 %), soluble sugar (76 %), pod yield (82.3 %) and seed protein (68 %) over infected plants. Moreover, biotic stress-induced oxidative stress biomarkers (proline, malondialdehyde, electrolyte leakage) and reactive oxygen species were significantly (p ≤ 0.05) lowered in fungal-infected greengram following fungicides and bioagents application. Furthermore, combined inoculation of bioagents significantly (p ≤ 0.05) reduced percent disease index (PDI) and increased % protection in biotic-stressed greengram. Besides, bioagents potentially increased disease resistance in M. phaseolina-infected greengram plants by modulating defense responses such as activities of β-1, 3 glucanase, catalase, phenylalanine ammonia lyase, polyphenol oxidase, peroxidase, and superoxide dismutase which significantly protected plants from fungal pathogen. Soil populations of pathogen and bioagents declined and considerably increased, respectively, at harvest. Application of antagonistic bacterial and fungal strains enhances greengram resistance to M. phaseolina while concurrently offering agriculture an environmentally friendly choice. Also, these strains can be applied as biocontrol agents in disease suppression over chemical fungicides.

A novel K57-specific Klebsiella pneumoniae phage disinfects milk and inhibits biofilm formation

Klebsiella pneumoniae (K. pneumoniae), especially serotype K57, is a major pathogen in dairy bovine mastitis, causing significant economic losses and posing a public health risk through contaminated dairy products. In addition, K57 is highly virulent among serotypes infecting humans, leading to serious conditions such as pyogenic liver abscess. In this study, we isolated and identified a bacteriophage called vB_Kp_Z57 that targets K57 serotype K. pneumoniae isolates from human and bovine sources. The phage has a latent period of 4 min and can produce approximately 233 phages per infected bacterial cell. vB_Kp_Z57 is stable over a temperature range of 4 °C to 60 °C and a pH range of 5–11. Sequencing and assembly of the phage genome revealed DNA molecules consisting of 40,316 base pairs. After genome annotation with bioinformatics software, it was discovered that the phage genome encodes 48 proteins, with no detected lysogeny genes, toxin genes, or tRNA-related genes. vB_Kp_Z57 was classified as a member of genus Przondovirus, family Autographiviridae. The results of bacteriostatic experiments show that vB_Kp_Z57 has a good effect on inhibiting bacterial growth. Notably, vB_Kp_Z57 significantly inhibited the growth of its host bacteria in milk at refrigerated temperatures of 4 °C and room temperature. Biofilm inhibition experiments revealed that phage vB_Kp_Z57 significantly reduced OD595 values in all phage-treated groups, indicating its potential as an effective biocontrol agent against K. pneumoniae. Our findings show the promise of vB_Kp_Z57 as a novel and environmentally friendly solution to address the challenges associated with K. pneumoniae in dairy production.

The Potential of Microorganisms for the Control of Grape Downy Mildew-A Review.

Plasmopara viticola (Berk.et Curtis) Berl. Et de Toni is the pathogen that causes grape downy mildew, which is an airborne disease that severely affects grape yield and causes huge economic losses. The usage of effective control methods can reduce the damage to plants induced by grape downy mildew. Biocontrol has been widely used to control plant diseases due to its advantages of environmental friendliness and sustainability. However, to date, only a few comprehensive reviews on the biocontrol of grape downy mildew have been reported. In this review, we summarize the biological characteristics of P. viticola and its infection cycle, followed by a detailed overview of current biocontrol agents, including bacteria and fungi that could be used to control grape downy mildew, and their control effects. Furthermore, potential control mechanisms of biocontrol agents against grape downy mildew are discussed. Lastly, suggestions for future research on the biocontrol of grape downy mildew are provided. This review provides the basis for the application of grape downy mildew biocontrol.

Bacteria as potential biocontrol agents for managing purple witchweed (Striga hermonthica) in grain sorghum

Abstract Purple witchweed [Striga hermonthica (Delile) Benth.], a highly destructive parasitic weed, poses a significant threat to sorghum [Sorghum bicolor (L.) Moench] cultivation. This hemiparasitic plant intrudes its root system into the host plant, leading to substantial yield losses, particularly in susceptible genotypes. In the pursuit of eco-friendly solutions, the biocontrol approach has gained attention as a potential management strategy for Striga. In this study, 13 bacterial strains belonging to the genera Bacillus, Gluconobacter, Pseudomonas, and Streptomyces were investigated in vitro for their efficiency in controlling the early-stage development of Striga. Among the tested strains, Streptomyces morookaensis NRRL B-12429 demonstrated significant inhibition of Striga seed germination and radicle elongation at 54.36% and 61.84%, respectively, when applied to preconditioned seeds with a synthetic germination stimulant. The effect of S. morookaensis on the inhibition of Striga seed germination was more pronounced in the presence of the host plant, sorghum, at 62.35%. However, biopriming of sorghum seeds with S. morookaensis did not enhance the inhibitory effects on Striga seed germination but resulted in a greater reduction in radicle elongation at 74.64% compared with non-primed seeds. Additionally, the biopriming with S. morookaensis promoted the growth of shoots and roots of germinating sorghum, regardless of the presence of Striga seeds. These findings highlight the potential of S. morookaensis strain NRRL B-12429 as a viable candidate for biocontrol agent applications in sorghum cultivation. Further exploration and investigation of its biocontrol capabilities can provide valuable insights for sustainable management practices against Striga infestations.

Biocontrol of Hemileia vastatrix, the Causal Agent of Coffee Leaf Rust, by Paenibacillus sp. NMA1017.

Coffee leaf rust (CLR), caused by Hemileia vastatrix, is considered a highly important phytosanitary problem in Mexico. Currently, there are few microorganisms used as biocontrol alternatives to chemical control of CLR in organic coffee fields in Mexico. This study evaluates the use of Paenibacillus sp. NMA1017 as a biocontrol agent to inhibit the development of H. vastatrix in in vitro and in vivo (greenhouse) experiments. Hemileia vastatrix urediniospores were placed on water agar plates, and then Paenibacillus sp. NMA1017 was inoculated simultaneously or 8 h later. Urediniospores germination rate was reduced by 94% when the NMA1017 strain was inoculated simultaneously with the urediniospores and reduced by 38% when NMA1017 was inoculated 8 h later. Experiments with 8-month-old Bourbon coffee plants that were infected with H. vastatrix showed that disease incidence was reduced by 38, 90, and 50% when NMA1017 was applied 8 days before, simultaneously, or 8 days after the application of H. vastatrix, respectively. Paenibacillus sp. NMA1017 also reduced the severity of CLR on the leaves by up to 62%. The germination urediniospores of other rust pathogens such as Puccinia sorghi (maize leaf rust), Puccinia triticina (wheat leaf rust), Puccinia graminis f. sp. tritici (black stem rust of wheat), Uromyces striatus (alfalfa leaf rust), and Phragmidium sp. (rosebush leaf rust) were also inhibited. Use of the potential biocontrol agent Paenibacillus sp. NMA1017 might help reduce the application of chemical fungicides for the control of CLR, making coffee a more sustainable crop and providing management options for organic coffee growers.

Endophytic fungi from &lt;i&gt;Dichrocephala integrifolia&lt;/i&gt;: Diversity, antifungal properties, enzymatic activities, and plant growth promotion

Dichrocephala integrifolia is a wild medicinal plant utilised in traditional healing and Ayurveda to cope up with several health issues by various groups of people around the world. Medicinal plants are associated with diverse fungal endophytes with potential bioactive properties. In this investigation, 26 fungal endophytes were isolated from D. integrifolia and three sterile forms using the Petri plate culture method. The endophytic isolation rate was highest for inflorescence (35.97%) and lowest for stem (15.61%). The highest colonization frequency was shown by F. solani (8.37%) and the lowest by Sterile morphotype 3 (1.36%). Out of the four plant parts, inflorescence was found to be highly infected, displaying an infection rate of 93.43% and the least infection occurred in the stem with 45%. The maximum number of isolated endophytic fungi belongs to the class Sordariomycetes, with a relative occurrence (%) of 71.72%. The Simpson’s diversity index reveals that the leaf endophytes were more diverse (0.94). Qualitative antifungal activity of the sporulating isolates against Curvularia lunata has shown that the maximum number of endophytes possessed Class 3 antagonism. Four isolates were selected based on screening of their antagonistic activity and their antifungal inhibition was calculated against nine fungal phytopathogens. Maximum inhibition (100%) was shown by Trichoderma sp. 2 (S2B2) against Alternaria alternata, A. brassicicola, Colletotrichum capsici, C. lunata, and Ustilaginoidea virens and least inhibition by Gliocladium sp. 1 (19.78%) against C. lunata. The four isolates were found to produce protease, lipase, amylase and cellulase enzymes. The isolates produced ammonia and hydrogen cyanide, but none of the isolates could solubilize phosphate. Potent biocontrol agents are much needed to replace synthetic chemicals and restore soil microflora.

New Trichoderma Strains Suppress Blue Mold in Oranges by Damaging the Cell Membrane of Penicillium italicum and Enhancing Both Enzymatic and Non-Enzymatic Defense Mechanisms in Orange Fruits

Blue mold disease, caused by Penicillium italicum (P. italicum), presents a significant challenge to orange fruits (Citrus sinensis L.) and other citrus crops globally. Biological control, particularly Trichoderma species, offers a promising alternative to synthetic fungicides. Therefore, this study aimed to isolate, identify, and evaluate the antagonistic activities of two Trichoderma isolates against P. italicum. These isolates were molecularly identified and assigned accession numbers PP002254 and PP002272, respectively. Both isolates demonstrated significant antifungal activity in dual culture assays. Moreover, the culture filtrates (CFs) of Trichoderma longibrachiatum PP002254 and Trichoderma harzianum PP002272 suppressed the mycelial growth of P. italicum by 77.22% and 71.66%, respectively. Additionally, CFs reduced the severity of blue mold on orange fruits by 26.85% and 53.81%, compared to 100% in the control group. Scanning electron microscopy revealed that treated P. italicum hyphae were shrunken and disfigured. Enzyme activities (catalase, peroxidase, polyphenol oxidase, and phenylalanine ammonia-lyase) in treated oranges increased, along with total soluble phenolics and flavonoids. Conversely, malondialdehyde (MDA) levels decreased in treated fruits. These findings suggest that T. longibrachiatum PP002254 and T. harzianum PP002272 could be effective biocontrol agents for managing blue mold and other citrus postharvest diseases.

Annotated checklist of chalcidid (Hymenoptera: Chalcididae) fauna of the Chhattisgarh region

Chalcidid wasps are a diverse group of insects belonging to the order Hymenoptera and family Chalcididae. They are natural biocontrol agents for many agricultural and forestry pests. The present study was undertaken from 2021 to 2023. The objective of this study was to document the Chalcididae diversity from the Chhattisgarh region. Based on this study, the first checklist of chalcidid wasps from Chhattisgarh state is prepared. In total, 71 species of chalcidid wasps were collected from 11 genera falling under four subfamilies. The chalcidid wasps identified belonged to the subfamilies, Chalcidinae, Dirhininae, Epitraninae and Haltichellinae of which, subfamily Haltichellinae was found to be most predominant. The results of this preliminary study shed light on the hidden biodiversity of chalcidid wasps.

An asymptomatic geminivirus activates autophagy and enhances plant defenses against diverse pathogens

Plant viral diseases cause great losses in agricultural production. Virus cross-protection is a strategy in which a mild virus is employed to shield plants against subsequent infections by severe viral strains. However, this approach is restricted to protection against the same viruses. In this study, we observed that pre-inoculation with apple geminivirus (AGV) reduced the accumulation of secondarily infected heterologous viruses, such as cucumber mosaic virus, potato virus X, and tobacco mosaic virus in Nicotiana benthamiana, tomato, and pepper plants. Transcriptional expression analysis showed that autophagy-related genes were transcriptionally up-regulated upon AGV inoculation at an early stage of infection. Accordingly, autophagic activity was observed to be elevated following AGV infection. Interestingly, AGV accumulation was reduced in autophagy-deficient plants, suggesting that autophagy activation promotes AGV infection in the plant. Moreover, pre-inoculation with AGV provided cross-protection against infection with a phytopathogenic bacterium (Pseudomonas syringae) and fungus (Botrytis cinerea) in Nicotiana species. In summary, our study showed that AGV, an asymptomatic virus, could protect plants against severe viral, fungal, and bacterial diseases to some extent through the activation of autophagy pathways, highlighting its potential as a biocontrol agent for managing a wide range of plant crop diseases in the field.

Genomic Characterization and Establishment of a Genetic Manipulation System for Trichoderma sp. (Harzianum Clade) LZ117.

Trichoderma species have been reported as masters in producing cellulolytic enzymes for the biodegradation of lignocellulolytic biomass and biocontrol agents against plant pathogens and pests. In our previous study, a novel Trichoderma strain LZ117, which shows potent capability in cellulase production, was isolated. Herein, we conducted multilocus phylogenetic analyses based on DNA barcodes and performed time-scaled phylogenomic analyses using the whole genome sequences of the strain, annotated by integrating transcriptome data. Our results suggest that this strain represents a new species closely related to T. atrobrunneum (Harzianum clade). Genes encoding carbohydrate-active enzymes (CAZymes), transporters, and secondary metabolites were annotated and predicted secretome in Trichoderma sp. LZ117 was also presented. Furthermore, genetic manipulation of this strain was successfully achieved using PEG-mediated protoplast transformation. A putative transporter gene encoding maltose permease (Mal1) was overexpressed, which proved that this transporter does not affect cellulase production. Moreover, overexpressing the native Cre1 homolog in LZ117 demonstrated a more pronounced impact of glucose-caused carbon catabolite repression (CCR), suggesting the importance of Cre1-mediated CCR in cellulase production of Trichoderma sp. LZ117. The results of this study will benefit further exploration of the strain LZ117 and related species for their applications in bioproduction.

Patulin inhibition of specific apple microbiome members uncovers Hanseniaspora uvarum as a potential biocontrol agent.

Penicillium expansum is a major postharvest pathogen of apples, causing loss in fruits through tissue damage, as well as in apple products due to contamination with the mycotoxin patulin. During infections, patulin is a cultivar-dependent virulence factor that facilitates apple lesion development. Patulin also has characterized antimicrobial activity and is important for inhibiting other competitive phytopathogens, but the role of this inhibitory activity has not been investigated in the context of the apple microbiome. In our current study, we isolated 68 apple microbiota and characterized their susceptibility to P. expansum extracts. We found Gram-negative bacteria and Basidiomycete yeast to demonstrate largely patulin-specific growth inhibition compared to Gram-positive and Ascomycete isolates. From co-cultures, we identified a Hanseniaspora and Gluconobacter pairing that reduced P. expansum biomass and found that Hanseniaspora uvarum alone is sufficient to reduce apple disease progression in vivo. We investigated possible mechanisms of H. uvarum biocontrol activity and found modest inhibition on apple puree plates, as well as a trend toward lower patulin levels at the wound site. Active biocontrol activity required live yeast, which also were effective in controlling Botrytis cinerea apple infections. Lastly, we explored the breadth of H. uvarum biocontrol activity with over 30 H. uvarum isolates and found consistent inhibition of P. expansum apple disease.

Biocontrol potential of Bacillus velezensis SEC-024A against southern blight of industrial hemp

Agroathelia rolfsii is the causative agent of southern blight of industrial hemp, which is a soil-borne infection resulting in grave economic losses in industrial hemp (Cannabis sativa L.). Chemical pesticides are currently the primary means of controlling this disease, but their use is harmful to human health as well as to the environment. Urgent action is needed to screen eco-friendly biocontrol agents against this pathogen. An endophytic bacterium SEC-024A was isolated from the leaf tissue of industrial hemp and exhibited a significant antagonistic effect on A. rolfsii, with a plate inhibition rate of 80.5 % and a pot control effect of 74.1 %. Through whole genome sequencing, SEC-024A was identified to be Bacillus velezensis. It contained multiple gene clusters involved in secondary metabolite biosynthesis and the genes that promote plant growth, colonization, and pathogen defense. Bacillus velezensis SEC-024A produces volatile organic compounds (VOCs) that serve as its primary fungistatic agents. Nine VOCs produced by SEC-024A were detected through gas chromatography-mass spectrometry. These VOCs were individually tested for their antifungal effects. Four volatiles, namely isobutyric acid, tiglic acid, trans-2-octenal, and 2-decanone, exhibited notable inhibitory effects on A. rolfsii with EC50 values of 0.012, 0.038, 0.009, and 0.046 μL/mL, respectively. Additionally, trans-2-octenal demonstrated broad-spectrum antifungal activity against various soil-borne pathogens. Together, this study suggests that B. velezensis SEC-024A holds promise as a novel microbial agent for improving plant growth and controlling diseases.