Effective Biological Control Research Articles

Artificial diets can manipulate the reproductive development status of predatory ladybird beetles and hold promise for their shelf-life management

With the development of biological control methods, the predatory ladybird beetle Harmonia axyridis Pallas (Coleoptera: Coccinellidae) has been widely used for pest control in agricultural production. Appropriate shelf-life management strategies could synchronize H. axyridis production with pest outbreaks, finally improving the effectiveness of biological control. Herein, we preliminarily explored whether an artificial diet could optimize the shelf-life management of H. axyridis. We compared the survival rate, nutrition accumulation, reproductive development, juvenile hormone (JH) related-gene expression levels, and stress resistance gene expression levels between aphid-fed and artificial diet-fed H. axyridis females. The results revealed that H. axyridis females maintained a high survival rate after being fed an artificial diet for 60 days, whereas the survival rate of aphid-fed females decreased. Continuous feeding of the artificial diet caused H. axyridis females to enter a diapause-like state, which was characterized by low JH levels, high triglycerides and trehalose accumulation, ovarian development inhibition, decreased Vgs expression levels, and increased stress resistance gene expression levels. This diapause-like state could be promptly recovered upon transferring to an aphid diet. These results indicate that the artificial diet could manipulate the reproductive development status of H. axyridis and lay the foundation for its shelf-life management.

Begomovirus Transmission to Tomato Plants Is Not Hampered by Plant Defenses Induced by Dicyphus hesperus Knight.

Plants can respond to insect infestation and virus infection by inducing plant defenses, generally mediated by phytohormones. Moreover, plant defenses alter host quality for insect vectors with consequences for the spread of viruses. In agricultural settings, other organisms commonly interact with plants, thereby inducing plant defenses that could affect plant-virus-vector interactions. For example, plant defenses induced by omnivorous insects can modulate insect behavior. This study focused on tomato yellow leaf curl virus (TYLCV), a plant virus of the family Geminiviridae and genus Begomovirus. It is transmitted in a persistent circulative manner by the whitefly Bemisia tabaci Gennadius (Hemiptera: Aleyrodidae), posing a global threat to tomato production. Mirids (Hemiptera: Miridae) are effective biological control agents of B. tabaci, but there is a possibility that their omnivorous nature could also interfere with the process of virus transmission. To test this hypothesis, this study first addressed to what extent the mirid bug Dicyphus hesperus Knight induces plant defenses in tomato. Subsequently, the impact of this plant-omnivore interaction on the transmission of TYLCV was evaluated. Controlled cage experiments were performed in a greenhouse setting to evaluate the impact of mirids on virus transmission and vector acquisition by B. tabaci. While we observed a reduced number of whiteflies settling on plants exposed to D. hesperus, the plant defenses induced by the mirid bug did not affect TYLCV transmission and accumulation. Additionally, whiteflies were able to acquire comparable amounts of TYLCV on mirid-exposed plants and control plants. Overall, the induction of plant defenses by D. hesperus did not influence TYLCV transmission by whiteflies on tomato.

Inhibition of Verticillium Wilt in Cotton through the Application of Pseudomonas aeruginosa ZL6 Derived from Fermentation Residue of Kitchen Waste.

To isolate and analyze bacteria with Verticillium wilt-resistant properties from the fermentation residue of kitchen wastes, as well as explore their potential for new applications of the residue. A total of six bacterial strains exhibiting Verticillium wilt-resistant capabilities were isolated from the biogas residue of kitchen waste fermentation. Using a polyphasic approach, strain ZL6, which displayed the highest antagonistic activity against cotton Verticillium wilt, was identified as belonging to the Pseudomonas aeruginosa. Bioassay results demonstrated that this strain possessed robust antagonistic abilities, effectively inhibiting V. dahliae spore germination and mycelial growth. Furthermore, P. aeruginosa ZL6 exhibited high temperature resistance (42°C), nitrogen fixation, and phosphorus removal activities. Pot experiments revealed that P. aeruginosa ZL6 fermentation broth treatment achieved a 47.72% biological control effect compared to the control group. Through activity tracking and protein mass spectrometry identification, a neutral metalloproteinase (Nml) was hypothesized as the main virulence factor. The mutant strain ZL6ΔNml exhibited a significant reduction in its ability to inhibit cotton Verticillium wilt compared to the strain P. aeruginosa ZL6. While the inhibitory activities could be partially restored by a complementation of nml gene in the mutant strain ZL6CMΔNml. This research provides a theoretical foundation for the future development and application of biogas residue as biocontrol agents against Verticillium wilt and as biological preservatives for agricultural products. Additionally, this study presents a novel approach for mitigating the substantial amount of biogas residue generated from kitchen waste fermentation.

Impact of Climate Change on Pest Biology, Behaviour and Their Distributions

Increased temperatures, rising CO2 levels, and changing precipitation patterns are influencing agricultural production. These changes can lead to the expansion of insect pests geographic distribution, increased survival during overwintering, more generations per year, altered synchrony with plants, changed interspecific interactions, higher risk of invasion by migratory pests, increased incidence of insect-transmitted plant diseases, and reduced effectiveness of biological control. There is a serious risk of economic losses in crop production. Human food security is challenged due to potential disruptions in agricultural systems. Climate change necessitates adaptive management strategies to address evolving pest dynamics. Future research should focus on developing modified integrated pest management tactics to cope with changing pest statuses. Monitoring climate and pest populations is crucial for understanding and predicting the impact of climate change on insect pests. This review examines the climate change effect on insect pest biology and behavior and explores the potential use of modern pest monitoring technologies, modeling prediction tools can aid in forecasting and planning for potential pest outbreaks. Integrated pest management (IPM) approaches should be adapted to incorporate the changing dynamics influenced by climate change. This may involve a combination of chemical, biological, cultural, and mechanical control methods.

Fungal plant pathogen Colletotrichum shisoi identified as a potential biological control agent of invasive Perilla frutescens in the United States

ABSTRACT Perilla frutescens (L.) Britton, an annual plant in the mint family, is considered invasive in the United States where it excludes native vegetation in natural areas and can cause respiratory illness in livestock. An effective biological control agent would improve perilla management, but none have been developed or proposed. This study investigated the biocontrol potential of a fungal pathogen, Colletotrichum shisoi, recently found infecting perilla in North America. The growth rates and morphology of seven Colletotrichum shisoi isolates were recorded. A virulence assay was performed on two perilla accessions, and the number and size of foliar lesions produced were used to select a single isolate, Colletotrichum shisoi isolate 21-072, exhibiting consistently high virulence for further evaluation of biocontrol efficacy and host specificity. Treatment with 0.5 mL of 1 × 107 spores/mL H2O caused a 52% reduction in shoot height and 84% reduction in root length of invasive perilla seedlings compared to a control treatment (P < 0.001). Seedlings treated with 1 × 108 spores/mL had a 100% mortality rate. Formulation of inoculum in a 1:1 water:oil emulsion reduced dew period requirements for causing >50% plant mortality from 48 to 24 h, but oil emulsions exhibited a mild phytotoxic effect on seedlings. Four additional perilla accessions were susceptible to C. shisoi, and none of 34 other species included in host range experiments developed disease symptoms following inoculation. These results suggest C. shisoi 21-072 could be an effective antagonist of perilla and likely poses a low risk of non-target impacts.

Effect of volatile compounds produced by Weissella cibaria BWL4 on Botrytis cinerea infection in fruit and complete genome sequence analysis of BWL4

Botrytis cinerea, one of the most harmful pathogenic fungi, causes significant postharvest decay in fruit. In this study, the effects of Weissella cibaria BWL4 on B. cinerea and the gray mold in grape and kiwifruit were evaluated. Additionally, the genome of an antagonistic bacterium BWL4 was sequenced. Results showed that the application of BWL4 inhibited B. cinerea infection in grape and kiwifruit. BWL4 produced antifungal volatile substances against B. cinerea. A gas chromatography–mass spectrometry of solid phase microextraction samples (SPME-GC-MS) analysis showed that BWL4 produced 54 volatiles. Among these volatiles, furfural, 2,6-dimethyl-pyrazine, dimethyl trisulfide, 2-pentyl furan, 2-ethyl hexanol, hexamethyl-cyclotrisiloxane, dodecamethyl-cyclohexasiloxane, 2,4-di-tert-butylphenol, and butylated hydroxytoluene, exhibited antifungal activity against B. cinerea, with MICs of 0.8 g L−1, 0.2 g L−1, 0.9 g L−1, 5 g L−1, 0.4 g L−1, 8 g L−1, 8 g L−1, 80 mg L−1, and 2 g L−1, respectively. In vivo assay showed that these compounds inhibited the disease development in kiwifruit. These compounds except dimethyl trisulfide can also inhibit gray mold in grape. The genome of W. cibaria BWL4 consists of a circular chromosome of 2.42 Mb and 6 plasmids. The genome size is 2.53 Mb, encoding 2327 putative genes. Several genes involved in the biosynthesis of some antifungal volatiles and 17 genes associated with biocontrol were identified. These results suggested that W. cibaria BWL4 can be considered as an effective biological control agent for the postharvest management of B. cinerea infection.

Биологична активност на 20 изолата на ентомопатогенни гъби от род Beauveria

Enzymes such as chitinase, proteinase and lipase are involved in the breakdown of insect cuticles. Their pathogenicity, virulence and specificity largely depend on the secretion of these enzymes by different microorganisms. Thus, the level of enzyme activity may also represent a diagnostic tool for selecting an effective biological control agent. In the present study, 20 different isolates of entomopathogenic fungi of the genus Beauveria were used and their lipolytic, chitinolytic and proteolytic activities were determined. Enzyme activities are assessed qualitatively according to the presence of a zone of lightening due to their degrading action and quantitatively expressed as a ratio between the zone of enzyme activity and the diameter of the colony. PCR analysis with gene-specific primers located in the sequence of the Beauveria bassiana chitinase - chitinase (chit1) gene (ACC AY145440) was also carried out. Amplifications of fragments with an expected length of 1225 bp were detected in the majority of Beauveria isolates studied. The studied isolates of the genus Beauveria differ in their lipolytic, proteolytic and chitin-degrading abilities. The highest total activity was found in four of them 336, 340, 487 and 501 and could be related to their virulence. The information on the enzymatic activities of the studied isolates will be necessary for the development of an efficient preparation for the biological control of the enemies.

Genomic and biological control of Sclerotinia sclerotiorum using an extracellular extract from Bacillus velezensis 20507.

Sclerotinia sclerotiorum is a known pathogen that harms crops and vegetables. Unfortunately, there is a lack of effective biological control measures for this pathogen. Bacillus velezensis 20507 has a strong antagonistic effect on S. Sclerotiorum; however, the biological basis of its antifungal effect is not fully understood. In this study, the broad-spectrum antagonistic microorganisms of B. velezensis 20507 were investigated, and the active antifungal ingredients in this strain were isolated, purified, identified and thermal stability experiments were carried out to explore its antifungal mechanism. The B. velezensis 20507 genome comprised one circular chromosome with a length of 4,043,341 bp, including 3,879 genes, 185 tandem repeats, 87 tRNAs, and 27 rRNAs. Comparative genomic analysis revealed that our sequenced strain had the closest genetic relationship with Bacillus velezensis (GenBank ID: NC 009725.2); however, there were significant differences in the positions of genes within the two genomes. It is predicted that B. velezensis 20507 encode 12 secondary metabolites, including difficidin, macrolactin H, fengycin, surfactin, bacillibactin, bacillothiazole A-N, butirosin a/b, and bacillaene. Results showed that B. velezensis 20507 produced various antagonistic effects on six plant pathogen strains: Exserohilum turcicum, Pyricularia oryzae, Fusarium graminearum, Sclerotinia sclerotiorum, Fusarium oxysporum, and Fusarium verticillioides. Acid precipitation followed by 80% methanol leaching is an effective method for isolating the antifungal component ME80 in B. velezensis 20507, which can damage the membranes of S. sclerotiorum hyphae and has good heat resistance. Using high-performance liquid chromatography, and Mass Spectrometry analysis, it is believed that fengycin C72H110N12O20 is the main active antifungal substance. This study provides new resources for the biological control of S. Sclerotiorum in soybeans and a theoretical basis for further clarification of the mechanism of action of B. velezensis 20507.

The landscape of fear in cattle farms? How the presence of barn swallow influences the activity of pest flies

Abstract Insectivorous birds are key suppliers of crucial ecosystem services, especially important for pest control in agricultural systems, and are globally decreasing at a fast rate. Among them, the declining barn swallow Hirundo rustica is particularly linked to cattle farms, where it is impacted by changes in livestock husbandry practices. The presence of swallows in cattle sheds might contribute to control insect pests, but such an effect had never been explored. With this work, we evaluated the potential control effect exerted by barn swallow on economically relevant pest insects, filth flies. To assess whether swallows inside the cattle sheds influence the activity rate of pest flies (especially Musca domestica and Stomoxys calcitrans), we assessed the relative abundance of swallows and flies over 16 weeks in nine different cattle farms in Non valley (Trentino, North‐east Italy), either occupied or not by nesting swallows. From April 2022 to August 2022, we collected weekly data on the activity rate of flies by placing eight white spot cards inside each of nine cattle sheds and counting the deposition of faecal and regurgitation spots by flies. Simultaneously, we collected weekly data regarding swallow occurrence, brood size, number and age of chicks. We applied generalised linear mixed effect models on the activity rate of flies, with spot card and farm as nested random effects, to account for potential random variability between cards and farms. We then assessed through model selection the effect of number of cows, number of swallows, and temperature or week alternatively. Temperature and week were modelled also in interaction with the number of swallows. Fly activity rate followed a seasonal trend with a peak in July, which was markedly lower where swallows were present. Moreover, the positive relationship between flies and temperature was lessened when swallows were abundant. Synthesis and applications. Barn swallows are effective biological control agents for lowering pest fly activity in livestock farms. Farmers were actively involved in swallow conservation within their sheds, also with a shared memorandum‐of‐intent. Harnessing the ecosystem services supplied by farmland birds could bring positive outcomes for people and biodiversity.

The Use of Microbiological Quality Control Techniques Adopted in a Human Tissue Bank

The Brazilian Ministry of Health’s Consolidation Ordinance No. 4 of September 28, 2017, provides the requisites for the evaluation of services for human tissue banks (bancos de tecidos humanos [BTH]) in terms of the microbiological control of environments (ISO 5 classification processing room) and products (culture for aerobic, anaerobic, and fungal pathogens), with a view to guaranteeing harmlessness in tissue transplants. Objectives: To highlight the techniques for microbiological control of distributed tissues adopted by a BTH over 5 years, as well as the results obtained from the application and use of these techniques. Methods: Using standard operating procedures (SOPs), the methodology, and frequency of collections, tests and room cleaning were established in advance. Microbiological cultures were taken from the tissues during all the collections and processing. During tissue processing, fingerprint samples on the processor’s gloves and in the processing room were seeded on blood agar plates. The processing room was cleaned weekly with biguanide and quaternary ammonium. The processing bench was sanitized with sterile 70% alcohol and the microbiological environmental control was carried out every 6 months by a qualified outsourced company. Results: In the period analyzed, the techniques proved effective in 46 (96%) cases, with contamination identified in only two (4%) processed and collected samples. The effectiveness and results were documented. Conclusion: Effective biological quality control methods are legitimately required, but they can be improved, raising the need to develop safe protocols for the quality of services and the safety of transplanted tissues.

Evaluation of the control efficacy of antagonistic bacteria from V-Ti magnetite mine tailings on kiwifruit brown spots in pot and field experiments.

Seemingly barren heavy-metal-polluted vanadium (V) and titanium (Ti) magnetite mine tailings contain various functional microbes, yet it is unclear whether this includes microbial resources relevant to the biological control of plant diseases. Kiwifruit brown leaf spot disease, caused by Corynespora cassiicola, can seriously reduce kiwifruit yield. To discover effective control measures for kiwifruit leaf spot, 18 bacteria strains among 136 tailing-isolated bacteria from V-Ti magnetite mine tailings were identified as inhibiting C. cassiicola by the confrontation plate method, indicating that antagonistic bacteria surviving in the V-Ti magnetite mine tailings were present at a low level. The 18 antagonistic strains could be divided into two BOX-A1R clusters. The 13 representative strains that were selected for phylogenetic tree construction based on their 16S rRNA sequences belonged to the Bacillus genus. Five predominant strains exhibited different toxin-production times and intensities, with four of them initiating toxin production at 32 h. Among them, Bacillus sp. KT-10 displayed the highest bacteriostatic rate (100%), with a 37.5% growth inhibition rate and an antagonistic band of 3.2 cm against C. cassiicola. Bacillus sp. KT10 also showed a significant inhibitory effect against the expansion speed of kiwifruit brown spots in the pot. The relative control effect was 78.48 and 83.89% at 7 days after the first and last spraying of KT-10 dilution, respectively, confirming a good effect of KT-10 on kiwifruit brown leaf spots in the field. This study demonstrated for the first time that there are some antagonistic bacteria to pathogenic C. cassiicola in V-Ti magnetite mine tailings, and Bacillus sp. KT10 was found to have a good control effect on kiwifruit brown leaf spots in pots and fields, which provided an effective biological control measurement for kiwifruit brown leaf spots.

Antifungal potential of lipopeptides produced by the Bacillus siamensis Sh420 strain against Fusarium graminearum.

Biological control is a more sustainable and environmentally friendly alternative to chemical fungicides for controlling Fusarium spp. infestations. In this work, Bacillus siamensis Sh420 isolated from wheat rhizosphere showed a high antifungal activity against Fusarium graminearum as a secure substitute for fungicides. Sh420 was identified as B. siamensis using phenotypic evaluation and 16S rDNA gene sequence analysis. An in vitro antagonistic study showed that Sh420's lipopeptide (LP) extract exhibited strong antifungal properties and effectively combated F. graminearum. Meanwhile, lipopeptides have the ability to decrease ergosterol content, which has an impact on the overall structure and stability of the plasma membrane. The PCR-based screening revealed the presence of antifungal LP biosynthetic genes in this strain's genomic DNA. In the crude LP extract of Sh420, we were able to discover several LPs such as bacillomycin, iturins, fengycin, and surfactins using ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry. Microscopic investigations (fluorescent/transmission electron microscopy) revealed deformities and alterations in the morphology of the phytopathogen upon interaction with LPs. Sh420 LPs have been shown in grape tests to be effective against F. graminearum infection and to stimulate antioxidant activity in fruits by avoiding rust and gray lesions. The overall findings of this study highlight the potential of Sh420 lipopeptides as an effective biological control agent against F. graminearum infestations.IMPORTANCEThis study addresses the potential of lipopeptide (LP) extracts obtained from the strain identified as Bacillus siamensis Sh420. This Sh420 isolate acts as a crucial player in providing a sustainable and environmentally friendly alternative to chemical fungicides for suppressing Fusarium graminearum phytopathogen. Moreover, these LPs can reduce ergosterol content in the phytopathogen influencing the overall structure and stability of its plasma membrane. PCR screening provided confirmation regarding the existence of genes responsible for biosynthesizing antifungal LPs in the genomic DNA of Sh420. Several antibiotic lipopeptide compounds were identified from this bacterial crude extract using ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry. Microscopic investigations revealed deformities and alterations in the morphology of F. graminearum upon interaction with LPs. Furthermore, studies on fruit demonstrated the efficacy of Sh420 LPs in mitigating F. graminearum infection and stimulating antioxidant activity in fruits, preventing rust and gray lesions.

Native Entomopathogenic Fungi Isolated from Rhynchophorus palmarum (Linnaeus, 1758) in Northeast Brazil.

Both palm weevils, the South American (Rhynchophorus palmarum) (SAPW) and the red palm weevil (R. ferrugineus, RPW), are present in South America, affecting commercial, ornamental, and native palms. These pests oviposit and thrive on selected Arecaceae. R. palmarum mainly infests coconut (Cocos nucifera), oil palms (Elaeis guineensis), and other ornamental and native palms in America, causing a significant social impact on growers. The weevils fulfill a significant ectosymbiotic macro- and microorganism role in the first period of larval development, worsening the damage which, during this period, is not yet apparent. Palm protection in the Brazilian context suggests the use of indigenous agents for microbiological biocontrol. This research identifies three Brazilian Beauveria bassiana isolates: CVAD01, CVAD02, and CVAD06. The results suggest that the strain's impact on R. palmarum can also be compared with that of the commercial strain Beauveria bassiana. Phylogenetic analysis allowed the delimitation of species of Beauveria (Hypocreales). Pathogenicity tests caused significant mortality in R. palmarum. The isolates CVAD01, CVAD02, and CVADO6 showed high pathogenicity between 7 and 21 days, with mortality rates between 90 and 100%, suggesting that they may be effective biological control agents of R. palmarum in the field when used, within available means, to mitigate the impact of R. palmarum and R. ferrugineus in South America.

Isolation and Characterization of fungal pathogens associated with Carica papaya L. and their biocontrol with Trichoderma sp.

Objective: In this work, we undertook the task of isolating and identifying Fusarium obtained from papaya fruits grown in the Veracruz region and carried out antibiotic tests to find a beneficial fungus that could exert biological control. Methodology: Juvenile fruits with necrosis and rot were collected from papaya plants of the Maradol variety, from here the pathogenic fungi were obtained, which were morphologically and molecularly characterized using the ITS gene. Subsequently, the isolated pathogenic fungi were confronted with the Trichoderma sp fungus. Results: Ten isolates were obtained, of which four were Fusarium solani. From the confrontations, a 66% to 100% percentage of inhibition in these pathogenic fungi was obtained. Limitations of the study: It is proposed that future evaluations carry out long-term follow-ups to evaluate the persistence and effectiveness of biological control. Findings/conclusions: Timely identification of pathogens could represent a biological control strategy in disease management programs.

Detecting and identifying pathogens and antagonistic bacteria associated with Ginkgo biloba leaf spot disease.

Leaf spot disease severely impacts Ginkgo biloba (G. biloba) yield and quality. While microbial agents offer effective and non-toxic biological control for plant diseases, research on controlling leaf spot disease in G. biloba is notably scarce. The pathogenic fungi were isolated and purified from diseased and healthy leaves of G. biloba, Subsequent examinations included morphological observations and molecular identification via PCR techniques. A phylogenetic tree was constructed to facilitate the analysis of these pathogenic fungi, and Koch's postulates were subsequently employed to reaffirm their pathogenic nature. The antagonistic experiment was employed to select biocontrol bacteria, and subsequently, the isolated biocontrol bacteria and pathogenic fungi were inoculated onto healthy leaves to assess the inhibitory effects of the biocontrol bacteria. Two pathologies responsible for the leaf spot disease on G. biloba were identified as Botryosphaeria dothidea and Neofusicoccum parvum via the analysis of phylogenetic tree and the application of Koch's Postulates. Additionally, we isolated two strains of biocontrol bacteria, namely Bacillus velezensis and Bacillus amyloliquefaciens. Their average inhibitory zones were measured at 4.78 cm and 3.46 cm, respectively. The inhibition zone of B. velezensis against N. parvum was 4 cm. B. velezensis showed a stronger inhibitory effect compared to B. amyloliquefaciens on the development of lesions caused by B. dothidea via leaf culture experiment. This research reports, for the first time, the presence of B. dothidea as a pathogenic fungus affecting G. biloba. Moreover, the biocontrol bacteria, B. velezensis and B. amyloliquefaciens, exhibited the capability to effectively inhibit the growth and reproduction of B. dothidea, indicating their promising potential as environmentally friendly biocontrol resources.

Evaluating the climatic suitability of Engytatus passionarius Minghetti et al. (Heteroptera, Miridae) as a biological control agent of the invasive stinking passion flower Passiflora foetida L. in Australia through ecological niche models

Passiflora foetida is a climbing vine, native to the Neotropical Region that is causing major economic and ecological damage in Australia, where it is rapidly spreading. Traditional control options, such as cutting, manual uprooting, and herbicide applications are only effective for local management. Currently, the plant bug Engytatus passionarius is the most promising biological control agent. Specificity tests performed in its native range in Argentina suggest it is highly specific to the plant, and it has not been observed in the field associated with other plants. As climate determines the establishment of insects, knowing if the environmental conditions suit their requirements is key to introducing a species in a region. Also, an overlap between the climatic niches of species is an indicator of similar requirements. To explore the possibilities of a successful establishment of E. passionarius in Australia, ecological niche models (ENM) were built for the plant bug and for the vine and their overlap was measured. The ENM projected to Australia recognized suitable environmental conditions for the establishment of E. passionarius in several regions where P. foetida is present, both for current and future scenarios. Moreover, the niche of the plant bug is almost completely overlapped with that of the vine. All the aforementioned evidence seems to indicate that E. passionarius has a good chance to become an effective biological control agent of P. foetida.

STEINERNEMA ADAMSI N. SP. (RHABDITIDA: STEINERNEMATIDAE), A NEW ENTOMOPATHOGENIC NEMATODE FROM THAILAND.

A new species of entomopathogenic nematode, Steinernema adamsi n. sp., was recovered from the soil of a longan tree (Dimocarpus sp.) in Mueang Lamphun District, Thailand, using baiting techniques. Upon analysis of the nematode's morphological traits, we found it to be a new species of Steinernema and a member of the Longicaudatum clade. Molecular analyses of the ITS rDNA and D2D3 of 28S rDNA sequences further confirmed that S. adamsi n. sp. is a new species of the Longicaudatum clade, which is closely related to Steinernema guangdongense and Steinernema longicaudam. Using morphometric analysis, the infective juveniles measure between 774.69 and 956.96 μm, males have a size range of 905.44 to 1,281.98 μm, and females are within the range of 1,628.21 to 2,803.64 μm. We also identified the symbiotic bacteria associated with the nematode based on 16S sequences as Xenorhabdus spp. closely related toXenorhabdus griffiniae. Furthermore, we have successfully assessed a cryopreservation method for the long-term preservation of S. adamsi n. sp. Successful cryopreservation of this new species will allow for the longer preservation of its traits and will be valuable for its future use. The discovery of this new species has significant implications for the development of effective biological control agents in Thailand, and our work contributes to our understanding of the diversity and evolution of entomopathogenic nematodes.

Biological Control Effect of Antagonistic Bacteria on Potato Black Scurf Disease Caused by Rhizoctonia solani

Potato black scurf, caused by Rhizoctonia solani, is a severe soil-borne disease that affects the quality and production of potatoes worldwide. In our study, we used Paenibacillus polymyxa YF and Bacillus amyloliquefaciens HT to determine the antagonistic ability of R. solani, with a particular focus on the antagonistic action of P. polymyxa YF to R. solani and its biocontrol effect on potato black scurf. In fermentation, filtrate assay, 50% filtrate of P. polymyxa YF and B. amyloliquefaciens HT inhibited the growth of R. solani by 85.55% and 82.86%, respectively. Microscopic observations showed notable morphological changes with mycelial collapse, atrophy, and deformation following treatment with the antagonistic filtrates. Moreover, cell membrane permeability results showed increased conductivity in bacteria-treated samples compared to the control. P. polymyxa YF exhibited stable colonization on potato plants and secreted various extracellular enzymes (protease, amylase, and cellulase), along with the synthetic substances with growth-promoting effects, such as siderophores and Indole-3-acetic acid (IAA). Whether it is in the excised tissue inoculation or potted experiment, the negative control showed the highest rank of disease symptoms. In the pot experiment, after YF treatment, physiological parameters showed remarkable changes in plant height, root length, stem thickness, and dry and fresh weight. Compared to blank control, the activities of the four resistant enzymes increased significantly in the P. polymyxa YF treatment group. The upregulation in the P. polymyxa YF group was 4.04, 0.54, 0.46, and 3.10 times, respectively. PCR analysis identified genes in both bacterial strains coding for antimicrobial lipopeptides, including fenB, ituC, and srfAA, which are associated with fengycin, iturin, and surfactin synthesis. These findings demonstrated that P. polymyxa YF has a prominent antagonistic effect on R. solani, suggesting its potential as an effective biological control agent for controlling potato black scurf.

Identification of neuropeptides and their G protein-coupled receptors in the predatory stink bug, Arma custos (Hemiptera: Pentatomidae).

The predatory stink bug Arma custos has been selected as an effective biological control agent and has been successfully massly bred and released into fields for the control of a diverse insect pests. As a zoophytophagous generalist, A. custos relies on a complex neuropeptide signaling system to prey on distinct food and adapt to different environments. However, information about neuropeptide signaling genes in A. custos has not been reported to date. In the present study, a total of 57 neuropeptide precursor transcripts and 41 potential neuropeptide G protein-coupled receptor (GPCR) transcripts were found mainly using our sequenced transcriptome data. Furthermore, a number of neuropeptides and their GPCR receptors that were enriched in guts and salivary glands of A. custos were identified, which might play critical roles in feeding and digestion. Our study provides basic information for an in-depth understanding of biological and ecological characteristics of the predatory bug and would aid in the development of better pest management strategies based on the effective utilization and protection of beneficial natural enemies.

Genome-wide transcriptome profiling reveals molecular response pathways of Trichoderma harzianum in response to salt stress.

Trichoderma harzianum exhibits a strong biological control effect on many important plant pathogens, such as Fusarium oxysporum, Botrytis cinerea, and Meloidogyne. However, its biocontrol effectiveness is weakened or reduced under salt stress. The aim of this study was to investigate the molecular response of T. harzianum to salt stress at the whole-genome level. Here, we present a 44.47 Mb near-complete genome assembly of the T. harzianum qt40003 strain for the first time, which was assembled de novo with 7.59 Gb Nanopore sequencing long reads (~170-fold) and 5.2 Gb Illumina short reads (~116-fold). The assembled qt40003 genome contains 12 contigs, with a contig N50 of 4.81 Mb, in which four of the 12 contigs were entirely reconstructed in a single chromosome from telomere to telomere. The qt40003 genome contains 4.27 Mb of repeat sequences and 12,238 protein-coding genes with a BUSCO completeness of 97.5%, indicating the high accuracy and completeness of our gene annotations. Genome-wide transcriptomic analysis was used to investigate gene expression changes related to salt stress in qt40003 at 0, 2% (T2), and 4% (T4) sodium chloride concentrations. A total of 2,937 and 3,527 differentially expressed genes (DEGs) were obtained under T2 and T4 conditions, respectively. GO enrichment analysis showed that the T2-treatment DEGs were highly enriched in detoxification (p < 0.001), while the T4 DEGs were mainly enriched in cell components, mostly in cellular detoxification, cell surface, and cell wall. KEGG metabolic pathway analysis showed that 91 and 173 DEGs were significantly enriched in the T2 and T4 treatments, respectively (p < 0.01), mainly in the glutathione metabolism pathway. We further experimentally analyzed the differentially expressed glutathione transferase genes in the glutathione metabolic pathway, most of which were downregulated (13/15). In addition, we screened 13 genes related to active oxygen clearance, including six upregulated and seven downregulated genes, alongside five fungal hydrophobic proteins, of which two genes were highly expressed. Our study provides high-quality genome information for the use of T. harzianum for biological control and offers significant insights into the molecular responses of T. harzianum under salt-stress conditions.