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Pine wilt disease poses a significant threat to pine forests worldwide, necessitating efficient and accurate detection of dead pine wood for effective disease control and forest management. Traditional deep learning methods based on supervised closed-set paradigms often struggle to address unknown interfering objects, causing false positives, missed detection, and increased annotation burdens. To overcome these challenges, we propose SS-OPDet, a semi-supervised open-set detection framework that leverages a small amount of labeled data along with abundant unlabeled data. SS-OPDet integrates a Weighted Multi-scale Feature Fusion module to dynamically integrate global- and local-scale features, thereby significantly improving representational accuracy for dead pine wood. Additionally, a Dynamic Confidence Pseudo-Label Generation strategy categorizes predictions by confidence level, effectively reducing training noise and maximizing the use of reliable unlabeled data. Experimental results from 7733 UAV images demonstrate that SS-OPDet achieves an average precision (APK) of 84.73%, a recall (RK) of 94.48%, an Absolute Open-Set Error (AOSE) of 271 and a Wilderness Impact (WI) of 0.0917%. Cross-region validation further confirms the robustness and generalization capability of the proposed framework. The proposed method offers a cost-effective and accurate solution for timely detection of pine wilt disease, providing substantial benefits to forest monitoring and management.

Monochamus alternatus larvae, as concealed trunk-boring pests, evade conventional insecticide contact due to their cryptic feeding niche. To overcome this limitation, previous studies have engineered strains of the naturally entomopathogenic bacterium Yersinia entomophaga. The lethality of these strains against M. alternatus was enhanced by incorporating extracellular secretion systems and enriching insecticidal proteins within the larval midgut. However, plasmid loss occurs during serial subculturing. Here, we established an engineered strain that expresses the red fluorescent protein gene mCherry to explore the applicability of bacterial conjugation transfer to Yersinia. We then constructed a chromosomally integrated strain (CSLH88-pCHSW) that incorporates extracellular secretion systems. The results of stability assays demonstrated 100% retention of the mCherry and Cry3Aa-T-HasA genes over 78 generations. SDS-PAGE and Western blot analyses confirmed the extracellular secretion of the Cry3Aa-T protein in the CSLH88-pCHSW strain. Bioassays revealed that the CSLH88-pCHSW strain was significantly more virulent against M. alternatus larvae than both the wild-type strain (CSLH88) and the plasmid-transformed strain (CSLH88-pCHKW), and exhibited markedly faster insecticidal kinetics. Our study reveals the application of bacterial conjugation transfer technology for constructing biocontrol strains. This genomically stabilized Yersinia strain eliminates the risks of failure associated with plasmid loss in the field, enabling the sustainable control of M. alternatus.

Herbivore-induced plant volatiles (HIPVs) play a pivotal role in mediating tritrophic interactions between plants, herbivores, and their natural enemies. Paracarophenax alternatus, a parasitic mite targeting the egg stage of Monochamus alternatus, has emerged as a promising biocontrol agent. However, its ability to detect Pinus massoniana-derived HIPVs for host insect localization remains unclear. G-protein-coupled receptors (GPCRs) may play a role in mediating the perception of HIPVs and associated chemosensory signaling pathways in mites. In this study, a total of 85 GPCRs were identified from P. alternatus. All GPCRs exhibited conserved transmembrane domains and stage-specific expression patterns, with 21 receptors significantly upregulated in viviparous mites. Combined with two previously identified odorant receptors (ORs), six candidate chemosensory receptors were selected for molecular dynamics simulations to validate their binding stability with key volatile compounds. The results demonstrate that specific GPCRs likely facilitate HIPV detection in mites, enabling precise host localization within dynamic ecological niches. Our findings provide critical insights into the molecular basis of mite–host interactions and establish a framework for optimizing P. alternatus-based biocontrol strategies against pine wilt disease vectors.

In this study, we focused on the screening and identification of reference genes for Paracarophenax alternatus Xu and Zhang. The laboratory population was used as the laboratory population, and samples were collected from mites at four different stages, including physogastry, viviparous, 5 d viviparous and phoresy. Then, the expression levels of seven candidate reference genes (α-tubulin, β-tubulin, RPS18, RPL13, GAPDH, EF1A, SDHA) were detected through qRT-PCR. Melting curves showed good gene specificity, and the amplification efficiency ranged from 90% to 102%. ΔCt analysis indicated that GAPDH was the most stable reference gene. The GeNorm software determined that the optimal number of reference genes was two, with GAPDH and RPS18 forming the most stable combination, and NormFinder identified RPS18 as the most stable reference gene. Although the BestKeeper software suggested that EF1A was the most stable, its p-value exceeded 0.05, rendering it unsuitable for use as a reference gene. Finally, through the RefFinder network tool, the most stable reference genes were identified as GAPDH and RPS18.

Background Monochamus alternatus Hope is one of the insect vectors of pinewood nematode ( Bursaphelenchus xylophilus ), which causes the destructive pine wilt disease. The microorganisms within the ecosystem, comprising plants, their environment, and insect vectors, form complex networks. This study presents a systematic analysis of the bacterial microbiota in the M. alternatus midgut and its habitat niche. Methods Total DNA was extracted from 20 types of samples (with three replicates each) from M. alternatus and various tissues of healthy and infected P. massoniana (pines). 16S rDNA amplicon sequencing was conducted to determine the composition and diversity of the bacterial microbiota in each sample. Moreover, the relative abundances of bacteria in the midgut of M. alternatus larvae were verified by counting the colony-forming units. Results Pinewood nematode infection increased the microbial diversity in pines. Bradyrhizobium , Burkholderia , Dyella , Mycobacterium , and Mucilaginibacter were the dominant bacterial genera in the soil and infected pines. These results indicate that the bacterial community in infected pines may be associated with the soil microbiota. Interestingly, the abundance of the genus Gryllotalpicola was highest in the bark of infected pines. The genus Cellulomonas was not found in the midgut of M. alternatus , but it peaked in the phloem of infected pines, followed by the phloem of heathy pines. Moreover, the genus Serratia was not only present in the habitat niche, but it was also enriched in the M. alternatus midgut. The colony-forming unit assays showed that the relative abundance of Serratia sp. peaked in the midgut of instar II larvae (81%). Conclusions Overall, the results indicate that the bacterial microbiota in the soil and in infected pines are correlated. The Gryllotalpicola sp. and Cellulomonas sp. are potential microbial markers of pine wilt disease. Additionally, Serratia sp. could be an ideal agent for expressing insecticidal protein in the insect midgut by genetic engineering, which represents a new use of microbes to control M. alternatus .

Background Pine wilt disease (PWD) is a destructive disease that endangers pine trees, resulting in the wilting, with yellowing and browning of the needles, and eventually the death of the trees. Previous studies showed that the Avr9/Cf-9 rapidly elicited ( PmACRE1 ) gene was downregulated by Bursaphelenchus xylophilus infection, suggesting a correlation between PmACRE1 expression and pine tolerance. Here, we used the expression of PmACRE1 in Arabidopsis thaliana to evaluate the role of PmACRE1 in the regulation of host defence against B. xylophilus infection. Results Our results showed that the transformation of PmACRE1 into A. thaliana enhanced plant resistance to the pine wood nematode (PWN); that is, the leaves of the transgenic line remained healthy for a longer period than those of the blank vector group. Ascorbate peroxidase (APX) activity and total phenolic acid and total flavonoid contents were higher in the transgenic line than in the control line. Widely targeted metabolomics analysis of the global secondary metabolites in the transgenic line and the vector control line showed that the contents of 30 compounds were significantly different between these two lines; specifically, the levels of crotaline, neohesperidin, nobiletin, vestitol, and 11 other compounds were significantly increased in the transgenic line. The studies also showed that the ACRE1 protein interacted with serine hydroxymethyltransferase, catalase domain-containing protein, myrosinase, dihydrolipoyl dehydrogenase, ketol-acid reductoisomerase, geranylgeranyl diphosphate reductase, S-adenosylmethionine synthase, glutamine synthetase, and others to comprehensively regulate plant resistance. Conclusions Taken together, these results indicate that PmACRE1 has a potential role in the regulation of plant defence against PWNs.

Lymantria xylina is a major pest in coastal casuarina shelterbelts and a species subject to quarantine regulations by countries to which it is non-native. Phototaxis is fundamental to the insect’s surveillance and risk assessment analysis, and it exhibits pronounced sexual dimorphism in compound eye structure. This dimorphism was investigated using scanning and transmission electron microscopy. Males displayed significantly larger compound eyes, characterized by greater ommatidial areas and a higher total number of facets per eye compared to females. From the distal to proximal end, the ommatidium consists of the cornea, primary and secondary pigment cells, crystalline cones, retinula cells, a rhabdom bundle, and basal retinal cells (in a “7 + 1” arrangement). The internal ultrastructure of the ommatidia is similar in both sexes. However, males possess significantly thinner cornea and extremely elongated crystalline cones. Based on external morphology, both sexes generally exhibit a parallel-symmetrical compound eye form, minimizing optical asymmetry to optimize nocturnal vision. These differences are attributed to the distinct visual demands of males for mate-searching in low-light environments, while females, being more stationary, have reduced visual needs. Paraffin sections of Lymantria xylina compound eyes further revealed that, during dark adaptation, pigment granules aggregated within the crystalline cone region to enhance low-light capture. Conversely, following intense light stimulation, these granules translocated to the perinuclear region of photoreceptor cells, forming a light-shielding configuration.

Lymantria xylina Swinhoe (Lepidoptera: Erebidae) is considered a potentially internationally invasive forest pest with limited knowledge about its phototactic behavior. This study investigated the phototaxis of L. xylina males and females using various insecticidal lamps in the field. The results showed that all lamps attracted both males and females, but females were captured in a very low numbers, with a female-to-male ratio of 1:322. The insecticidal lamp with a peak wavelength of 363 nm was most effective for male trapping. Males exhibited a distinct light-trapping rhythm, peaking around midnight (23:00–0:00), with 29.5% capture, while females were most active at dusk (19:00–20:00), with 44.4% capture. Light-trapped females were smaller and lighter than indoor-emerged females and had lower egg-carrying capacity. Females, when laying eggs, did not exhibit phototactic behavior. Only 14.6% of non-ovipositing females showed phototactic behavior, and only 0.6% flew directly toward the lamp. These findings suggest that while non-ovipositing females can exhibit phototaxis, only a small proportion will, potentially reducing the risk of long-distance dispersal of L. xylina via ocean-going freighters. The results showed that the non-ovipositing females could fly under phototaxis, but their phototaxis was limited. This study provides a basis for the risk assessment of the long-distance dispersal of L. xylina via ocean-going freighters through female moth phototactic flight.

The rapid spread of pine wilt disease has become a major crisis with regard to forest ecological security. Against the background of China’s concerted effort to achieve carbon neutrality by 2060, balancing economic and environmental costs during the environmentally sound treatment of nematode-infected pine wood is an issue worthy of attention. In this study, we constructed an evolutionary game model of the central government, local governments, and infected wood management-related enterprises and analyzed the evolutionary process of the three parties with stable strategies based on a full consideration of actual circumstances. To verify the theoretical results, we conducted numerical simulations using MATLAB. The results of the study are as follows. 1) The central government plays a decisive role in how infected wood is handled. The greater the degree of regulation is, the greater the local government favors low carbon emissions and the more companies will choose the pulverization method. 2) Local governments bear the main responsibility in the processing of infected wood, and only when local governments are sufficiently penalized will enterprises choose to pulverize the wood and dispose of it. 3) Policy subsidies are an effective measure that will help the three parties in the model reach a balance more quickly. Overall, this study clarified the realization path of low-carbon treatment of nematode-infected pine wood from a policy perspective. The study results should help promote the low-carbon treatment of nematode-infected pine wood and provide a basis for the formulation of relevant forestry policies in the context of carbon neutrality.

The red palm weevil (RPW), Rhynchophorus ferrugineus (Oliver) is an important pest of palms that causes significant damage by boring into and feeding within palm stem tissues. Here, we studied the proteolytic process of Cry3Aa in the RPW to understand the mechanism of Cry toxicity. The bioassays showed that Cry3Aa toxin is weakly toxic to the RPW. Proteolytic activation assays indicated the Cry3Aa protein is digested into smaller fragments than the 55-kDa activated fragments under different conditions. In particular, at higher mass ratios of gut protease and Cry3Aa protein (5:1, 2:1, and 1:1, respectively), and at 36.9°C for 16 h in a solution of pH 8.6, the Cry3Aa protoxin is over-digested by the gut proteases of weevil larvae. Moreover, the zymogram analysis of the gut proteases revealed the RPW larvae harbors intestinal digestive enzymes mainly composed of serine proteases.This study describes the proteolytic activation process of Cry3Aa in the midgut of RPW larvae.