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Wheat straw (WS) is a potential biomass for production of monomeric sugars. However, the enzymatic hydrolysis ratio of cellulose in WS is relatively low due to the presence of lignin and hemicellulose. To enhance the enzymatic conversion of WS, we tested the impact of three different pretreatments, e.g. sulfuric acid (H 2 SO 4 ), sodium hydroxide (NaOH), and hot water pretreatments to the enzymatic digestions. Among the three pretreatments, the highest cellulose conversion rate was obtained with the 4% NaOH pretreatment at 121 °C (87.2%). In addition, NaOH pretreatment was mainly effective in removing lignin, whereas the H 2 SO 4 pretreatment efficiently removed hemicellulose. To investigate results of pretreated process for enhancement of enzyme-hydolysis to the WS, we used scanning electron microscopy, X-ray diffraction, and Fourier transform infrared spectroscopy to analyze structural changes of raw and treated materials. The structural analysis indicated that after H 2 SO 4 and NaOH pretreatments, most of the amorphous cellulose and partial crystalline cellulose were hydrolyzed during enzymatic hydrolysis. The findings of the present study indicate that WS could be ideal materials for production of monomeric sugars with proper pretreatments and effective enzymatic base hydrolysis.

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.

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 .

The appressorium is a specialised infection structure formed by Beauveria bassiana during host invasion. This study used sulforaphane to regulate the formation rate of B. bassiana appressoria, evaluated the correlation between appressorium formation and fungal pathogenicity, and explored its impact on insect cuticular metabolism. The results showed that sulforaphane significantly modulated appressorium formation. Spore suspensions with varying appressorium formation rates were injected into Opisina arenosella and Bombyx mori larvae. As the appressorium formation rate increased, B. bassiana exhibited enhanced pathogenicity, leading to accelerated larval mortality. A significant positive correlation (p ≤ 0.05) was observed between appressorium formation and pathogenicity. LC-MS analysis revealed that, prior to appressorium development, larvae activated defence mechanisms involving secondary metabolites, hormone signalling, and toxin metabolism pathways. Following appressorium formation, 61 unique cuticular compounds were identified, along with activation of host lipid metabolism (notably glycerophospholipid degradation), programmed cell death pathways (ferroptosis, necroptosis), and enhanced energy metabolism via the citric acid cycle—collectively indicating disruption of the epidermal defence barrier. Overall, appressorium development by B. bassiana significantly reshapes the metabolic landscape of the larval cuticle, thereby enhancing fungal virulence. This study provides a theoretical foundation for understanding the pathogenic mechanisms of B. bassiana.

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.

The casuarina moth, Lymantria xylina , is a serious pest threatening subtropical regions through severe defoliation and strong invasive potential. Despite its economic impact and high invasion risk, a high-quality reference genome remains lacking. To bridge this knowledge gap, we generated a chromosome-level genome assembly for L. xylina combining Illumina short-reads, Oxford Nanopore long-reads, and high-throughput chromatin conformation capture (Hi-C) scaffolding data. Following long-reads based assembly and Hi-C scaffolding, the final genome assembly totals 977.74 Mb, with 930.50 Mb (95.17%) of sequences anchored onto 31 pseudo-chromosomes, achieving a scaffold N50 of 34.15 Mb. The genome assembly, featuring fully assembled telomeres on all 31 pseudo-chromosomes, demonstrates 94.5% Benchmarking Universal Single-Copy Orthologs (BUSCO) completeness and high accuracy with consensus quality value of 31.72. Repetitive elements constitute 77.18% of the genome, and 18,484 protein-coding genes were predicted, with 95.21% functionally annotated. This high-quality genome assembly provides a critical foundation for elucidating interaction mechanisms with host plants and natural enemies (nucleopolyhedrovirus, Beauveria bassiana ), for developing enhanced pest management and control strategies.

Dendrolimus houi is one of the most common caterpillars infesting Gymnosperm trees, and widely distributed in several countries in Southeast Asia, and exists soley or coexists with several congeners and some Lasiocampidae species in various forest habitats. However, natural hybrids occasionally occur among some closely related species in the same habitat, and host preference, extreme climate stress, and geographic isolation probably lead to their uncertain taxonomic consensus. The mitochondrial DNA (mtDNA) of D. houi was extracted and sequenced by using high-throughput technology, and the mitogenome composition and characteristics were compared and analyzed of these species, then the phylogenetic relationship was constructed using the maximum likelihood method (ML) and the Bayesian method (BI) based on their 13 protein-coding genes (PCGs) dataset, which were combined and made available to download which were combined and made available to download among global Lasiocampidae species data. Mitogenome of D. houi was 15,373 bp in length, with 37 genes, including 13 PCGs, 22 tRNA genes (tRNAs) and 2 rRNA genes (rRNAs). The positions and sequences of genes were consistent with those of most known Lasiocampidae species. The nucleotide composition was highly A+T biased, accounting for ~80% of the whole mitogenome. All start codons of PCGs belonged to typical start codons ATN except for COI which used CGA, and most stop codons ended with standard TAA or TAG, while COI, COII, ND4 ended with incomplete T. Only tRNASer (AGN) lacked DHU arm, while the remainder formed a typical \"clover-shaped\" secondary structure. For Lasiocampidae species, their complete mitochondrial genomes ranged from 15,281 to 15,570 bp in length, and all first genes started from trnM in the same direction. And base composition was biased toward A and T. Finally, both two methods (ML and BI) separately revealed that the same phylogenetic relationship of D. spp. as (((D. punctatus + D. tabulaeformis) + D. spectabilis) + D. superans) + (D. kikuchii of Hunan population + D. houi) as in previous research, but results were different in that D. kikuchii from a Yunnan population was included, indicating that different geographical populations of insects have differentiated. And the phylogenetic relationship among Lasiocampidae species was (((Dendrolimus) + Kunugia) + Euthrix) + Trabala). This provides a better theoretical basis for Lasiocampidae evolution and classification for future research directions.

Rhynchophorus ferrugineus is a quarantine pest that mainly damages plants in tropical regions, which are essential economic resources. Cry3Aa has been used to control coleopteran pests and is known to be toxic to R. ferrugineus . The binding of the Cry toxin to specific receptors on the target insect plays a crucial role in the toxicological mechanism of Cry toxins. However, in the case of R. ferrugineus , the nature and identity of the receptor proteins involved remain unknown. In the present study, pull-down assays and mass spectrometry were used to identify two proteins of aminopeptidase N proteins (RfAPN2a and RfAPN2b) in the larval midguts of R. ferrugineus . Cry3Aa was able to bind to RfAPN2a ( Kd = 108.5 n M ) and RfAPN2b ( Kd = 68.2 n M ), as well as midgut brush border membrane vesicles ( Kd = 482.5 n M ). In silico analysis of both RfAPN proteins included the signal peptide and anchored sites for glycosyl phosphatidyl inositol. In addition, RfAPN2a and RfAPN2b were expressed in the human embryonic kidney 293T cell line, and cytotoxicity assays showed that the transgenic cells were not susceptible to activated Cry3Aa. Our results show that RfAPN2a and RfAPN2b are Cry3Aa-binding proteins involved in the Cry3Aa toxicity of R. ferrugineus . This study deepens our understanding of the action mechanism of Cry3Aa in R. ferrugineus larvae.

Aedes aegypti, a crucial vector mosquito that transmits many diseases that cause millions of deaths worldwide, can be controlled with Bacillus thuringiensis subsp. israelensis (Bti). The larvicidal activity of Bti against Ae. aegypti is due primarily to Cry4Aa, Cry4Ba, and Cry11Aa, and Cyt1Aa, a protein that synergizes the activity of the Cry proteins. Interestingly, Galectins-6 and Galectins-14, members of a family of β-galactoside-binding proteins that play a role in immune responses insects, have been shown to decrease the activity of Bti toxins. The activity of other Galectins, particularly Galectin-8A, against the Cry proteins is not known. Toward this end, we cloned the gene coding for galactin-8A and expressed the recombinant protein and purified protein. The bioassay results indicated that Galectin-8A can also reduce the toxicity of Cry11Aa, but it was much stronger than Galectin-6. To investigate the interactions among Galectin-8A, Cry11Aa, and toxin receptors, Octet Red System analysis, Western blot, far-Western blot, and ELISA assay were also performed. The Octet Red System result showed that Galectin-8A could also bind to BBMVs of Ae. aegypti, with a lower kDa value than that of Galectin-6, indicating that Galectin-8A had a stronger binding affinity to BBMVs than Galectin-6. Western blot, far-Western blot, and ELISA assay analyses also demonstrated that Galectin-8A bound to Ae. aegypti receptor ALP1 and APN2, consistent with the protein docking simulation results. These findings support the conclusion that Galectin-8A blocks with ALP1 and APN2 more effectively than Galectin-6, which may subsequently reduce the toxicity of Cry11Aa in Ae. aegypti.

Background Quantitative real-time reverse transcription-polymerase chain reaction has been widely used in gene expression analysis, however, to have reliable and accurate results, reference genes are necessary to normalize gene expression under different experimental conditions. Several reliable reference genes have been reported in plants of Traditional Chinese Medicine, but none have been identified for Euscaphis konishii Hayata. Results In this study, 12 candidate reference genes, including 3 common housekeeping genes and 9 novel genes based on E. konishii Hayata transcriptome data were selected and analyzed in different tissues (root, branch, leaf, capsule and seed), capsule and seed development stages. Expression stability was calculated using geNorm and NormFinder, the minimal number of reference genes required for accurate normalization was calculated by Vn/Vn + 1 using geNorm. EkEEF - 5A - 1 and EkADF2 were the two most stable reference genes for all samples, while EkGSTU1 and EkGAPDH were the most stable reference genes for tissue samples. For seed development stages, EkGAPDH and EkEEF - 5A - 1 were the most stable genes, whereas EkGSTU1 and EkGAPDH were identified as the two most stable genes in the capsule development stages. Two reference genes were sufficient to normalize gene expression across all sample sets. Conclusion Results of this study revealed that suitable reference genes should be selected for different experimental samples, and not all the common reference genes are suitable for different tissue samples and/or experimental conditions. In this study, we present the first data of reference genes selection for E. konishii Hayata based on transcriptome data, our data will facilitate further studies in molecular biology and gene function on E. konishii Hayata and other closely related species.