PEST MANAGEMENT SCIENCE
·
2024
·
IF 3.8
Xiangying Zou,Jiayu Jia,Tengfei Zhu,Shouping Cai,Yiman He,Shunde Su,Yu Fang,Jian Li,Guifang Lin,Jun Su
BACKGROUND Pine wood nematode (PWN; Bursaphelenchus xylophilus ) is the causative agent of pine wilt disease (PWD), which is considered the most dangerous biohazard to conifer trees globally. The transmission of PWN relies on insect vectors, particularly the Japanese pine sawyer (JPS; Monochamus alternatus ). However, the molecular mechanism underlying PWN-JPS assembly remains largely unknown. RESULTS Here, we found that both geographical and gender could significantly affect the PCA (PWN carrying amount) of JPS; thus, JPS transcriptomes from diverse locations and genders were explored regard to PWN loading. Due to the shortage of genomes, we developed a full-length reference transcriptome for analyzing next-generation sequencing data. A comparative genomic study was performed, and 11 248 potential PWN-carrying associate genes (β) were nominated in JPS by using the reported genomes of PWN and non-PWN carrier insect species. Then, 151 differentially expressed transcripts (DETs), 28 of them overlapped with β, correlated with the PCA of JPS were nominated by RNA-Seq, and found that fatty acid β-oxidation might be the key factor that affected the PCA of JPS. Furthermore, JPS fatty acid β-oxidation rates were experimentally decreased using the inhibitor Etomoxir, leading to an increased PCA of JPS. Meanwhile, silencing MaCPT1 in JPS by RNA interference led to a decreased fatty acid β-oxidation rate and increased PCA of JPS. CONCLUSIONS In conclusion, MaCPT1 was able to decrease the PWN-JPS assembly formation through the fatty acid β-oxidation of JPS. These results provide new insights for exploring the impact of PWN invasion on JPS. © 2024 Society of Chemical Industry.
研究领域:
植物病理学
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昆虫学
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分子生物学
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基因组学
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转录组学
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产品关联
DOI
PubMed
高影响因子
Jove-Journal of Visualized Experiments
·
2022
·
IF 1.424
Xiaofang Liu,Xiang Zhou,Lifeng Zhou,Jiafu Hu,Kai Guo
The pinewood nematode, Bursaphelenchus xylophilus, is one of the most destructive invasive species worldwide, causing the wilting and eventual death of pine trees. Despite the recognition of their economic and environmental significance, it has thus far been impossible to study the detailed gene functions of plant-parasitic nematodes (PPNs) using conventional forward genetics and transgenic methods. However, as a reverse genetics technology, RNA interference (RNAi) facilitates the study of the functional genes of nematodes, including B. xylophilus. This paper outlines a new protocol for RNAi of the ppm-1 gene in B. xylophilus, which has been reported to play crucial roles in the development and reproduction of other pathogenic nematodes. For RNAi, the T7 promoter was linked to the 5′-terminal of the target fragment by polymerase chain reaction (PCR), and double-stranded RNA (dsRNA) was synthesized by in vitro transcription. Subsequently, dsRNA delivery was accomplished by soaking the nematodes in a dsRNA solution mixed with synthetic neurostimulants. Synchronized juveniles of B. xylophilus (approximately 20,000 individuals) were washed and soaked in dsRNA (0.8 µg/mL) in the soaking buffer for 24 h in the dark at 25 °C.The same quantity of nematodes was placed in a soaking buffer without dsRNA as a control. Meanwhile, another identical quantity of nematodes was placed in a soaking buffer with green fluorescent protein (gfp) gene dsRNA as a control. After soaking, the expression level of the target transcripts was determined using real-time quantitative PCR. The effects of RNAi were then confirmed using microscopic observation of the phenotypes and a comparison of the body size of the adults among the groups. The current protocol can help advance research to better understand the functions of the genes of B. xylophilus and other parasitic nematodes toward developing control strategies through genetic engineering.
研究领域:
分子生物学
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植物病理学
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线虫学
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遗传学
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产品关联
DOI
PubMed
高影响因子
PLANT DISEASE
·
2026
·
IF 2.8
Sai-Nan Li,Bao-Ming Shen,Jin-Xiu Yu,Yun Tan,Ai-Rong Shen,Li-Na Liu,Zhu-Ming Tan
Pine wilt disease, caused by the pinewood nematode (PWN) Bursaphelenchus xylophilus, devastates pine forests worldwide. Although the medicinal fungus Wolfiporia hoelen exhibits nematicidal activity against PWN, its specific bioactive constituents remain unidentified. This study investigated the nematicidal activity of volatile organic compounds (VOCs) released from W. hoelen against PWN. Results indicated that VOCs from W. hoelen exhibit significant inhibitory efficacy against second-stage juveniles (J2s) of PWN, achieving 90% mortality after 72 h of exposure. In contrast, ovicidal activity was limited (6% hatching inhibition after 24 h). To analyze the VOCs produced by W. hoelen via solid-phase microextraction gas chromatography–mass spectrometry (SPME-GC-MS), we identified six major and commercially available VOCs: (Z)-5-octen-1-ol, 1-octen-3-ol, 3-octanol, 3-octanone, phenylglyoxal, and phenylacetaldehyde. Five compounds (excluding phenylglyoxal) exhibited nematicidal activity against PWN J2s through both contact and fumigation. These findings demonstrate the efficacy of W. hoelen and its VOCs as promising candidates for management of PWN.
✓ 已验证
DOI
PubMed
高影响因子
PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY
·
2021
·
IF 3.963
Jie Chen,Xin Hao,Buyong Wang,Ling Ma
Bursaphelenchus xylophilus is one of the most dangerous forest pathogens in the world, causing devastating pine forest deaths with considerable economic losses. In this study, we investigated the B. xylophilus RNA sequence responses of two different concentrations of levamisole hydrochloride (LH). We observed that body-wall muscle twitching, paralysis and, ultimately, death. 2.5 mg/ml and 3.5 mg/ml LH have toxicological effects on B. xylophilus , with mortality increasing significantly with concentration ( p < 0.05). RNA sequencing, differential gene expression analysis, and cluster analysis were performed, and 336, 384, 6 genes with significant variance in expression were identified. Gene Ontology annotation and Kyoto Encyclopedia of Genes and Genomes pathway analyses of the 12 intersecting genes revealed that these genes are mostly involved in metabolism of xenobiotics and have essential roles in drug sensitivity. Through the trend analysis of DEGs, it was divided into 8 modules, and the significant modules were selected to construct the co-expression network as the central genes of the drug metabolism-cytochrome P450 pathway (ko00982) and metabolism of xenobiotics by cytochrome P450 (ko00980). Eight highly related genes were identified, including cuticle collagen, cystathionine beta-synthase, endochitinase, pyruvate dehydrogenase E1 component subunit beta , aldehyde dehydrogenase , lipase , and zinc metalloproteinase . The expression levels of these genes were upregulated significantly at low concentrations and were significantly related to the resistance of B. xylophilus to LH. This study shows that B. xylophilus gene family expansions occurred in xenobiotic detoxification pathways through gene expression and potential horizontal correlated gene transfer with LH and helps to elucidate LH lethality and the evolutionary mechanisms underlying the adaptations of B. xylophilus to the environment. These results contributing to our understanding of B. xylophilus under LH and provide a data platform to providing a basis for its control.
研究领域:
分子生物学
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基因组学
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转录组学
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线虫学
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药物基因组学
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DOI
PubMed
高影响因子
MOLECULES
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2016
·Vol. 21
·期 10
·1276
·
IF 2.465
Gai Liu,Daowan Lai,Qi Zhi Liu,Ligang Zhou,Zhi Long Liu
During a screening program for new agrochemicals from Chinese medicinal herbs, the ethanol extract ofNotopterygium incisumrhizomes was found to possess strong nematicidal activity against the two species of nematodes,Bursaphelenchus xylophilusandMeloidogyne incognita. Based on bioactivity-guided fractionation, the four constituents were isolated from the ethanol extract and identified as columbianetin, falcarindiol, falcarinol, and isoimperatorin. Among the four isolated constituents, two acetylenic compounds, falcarindiol and falcarinol (2.20–12.60 μg/mL and 1.06–4.96 μg/mL, respectively) exhibited stronger nematicidal activity than two furanocoumarins, columbianetin, and isoimperatorin (21.83–103.44 μg/mL and 17.21–30.91 μg/mL, respectively) against the two species of nematodes,B. xylophilusandM. incognita. The four isolated constituents also displayed phototoxic activity against the nematodes. The results indicate that the ethanol extract ofN.incisumand its four isolated constituents have potential for development into natural nematicides for control of plant-parasitic nematodes.
研究领域:
天然产物化学
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线虫学
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农用化学品研究
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植物化学
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害虫防治
✓ 已验证
DOI
高影响因子
ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY
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2024
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IF 6.2
Buyong Wang,Rongrong Wen,Xuenan Mao,Jie Chen,Xin Hao
Bursaphelenchus xylophilus is a pathogenic nematode responsible for pine wilt disease, which can cause the demise of pine trees and discoloration of trunks. As rotenone is an important botanical pesticide, its impact on B. xylophilus was investigated through RNA-seq to understand the response mechanism of nematode. The bioassay results yielded the 12-h LC 30 (1.35 mg L −1 ) and LC 50 (2.60 mg L −1 ) values for rotenone. Differential gene expression analysis identified 172 and 614 differentially expressed genes (DEGs) in B. xylophilus exposed to two different concentrations of rotenone (1.35 and 2.60 mg L −1 ). To validate these findings, the expression patterns of 10 DEGs were confirmed through RT-qPCR. Additionally, all DEGs were categorized into eight gene expression profiles using STEM. Notably, the gene ontology (GO) processes of "single-organism process," "metabolic process," and "catalytic activity" were prominently enriched in rotenone-treated samples, suggesting a role for metabolic and catalytic pathways in the nematode's response to rotenone stress. KEGG pathways related to "carbon metabolism," "drug metabolism-cytochrome P450," and "metabolism of xenobiotics by cytochrome P450" were similarly enriched, indicating potential mechanisms for detoxification resistance and oxidative stress resistance. The analysis pointed to the pivotal roles of detoxification- and oxidoreduction-related genes, as well as signal transduction-related genes, in enabling B. xylophilus to adapt to rotenone exposure. These insights could markedly enhance our understanding of nematode resistance mechanisms and potentially inform the development of more effective rotenone-based strategies for controlling B. xylophilus .
研究领域:
植物保护
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分子生物学
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农药科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
LANGMUIR
·
2025
·
IF 3.9
Fei Teng,Zhaogui Wang,Mingyu Xue,Hui Li,Quanmin Wen,Peng Xu,Dejun Hao
Pine wilt disease is caused by Bursaphelenchus xylophilus invasion and has a great impact on global pine resources. Injection of emamectin benzoate (EB) into pine trunks is an effective way to control B. xylophilus. However, EB has limited aqueous solubility, easily photodissociates, and its long-term use causes resistance problems. In this study, the metal–organic framework material ZIF-8 was prepared, and EB was loaded onto ZIF-8 through physical adsorption, resulting in a pH-responsive EB@ZIF-8 nanopesticide with a high drug loading rate (36.04%). Simulated release experiments of EB@ZIF-8 under different pH conditions showed a cumulative release of 64.43% within 10 h at pH 5.0, which increased as the pH decreased, compared to conditions at pH 7.0 and 9.0. EB@ZIF-8 showed positive insecticidal activity in a laboratory biological test (LC50 = 52.503 mg/L). In addition, 7 days after trunk injection in Pinus massoniana, EB residuals were detected in all parts of the branches, with the lowest residue level being 73.39 mg/kg. The proportion of the EB residue in the lower, middle, and upper branches of P. massoniana gradually increased, reaching 43.92% at 28 days after trunk injection, indicating that EB was distributed evenly in the tree. Therefore, EB@ZIF-8 had good morphology, structure, and pH-responsive release performance. EB@ZIF-8 also exhibited a high nematocidal ability due to its smaller particle size and rapid transport in P. massoniana. This study provides a new approach for the development of highly effective EB preparations, improvement of pesticide utilization, and B. xylophilus control.
研究领域:
植物保护
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材料科学
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农业科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Biology-Basel
·
2021
·Vol. 10
·期 8
·785
·
IF 5.079
Long Pan,Rong Cui,Yongxia Li,Wei Zhang,Jianwei Bai,Juewen Li,Xingyao Zhang
Simple SummaryPine wilt disease caused by the nematodeBursaphelenchus xylophiluscauses significant harm to China’s forests, but there are currently no effective prevention and control measures. Additionally, this devastating disease is currently spreading northward. We determined that third-stage dispersal juveniles ofB. xylophiluscan resist low-temperature stress by cryptobiosis, allowing these nematodes to tolerate a greater range of temperatures. These results facilitate the prediction of potential areas at risk forB. xylophilusin the mid-temperature and cold temperature zones of China.Nematodes can enter cryptobiosis by dehydration as an adaptation to low-temperature environments and recover from cryptobiosis by rehydration after environmental improvement. In this work, the survival ofBursaphelenchusxylophilusthird-stage dispersal juveniles was studied in response to low-temperature treatment. The average survival rates were 1.7% after −80 °C treatment for 30 d and 82.2% after −20 °C treatment for 30 d. The changes of water content and inorganic salt ions that occur in pine trees during winter gradually alter the osmotic pressure in the liquid environment to dehydrateB. xylophilusjuveniles, resulting in improved survival after low-temperature treatment. The survival rate at −20 °C improved to 92.1% when the juveniles entered cryptobiosis by osmotic regulation. The results of this study demonstrate thatB. xylophilusthird-stage dispersal juveniles can resist low-temperature stress through cryptobiosis, providing the theoretical basis for the identification of areas potentially vulnerable toB. xylophilusin the mid-temperature and cold temperature zones of China.
✓ 已验证
DOI
高影响因子
Genes
·
2022
·Vol. 13
·期 11
·1956
·
IF 4.141
Jie Chen,Xin Hao,Ruina Tan,Yang Li,Bowen Wang,Jialiang Pan,Wei Ma,Ling Ma
Bursaphelenchus xylophilus(PWN) causes pine wilt disease (PWD), which is one of the most devastating pine diseases worldwide. Cytochrome P450 (CYP) catalyzes the biosynthetic metabolism of terpenoids and plays an important role in the modification of secondary metabolites in all living organisms. We investigated the molecular characteristics and biological functions ofBx-cyp29A3inB. xylophilus. The bioinformatics analysis results indicated thatBx-cyp29A3has a transmembrane domain and could dock with L(−)-carvone. The gene expression pattern indicated thatBx-cyp29A3was expressed in 0.2, 0.4, 0.6, 0.8, and 1.0 mg/mL L(−)-carvone solutions. TheBx-cyp29A3expression increased in a dose-dependent manner and peaked at 24 h of exposure when the L(−)-carvone solution concentration was 0.8 mg/mL. However, the gene expression peaked at 0.6 mg/mL after 36 h. Furthermore, RNA interference (RNAi) indicated thatBx-cyp29A3played an essential role in the response to L(−)-carvone. The mortality rates of theBx-cyp29A3knockdown groups were higher than those of the control groups in the 0.4, 0.6, 0.8, and 1.0 mg/mL carvone solutions after 24 h of exposure or 36 h of exposure. In summary, bioinformatics provided the structural characteristics and conserved sequence properties ofBx-cyp29A3and its encoded protein, which provided a target gene for the study of the P450 family ofB. xylophilus. Gene silencing experiments clarified the function ofBx-cyp29A3in the immune defense ofB. xylophilus.This study provides a basis for the screening of new molecular targets for the prevention and management ofB. xylophilus.Keywords:cytochrome P450;L(−)-carvone;Bursaphelenchus xylophilus;RNAi
研究领域:
分子生物学
·
植物病理学
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线虫学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Forests
·
2026
·Vol. 17
·期 6
·668
·
IF 3.1
Xianglian Jin,Zhen Zhang,Jing Ning,Ziying Liu,Chen Zhang,Ling Ma,Yanfen Zhang
Pine wilt disease poses a serious threat to pine forest health, and efficient biocontrol agents are urgently needed.Bacillus velezensisYG4-2 is a pine endophytic bacterium with strong nematicidal activity againstBursaphelenchus xylophilus, but the effect of oxygen conditions on its activity remains unclear. In this study, we compared the growth, nematicidal activity, and extracellular metabolite profiles of YG4-2 under aerobic and anaerobic conditions. Aerobic cultivation significantly increased bacterial biomass and enhanced the nematicidal activity of both bacterial cultures and cell-free supernatants. LC-MS/MS-based untargeted metabolomics revealed clear metabolic differences between the two oxygen conditions, with 19 metabolites significantly upregulated under aerobic conditions. Activity validation showed that kynurenic acid exhibited the strongest nematicidal activity among the tested candidate metabolites. Targeted LC-MS/MS further confirmed the accumulation of kynurenic acid in aerobic culture supernatants, with an estimated concentration of 0.38 ng/mL, and no obvious signal was detected under anaerobic conditions. These results suggest that oxygen enhances the nematicidal activity of YG4-2 by promoting growth and regulating extracellular metabolite production, with kynurenic acid as a potential oxygen-responsive active metabolite.
✓ 已验证
DOI
高影响因子
FOREST PATHOLOGY
·
2019
·Vol. 49
·期 2
·e12467
·
IF 1.434
Buyong Wang,Rongrong Wen,Ling Ma,Yan Ma,T. Hua Zhou
The Forkhead box O transcription factor DAF-16 is an important downstream transcription factor in the well-conserved insulin signalling (IIS) pathway in Caenorhabditis elegans . The nuclear localization of DAF-16 can activate a series of genes that mediate oxidative stress, heat shock, xenobiotic responses, innate immunity, metabolism and autophagy. In this study, a Bxdaf-16-2b gene encoding 510 amino acids was cloned from Bursaphelenchus xylophilus . Sequence analysis showed that the BxDAF-16-2b protein shared the highest similarity value (92%) with the DAF16-2 (accession no. ADD84716.1 ) of B. xylophilus . Using in situ hybridization, Bxdaf-16-2b was shown to be constitutively expressed in the circum-pharyngeal nerve rings and intestines of males, and in the head neurons and the ovaries of females. Quantitative RT-PCR showed that the highest gene expression was found in females, followed by juveniles, males and eggs. Bxdaf-16-2b expression was up-regulated after treatment with the botanical pesticides rotenone and matrine. RNA interference showed that the expression of Bxdaf-16-2b gene was reduced by dsRNA, and the silencing of Bxdaf-16-2b suppressed the expression of several IIS pathway genes. Bursaphelenchus xylophilus showed increased mortality at sublethal doses of rotenone and matrine when subjected to Bxdaf-16-2b gene silencing. Thus, Bxdaf-16-2b may participate in the stress response of B. xylophilus and play important roles in the immune system. This study paves the way towards design of new drug targets for non-polluting control of nematodes.
研究领域:
分子生物学
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遗传学
·
植物保护
✓ 已验证
产品关联
DOI
高影响因子
JOURNAL OF ASIAN NATURAL PRODUCTS RESEARCH
·
2019
·
IF 1.17
Yue-E Tian,Di Sun,Jin-Ming Yang,Zhi-Ping Che,Sheng-Ming Liu,Xiao-Min Lin,Jia Jiang,Gen-Qiang Chen
Sixteen sulfonate derivatives of maltol were synthesized and screened in vitro for their anti-oomycete and nematicidal activity against Phytophthora capsici and Bursaphelenchus xylophilus, respectively. Among all the compounds, 3e, 3m, and 3p exhibited the most promising and pronounced anti-oomycete activity against P. capsici than zoxamide, and the EC50 values of 25.42, 18.44, 23.69, and 27.99 mg/L, respectively; compounds 3e, 3m, 3n, and 3p exhibited potent nematicidal activity with LC50 values ranging from 1 to 2 mg/L, especially 3m and 3n showed the best promising and pronounced nematicidal activity, with LC50 values of 1.1762 and 1.2384 mg/L, respectively.
研究领域:
农业化学
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植物病理学
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线虫学
·
药物化学
✓ 已验证
DOI
高影响因子
危险品化学品经营许可证（带存储）