Pesticide Biochemistry and Physiology最新文献_第10页

The function of HgLac in Heterodera glycines and its potential as a control target HgLac在异线虫甘氨酸中的作用及其作为控制靶点的潜力

IF 4.2 1区农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Pesticide Biochemistry and Physiology

Pub Date : 2024-11-27 DOI: 10.1016/j.pestbp.2024.106225

Dongya Wang , Fangcao Wu , Xiongbiao Xu , Deliang Peng , Yuxi Duan , Huan Peng , Haiyan Wu

The soybean cyst nematode (SCN; Heterodera glycines) is one of the most devastating pathogens for soybean production. The second stage juvenile (J2) invades the host root, develops and form white females which then become brown cysts enter the soil. The brown cyst wall plays a key role in protecting inside eggs from adverse environmental conditions. However, the function of cyst wall tanning (sclerotization and pigmentation) in nematodes is not clear. A browning-related gene discovered from the whole-genome sequencing was cloned and characterized in this study, the gene was confirmed to be the laccase gene and was named HgLac. HgLac mRNA and HgLac protein was detected in the epidermis of juveniles using in situ hybridization and immunolocalization techniques. The HgLac expression level was greater in fourth-stage juveniles (J4s) than in the other stages. Knockdown of HgLac by in vitro RNA interference (RNAi) significantly decreased the infectivity, development and reproduction of J2s but had no effect on cyst wall tanning. Further research revealed that HgLac expression in nematodes was significantly suppressed by 35.41–59.17 % through in planta RNAi, 52.96–58.19 % females could not tan successfully, and the female wall was very soft and fragile, with a low egg hatching rate (1.33 %), which was significantly lower than that of normal females (68.85 %). These results indicate that HgLac plays a key role in cyst wall tanning and suppressing the development and reproduction of the SCN, which provides new ideas for the use of this gene as a target to control SCN.

大豆囊肿线虫(SCN；异源线虫（Heterodera glycines）是大豆生产中最具破坏性的病原体之一。第二阶段幼虫（J2）侵入寄主根部，发育形成白色雌虫，然后变成棕色囊进入土壤。棕色囊肿壁在保护卵子免受不利环境条件的影响方面起着关键作用。然而，线虫囊壁晒黑（硬化和色素沉着）的功能尚不清楚。本研究克隆并鉴定了从全基因组测序中发现的一个褐变相关基因，确认该基因为漆酶基因，并命名为HgLac。采用原位杂交和免疫定位技术检测幼鱼表皮中HgLac mRNA和HgLac蛋白的表达。第4期幼鱼（J4s）的HgLac表达量高于其他各期。体外RNA干扰（RNAi）敲除HgLac可显著降低J2s的感染性、发育和繁殖，但对囊壁鞣制无影响。进一步研究发现，HgLac在线虫中的表达被植物RNAi显著抑制了35.41 ~ 59.17%,52.96 ~ 58.19%的雌虫不能成功鞣制，雌虫壁非常柔软脆弱，卵孵化率较低（1.33%），显著低于正常雌虫（68.85%）。这些结果表明，HgLac在囊肿壁晒黑和抑制SCN的发育和繁殖中起着关键作用，这为利用该基因作为控制SCN的靶点提供了新的思路。

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引用次数: 0

Vitamin E alleviates zebrafish intestinal damage and microbial disturbances caused by pyraclostrobin

IF 4.2 1区农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Pesticide Biochemistry and Physiology

Pub Date : 2024-11-26 DOI: 10.1016/j.pestbp.2024.106221

Mengna Zhang , Feng Zhao , Mengyu Guo , Manman Duan , Yao Xie , Lihong Qiu

Pyraclostrobin (PY) is highly toxic to aquatic organisms, and its increased residues in aquatic environments may have harmful effects on the intestine of aquatic creatures. Previous research demonstrated that vitamin E (VE) alleviated the acute toxicity of PY to zebrafish. This study further explored the mitigation effect of VE on PY-induced intestinal toxicity in fish and the underlying mechanisms by exposing adult zebrafish to PY (10, 20 μg/L) with or without 4 μM VE supplementation for 21 days. The results showed that VE alleviated the gut histopathological lesions caused by PY. VE co-exposure also improved PY-induced intestinal inflammation and restored the expression level of genes encoding intestinal tight junction protein. Furthermore, VE restored the anti-oxidation level inhibited by PY and reduced pro-apoptotic cytokine level and apoptotic enzyme activity increased by PY. 16S rRNA high-throughput sequencing showed that VE improved the zebrafish intestinal flora imbalance caused by 20 μg/L PY, increased the relative abundance of beneficial bacterium Cetobacterium, and reduced the relative abundance of pathogenic bacteria. In conclusion, VE alleviated PY-induced intestinal toxicity via repairing the damaged intestinal mucosal barrier, inhibiting inflammation, reducing oxidative stress and apoptosis, and improving the intestinal microbial disorder in zebrafish.

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引用次数: 0

Exploration of the transfluthrin effects on fertility and pregnancy outcomes: An in-vivo study in rat 探讨氟氯氰菊酯对生育力和妊娠结果的影响：大鼠体内研究

IF 4.2 1区农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Pesticide Biochemistry and Physiology

Pub Date : 2024-11-22 DOI: 10.1016/j.pestbp.2024.106220

Akhilesh Mishra , Gayatri Dewangan , Megh Singh Dhakad , Subash Chandra Sonkar , Jasbir Dalal , Sunita Pradhan , Divakar Sharma , Vandana Roy , Bidhan Chandra Koner

Transfluthrin (TFL) is a rapid-acting pyrethroid insecticide, commonly used worldwide, however its overuse and or misuse has been associated with adverse effects on human health. This study was designed to investigate the impact of TFL on fertility and pregnancy in Sprague–Dawley rats exposed orally and by inhalation. In this study, 48 adult Sprague–Dawley rats of 6–8 months age group were randomly selected with equal numbers of males and females. Each group was housed separately in a standard laboratory condition and their daily consumption of food and water, along with weight of each group were monitored. The blood was collected from each rat on 0 (zero) and 85th day of oral and inhalation exposure to TFL to determine the hormone levels by ELISA. The findings revealed that TFL exposure through oral and inhalation route reduced hormones level in serum such as testosterone, follicle-stimulating hormone, luteinizing hormone, progesterone, estrogen, and prolactin in both male and female rats. In conclusion, current research suggest the negative effects of TFL exposure on fertility, adversely affecting pregnancy outcomes, reducing the mating rate, and lowering the number of pups. It also stresses the significance of controlling its use to prevent any negative consequences.

高效氟氯氰菊酯（TFL）是一种速效拟除虫菊酯杀虫剂，在全球范围内被普遍使用，但其过度使用或滥用会对人类健康造成不良影响。本研究旨在调查口服和吸入 TFL 对 Sprague-Dawley 大鼠生育和怀孕的影响。本研究随机挑选了 48 只 6-8 个月大的成年 Sprague-Dawley 大鼠，雌雄数量相等。每组大鼠在标准的实验室条件下分别饲养，并监测其每天的食物和水消耗量以及体重。在口服和吸入 TFL 的第 0 天（零天）和第 85 天，采集每只大鼠的血液，用 ELISA 方法测定激素水平。研究结果显示，通过口服和吸入途径接触 TFL 会降低雌雄大鼠血清中的激素水平，如睾酮、卵泡刺激素、黄体生成素、孕酮、雌激素和催乳素。总之，目前的研究表明，接触 TFL 对生育能力有负面影响，会对妊娠结果产生不利影响，降低交配率，并减少幼崽数量。研究还强调了控制其使用以防止任何负面影响的重要性。

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引用次数: 0

Knockdown of the β-N-acetylhexosaminidase genes by RNA interference inhibited the molting and increased the mortality of the white-backed planthopper, Sogatella furcifera 用RNA干扰法敲低白背飞虱的β- n -乙酰己糖氨酸酶基因，抑制了飞虱的脱毛，提高了飞虱的死亡率

IF 4.2 1区农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Pesticide Biochemistry and Physiology

Pub Date : 2024-11-22 DOI: 10.1016/j.pestbp.2024.106216

Pan-Pan Guo , Xi-Bin Yang , Hong Yang , Cao Zhou , Gui-Yun Long , Dao-Chao Jin

β-N-Acetylglucosaminidases and/or β-N-acetylhexosaminidases (NAGs / Hexes) are crucial exonucleases, playing a crucial role in the insect molting process. SfHex3 and SfHex4 contain conserved catalytic domains of GH20 and GH20b, clustered into NAG2 and NAG1 group, respectively. SfHex3 and SfHex4 were mainly highly expressed in the 4th–5th instar nymphs, as well as in the integument and ovary. The expression level of SfHex3 gradually decreased in male and female adults, and SfHex4 on the first day of female was significantly higher than that on the first day of male. In addition, RNA interference (RNAi) results demonstrated that the downregulation of SfHex3 and SfHex4 expression in 5th-instar nymphs resulted in failed molting, and a high mortality. Furthermore, after RNAi with SfHex3 and SfHex4, the transcript levels on key genes of the chitin metabolism pathway (SfCHS1, SfCHS1a, SfCHS1b, SfTRE1, SfTRE2, SfCht5, and SfCht7) were significantly decreased compared to the control group. Meanwhile the expression levels of SfHex3 and SfHex4 were up-regulated after 6 h and 12 h of 20E treatment. And the transcription levels of SfHex3 and SfHex4 were significantly inhibited at nitenpyram LC₂₀, LC₅₀, and LC₉₀ after 96 h of treatment, in 3rd nymphs of Sogatella furcifera. In conclusion, SfHex3 and SfHex4 play important roles in the nymphal development of S. furcifera, contributing to the molting process from nymph to adult. This study not only enhances our understanding of the nitenpyram in pest control, but also provides a foundation for the development of new control strategies using RNAi to targeting SfHex3 and SfHex4.

β- n -乙酰氨基葡萄糖酶和/或β- n -乙酰己糖氨酸酶（nag / Hexes）是重要的外切酶，在昆虫的蜕皮过程中起着至关重要的作用。SfHex3和SfHex4含有GH20和GH20b的保守催化结构域，分别聚集在NAG2和NAG1基团中。SfHex3和SfHex4主要在4 ~ 5龄若虫、被皮和子房中高表达。SfHex3在雄性和雌性成虫体内的表达量逐渐降低，雌性成虫第一天SfHex4的表达量显著高于雄性成虫第一天。此外，RNA干扰（RNAi）结果表明，下调SfHex3和SfHex4在5龄若虫体内的表达可导致脱皮失败和高死亡率。此外，SfHex3和SfHex4 RNAi后，几丁质代谢途径关键基因（SfCHS1、SfCHS1a、SfCHS1b、SfTRE1、SfTRE2、SfCht5和SfCht7）的转录水平与对照组相比显著降低。同时，在20E处理6 h和12 h后，SfHex3和SfHex4的表达水平上调。SfHex3和SfHex4的转录水平在处理96 h后，在nitenpyram LC20、LC50和LC90处被显著抑制。综上所述，SfHex3和SfHex4在蜜蛾若虫发育过程中起着重要的作用，参与了蜜蛾从若虫到成虫的蜕皮过程。该研究不仅加深了我们对nitenpyram在害虫防治中的认识，而且为利用RNAi靶向SfHex3和SfHex4开发新的防治策略提供了基础。

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引用次数: 0

Distribution, frequency and molecular basis of penoxsulam, metamifop and florpyrauxifen-benzyl resistance in Echinocloa spp. from rice fields across Jiangsu Province, China 江苏省稻田棘球绦虫对培诺舒兰、甲虫脒和氟虫腈的抗性分布、频率和分子基础

IF 4.2 1区农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Pesticide Biochemistry and Physiology

Pub Date : 2024-11-22 DOI: 10.1016/j.pestbp.2024.106218

Wei Deng , Hanqi Yin , Zhixun Ge , Sai Yao , Jiawen Wu , Axiu Zhu , Qian Yang , Shuzhong Yuan

Barnyard grasses (Echinochloa spp.) are the most troublesome weeds in rice ecosystems of Jiangsu Province, China, and long-term use of herbicides inevitably cause the development of resistance. In this study, 122 field-sampled populations were tested for resistance to penoxsulam (PEN), metamifop (MET), florpyrauxifen-benzyl, (FPB), propanil, and tripyrasulfone. Field-dose assays revealed that 51, 7, and 6 populations have either developed or were developing resistance to penoxsulam, metamifop and florpyrauxifen-benzyl, and all 122 populations were still sensitive to propanil and tripyrasulfone. Dose-response assays identified that 41.8 % of populations exhibited 6- to 830-fold resistance to PEN; 5.7 % of populations displayed 2.2- to 13.9-fold resistance to MET; and 4.9 % of populations showed 2.4- to 11.1-fold resistance to FPB. Trp-574-Leu mutations in EcALS1 or EcALS3 were found in 18.0 % of populations. Trp-2027-Cys, Cys-2088-Arg, and Gly-2096-Ala mutations in EcACC1, EcACC3, or EcACC5 were detected in 4.1 % of populations, and no known resistance-associated target-site gene mutations were confirmed in the rest of herbicide-resistant populations. Furthermore, the P450 inhibitor malathion and GST inhibitor NBD-Cl significantly reduced the resistance to three herbicides in 38.5 % of populations, especially those without the target-site gene mutations, indicating both target-site mutations and P450s and/or GSTs-mediated metabolisms play important roles in herbicide resistance evolution in Echinochloa spp. These findings are helpful to formulate an appropriate strategy for effective and sustainable control of Echinochloa spp. by mixtures of herbicide with different modes of action and mitigate weed resistance evolution.

稗草（Echinochloa spp.）是江苏省水稻生态系统中最麻烦的杂草，长期使用除草剂不可避免地会产生抗性。在这项研究中，对122个野外取样的种群进行了对培诺舒南（PEN）、甲氨磷（MET）、氟吡虫啉苄（FPB）、丙烯和三嘧磺酮的抗性检测。现场剂量测定结果显示，51、7和6只种群对培诺舒朗、甲氧虫乐和氟吡虫啉苯产生或正在产生抗性，122只种群对丙烯和三嘧磺酮仍敏感。剂量反应试验表明，41.8%的种群对PEN表现出6至830倍的抗性；5.7%的种群对MET的抗性为2.2 ~ 13.9倍；4.9%的种群对FPB的抗性为2.4 ~ 11.1倍。在18.0%的人群中发现了EcALS1或EcALS3的Trp-574-Leu突变。在4.1%的EcACC1、EcACC3或EcACC5群体中检测到Trp-2027-Cys、Cys-2088-Arg和Gly-2096-Ala突变，其余除草剂抗性群体中未发现已知的抗性相关靶位基因突变。此外，P450抑制剂马拉硫磷和GST抑制剂NBD-Cl在38.5%的群体中显著降低了对三种除草剂的抗性，特别是那些没有靶位基因突变的群体。研究结果表明，靶位点突变和p450和/或gsts介导的代谢在紫锥藻的抗性进化中起重要作用，有助于制定不同作用模式的除草剂混合防治策略，减缓紫锥藻的抗性进化。

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引用次数: 0

Interaction between imidacloprid residues in maize rhizospheric soil and soil nematode community

IF 4.2 1区农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Pesticide Biochemistry and Physiology

Pub Date : 2024-11-21 DOI: 10.1016/j.pestbp.2024.106219

Jiaming Zhang , Yue Wu , Jiayi Liu, Xinru Wang, Weijia Pang, Juan Hua, Shihong Luo

Although imidacloprid has been shown to present potential risks to non-target invertebrates and vertebrates, researches exploring this risk from the perspective of the underground ecosystem remains incomplete. In this study, we determined that the presence of imidacloprid significantly reduced the abundance and diversity of soil nematodes in maize rhizospheric soil. Furthermore, imidacloprid also exerted negative effects on the body length, reproduction, locomotion, lipid accumulation, lipofuscin accumulation, and acetylcholinesterase activity in the model organism Caenorhabditis elegans. These toxic phenotypes are correlated with the upregulation of fat-2, fat-6, hsp-16.41, and hsp-16.2, along with the downregulation of ace-1, ace-2, and ace-3. In response to these toxic effects of imidacloprid, nematodes also developed corresponding adaptive mechanisms. UPLC-MS/MS analysis revealed that nematodes could convert imidacloprid to imidacloprid-guanidine and imidacloprid-urea to reduce the toxicity of imidacloprid. Moreover, C. elegans and Meloidogyne incognita exhibited repellent behavior towards imidacloprid-treated area, even the concentration of imidacloprid is only 0.4 μg/mL. This study revealed the interaction between imidacloprid and nematodes, providing a basis for understanding the potential risks of non-target soil nematodes after application of imidacloprid in sustainable agriculture and the resistance mechanism of nematodes to nematocidal pesticide.

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引用次数: 0

Effect of various physical and chemical stress conditions on the infectivity and survival of Heterorhabditis indica and Steinernema feltiae: Relationship with lipid oxidative stress 不同物理和化学胁迫条件对印度异habditis and steinnema feltiae感染和存活的影响：与脂质氧化应激的关系

IF 4.2 1区农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Pesticide Biochemistry and Physiology

Pub Date : 2024-11-21 DOI: 10.1016/j.pestbp.2024.106217

Lenin González-Paz , Carla Lossada , Dayrana Mora , Edgar Portillo , Ernesto San-Blas

Entomopathogenic nematodes (EPNs) of the genera Heterorhabditis and Steinernema represent an alternative for the biological control of insects. The limited half-life of EPNs is still one of the most concerning issues in their commercialization. Lipid peroxidation (LPO) caused by reactive oxygen species (ROS) may be one of the most important causes of loss of infectivity and survival of EPNs when exposed to various physicochemical stress conditions (temperature, pH, hypoxia and osmotic pressure). Because LPO generates free radicals (FRs), it can trigger membrane peroxidation and lipid energy reserves of EPNs. However, in EPNs there is no data on the role of LPO on their physiology, making strategies for the conservation of derived biopreparations difficult. In this sense, the influence of LPO on the species of EPNs S. feltiae and H. indica under various conditions of physicochemical stress was studied. In both EPNs, the proposed stress conditions altered infectivity and survival over time, generating ROS associated with LPO with a variable tolerance depending on the species, type and time of exposure to stress. A relationship was observed between the LPO induced by stress conditions and infectivity-survival.

虫病线虫（Heterorhabditis和Steinernema）是昆虫生物防治的一种选择。epn的有限半衰期仍然是其商业化中最受关注的问题之一。活性氧（ROS）引起的脂质过氧化（LPO）可能是epn暴露于各种物理化学胁迫条件（温度、pH、缺氧和渗透压）时丧失感染性和存活的最重要原因之一。由于LPO产生自由基（FRs），它可以触发epn的膜过氧化和脂质能量储备。然而，在epn中，没有关于LPO在其生理上的作用的数据，这使得保护衍生生物修复的策略变得困难。在此基础上，研究了LPO在不同物化胁迫条件下对EPNs S. feltiae和H. indica的影响。在这两种epn中，随着时间的推移，所提出的应激条件改变了传染性和存活率，产生与LPO相关的ROS，其耐受性取决于暴露于应激的物种、类型和时间。应激诱导的LPO与感染存活率之间存在一定的关系。

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引用次数: 0

Fomesafen induced ecotoxicological responses, bioaccumulation, and implications for soil ecosystems remediation in three ecotypes earthworms 三种生态型蚯蚓的福美沙芬诱导生态毒理学反应、生物累积以及对土壤生态系统修复的影响

IF 4.2 1区农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Pesticide Biochemistry and Physiology

Pub Date : 2024-11-19 DOI: 10.1016/j.pestbp.2024.106215

Yan Ge, Siqi Wang, Siqi Miao, Dongxing Zhou

Fomesafen (FSA) is a herbicide commonly used in soybean fields, but its long half-life in the soil can pose pollution risks to the soil ecosystem. Earthworms, which have an indicative function for soil health and play a vital role in maintaining soil ecological functions, have not been fully studied in terms of their susceptibility to FSA. This study examined the effects of different concentrations of FSA on three ecotypes of earthworms (Eisenia fetida (epigeic), Metaphire guillelmi (anecic), and Aporrectodea caliginosa (endogeic)) and found varying trade-off strategy of their growth and reproduction. When subjected to FSA stress, E. fetida prioritized reproduction, A. caliginosa prioritized growth, while M. guillelmi maintained consistent growth and reproduction, effectively allocating the acquired biomass to reproduction. The study also analyzed the oxidative stress response, gene expression, and an integrated biomarker response. The strongest induction performance of the three ecotypes of earthworms was in the expression of antioxidant enzymes such as CAT, POD, SOD, and P450 expression, while the performance was significantly different in AChE activity, AChE and HSP70 expression, which may lead to differences in toxicity and detoxification mechanisms of earthworms in different ecotypes. The partial least squares-path model (PLS-PM) also showed that the three ecotypes of earthworms exhibited different toxicity, detoxification and regulatory pathways under FSA stress. M. guillelmi had the highest IBRv2 index (28.67–42.90). At the same time, the FSA bioaccumulation tended to be stable with increasing concentration, and its main detoxification ability (GST activity and P450 expression) could still maintain the same level as CK at high concentrations, indicating that it is very likely to be a potential earthworm type for FSA remediation. This study firstly provides a comprehensive assessment of FSA's toxic effects on different ecotypes of earthworms and highlights the importance of identifying potential resistant species for soil remediation.

福美双（FSA）是一种常用于大豆田的除草剂，但它在土壤中的半衰期很长，会对土壤生态系统造成污染风险。蚯蚓对土壤健康具有指示性功能，在维护土壤生态功能方面发挥着重要作用，但人们尚未充分研究蚯蚓对 FSA 的易感性。本研究考察了不同浓度的 FSA 对三种生态型蚯蚓（Eisenia fetida（外生型）、Metaphire guillelmi（内生型）和 Aporrectodea caliginosa（内生型））的影响，发现它们的生长和繁殖有不同的权衡策略。当受到 FSA 胁迫时，E. fetida 优先考虑繁殖，A. caliginosa 优先考虑生长，而 M. guillelmi 则保持一致的生长和繁殖，有效地将获得的生物量分配给繁殖。研究还分析了氧化应激反应、基因表达和综合生物标志物反应。三种生态型蚯蚓的诱导表现最强的是CAT、POD、SOD等抗氧化酶的表达和P450的表达，而在AChE活性、AChE和HSP70的表达上表现有显著差异，这可能导致不同生态型蚯蚓毒性和解毒机制的差异。偏最小二乘路径模型（PLS-PM）也表明，三种生态型的蚯蚓在FSA胁迫下表现出不同的毒性、解毒和调节途径。M. guillelmi 的 IBRv2 指数最高（28.67-42.90）。同时，其FSA生物累积量随浓度增加而趋于稳定，其主要解毒能力（GST活性和P450表达）在高浓度下仍能保持与CK相同的水平，这表明它很有可能成为一种潜在的FSA修复蚯蚓类型。这项研究首次全面评估了 FSA 对不同生态型蚯蚓的毒性影响，并强调了识别潜在抗性物种对土壤修复的重要性。

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引用次数: 0

Identification and characterization of ATP-binding cassette transporters involved in chlorantraniliprole tolerance of model insect Drosophila melanogaster and agricultural pest Spodoptera frugiperda 模式昆虫黑腹果蝇和农业害虫弗罗里珀虫耐受氯酯的 ATP 结合盒转运体的鉴定和特征描述

IF 4.2 1区农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Pesticide Biochemistry and Physiology

Pub Date : 2024-11-17 DOI: 10.1016/j.pestbp.2024.106212

Lixiang Wang , Bin Wen , Shaoyi Guo , Yujie Han , Zhongyuan Deng , Qian Ding , Xianchun Li

ATP-binding cassette (ABC) transporter family is one of the largest transporter families, which plays an important role in insecticide tolerance. In this study, we found that the ABC transporter inhibitor verapamil could significantly enhance the toxicity of chlorantraniliprole (CHL) to the model insect Drosophila melanogaster. Forty-six ABC transporter genes of D. melanogaster were knocked down through the daughterless-GAL4 (Da-GAL4) strain. The subsequent bioassay result showed that D. melanogaster with DmCG5772, DmCG1494, and DmCG5853 genes silencing significantly increased mortality after CHL treatment. Based on the genome of the fall armyworm (FAW), three genes with the best hits were identified, and SfABCA1 (XM_035576510.2) and SfABCG10 (XM_035577893.2) were successfully cloned. Spatiotemporal expression pattern analysis showed that SfABCA1 and SfABCG10 were both highly expressed in adult and pupal stages. Hemolymph was also a tissue with high expression of these two genes. LC₁₀ dose of CHL could induce the expression levels of SfABCA1 and SfABCG10, with SfABCG10 upregulated 8-fold after 48 h of CHL treatment. Furthermore, overexpression of SfABCA1 and SfABCG10 increased the viability of Sf9 cell under CHL treatment. Our findings indicate that SfABCA1 and SfABCG10 might associate with the tolerance of CHL in S. frugiperda. These results are not only helpful in understanding the role of ABC transporters in CHL tolerance of other agricultural pests, but also lay a theoretical foundation for delaying the development of CHL resistance in pest management.

ATP结合盒（ABC）转运体家族是最大的转运体家族之一，在杀虫剂耐受性方面发挥着重要作用。本研究发现，ABC转运体抑制剂维拉帕米能显著增强氯虫苯甲酰胺（CHL）对模式昆虫黑腹果蝇的毒性。通过无子-GAL4（Da-GAL4）菌株敲除了黑腹果蝇的46个ABC转运体基因。随后的生物测定结果表明，沉默DmCG5772、DmCG1494和DmCG5853基因的黑腹果蝇在CHL处理后死亡率显著增加。根据秋军虫（FAW）的基因组，确定了三个命中率最高的基因，并成功克隆了SfABCA1（XM_035576510.2）和SfABCG10（XM_035577893.2）。时空表达模式分析显示，SfABCA1和SfABCG10在成虫和蛹期均有高表达。血淋巴也是这两个基因高表达的组织。LC10剂量的CHL可诱导SfABCA1和SfABCG10的表达水平，其中SfABCG10在CHL处理48小时后上调了8倍。此外，SfABCA1和SfABCG10的过表达提高了Sf9细胞在CHL处理下的存活率。我们的研究结果表明，SfABCA1和SfABCG10可能与节肢动物对CHL的耐受性有关。这些结果不仅有助于了解 ABC 转运体在其他农业害虫耐 CHL 能力中的作用，而且为在害虫管理中延缓 CHL 抗性的发展奠定了理论基础。

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引用次数: 0

Functional characterization of four antenna-enriched odorant binding proteins in Rhaphuma horsfieldi reveals the importance of RhorOBP1 in odorant recognition and insecticide resistance 马蹄金四种触角丰富的气味结合蛋白的功能表征揭示了 RhorOBP1 在气味识别和杀虫剂抗性中的重要性

IF 4.2 1区农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Pesticide Biochemistry and Physiology

Pub Date : 2024-11-17 DOI: 10.1016/j.pestbp.2024.106210

Yin-Lan Liang , Shun-Si Li , Ning-Na Yin, Shu-Lin Li, Yu-Yue Lu, Nai-Yong Liu

The cerambycid beetles are key players for the sustenance of biodiversity in the forest ecosystem, but in most cases are well known due to their harmfulness to agricultural and forest plants. Here, we characterized the odorant binding protein (OBP) gene family in Rhaphuma horsfieldi, emphasizing the roles of RhorOBP1 in odorant reception and insecticide sequestering. A homology-based search led to the identification of 35 RhorOBP genes with a major distribution in the Minus-C OBPs clade (21/35 genes). Expression profiles showed that RhorOBP1–RhorOBP4 had the abundant expression in antennae. Binding assays revealed that the four RhorOBPs exhibited diverse odorant response profiles tuned differentially to various classes of plant odorants, comprising walnut-derived host volatiles and ordinary floral scents. Two broadly tuned RhorOBP1 and RhorOBP2 exhibited different chain length-dependent binding properties to 10C12C alcohols, aldehydes or acetates. Compared with other three proteins, RhorOBP1 reduced the binding to ligands with high affinities at pH 5.0 (1.27–6.72-fold differences relative to pH 7.4). Molecular docking and point-mutation experiments confirmed that Ser107, Tyr118, Tyr119 and Phe120 situated in the binding pocket of RhorOBP1 were critical determinants for the recognition of 14, 15, 10 and 10 compounds, respectively. On the other hand, RhorOBP1 could strongly bind six insecticides, particularly chlorpyrifos (dissociation constant, K_i = 3.69 ± 0.74 μM). This study has provided insights into different binding properties of four antenna-enriched RhorOBPs in R. horsfieldi and identifies a dual role of RhorOBP1 in the binding of odorants and insecticides.

啮齿目甲虫是维持森林生态系统生物多样性的关键角色，但在大多数情况下，由于其对农业和森林植物的危害而广为人知。在这里，我们描述了马蹄金甲虫气味结合蛋白（OBP）基因家族的特征，强调了 RhorOBP1 在气味接收和杀虫剂封存中的作用。通过基于同源性的搜索，确定了 35 个 RhorOBP 基因，这些基因主要分布在 Minus-C OBPs 支系中（21/35 个基因）。表达谱显示，RhorOBP1-RhorOBP4在触角中的表达量最大。结合试验显示，四种 RhorOBPs 对不同类别的植物气味（包括核桃提取的宿主挥发物和普通花香）表现出不同的气味反应特征。两种广泛调谐的 RhorOBP1 和 RhorOBP2 对 10C12C 醇、醛或醋酸盐表现出不同的链长依赖性结合特性。与其他三种蛋白质相比，RhorOBP1 在 pH 值为 5.0 时与配体的结合亲和力较低（与 pH 值为 7.4 时相比，相差 1.27-6.72 倍）。分子对接和点突变实验证实，位于 RhorOBP1 结合口袋中的 Ser107、Tyr118、Tyr119 和 Phe120 分别是识别 14、15、10 和 10 种化合物的关键决定因素。另一方面，RhorOBP1 能强烈结合六种杀虫剂，尤其是毒死蜱（解离常数 Ki = 3.69 ± 0.74 μM）。这项研究揭示了马蹄蟾中四种富含触角的 RhorOBPs 的不同结合特性，并确定了 RhorOBP1 在气味剂和杀虫剂结合过程中的双重作用。

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引用次数: 0