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β-Caryophyllene wrapped by nanoliposomes efficiently increases the control effect on Bemisia tabaci MED 纳米脂质体包裹的 β-茶碱能有效提高对烟粉虱 MED 的防治效果
IF 4.2 1区 农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-08-14 DOI: 10.1016/j.pestbp.2024.106082

Bemisia tabaci poses a severe threat to plants, and the control of B. tabaci mainly relies on pesticides, which causes more and more rapidly increasing resistance. β-Caryophyllene is a promising ingredient for agricultural pest control, but its feature of poor water solubility need to be improved in practical applications. Nanotechnology can enhance the effectiveness and dispersion of volatile organic compounds (VOCs). In this study, a nanoliposome carrier was constructed by ethanol injection and ultrasonic dispersion method, and β-caryophyllene was wrapped inside it, thus solving the defect of poor solubility of β-caryophyllene. The size of the β-caryophyllene nanoliposomes (C-BT-NPs) was around 200 nm, with the absolute value of the zeta potential exceeding 30 mV and a PDI below 0.5. The stability was also maintained over a 14-d storage period. C-BT-NPs showed effective insecticidal activity against B. tabaci, with an LC50 of 1.51 g/L, outperforming thiamethoxam and offering efficient agricultural pest control. Furthermore, C-BT-NPs had minimal short-term impact on the growth of tomato plants, indicating that they are safety on plants. Therefore, the VOCs using nanoliposome preparation technology show promise in reducing reliance on conventional pesticides and present new approaches to managing agricultural pests.

烟粉虱(Bemisia tabaci)对植物构成严重威胁,防治烟粉虱主要依靠杀虫剂,但杀虫剂的抗药性越来越强。β-茶碱是一种很有前景的农业害虫防治成分,但其水溶性差的特点需要在实际应用中加以改进。纳米技术可以提高挥发性有机化合物(VOCs)的功效和分散性。本研究采用乙醇注射法和超声波分散法构建了一种纳米脂质体载体,并将 β-石竹烯包裹在其中,从而解决了 β-石竹烯溶解性差的缺陷。β-石竹烯纳米脂质体(C-BT-NPs)的尺寸约为 200 nm,zeta 电位的绝对值超过 30 mV,PDI 低于 0.5。在 14 天的储存期内也能保持稳定。C-BT-NPs 对烟粉虱具有有效的杀虫活性,半致死浓度为 1.51 g/L,优于噻虫嗪,可有效控制农业害虫。此外,C-BT-NPs 对番茄植物生长的短期影响极小,表明其对植物是安全的。因此,采用纳米脂质体制备技术的挥发性有机化合物有望减少对传统杀虫剂的依赖,为农业害虫防治提供了新方法。
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引用次数: 0
Inhibitory mechanism of 4-ethyl-1,2-dimethoxybenzene produced by Streptomyces albidoflavus strain ML27 against Colletotrichum gloeosporioides 白僵菌菌株 ML27 产生的 4-乙基-1,2-二甲氧基苯对球孢子菌的抑制机制
IF 4.2 1区 农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-08-13 DOI: 10.1016/j.pestbp.2024.106086

Actinomycetes have emerged as significant biocontrol resources due to their rich array of bioactive natural products. While much research has historically focused on secondary metabolites isolated from their fermentation broth, there remains a dearth of reports on their volatile organic compounds (VOCs). Here, strain ML27, isolated from soil, was identified as Streptomyces albidoflavus based on morphological features, physiological, biochemical, and molecular characteristics (16S rRNA, atpD, recA, and rpoB gene sequences). VOCs from S. albidoflavus strain ML27 were effectively captured using solid-phase microextraction (SPME) and tentatively identified through gas chromatography–mass spectrometry (GC/MS). Among these compounds, 4-ethyl-1,2-dimethoxybenzene exhibited broad-spectrum antifungal activity and demonstrated efficacy in controlling citrus anthracnose, with a control efficacy of 86.67%. Furthermore, the inhibitory mechanism of 4-ethyl-1,2-dimethoxybenzene against Colletotrichum gloeosporioides was revealed. Results indicated that 4-ethyl-1,2-dimethoxybenzene induced swelling, deformity, and breakage in C. gloeosporioides mycelia, and significantly inhibited spore germination. Transcriptome analysis revealed that 4-ethyl-1,2-dimethoxybenzene inhibited the growth and development of C. gloeosporioides primarily by disrupting energy metabolism and the integrity of the cell wall and membrane. Based on these results, it is promising to develop 4-ethyl-1,2-dimethoxybenzene as a novel biopesticide for controlling citrus anthracnose.

放线菌具有丰富的生物活性天然产物,已成为重要的生物控制资源。尽管许多研究历来侧重于从其发酵液中分离出的次生代谢物,但有关其挥发性有机化合物(VOCs)的报道仍然很少。根据形态特征、生理、生化和分子特征(16S rRNA、atpD、recA 和 rpoB 基因序列),本文将从土壤中分离出的菌株 ML27 鉴定为白化链霉菌。利用固相微萃取技术(SPME)有效捕获了白僵菌菌株 ML27 中的挥发性有机化合物,并通过气相色谱-质谱联用技术(GC/MS)进行了初步鉴定。在这些化合物中,4-乙基-1,2-二甲氧基苯具有广谱抗真菌活性,对柑橘炭疽病有一定的防治效果,防治效果达 86.67%。此外,还揭示了 4-乙基-1,2-二甲氧基苯对球孢子菌的抑制机理。结果表明,4-乙基-1,2-二甲氧基苯能诱导球孢子菌菌丝膨胀、变形和断裂,并显著抑制孢子萌发。转录组分析表明,4-乙基-1,2-二甲氧基苯主要通过破坏能量代谢以及细胞壁和细胞膜的完整性来抑制球孢子菌的生长和发育。基于这些结果,开发 4-乙基-1,2-二甲氧基苯作为新型生物农药来防治柑橘炭疽病是很有希望的。
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引用次数: 0
Plant-derived citronellol can significantly disrupt cell wall integrity maintenance of Colletotrichum camelliae 植物提取的香茅醇能极大地破坏山茶科 Colletotrichum camelliae 细胞壁完整性的维持
IF 4.2 1区 农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-08-13 DOI: 10.1016/j.pestbp.2024.106087

Anthracnose, a fungal disease, commonly infects tea plants and severely impacts the yield and quality of tea. One method for controlling anthracnose is the application of citronellol, a plant extract that exhibits broad-spectrum antimicrobial activity. Herein, the physiological and biochemical mechanism by which citronellol controls anthracnose caused by Colletotrichum camelliae was investigated. Citronellol exhibited excellent antifungal activity based on direct and indirect mycelial growth inhibition assays, with EC50 values of 76.88 mg/L and 29.79 μL/L air, respectively. Citronellol also exhibited good control effects on C. camelliae in semi-isolated leaf experiments. Optical and scanning electron microscopy revealed that citronellol caused C. camelliae mycelia to thin, fracture, fold and deform. Transmission electron microscopy revealed that the mycelial cell walls collapsed inward and separated, and the organelles became blurred after treatment with citronellol. The sensitivity of C. camelliae to calcofluor white staining was significantly enhanced by citronellol, while PI staining showed minimal fluorescence, and the relative conductivity of mycelia were not significantly different. Under citronellol treatment, the expression levels of β-1,3-glucanase, chitin synthase, and chitin deacetylase-related genes were significantly decreased, while the expression levels of chitinase genes were increased, leading to lower chitinase activity and increased β-1,3-glucanase activity. Therefore, citronellol disrupted the cell wall integrity of C. camelliae and inhibited normal mycelial growth.

炭疽病是一种真菌病害,通常会感染茶树,严重影响茶叶的产量和质量。防治炭疽病的方法之一是施用香茅醇,这是一种具有广谱抗菌活性的植物提取物。本文研究了香茅醇控制由山茶小孢子菌引起的炭疽病的生理生化机制。在直接和间接菌丝生长抑制试验中,香茅醇表现出优异的抗真菌活性,其 EC50 值分别为 76.88 mg/L 和 29.79 μL/L 空气。在半隔离叶片实验中,香茅醇对山茶菌也表现出良好的控制效果。光学显微镜和扫描电子显微镜显示,香茅醇会导致山茶菌菌丝变细、断裂、折叠和变形。透射电子显微镜显示,经香茅醇处理后,菌丝细胞壁向内塌陷、分离,细胞器变得模糊不清。在香茅醇的作用下,C. camelliae 对钙氟白染色的敏感性明显增强,而 PI 染色显示的荧光极少,菌丝体的相对电导率也无明显差异。在香茅醇处理下,β-1,3-葡聚糖酶、几丁质合成酶和几丁质脱乙酰酶相关基因的表达水平明显降低,而几丁质酶基因的表达水平升高,导致几丁质酶活性降低,β-1,3-葡聚糖酶活性升高。因此,香茅醇破坏了山茶科植物细胞壁的完整性,抑制了菌丝的正常生长。
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引用次数: 0
Activity of OSBPI fungicide fluoxapiprolin against plant-pathogenic oomycetes and its systemic translocation in plants OSBPI 杀菌剂氟唑菌酰胺对植物致病性卵菌的活性及其在植物体内的系统迁移
IF 4.2 1区 农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-08-13 DOI: 10.1016/j.pestbp.2024.106085

Fluoxapiprolin, a novel piperidinyl thiazole isoxazoline fungicide, was developed by Bayer Crop Science in 2012. Despite its well-documented inhibitory activity against plant pathogenic oomycetes such as Phytophthora capsici and Phytophthora infestans, limited information regarding its antifungal spectrum and protective and curative activity is available. Fluoxapiprolin exhibited strong inhibitory activity against Phytophthora spp. and several Pythium spp., with EC50 values ranging from 2.12 × 10−4 to 2.92 μg/mL. It was much more effective against P. capsici in inhibiting mycelial growth, sporangium production, and cystospore germination than at reducing zoospore release. Moreover, fluoxapiprolin displayed both protective and curative activity against P. capsici infection in pepper plants under greenhouse conditions, with systemic translocation capability confirmed by High Performance Liquid Chromatography (HPLC) analysis. The results demonstrated the strong inhibitory activity of fluoxapiprolin against economically important plant oomycete pathogens, including Phytophthora spp. and several Pythium spp., and its certain translocation activity in pepper plants.

Fluoxapiprolin 是一种新型哌啶基噻唑异噁唑啉类杀菌剂,由拜耳作物科学公司于 2012 年开发。尽管其对植物病原卵菌(如疫霉属和疫霉属)的抑制活性已得到充分证实,但有关其抗真菌谱、保护和治疗活性的信息却十分有限。Fluoxapiprolin 对疫霉属(Phytophthora spp.)和几种疫霉属(Pythium spp.)具有很强的抑制活性,EC50 值从 2.12 × 10-4 到 2.92 μg/mL 不等。在抑制菌丝生长、孢子囊产生和子囊孢子萌发方面,氟唑菌酰胺比减少子囊孢子释放更有效。此外,在温室条件下,氟唑虫酰胺对辣椒植株的蘑菇伞菌感染具有保护和治疗活性,高效液相色谱(HPLC)分析证实了其系统转移能力。研究结果表明,氟唑禾草灵对具有重要经济价值的植物卵菌病原体(包括疫霉属和几种疫霉属)具有很强的抑制活性,并且在辣椒植株中具有一定的转座活性。
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引用次数: 0
Identification and CRISPR-Cas9 validation of a novel β-adrenergic-like octopamine receptor mutation associated with amitraz resistance in Varroa destructor 鉴定并用 CRISPR-Cas9 验证与 Varroa destructor 抗阿米曲拉相关的新型 β-肾上腺素能样章胺受体突变
IF 4.2 1区 农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-08-08 DOI: 10.1016/j.pestbp.2024.106080

Varroa destructor is widely recognized as a significant contributor to colony collapse disorder. Chemical acaricides, such as amitraz, have been extensively used for Varroa control due to their selectivity within beehives. However, the increasing number of cases of amitraz resistance across global V. destructor populations poses a significant challenge. In this study, we conducted a comprehensive molecular screening of the β-adrenergic-like octopamine receptor (Octβ2R), the target-site of amitraz, across 66 Turkish and 63 Belgian V. destructor populations. Although previously reported amitraz resistance mutations were not detected, the screening revealed a novel Y337F mutation located within transmembrane 7 (TM7) of Octβ2R in Turkish Varroa populations. Notably, this mutation was identified in the last residue of the highly conserved NPxxY motif associated with the activation of G-protein coupled receptors (GPCR). Among the 66 Varroa samples from Türkiye, twenty harbored the Y337F mutation, with eight samples exhibiting fixation of the mutation. Subsequent bioassays revealed over 8-fold resistance to amitraz in populations that contain the Y337F mutation. Genotyping of mites after exposure to 10 mg a.i./l amitraz demonstrated that all surviving mites were homozygous for the Y337F mutation, whereas dead mites carried susceptible alleles, providing genetic linkage between mutation and phenotype. Further, we used CRISPR-Cas9 editing to introduce the Y337F mutation in the orthologous Octβ2R of the model organism Tetranychus urticae. Crispants exhibited over threefold resistance to amitraz. In conclusion, this study identified and validated a novel amitraz resistance mutation. Additional research is required to further evaluate the phenotypic strength of Y337F in the context of operational resistance with current treatment strategies.

人们普遍认为破坏性瓦罗虫是造成蜂群崩溃紊乱的一个重要因素。化学杀螨剂(如双甲脒)因其在蜂箱内的选择性而被广泛用于控制瓦虫。然而,全球破坏螨种群对双甲脒产生抗药性的情况越来越多,这给我们带来了巨大的挑战。在这项研究中,我们对 66 个土耳其和 63 个比利时的破坏者种群中阿米曲拉的靶位点--β-肾上腺素能样章胺受体(Octβ2R)进行了全面的分子筛选。虽然没有检测到以前报告过的阿米曲唑抗性突变,但这次筛选在土耳其的 Varroa 群体中发现了一个位于 Octβ2R 跨膜 7 (TM7) 内的新型 Y337F 突变。值得注意的是,这一突变是在与激活 G 蛋白偶联受体(GPCR)相关的高度保守的 NPxxY 矩阵的最后一个残基上发现的。在来自土耳其的 66 个 Varroa 样本中,有 20 个携带 Y337F 突变,其中 8 个样本显示出突变固定。随后的生物测定显示,含有 Y337F 突变的种群对双甲脒的抗性超过 8 倍。对暴露于 10 mg a.i./l 阿米曲拉后的螨虫进行基因分型,结果表明所有存活的螨虫都是 Y337F 突变的同源基因,而死亡的螨虫则携带易感等位基因,这提供了突变与表型之间的遗传联系。此外,我们还利用 CRISPR-Cas9 编辑技术在模式生物 Tetranychus urticae 的同源 Octβ2R 中引入了 Y337F 突变。Crispants 对双甲脒的抗性超过三倍。总之,这项研究发现并验证了一种新型的双甲脒抗性突变。还需要进行更多的研究,以进一步评估 Y337F 在当前治疗策略下的抗药性表型强度。
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引用次数: 0
Effects of the novel acaricide acynonapyr on the calcium-activated potassium channel 新型杀螨剂炔草酯对钙激活钾通道的影响
IF 4.2 1区 农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-08-08 DOI: 10.1016/j.pestbp.2024.106074

Resistance to insecticides and acaricides is a major impediment to effectively controlling insect pests worldwide. These pests include the two-spotted spider mite Tetranychus urticae (T. urticae), which exists globally. This polyphagous herbivore causes major agricultural problems and can develop resistance to the agents above. Therefore, the continuous development of acaricides with new modes of action is important to circumvent the resistance of insects to pesticides. Acynonapyr is a novel class of acaricides containing an azabicyclo ring. In this study, we determined the activity of acynonapyr and its analogs on calcium-activated potassium (KCa2) channels in two-spotted spider mites using electrophysiological techniques (patch-clamp). We also examined their acaricidal efficacy against mites in the laboratory. The acynonapyr and analogs blocked T. urticae KCa2 (TurKCa2) channels in a concentration-dependent manner. A comparison of acaricidal activity against T. urticae with inhibitory activity against TurKCa2 revealed that TurKCa2 channels are the primary toxicological targets. Finally, we examined the effect of acynonapyr on Homo sapiens KCa2 (HsaKCa2.2) channels and demonstrated that the compound at 10 μM had a limited effect on the activity of this channel.

对杀虫剂和杀螨剂的抗药性是有效控制全球害虫的主要障碍。这些害虫包括存在于全球的双斑蜘蛛螨(Tetranychus urticae,T. urticae)。这种多食性食草动物会造成严重的农业问题,而且会对上述药剂产生抗药性。因此,不断开发具有新作用模式的杀螨剂对于避免昆虫对杀虫剂产生抗药性非常重要。炔草酯是一类含有氮杂双环的新型杀螨剂。在这项研究中,我们利用电生理技术(膜片钳)测定了炔草酯及其类似物对二斑蛛螨体内钙激活钾(KCa2)通道的活性。我们还在实验室中检测了它们对螨虫的杀螨功效。炔草酯及其类似物以浓度依赖性的方式阻断了T. urticae KCa2(TurKCa2)通道。通过比较对荨麻螨的杀螨活性和对 TurKCa2 的抑制活性,我们发现 TurKCa2 通道是主要的毒性靶标。最后,我们研究了炔草酯对智人 KCa2(HsaKCa2.2)通道的影响,结果表明 10 μM 的化合物对该通道的活性影响有限。
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引用次数: 0
Harnessing Lecanicillium attenuatum: A novel strategy for combatting Nilaparvata lugens in rice fields 利用Lecanicillium attenuatum:对抗稻田中Nilaparvata lugens的新策略
IF 4.2 1区 农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-08-08 DOI: 10.1016/j.pestbp.2024.106078

Nilaparvata lugens is a notorious rice pest causing significant annual yield and economic losses. The use of entomopathogenic fungi offers a promising and eco-friendly approach to sustainable pest management programs. However, research in this area is currently limited to a few specific types of insects and other arthropods. This study aimed to analyze the biocontrol potential of Lecanicillium attenuatum against N. lugens. Bioassays showed that L. attenuatum 3166 induced >80% mortality in N. lugens following 7 d exposure. Greenhouse and field investigations demonstrated that L. attenuatum 3166 application leads to a substantial reduction in N. lugens populations. Under greenhouse conditions, fluorescence was detected in GFP-labeled L. attenuatum 3166 hyphae enveloping the bodies of N. lugens. In field trials, L. attenuatum 3166 treatment exhibited a control efficacy of up to 68.94% at 14 d post-application, which was comparable to that of the commercial entomopathogenic fungal agent. Genomic sequencing of L. attenuatum 3166 revealed a comprehensive array of genes implicated in its infestation and lethality. Further, the transcriptome sequencing analysis highlighted the elevated expression levels of genes encoding proteases, chitinases, cutinases, and phospholipases. Our findings highlight the potential of L. attenuatum 3166 as an effective biological control agent against N. lugens.

Nilaparvata lugens 是一种臭名昭著的水稻害虫,每年都会造成巨大的产量和经济损失。使用昆虫病原真菌为可持续虫害管理计划提供了一种前景广阔的生态友好型方法。然而,该领域的研究目前仅限于几种特定类型的昆虫和其他节肢动物。本研究旨在分析 Lecanicillium attenuatum 对 N. lugens 的生物防治潜力。生物测定显示,L. attenuatum 3166 在接触 N. lugens 7 d 后可诱导其死亡 80%。温室和田间调查表明,施用 L. attenuatum 3166 能大幅减少 N. lugens 的数量。在温室条件下,GFP 标记的 L. attenuatum 3166菌丝包裹着 N. lugens 的身体,可以检测到荧光。在田间试验中,施用 L. attenuatum 3166 后 14 d 的防治效果高达 68.94%,与商业昆虫病原真菌制剂的防治效果相当。对 L. attenuatum 3166 的基因组测序发现了一系列与其侵染和致死有关的基因。此外,转录组测序分析突出表明,编码蛋白酶、几丁质酶、角质酶和磷脂酶的基因表达水平升高。我们的研究结果凸显了 L. attenuatum 3166 作为一种有效的生物防治剂防治 N. lugens 的潜力。
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引用次数: 0
Biocontrol activity and potential mechanism of Bacillus cereus G5 against Meloidogyne graminicola 蜡样芽孢杆菌 G5 对禾谷圆线虫的生物防治活性和潜在机制
IF 4.2 1区 农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-08-08 DOI: 10.1016/j.pestbp.2024.106079

Root-knot nematodes (Meloidogyne spp.) are highly destructive pests that cause significant yield losses annually. Biological control of nematodes has emerged as a potential alternative in sustainable agriculture. In this study, we originally isolated Bacillus cereus G5 from the rhizosphere soil of rice (Oryza sativa). Treatment with the fermentation supernatant of G5 in vitro demonstrated high toxicity to second-stage juveniles (J2) of Meloidogyne graminicola and remarkably inhibited egg hatching. Moreover, G5 steadily colonized rhizosphere soil and rice seedlings, and exhibited excellent biocontrol efficacy against M. graminicola under greenhouse conditions. Notably, the volatile organic compounds (VOCs) produced by G5 displayed high fumigant activity against M. graminicola. The G5 VOCs efficiently reduced the gall index and nematode population in rice roots, while also promoting rice growth in double-layered pot tests. Additionally, the expression of defense genes involved in the salicylic acid (OsNPR1, OsWRKY45, OsPAL1), jasmonic acid (OsJaMYB, OsAOS2) and ethylene (OsACS1) signalling pathways was significantly upregulated in rice seedlings treated with G5 VOCs. This suggests that G5 VOCs contribute to eliciting plant defense responses. Furthermore, we identified 14 major VOCs produced by G5 using solid-phase micro-extraction gas chromatography and mass spectrometry (SPEM–GC–MS). Notably, allomatrine, morantel, 1-octen-3-ol and 3-methyl-2-butanol displayed strong contact nematicidal activity. Among these, only 1-octen-3-ol demonstrated fumigant activity against J2s of M. graminicola, with an LC50 value of 758.95 mg/L at 24 h. Overall, these results indicated that the B. cereus G5 and its synthetic VOCs possess high potential as biocontrol agents for managing root-knot nematodes.

根结线虫(Meloidogyne spp.)线虫的生物防治已成为可持续农业的一种潜在替代方法。在这项研究中,我们最初从水稻(Oryza sativa)的根瘤土壤中分离出蜡样芽孢杆菌 G5。在体外用 G5 的发酵上清液处理后,发现其对禾谷圆线虫第二期幼虫(J2)具有很高的毒性,并能显著抑制卵的孵化。此外,G5 还能稳定地定植于根瘤土壤和水稻秧苗,在温室条件下对禾谷圆线虫有很好的生物防治效果。值得注意的是,G5 产生的挥发性有机化合物(VOCs)对禾谷介壳虫具有很高的熏蒸活性。在双层盆栽试验中,G5 VOCs 能有效降低水稻根部的虫瘿指数和线虫数量,同时还能促进水稻生长。此外,参与水杨酸(OsNPR1、OsWRKY45、OsPAL1)、茉莉酸(OsJaMYB、OsAOS2)和乙烯(OsACS1)信号通路的防御基因的表达在使用 G5 VOCs 处理的水稻幼苗中显著上调。这表明 G5 VOCs 有助于激发植物防御反应。此外,我们还利用固相微萃取气相色谱和质谱法(SPEM-GC-MS)鉴定了 G5 产生的 14 种主要挥发性有机化合物。值得注意的是,异马菌素、莫兰特尔、1-辛烯-3-醇和 3-甲基-2-丁醇具有很强的接触杀线虫活性。总之,这些结果表明 B. cereus G5 及其合成挥发性有机化合物具有很高的生物防治根结线虫的潜力。
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引用次数: 0
Validation of shikimate dehydrogenase as the herbicidal target of drupacine and screening of target-based compounds with high herbicidal activity 验证莽草酸脱氢酶是 drupacine 的除草靶标并筛选具有高除草活性的靶标化合物
IF 4.2 1区 农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-08-05 DOI: 10.1016/j.pestbp.2024.106077

The discovery of new targets and lead compounds is the key to developing new pesticides. The herbicidal target of drupacine has been identified as shikimate dehydrogenase (SkDH). However, the mechanism of interaction between them remains unclear. This study found that drupacine specifically binds to SkDH with a dissociation equilibrium constant (KD) of 8.88 μM and a Kd value of 2.15 μM, as confirmed by surface plasmon resonance and microscale thermophoresis. Site-directed mutagenesis coupled with fluorescence quenching analysis indicated that residue THR431 was the key amino acid site for drupacine binding to SkDH. Nine compounds with the best binding ability to SkDH were identified by virtual screening from about 120,000 compounds. Among them, compound 8 showed the highest inhibition rate with values of 41.95% against SkDH, also exhibiting the strongest herbicidal activity. This research identifies a novel potential target SkDH and a candidate lead compound with high herbicidal activity for developing new herbicides.

发现新的靶标和先导化合物是开发新农药的关键。目前已确定 drupacine 的除草靶标是莽草酸脱氢酶(SkDH)。然而,它们之间的相互作用机制仍不清楚。本研究发现,经表面等离子体共振和微尺度热泳证实, drupacine 能与 SkDH 特异性结合,其解离平衡常数(K)为 8.88 μM,K 值为 2.15 μM。定点突变和荧光淬灭分析表明,残基 THR431 是 drupacine 与 SkDH 结合的关键氨基酸位点。通过虚拟筛选,从约 120,000 个化合物中确定了 9 个与 SkDH 结合能力最强的化合物。这项研究发现了一个新的潜在靶标 SkDH 和一个具有高除草活性的候选先导化合物,可用于开发新的除草剂。
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引用次数: 0
Expressional respones of hsp70 genes against abiotic and entomopathgenic stresses in four different noctuid larval species (Lepidoptera: Noctidae) 四种不同夜蛾幼虫(鳞翅目:夜蛾科)中 hsp70 基因对非生物胁迫和昆虫病原胁迫的表达反应
IF 4.2 1区 农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-08-05 DOI: 10.1016/j.pestbp.2024.106059

Heat shock proteins (Hsps) are stress response proteins. In a previous study, host larval Hsp70s were identified as the structural proteins of virions of Heliothis virescens ascovirus 3h (HvAV-3h), an insect virus that mainly infects noctuid larvae. To investigate the response of hsp70s of healthy Mythimna separata, Spodoptera exigua, Spodoptera frugiperda, and Spodoptera litura larvae to various abiotic or entomopathogenic stresses, quantitative PCR was used to detect larval hsp70s expression patterns. Results showed distinct expression patterns of hsp70s in response to different abiotic stresses. Notably, Mshsp70 expression pattern resembled Slhsp70 under most treatments. In healthy larvae, no tissue tropism was observed concerning the relative expression of Mshsp70, Sfhsp70, and Slhsp70. After infection with HvAV-3h, the expression of hsp70s in all dissected tissues of all tested larval species increased. Significant differences were found in the fat bodies of M. separata, S. exigua, and S. litura as well as in the hemolymph of S. exigua and S. litura. Subsequent silencing of Slhsp70, resulted in a significant decrease in DNA replication levels of HvAV-3h in S. litura larvae at 24 and 72 h post RNA interference, indicating that Slhsp70 is necessary for DNA replication in HvAV-3h. These data can provide references for the studying on the stress response of noctuid larvae to different environmental factors.

热休克蛋白(Hsps)是一种应激反应蛋白。之前的一项研究发现,宿主幼虫的 Hsp70s 是 Heliothis virescens ascovens virus 3h (HvAV-3h)病毒的结构蛋白,HvAV-3h 是一种主要感染夜蛾幼虫的昆虫病毒。为了研究健康的、、和幼虫对各种非生物或昆虫病原胁迫的反应,使用定量 PCR 检测幼虫的表达模式。结果表明,幼虫在不同的非生物胁迫下有不同的表达模式。值得注意的是,大多数处理下的表达模式相似。在健康幼虫体内,没有观察到Ⅳ、Ⅴ和Ⅵ的相对表达有组织趋向性。感染 HvAV-3h 后,所有受试幼虫的所有解剖组织中的、、和的表达量都有所增加。在Ⅳ、Ⅴ和Ⅳ的脂肪体以及Ⅴ和Ⅳ的血淋巴中发现了显著差异。RNA干扰后24小时和72小时,HvAV-3h在幼虫体内的DNA复制水平显著下降,这表明HvAV-3h的DNA复制是必需的。这些数据可为研究夜蛾幼虫对不同环境因素的应激反应提供参考。
{"title":"Expressional respones of hsp70 genes against abiotic and entomopathgenic stresses in four different noctuid larval species (Lepidoptera: Noctidae)","authors":"","doi":"10.1016/j.pestbp.2024.106059","DOIUrl":"10.1016/j.pestbp.2024.106059","url":null,"abstract":"<div><p>Heat shock proteins (Hsps) are stress response proteins. In a previous study, host larval Hsp70s were identified as the structural proteins of virions of Heliothis virescens ascovirus 3h (HvAV-3h), an insect virus that mainly infects noctuid larvae. To investigate the response of <em>hsp70s</em> of healthy <em>Mythimna separata</em>, <em>Spodoptera exigua</em>, <em>Spodoptera frugiperda</em>, and <em>Spodoptera litura</em> larvae to various abiotic or entomopathogenic stresses, quantitative PCR was used to detect larval <em>hsp70s</em> expression patterns. Results showed distinct expression patterns of <em>hsp70s</em> in response to different abiotic stresses. Notably, <em>Mshsp70</em> expression pattern resembled <em>Slhsp70</em> under most treatments. In healthy larvae, no tissue tropism was observed concerning the relative expression of <em>Mshsp70</em>, <em>Sfhsp70</em>, and <em>Slhsp70</em>. After infection with HvAV-3h, the expression of <em>hsp70s</em> in all dissected tissues of all tested larval species increased. Significant differences were found in the fat bodies of <em>M. separata</em>, <em>S. exigua</em>, and <em>S. litura</em> as well as in the hemolymph of <em>S. exigua</em> and <em>S. litura</em>. Subsequent silencing of <em>Slhsp70</em>, resulted in a significant decrease in DNA replication levels of HvAV-3h in <em>S. litura</em> larvae at 24 and 72 h post RNA interference, indicating that <em>Slhsp70</em> is necessary for DNA replication in HvAV-3h. These data can provide references for the studying on the stress response of noctuid larvae to different environmental factors.</p></div>","PeriodicalId":19828,"journal":{"name":"Pesticide Biochemistry and Physiology","volume":null,"pages":null},"PeriodicalIF":4.2,"publicationDate":"2024-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141941322","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
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Pesticide Biochemistry and Physiology
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