Isolation and identification of acaricidal compounds in Inula japonica (Asteraceae)

We identified acaricidal compounds in Inula japonica Thunberg (Asteraceae) that were effective against carmine spider mite, Tetranychus cinnabarinus (Boisduval). Petroleum-ether extracts from I. japonica were toxic to T. cinnabarinus, with a 50% lethal concentration (LC50) value of 1.18 mg/ml. Silica gel column chromatography was used to separate the acaricidal components. Seventeen of 38 fraction groups had mite mortality rates >50%, nine fraction groups had rates >60%, and three fraction groups had rates >80% at 2 mg/ml. The major volatile compounds in the bioactive fraction groups were identified by gas chromatography-mass spectroscopy, and beta-sitosterol (1), stigmasterol (2), lupeol (3), and alpha-amyrin (4) were identified and determined to have acaricidal activity against T. cinnabarinus in vitro.     

Acaricidal activity of Juglans regia leaf extracts on Tetranychus viennensis and Tetranychus cinnabarinus (Acari: Tetranychidae)

Leaf extracts of the walnut, Juglans regia L., were evaluated under laboratory conditions to determine their acaricidal activity on the mites Tetranychus cinnabarinus (Boisduval) and Tetranychus viennensis Zacher (Acari: Tetranychidae). Extracts had both contact and systemic toxicity to these mites. The four solvents tested for preparing crude extracts were petroleum ether, chloroform, ethyl acetate, and methanol. Methanol was the most efficient solvent, with an extraction rate from 17.06 + 0.80 to 20.27 +/- 0.28%. Petroleum ether was the least effective solvent, with extraction rates from 2.30 +/- 0.13 to 2.71 +/- 0.13%. However, the crude extracts with petroleum ether resulted in the highest mite mortality (79.04 +/- 0.52%) in a slide dip bioassay. Mites mortalities from the concentrated extracts prepared by chloroform, ethyl acetate, methanol, or distilled water were significantly lower than petroleum ether. The mean lethal concentrations (LC50) of the extracts from petroleum ether, chloroform, ethyl acetate, methanol, and distilled water to the two mite species were 0.73 +/- 0.04, 1.66 +/- 0.28, 4.96 +/- 0.35, 7.45 +/- 0.67, and 9.91 +/- 0.32 mg/ml, respectively. After liquid chromatography and thin-layer chromatography, the concentrated extracts of petroleum ether were separated into eight fractions and tested for acaricidal activity. Fraction 6 produced significantly higher mite mortality rates than the other groups, killing approximately 90% of both species.     

Acaricidal activities of extracts of Kochia scoparia against Tetranychus urticae, Tetranychus cinnabarinus, and Tetranychus viennensis (Acari: Tetranychidae)

Extracts of an annual herbaceous plant, Kochia scoparia (L.) Schrad (Macrophomina), were bioassayed to determine their acaricidal activities against Tetranychus urticae Koch, Tetranychus cinnabarinus (Boisduval), and Tetranychus viennensis Zacher (Acari: Tetranychidae) in the laboratory. Extracts had both contact and systemic toxicity to these mites. Three solvents were tested for preparing crude extracts: petroleum ether, chloroform, and methanol. Methanol was the most effective solvent, extracting 3.11-4.53% of the acaricide. Petroleum ether was the least effective solvent, extracting 1.25-1.54%. However, extracts with chloroform resulted in the highest mite mortality (78.86%), and ultrasound-assisted extraction required the least time (10 min). Concentrated extracts were prepared using chloroform, methyl acetate, or distilled water as a solvent. Mite mortalities from the concentrated extracts by methyl acetate or distilled water were significantly lower than those by chloroform. The mean lethal concentrations (LC50) of the extracts by chloroform, methyl acetate, and distilled water to the mites were 0.71 +/- 0.06, 2.08 +/- 0.16 and 8.75 +/- 0.062 mg/ml, respectively. After liquid chromatography and thin layer chromatography, the concentrated extracts by chloroform were separated into seven groups of isolated fractions and tested for acaricidal activity.     

木鳖子提取物对朱砂叶螨的触杀活性

测定了木鳖子提取物对朱砂叶螨的触杀活性。用甲醇、氯仿、石油醚3种不同极性的溶剂提取,石油醚提取率最高为30.79%,且对朱砂叶螨成螨和卵的触杀活性均高于其他两种溶剂的提取物,24 h校正死亡率分别为77.52%和72.04%。用甲醇对石油醚提取物萃取,发现甲醇萃取物活性明显高于石油醚部分,对成螨和卵处理24 h后的校正死亡率分别为89.60%和74.65%,产卵抑制率为62.74%,驱避率为58.23%。柱层析对甲醇萃取物进行分离得到10种组分,组分5活性最高,浓度为2 mg/mL时,24 h校正死亡率为86.15%。用薄层层析和气相色谱质谱联用仪分别检测组分5纯度和主成分,初步确定活性成分为α-菠菜甾醇。     

姜黄素类化合物对朱砂叶螨的生物活性

分别用玻片浸渍法和叶片浸渍法测定了从姜黄中分离的姜黄素(curcumin,CCM)、去甲氧基姜黄素(demethoxycurcumin,DMC)和双去甲氧基姜黄素(bisdemethoxycurcumin,BDMC) 3种天然姜黄素类化合物对朱砂叶螨Tetranychus cinnabarinus Boisduval的成螨、若螨、卵的触杀活性以及对成螨产卵的抑制作用。结果表明：3种化合物对朱砂叶螨的生物活性大小依次为BDMC>DMC>CCM。3种化合物对朱砂叶螨成螨触杀活性最高者为BDMC,处理24 h和48 h其LC₅₀分别为1.18和0.51 mg/mL。对若螨触杀活性的大体趋势与对成螨的相同,其中处理48 h,BDMC对若螨的LC₅₀最小,为2.48 mg/mL。3种化合物对朱砂叶螨卵的触杀毒力也同样表现为BDMC>DMC>CCM。3种单体化合物都表现出一定的对朱砂叶螨雌成螨的产卵抑制作用。对姜黄素类化合物构效关系的初步研究,明确了甲氧基在姜黄素模板上对杀螨活性的贡献,对于开发具经济价值的叶螨类杀螨剂或者筛选先导化合物模板,具有一定的参考价值。     

Bioactivity of the metabolites from endophytic actinomycete Fq24 against Tetranychus cinnabarinus

An endophytic actinomycete strain Fq24 was isolated from healthy tomato plants. The acaricidal substances in the metabolites from Fq24 were collected and identified by the methods of extraction, column chromatography, and gas chromatography-mass spectrometry (GC-MS), and their bioactivities against Tetranychus cinnabarinus were measured with slide-dip and leaf-residue methods. Among the extracts, petroleum ether extract had high bioactivity in contact toxicity and oviposition deterrent against T. cinnabarinus. Its lethal concentration of 50% (LC50) was 52.57 mg x L(-1), and its oviposition deterrent concentration of 50% (ODC50) was 43.18 mg x L(-1). The identification with GC-MS showed that the main chemical component of fraction S11 was methyl hexadecanoate, whose molecular formula was C17H34O2, being one of the substances with acaricidal activity in the metabolites from Fq24. The 24 h corrected mortality rate of female mite at 5 mg x mL(-1) of methyl hexadecanoate was 78.3%, and the oviposition deterrent rate was 81.6%.     

东莨菪内酯与双脱甲氧基姜黄素对朱砂叶螨毒力的温度效应

为明确植物性杀螨活性物质东莨菪内酯与双脱甲氧基姜黄素对朱砂叶螨Tetranychus cinnabarinus雌成螨毒力的温度效应, 采用玻片浸渍法测定了两者不同温度下的杀螨活性。结果表明： 在8～26℃的温度范围内, 东莨菪内酯和双脱甲氧基姜黄素对朱砂叶螨雌成螨的毒力呈正温度系数; 在26～34℃的温度范围内两者对朱砂叶螨雌成螨的毒力呈负温度系数。其中26℃下东莨菪内酯和双脱甲氧基姜黄素表现出较好杀螨活性, 处理后48 h的致死中浓度(LC₅₀)分别为0.1884和0.3376 mg/mL; 23℃下的毒力次之。致死中浓度(y)与温度(x)关系的拟合方程为： 东莨菪内酯: y₁= 0.006x₁²-0.278x₁+3.403; 双脱甲氧基姜黄素: y₂=0.007x₂²-0.354x₂+4.826。对y求最小值得出, 东莨菪内酯和双脱甲氧基姜黄素对朱砂叶螨雌成螨的最高毒力温度分别为23.2℃和25.3℃, LC₅₀分别为0.1828和0.3504 mg/mL。据此认为, 在一定的温度范围内, 随着温度的升高, 这两种植物性杀螨活性物质对朱砂叶螨的毒力与温度先呈正相关, 到达最佳毒力温度后再呈负相关。     

东莨菪内酯对朱砂叶螨的驱避和产卵抑制活性

本研究采用叶碟浸渍法测定了东莨菪内酯对朱砂叶螨Tetranychus cinnabarinus(Biosduvals)雌成螨的触杀活性,采用选择性的半叶法测定了驱避活性,同时采用非选择性的全叶法测定了产卵抑制活性。结果表明,东莨菪内酯对朱砂叶螨雌成螨具有较好的触杀活性,采用叶碟浸渍法处理48h后的LC50为0.297mg·mL-1,LC30为0.105mg·mL-1,LC10为0.023mg·mL-1。采用LC50、LC30和LC103个浓度的东莨菪内酯处理雌成螨后,发现东莨菪内酯对雌成螨没有表现出明显的驱避活性,处理区和对照区24h和48h的着螨率均为50%左右;采用LC50和LC102个浓度处理雌成螨后表现出了一定的产卵抑制活性,平均产卵抑制率分别为23.02%和13.23%,而LC30对雌成螨的产卵量表现出了促进作用,平均产卵抑制率为-8.25%,这可能是由于毒物兴奋效应所致。本研究旨在为东莨菪内酯的进一步开发和应用奠定理论基础。     

生防放线菌Fq24代谢产物对朱砂叶螨的生物活性

菌株Fq24是从健康番茄植株分离得到的一株植物内生放线菌.为了研究它对朱砂叶螨生物活性的影响,通过萃取、柱层析、气相色谱-质谱(GC-MS)等技术对Fq24代谢产物中的杀螨活性物质进行了分离和结构鉴定,并采用玻片浸渍法和叶片残毒法测定了这些代谢产物的生物活性.结果表明:石油醚萃取物对朱砂叶螨具有较强的触杀和产卵忌避作用.触杀作用的LC50为52.57 mg·L-1,产卵忌避作用的ODC50为43.18 mg·L-1.经GC-MS分析,流份S11的主要化学成分为棕榈酸甲酯,分子式为C17H34O2,是代谢产物中的杀螨活性物质之一.质量浓度为5 mg·mL-1的棕榈酸甲酯对雌成螨的24h校正死亡率为78.3％,对雌成螨24h产卵驱避率为81.6％.     

朱砂叶螨体内感染的Wolbachia的wsp基因序列测定与分析

应用Wolbachia的wsp基因特异引物,通过PCR扩增法对我国朱砂叶螨Tetranychus cinnabarinus7个地理种群进行了检测。在采自黑龙江佳木斯、安徽安庆、江苏镇江和浙江慈溪的4个地理种群中扩增出了596bp左右的Wolbachia的wsp基因片段,而在河北威县、山东滨州和湖北赤壁3个地理种群中未发现这个Wolbachia特征基因片段,表明 Wolbachia在我国朱砂叶螨中的侵染较为普遍。通过对我国朱砂叶螨体内感染的 Wolbachia的wsp基因序列进行系统发育分析,得出它们全部与B大组的Ori组的Wolbachia株十分相近或完全相同,提示它们可能是相近或相同的株。     

Acaricidal properties of Artemisia absinthium and Tanacetum vulgare (Asteraceae) essential oils obtained by three methods of extraction

Essential oils of Artemisia absinthium L. and Tanacetum vulgare L. were extracted by three methods, a microwave assisted process (MAP), distillation in water (DW) and direct steam distillation (DSD), and tested for their relative toxicity as contact acaricides to the two spotted spider mite, Tetranychus urticae Koch. All three extracts of A. absinthium and of T. vulgare were lethal to the spider mite but to variable degrees. The LC50 obtained from the DSD oil of A. absinthium was significantly lower (0.04 mg/cm2) than that of the MAP (0.13 mg/cm2) and DW (0.13 mg/cm2) oil of this plant species. DSD and DW extracts of T. vulgare were more toxic (75.6 and 60.4% mite mortality, respectively, at 4% concentration) to the spider mite than the MAP extract (16.7% mite mortality at 4% concentration). Chromatographic analysis indicated differences in composition between the more toxic DSD oil of A. absinthium and the other two extracts of this plant, indicating that a sesquiterpene (C15H24) compound present in the DSD oil and absent in the other two may enhance the toxicity of the DSD oil. Chemical analysis of the T. vulgare extracts indicated that beta-thujone is by far the major compound of the oil (>87.6%) and probably contributes significantly to the acaricidal activity of the oil.     

双去甲氧基姜黄素对朱砂叶螨的生物活性及作用方式

在室温26℃±1℃、湿度60%～80%、光照14 h条件下,测定了天然植物活性成分双去甲氧基姜黄素对重要植食性害螨朱砂叶螨各螨态的触杀和熏蒸活性,对幼螨、若螨、成螨的驱避活性,以及对雌成螨的产卵抑制活性.采用玻片浸渍法测得双去甲氧基姜黄素对朱砂叶螨雌成螨48 h的LCso为0.433 mg·mL-1.在0.883 mg·mL-1(LC70)的浓度下,双去甲氧基姜黄素对朱砂叶螨不同螨态触杀活性的大小依次为幼螨＞若螨＞成螨＞卵,其中对幼螨24h和48 h的校正死亡率分别为60.0%和83.3%;对朱砂叶螨各螨态的熏蒸作用不明显,024h和48 h的校正死亡率均小于3%;对幼螨、若螨以及成螨均表现出较强的驱避作用,其中对幼螨的效果最好,不同处理时间的驱避率均在85%以上,其次是若螨,对成螨的驱避性相对较差,72 h的驱避率仅为47.8%.同时对雌成螨有明显的产卵抑制作用,处理后120 h产卵抑制率达到89.3%.表明双去甲氧基姜黄素对朱砂叶螨的主要作用方式为触杀、驱避和产卵抑制作用.     

Acaricidal activity of Aloe vera L. leaf extracts against Tetranychus cinnabarinus (Boisduval) (Acarina: Tetranychidae)

Four different extracts of Aloe vera L. leaves were evaluated for acaricidal activity against female adults of carmine spider mite, Tetranychus cinnabarinus (Boisduval), by slide-dip bioassay. At 72 h after treatment, the acetone extract showed the strongest acaricidal activity with LC₅₀ value of 90 ppm. The LC₅₀ values for ethyl acetate, water, and ethanol extracts were 113, 340, and 391 ppm, respectively. The acetone extract was fractionated using a silica gel column. Among the twenty-two fractions obtained the fifth, tenth, eleventh, twelfth, fifteenth, and seventeenth fractions showed strong acaricidal activity, causing 80.39 to 92.16% mortality at 72 h after treatment. The tenth and eleventh fractions had the strong activity, with LC₅₀ values of 44 ppm and 33 ppm, respectively. The results suggested that A. vera has a great potential for development as a botanical acaricide for T. cinnabarinus control.     

朱砂叶螨对三种杀螨剂的抗性选育与抗性风险评估

为评价朱砂叶螨Tetranychus cinnabarinus对3种杀螨剂的抗性风险,在实验室抗性品系选育基础上,应用数量遗传学中的域性状分析法,研究了朱砂叶螨北碚种群对甲氰菊酯、阿维菌素和哒螨灵3种杀螨剂的抗性现实遗传力,并对3种药剂在不同杀死率下抗性发展的速率进行了预测。结果表明：分别单一连续汰选16代后,朱砂叶螨对甲氰菊酯、阿维菌素的抗性倍数分别达26.54和4.51倍,对哒螨灵表现为敏感性降低（抗性倍数为1.16倍）;朱砂叶螨对甲氰菊酯、阿维菌素和哒螨灵的抗性现实遗传力分别为0.2472,0.1519和0.0160。在室内选择条件下,杀死率为50%～90%时,要获得10倍抗性,甲氰菊酯仅需要13～6代,阿维菌素需要约21～10代;哒螨灵需要约197～89代;在田间选择,三种药剂都将需要更长的时间。抗性筛选16代结果表明,抗性风险较高的是菊酯类的甲氰菊酯,其次是生物源农药阿维菌素,杂环类的哒螨灵抗性风险较小。试验结果可为朱砂叶螨抗性治理提供参考。     

阿维菌素与植物精油复配对朱砂叶螨杀螨的增效作用研究

为了得到对朱砂叶螨(Tetranychuscinnabarinus)具有较好杀螨活性的植物精油与阿维菌素复配配方,并为杀螨剂开发应用提供指导,本研究采用喷雾法测定了柠檬草、广藿香、山鸡椒、亚洲薄荷植物精油及阿维菌素对朱砂叶螨的毒力,分别采用共毒因子法和共毒系数法评价了山鸡椒和亚洲薄荷精油对阿维菌素的增效作用和复配最佳配比.结果表明,柠檬草精油和广藿香精油基本无杀螨活性,山鸡椒精油、亚洲薄荷精油处理朱砂叶螨24h后LC50分别为772.801mg/L和1040.187mg/L.阿维菌素与亚洲薄荷1∶272,1∶679复配具有增效作用.阿维菌素与亚洲薄荷1∶400复配时共毒系数最大,可达160.因此,阿维菌素与亚洲薄荷1∶400复配防治朱砂叶螨具有明显增效作用,这为杀螨剂的开发应用研究提供了参考.     

Various effects of ethanolic extract of Mentha pulegium on the two-spotted spider mite,Tetranychus urticae (Tetranychidae)

Ethanolic extract of Mentha pulegium was evaluated against the adults of important arthropod pest species, the two-spotted spider mite, Tetranychus urticae. Potential acaricidal impact of plant was determined by contact application. Phytotoxicity was recorded after 24 h. The value of LC₅₀ was 59,149 mg L^?1. Also, LT₅₀ values were 27.42, 25.56 and 19.79 h for 35,000, 50,000 and 100,000 mg L^?1, respectively. Also, results showed that this ethanolic extract had impact on repellency of T. urticae. All of the tested concentrations were similar in the repellency test. On the other hand, the used concentrations (1000, 4000, 6000 and 8000 mg L^?1) affected on the oviposition of mite females. These extract doses significantly decreased the egg laying on the treated surface. The current study indicated that this ethanolic extract can be used as a safe acaricide on T. urticae.     

The potential of Lagenaria rootstock to confer resistance to the carmine spider mite, Tetranychus cinnabarinus (Acari: Tetranychidae) in Cucurbitaceae

Antibiosis and resistance of six Cucurbita and two Lagenaria accessions to the carmine spider mite, Tetranychus cinnabarinus Boisduval, were evaluated in the laboratory. Significant differences among accessions were observed three days after the inoculation of detached leaf discs. The Lagenaria accessions, Slawi and Sus, proved to be the most resistant to mites, with average populations of mite eggs, 87 and 95%, respectively less than that of the susceptible C. pepo accession, Orangetti. The Cucurbita accessions, Tace, Brava, Tetsukabuto, Phoenix and TZ-148 had mite egg totals 4, 9, 13, 26 and 40%, respectively, less than those of accession Orangetti. The Sus accession of Lagenaria was resistant to T. cinnabarinus from the four-leaf stage until fruit set in laboratory and field tests. Grafting the susceptible Brava onto Sus rootstock increased the resistance of the scion to the same level as that of non-grafted Sus. Grafting the susceptible Cucumis melo Noy Yizre'el on resistant or susceptible rootstocks of Cucurbita and Lagenaria accessions did not affect its susceptibility to T. cinnabarinus. The results indicate that resistance to T. cinnabarinus can be transferred by grafting from Lagenaria stocks to Cucurbita scions but not in the opposite direction.     

Sensitivity of two greenhouse pests to vapours of essential oils

Vapours of essential oils extracted from cumin, Cuminum cyminum L., anise, Pimpinella anisum L., oregano, Origanum syriacum var bevanii (Holmes) Ietswaart and eucalyptus, Eucalyptus camaldulensis Dehn., were found to be toxic to two greenhouse pests, viz. the carmine spider mite, Tetranychus cinnabarinus (Boisd.) (Tetranychidae, Tetranychini) and cotton aphid, Aphis gossypii Glov. (Aphididae, Aphidini). In general, a minimum dose of 0.5 µl/l air and 2–3 days of exposure was required for 99% mortality using essential oils of the first three plants.     

Acaricidal Activity of Pla.nt Essential Oils against Tyrophagus putrescentiae (Acari: Acaridae)

The acaricidal activity of 54 commercially available plant essential oils against Tyrophagus putrescentiae adults was compared with those of benzyl benzoate and N, N-diethyl-m-toluamide (deet). Responses varied with test oils. In a test with an impregnated fabric disc bioassay at 12.7 μg/cm², over 80% mortality was observed with bay, citronella java, clove bud, clove leaf, lemongrass, nutmeg, oregano, pimento berry, thyme red, and thyme white oils. These oils were more toxic to T. putrescentiae adults than benzyl benzoate and deet. At 6.4 μg/cm², lemongrass and thyme white oils exhibited 76 and 84% mortality, respectively, whereas the acaricidal activity of the other essential oils was significantly decreased. In a fumigation test with adult mites, lemongrass and pimento berry oils were much more effective in closed containers than in open ones, indicating that the effects of these oils were largely due to action in the vapor phase. Plant essential oils described merit further study as potential T. putrescentiae control agents.     

迷迭香油和花椒油对朱砂叶螨的生物活性

【目的】明确室内条件下迷迭香油和花椒油对朱砂叶螨Tetranychus cinnabarinus (Boisduval)的生物活性。【方法】采用熏蒸法和驱避法测定迷迭香油和花椒油对5日龄朱砂叶螨雌成螨的熏蒸活性、产卵抑制活性和驱避活性;并采用EthoVision XT6分析亚致死浓度下两种精油对雌成螨行为的影响。【结果】分别将5日龄雌成螨熏蒸处理12, 24和48 h后,迷迭香油对朱砂叶螨的致死率均显著高于花椒油处理组（P＜0.05）,且随着熏蒸时间的延长或精油浓度的升高,其熏蒸活性均增强;不同浓度的迷迭香油处理朱砂叶螨雌成螨24 h后的产卵抑制率均显著高于花椒油处理组（P＜0.05）。迷迭香油和花椒油处理朱砂叶螨雌成螨24 h的LC₃₀分别为6.731和120.142 μL/L;迷迭香油对朱砂叶螨雌成螨的驱避活性明显高于花椒油处理组。两种精油亚致死浓度处理朱砂叶螨雌成螨后,其自发运动方式发生了明显变化,移动频率在LC₁₀ 时呈现最高;且迷迭香油对朱砂叶螨雌成螨自发运动能力的影响比花椒油处理的影响更为明显。【结论】迷迭香油对朱砂叶螨的生物活性明显高于花椒油,因此,迷迭香油对朱砂叶螨的生物防治具有较高的应用价值。本研究为绿色蔬菜植物源杀螨剂的研究提供了科学依据。