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。     

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。     

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.     

朱砂叶螨抗炔螨特品系相对适合度

为明确朱砂叶螨抗炔螨品系的相对适合度,在朱砂叶螨敏感品系(SS)的基础上使用炔螨特持续筛选34代获得了炔螨特抗性品系(PRR34)。通过统计2个品系的实验种群生命表,对其发育历期、繁殖力及种群参数差异进行了比较。结果表明,朱砂叶螨敏感品系经炔螨特持续筛选34代后(PPR34),其抗性倍数为37.78。与SS品系相比,PPR34卵孵化率、发育历期和世代周期均无显著差异,但产卵前期显著延长了37.07%,单雌产卵量减少14.61%,雌雄性(♀/♂)比增加了23.38%。采用Weibull分布拟合存活曲线模型为DeeveyⅠ型,SS品系和PRR34品系的参数c值分别为2.04和2.38。其种群趋势指数、内禀增长率(rm)、净生殖率(R0)分别为123.78、0.24和25.34,以及111.91、0.26和28.99;PRR34品系的相对适合度为1.14,与敏感品系相比存在一定优势。因此,与敏感品系相比,朱砂叶螨抗炔螨特品系单雌产卵量下降,但雌性比增加,种群适合度优势较高,可能导致抗性种群迅速增长并增加防控难度。     

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.     

抗阿维菌素朱砂叶螨的热激反应及热激蛋白

选用朱砂叶螨Tetranychus cinnabarinus阿维菌素抗性品系和敏感品系,测定了热预刺激后其在极限高温下的存活率,并应用SDS-PAGE技术研究了热激蛋白(HSPs)的种类及其含量。结果表明：非致死的热预刺激能显著提高朱砂叶螨耐极限温度的能力。两个品系在不同温度热激处理后,其蛋白质种类和含量发生了变化。正常情况下,朱砂叶螨敏感品系与阿维菌素抗性品系相比缺失8条条带;敏感品系热激后,增加了分子量分别为97.2,74.3,62.4,53.0和30.3 kDa的5条条带; 抗性品系热激后没有特异蛋白带的产生,但进一步高温胁迫后有些蛋白表达增强。此结果有助于解释朱砂叶螨抗性品系存在高温适合度优势现象。     

Biological parameters of the predatory mite Cheletomimus bakeri (Acari: Cheyletidae) feeding on Tetranychus cinnabarinus (Acari: Tetranychidae)

Developmental periods of egg, larva and nymphal stages and fecundity as well as predation of Cheletomimus bakeri (Acari: Cheyletidae) feeding on Tetranychus cinnabarinus (Acari: Tetranychidae) were evaluated at different temperatures (15, 20, 25, 30 and 35°C) at 65 ± 10% relative humidity and 16 L: 8D in the laboratory. The development periods of C. bakeri from egg through adult decreased significantly when the temperature was increased from 20°C to 35°C. Egg and total development periods of C. bakeri at 20, 25, 30 and 35°C were 13.86, 7.98, 5.07, 4.08 days and 58.66, 41.51, 21.21, 22.92 days, respectively. The highest numbers of total and daily egg production were found at 20°C and 30°C, respectively. Net reproductive rate (R₀ = 13.29), mean generation time (T = 88.30), gross reproductive rate (GRR = 17.46) and doubling time (DT = 23.66) were the highest at 20°C. The intrinsic rate of increase (r_m = 0.0592) and finite capacity for increase (λ = 1.061) for C. bakeri were the highest at 30°C. Predation of C. bakeri increased throughout the range of prey densities. The highest consumption number of C. bakeri feeding on T. cinnabarinus males per day was 4.63, 4.70 and 4.60 when confined to 40, 80 and 160 individuals, respectively. Our data suggest that C. bakeri does not appear to have much promise for the control of spider mites because of the characteristics of the predator such as slow development period, poor searching capacity and low intrinsic rate of increase.     

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.     

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。     

Effect of temperature on toxicities of scopoletin and bisdemethoxycurcumin against Tetranychus cinnabarinus ( Acari:Tetranychidae )

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°C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.     

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。     

Toxicity of ethanolic extracts from Lippia origanoides and Gliricidia sepium to Tetranychus cinnabarinus (Boisduval) (Acari: Tetranychidae)

Botanical compounds with insecticidal and acaricidal activities have been used in pest management with different levels of success. Toxicity of ethanolic extracts obtained from wild oregano (Lippia origanoides) and gliricidia (Gliricidia sepium) to Tetranychus cinnabarinus (Boisduval) were evaluated. Mite population was collected from black bean plants growing in Urachiche Municipality, Yaracuy State, Venezuela. Ethanolic extracts of wild oregano and gliricidia leaves were evaluated at different concentrations (5, 10, 15, and 20%) using the leaf disk immersion technique. The presence of alkaloids, flavonoids, phenols and tannins, essential oils and saponins was verified in the plant material used in our study. Tetranychus cinnabarinus oviposition decreased at a rate of 43.7% or 57% when 5% oregano or gliricidia extracts were used, respectively. Also, 10% oregano or gliricidia extracts caused 42.2% or 72.5% of mortality to T. cinnabarinus, respectively. Ethanolic extracts showed acaricidal effects on T. cinnabarinus, as evidenced by maximum mortality (96.6% and 100% caused by wild oregano and gliricidia, respectively) when used at a concentration of 20%. Our results showed that gliricidia and wild oregano are promising for the management of T. cinnabarinus, although their field efficacy remains to be evaluated.     

朱砂叶螨刺吸胁迫对玉米防御信号分子的诱导作用

【目的】探明朱砂叶螨Tetranychus cinnabarinus持续为害对玉米Zea mays叶片内茉莉酸(jasmonic acid,JA)、水杨酸(salicylic acid,SA)、乙烯(ethylene,ET)、一氧化氮(nitricoxide,NO)、脱落酸(abscisic acid,ABA)和过氧化氢(hydrogen peroxide,H2O2)6个防御信号分子的诱导作用。【方法】室内人工接螨(10,20和30头/叶),采用分光光度法(SP)、高效液相色谱法(HPLC)和酶联免疫法(ELISA),测定了朱砂叶螨持续为害0,24,48,72和96 h后,玉米幼苗叶片内6个信号分子的含量。【结果】朱砂叶螨持续刺吸为害玉米幼苗叶片后,JA,ABA和H2O23个信号分子含量在叶螨刺吸为害24 h内迅速上升,在24 h时达高峰值,叶螨密度为30头/叶时其含量分别为同期未接螨对照的4.13,3.84和3.20倍,24-48 h内迅速下降;此后,ABA和H2O2含量维持在较低水平,而JA含量在48-96 h内又上升至次高峰值。NO含量则在24-48 h内上升较快,48 h时达最高,叶螨密度为30头/叶时其含量为同期未接螨对照的5.09倍。SA和ET含量在96 h内均随刺吸时间的延长而增大,96 h时最高,叶螨密度为30头/叶时其含量分别为同期未接螨对照的5.17和2.99倍。叶螨密度为30头/叶时,6个信号分子含量均显著高于同期未接螨对照(P0.05)。【结论】朱砂叶螨为害对玉米叶片内JA,SA,ET,NO,ABA和H2O2均具有诱导作用,且6个信号分子在叶螨持续为害玉米叶片后循序被诱导。     

朱砂叶螨HSP90基因克隆及原核表达

采用RT-PCR及RACE技术克隆朱砂叶螨Tetranychus cinnabarinus的热激蛋白90(HSP90)基因, 并进行序列分析, 得到一条长2 595 bp的cDNA序列, 该序列开放阅读框（open reading frame, ORF）为2 169 bp, 编码722个氨基酸, 分子量约为83.45 kDa, 理论等电点为4.81, 3′非编码区（untranslated region, UTR）为249 bp, 5′UTR为177 bp。通过Antheprot分析发现5个HSP90家族的签名序列及胞质HSP90特征序列MEEVD。同源性分析表明, 朱砂叶螨HSP90编码区核苷酸序列和其他已知的HSP90, 尤其是节肢动物昆虫的HSP90, 具有很高的相似性。将鉴定正确的原核重组表达质粒pET43a-TcHSP90, 转化大肠杆菌Escherichia coli BL21(origami) 进行原核表达, 应用SDS-PAGE和Western blotting技术分离并检测融合蛋白, 结果表明构建的原核表达质粒可以在宿主菌中稳定、正确表达。朱砂叶螨TcHSP90基因的克隆、原核表达, 为进一步研究HSP90的性质和功能的研究提供有用的实验材料。     

东莨菪内酯与双脱甲氧基姜黄素对朱砂叶螨致死效应模拟

采用玻片浸渍法测定了植物源杀螨活性物质东莨菪内酯、双脱甲氧基姜黄素及两者联合作用最佳增效配比(东:双=7:6)对朱砂叶螨雌成螨的致死效应,并采用时间-剂量-死亡率(TDM)模型进行模拟.结果表明:所建TDM模型均通过Hosmer-Lemoshow拟合异质性检验,朱砂叶螨雌成螨对3种药剂浓度变化的敏感程度为东莨菪内酯＞两者最佳配比＞双脱甲氧基姜黄素,双脱甲氧基姜黄素、东莨菪内酯和两者最佳配比对朱砂叶螨雌成螨的致死率高峰期分别为处理后32、28和32 h;处理后48 h的LC50和LC90分别为0.3324、0.2035、0.2195mg·mL-1和2.1198、0.9521、1.1617 mg·mL-1;浓度为1.0和2.0 mg·mL-1处理下,LT50分别为7.4、6.0、6.1h和6.4、4.8、5.0h.因此,东莨菪内酯与双脱甲氧基姜黄素最佳配比和东莨菪内酯具有相近的杀螨动态及时间-剂量效应,表现出较好的杀螨活性和增效作用,具有一定的开发利用价值.     

Effects of simulated acid rain on the physiology of carmine spider mite, Tetranychus cinnabarinus (Boisduvals) (Acari: Tetranychidae)

Abstract:  The physiological effect of simulated acid rain sprayed on carmine spider mite Tetranychus cinnabarinus (Boisduvals) and host plant, were measured in a series of laboratory trials. We examined potential changes in three kinds of protective enzymes [peroxidase (POD), superoxide dismutase (SOD) and catalase (CAT)] and three hydrolases [acid phosphatase (ACP), alkaline phosphatase (ALP) and carboxylesterase (CarE)] in response to changes in pH values of simulated acid rain at different time of exposure. POD, SOD and CAT activities increased significantly with the increase in the acidity of the acid rain, reaching the highest levels at pH 4.0 or 3.0, and then declined. Changes in ACP activity were similar to those observed in the protective enzymes. The increasing extent of the activities of these four enzymes after 30 and 45 days treatment became smaller than that after 15 days treatment . ALP activities decreased as pH value declined. There were no significant changes in CarE activities after 15 and 45 days, but that in pH 4.0 and 3.0 decreased after 30 days. The enhanced anti-oxidation enzyme levels (POD, SOD and CAT) and ACP activities in pH 4.0 and 3.0 reduced the effects of these toxic products on mites, resulting in the strengthening of the defensive power, and increase in survival and reproductive power of the mites, thus leading to an increase in the density of mites on host plant. From these results, we inferred that POD, SOD, CAT and ACP might be relevant to population changes of mites under acid rain pressure.     

Population Dynamics of Tetranychus Kanzawai (Acari: Tetranychidae) on Hydrangea

The seasonal occurrence of Tetranychus kanzawai Kishida populations on hydrangea (Hydrangea macrophylla) was studied at two different localities in Ibaraki, Japan, during a three-year period. There were two types of seasonal population trends: one with a population peak from May to June, and the other with the spring peak in June and the autumn peak in September–October. Each year the populations on hydrangea plants abruptly declined just after the spring peak. Predators showed a delayed density-dependent numerical response. The population crashed even in the absence of predators, suggesting that the predators had nothing to do with the June decline. Furthermore, the rate of development from larva to adult and the fecundity in adult females on detached hydrangea leaves decreased markedly just prior to the abrupt decline in density in June. Consequently, seasonal changes in plant quality (perhaps influenced by secondary compounds) seem to contribute to the drastic decline of T. kanzawai density on hydrangea in June.     

Distribution of the endosymbiotic bacterium Cardinium in Chinese populations of the carmine spider mite Tetranychus cinnabarinus (Acari: Tetranychidae)

Abstract:  ' Candidatus Cardinium hertigii ( Cardinium )' from the Bacteroidetes group is a symbiont that has recently received attention as a reproductive manipulator of its arthropod hosts. Though the spatial distribution of symbionts depends heavily on that of their hosts, evidence has shown that dispersal limitations also have effect on some micro-organisms. In order to estimate the distribution of Cardinium in Chinese populations of the carmine spider mite, twenty seven geographical populations of Tetranychus cinnabarinus were collected and screened, using Cardinium -specific primers for the 16S ribosomal DNA. Six geographical populations collected between 30° and 36°N tested positive for the presence of Cardinium , whereas it could not be detected in populations collected between 19°–29°N and 37°–46°N, suggesting that the symbiont is not evenly distributed, comparing with a wider distribution of Wolbachia previously reported in the same populations. In addition, the infection rates of Cardinium seem to increase from the south to the north of China. All the six Cardinium -infected populations were found to be doubly infected with Wolbachia . Phylogenetic analysis based on the 16S rDNA gene sequence of Cardinium shows a close relationship between Cardinium in Chinese carmine spider mite and symbionts found in other species in the Tetranychidae.