Time-mortality relationships of larvae and adults of grain beetles exposed to extreme cold

Mortality effects of low temperature exposure to −16 °C were investigated on larvae and adults of the grain beetles Tribolium confusum Jacquelin du Val (Coleoptera: Tenebrionidae), Oryzaephilus surinamensis (L.) (Coleoptera: Silvanidae) and Trogoderma granarium (Everts) (Coleoptera: Dermestidae), in the laboratory. The main effects and interactions of exposure time (0.2, 0.5, 1, 2, 4, 8, 12, 24 and 48 h), developmental stage (larva and adult) and individual’s age (young and old) were examined. After 4 h of exposure 100% mortality has been achieved in most of the treatments. The only exceptions were that of old larvae of T. granarium and O. surinamensis, and adults of T. granarium which needed 48, 8, and 24 h, respectively. T. granarium was the most cold-hardy species irrespective of developmental stage, and age, followed in decreasing order by O. surinamensis and T. confusum. Younger adults were generally more susceptible to cold in the cases of O. surinamensis and T. confusum. On the contrary, older adults of T. granarium suffered higher mortality than younger ones. Larvae of T. granarium were generally more cold-tolerant than adults but the opposite pattern was observed in O. surinamensis and T. confusum. Main effects of exposure time, developmental stage and individual’s age on mortality proved to be significant for all species, with the exception of T. granarium where the effect of developmental stage proved to be insignificant. Lethal time values were estimated via probit analysis. Values of LT₅₀ ranged between 0.7 and 1.0 h for T. confusum, 0.7-1.9 h for O. surinamensis and 1.2-3.4 h for T. granarium. Our results are discussed with findings from relevant studies and the possibility of including extreme cooling to IPM programs against stored product pests is investigated.     

Efficacy of vaporised ethyl formate/carbon dioxide formulation against stored‐grain insects: effect of fumigant concentration,exposure time and two grain temperatures

BACKGROUND: The ethyl formate/carbon dioxide (CO₂) formulation Vapormate? is a rapid‐acting fumigant being developed for the control of stored‐grain insects. The effects have been investigated of concentration, exposure times of 1, 3, 24 and 72 h and two grain temperatures, 15 and 25 °C, on its efficacy against mixed‐stage cultures of Sitophilus oryzae (L.) Tribolium castaneum (Herbst) and strongly phosphine‐resistant Rhyzopertha dominica (F.) strain QRD569. RESULTS: High mortalities (≥92%) of mixed‐stage cultures of all three species were obtained when grain was fumigated with the formulation (193 g m^?3 ethyl formate) for 1 h. Complete control of R. dominica QRD569 and T. castaneum was achieved with 63 and 76 g m^?3 ethyl formate respectively, with exposure for 24 h, whereas mean mortality of S. oryzae was 86% under the same conditions. Mortalities of S. oryzae juvenile stages were significantly lower than adults under the conditions tested, which was due to pronounced tolerance of mid‐stage pupae to the fumigant. Reducing grain temperature from 25 to 15 °C had no effect on insect mortality. CONCLUSION: Ethyl formate/CO₂ formulation is highly effective against stored‐grain insects over a range of concentrations and exposure times. Efficacious fumigations were conducted in as little as 1 h, and a strongly phosphine‐resistant R. dominica strain was readily controlled with the fumigant. Copyright © 2009 CSIRO, Australia. Published by John Wiley & Sons, Ltd     

番茄黄化曲叶病毒的鉴定与群体进化分析

番茄黄化曲叶病(tomato yellow leaf curl disease,TYLCD)是番茄上的重要病害,可由多种双生病毒引起。为了明确北京地区番茄黄化曲叶病的病毒种类和病毒基因组变异情况,本研究收集了51个采自北京不同区县的病毒疑似样品,进行了双生病毒的检测及分离物的基因组分析。经双生病毒简并引物检测25个样品呈阳性;扩增获得10个代表性分离物的全基因组,大小均为2 781bp,共含有6个开放阅读框;经比对,10个分离物与Tomato yellow leaf curl virus-Israel(TYLCV-IL)同源率大于99%;基于我国TYLCV分离物全基因组序列的变异分析表明,TYLCV在进化上比较保守,整个基因组上的变异是不均匀的;系统发育树显示我国TYLCV分离物在进化树上可分为3个组群,分析了我国TYLCV群体进化情况。     

Toxicity nanoinsecticide based on clove essential oil against Tribolium castaneum (Herbst)

This study aimed to characterize nanoparticles loaded with clove (Syzygium aromaticum) essential oil-based polyethylene glycol (PEG) and to know their insecticidal activity against red flour beetle (Tribolium castaneum). The nanoparticles have irregular shapes in good dispersion. The nanoformulation could not enhance clove oil contact toxicity to T. castaneum, but could protect the oil from degradation and evaporation while simultaneously allowing sustained release, as indicated by the continued high toxicity for 16 weeks of storage.     

赤拟谷盗章鱼胺受体3(TcOctβR3)cDNA克隆、表达及功能

【目的】章鱼胺信号系统在调节昆虫行为和生理过程中具有至关重要的作用。赤拟谷盗(Tribolium castaneum)作为一种模式昆虫,被广泛用于解析昆虫生长发育及生理等调控机制的研究工作。本研究以赤拟谷盗为对象,旨在明确章鱼胺受体在调节赤拟谷盗行为和生理方面的功能。【方法】根据Gen Bank登录的相关序列信息(XP_008198078),利用RT-PCR技术克隆赤拟谷盗章鱼胺受体基因Tc OctβR3的c DNA序列。利用在线生物信息学分析软件预测该基因的开放阅读框、编码的氨基酸序列以及跨膜结构域等信息,基于邻接法构建该基因与其他昆虫相关序列的系统发育树,明确系统进化关系。分别提取赤拟谷盗各发育阶段(卵、幼虫、蛹和成虫)、不同组织(中枢神经系统、脂肪体、中肠、后肠、马氏管、精巢和卵巢)以及饥饿胁迫后的RNA,以赤拟谷盗核糖体蛋白S3(Tc RPS3)为内参基因,采用实时定量PCR技术分析该基因在赤拟谷盗不同发育阶段、不同组织以及在饥饿胁迫下的表达模式。运用哺乳动物异源表达系统在人胚胎肾细胞HEK293中瞬时表达Tc OctβR3,进而利用第二信使c AMP含量测定技术分析Tc OctβR3与配体的结合能力。最后,通过体外合成赤拟谷盗Tc OctβR3的双链RNA,利用RNA干扰以及轨迹球行为分析等技术探究该基因的生理功能。【结果】序列分析结果表明,赤拟谷盗Tc OctβR3开放阅读框全长1 305 bp,编码434个氨基酸,序列中含有G蛋白偶联受体典型的7个跨膜结构域。基于邻接法构建的系统发育树表明,该基因编码的蛋白质与小蜂甲(Aethina tumida)的OctβR3亲缘关系最近。实时定量PCR分析结果表明,Tc OctβR3在赤拟谷盗各发育阶段均有表达,尤其在低龄幼虫期转录水平最高,而在其他发育阶段表达量无显著差异;在赤拟谷盗不同组织中,Tc OctβR3在中枢神经系统的表达量显著高于其他组织;赤拟谷盗幼虫在经饥饿处理24 h的过程中,Tc OctβR3的表达量呈先下降后上升的趋势,且在处理6 h的表达量最低,为对照的0.47倍,在16 h表达量最高,为对照的1.80倍,最后恢复到正常水平。通过HEK293细胞异源表达Tc OctβR3后,c AMP测定结果表明章鱼胺(OA)呈浓度依赖性地激活Tc OctβR3,其EC50为8.68×10-7 mol·L-1,萘甲唑啉(NA)的激动活性最高,其有效中浓度EC50为8.56×10-8 mol·L-1。4种供试生物胺激动剂活性强弱为:萘甲唑啉酪胺(TA)章鱼胺多巴胺(DA)。进一步采用RNA干扰技术的分析结果表明,注射ds RNA能有效抑制Tc OctβR3的表达,沉默效率高达61.5%,但干扰该基因表达后不会影响赤拟谷盗成虫的爬行速度和产卵量。【结论】Tc OctβR3在赤拟谷盗中枢神经系统可能发挥重要作用,能调节幼虫对饥饿胁迫的响应。明确了Tc OctβR3的分子生物学特性,可为将来以其为靶标筛选高效激动剂和抑制剂提供理论依据。     

两种亚菊属植物挥发油及主要成分对赤拟谷盗幼虫的活性研究

本文旨在研究川甘亚菊和灌木亚菊挥发油及其主要成分(1,8-桉叶油醇、樟脑、马鞭草烯醇、冰片和桃金娘烯醇)对赤拟谷盗幼虫的活性作用。采用水蒸气蒸馏法提取挥发油并通过熏蒸、触杀和驱避等测试方法评价两种挥发油及其主要成分对赤拟谷盗幼虫的生物活性。结果表明,川甘亚菊挥发油、灌木亚菊挥发油和1,8-桉叶油醇均具有一定的熏蒸活性(LC₅₀分别为94.60、81.17 mg/L和18.42 mg/L)和触杀活性(LD₅₀分别为216.93、100.50μg/头和52.69μg/头)。同时,在高测试浓度(78.63 nL/cm~2和15.73 nL/cm~2)时,分别处理2 h和4 h后川甘亚菊和灌木亚菊挥发油以及桃金娘烯醇对赤拟谷盗幼虫均表现出较明显的驱避活性,冰片驱避活性接近于阳性对照避蚊胺。因此,川甘亚菊和灌木亚菊挥发油对赤拟谷盗的幼虫有一定的防治作用。     

高浓度CO2气调防治谷蠹及杂拟谷盗的研究

将谷蠹Rhyzopertha dominica(Fab.）成虫及杂拟谷盗Tribolium confusum Jacquelin du val卵、幼虫、肾和成虫分别暴露于35％CO2和11％O2，35％CO2和21％O2，75％CO2和11％O2，75％CO2和21％O2四种气调环境中12、24、36、48、60、72、84、96、108、120小时，结果表明谷蠹成虫对四种气调环境都很敏感，其LT50分别为34.6、42.0、18.6、17.8小时。杂拟谷盗成虫较耐35％CO2、11％O2及35％CO2、21％O2气调环境，处理120小时后，成虫死亡率低于12％。卵及幼虫对四种气调环境较为敏感，蛹对35％CO2、11％O2的耐受性比35％CO2、21％O2强，前者LT50为2248.3小时，后者为124.5小时，杂拟谷盗卵、幼虫、蛹及成虫对四种气调环境敏感顺序为：卵＞幼虫＞蛹＞成虫。     

甘蔗害虫综合防治研究

1981—1984年广东省开展甘蔗害虫综合防治研究、试验、示范。采取“以农业防治为基础,释放赤眼蜂防治蔗螟;科学用药,保护天敌,充分发挥天敌自然控制多种害虫的作用”。在10万亩大面积上,收到到了显著的经济效益、生态效益和环境保护效益。     

拟澳洲赤眼蜂和短管赤眼蜂在菜芯地的扩散动态

在不同生长期的菜芯地释放赤眼蜂防治小菜蛾,当蜂量为2000头／点时,拟澳洲赤眼蜂在田间的有效扩散距离(寄生率大于60％),苗期为3米,生长中期和收获期为4米。短管赤眼蜂在田间的有效扩散距离,苗期和生长中期为4米,收获期为5米,较拟澳洲赤眼蜂具有更强的扩散能力。     

电子束辐照对赤拟谷盗保护酶系的影响

研究了赤拟谷盗Tribolium castaneurn（Herbst）不同虫态（3-4龄幼虫、1-2龄蛹和羽化1天成虫）体内过氧化物酶（POD）、过氧化氢酶（CAT）、超氧歧化酶（SOD）活力在电子束辐照条件下的变化。结果表明,赤拟谷盗不同虫态的POD活力：羽化1 d成虫〉1-2龄蛹〉3-4龄幼虫;CAT活力：羽化1 d成虫〉1-2龄蛹〉3-4龄幼虫;SOD活力：3-4龄幼虫〉1-2龄蛹〉羽化1 d成虫。辐照处理后,保护酶活力随辐照剂量的增加呈先上升后下降的趋势,赤拟谷盗3种虫态POD活力均在500 Gy辐照剂量最高;3-4龄幼虫和1-2龄蛹CAT活力在420 Gy辐照剂量最高,羽化1 d成虫300 Gy最高;3-4龄幼虫和1-2龄蛹SOD活力在420 Gy辐照剂量最高,羽化1 d成虫500 Gy最高。本试验结果有助于阐明电子束辐照对储粮害虫的生理生化效应。