孟氏隐唇瓢虫和台毛艳瓢虫对茶椰圆蚧的捕食作用

本首次报道,孟氏隐唇瓢虫和志艳瓢虫是茶椰圆蚧的重要捕食性天敌。1998－1999年我们研究了这两种瓢虫的生活习性及其对茶椰圆蚧二龄若虫的捕食作用。这两种瓢虫成6虫对共椰圆蚧二龄若虫的功能反应均可用HollingⅡ型模型模拟。孟氏隐唇瓢虫的模型参数为：α'=1.1302,Th=0.0093,maxNa=107.6233;台毛艳主虫的模型参数为：α'=1.0005,Th=0.0126,maxNa=79.3741。温度对孟氏隐唇瓢虫捕食作用的影响大于对台毛艳瓢虫的影响,个体间相互干扰对捕食作用的影响,孟氏隐唇瓢虫大于台毛艳瓢虫孟氏隐唇瓢虫干扰反应模拟模型参数为：Q＝0.4045,m=0.4359;台毛艳瓢虫干扰反应模拟模型参数为：0.3335m=0.4042。     

孟氏隐唇瓢虫研究现状及其种质资源描述规范的建立

孟氏隐唇瓢虫Cryptolaemus montrouzieri Mulsant原产于澳大利亚,是粉蚧的重要捕食性天敌,作为生物防治重要的天敌资源被广泛引进世界各地.本文介绍了国内外近几十年来对孟氏隐唇瓢虫在生活史、行为、抗药性以及生物防治等方面的研究概况,并提出建立孟氏隐唇瓢虫种质资源描述规范的必要性,为充分发挥其控害潜能提供理论依据.     

孟氏隐唇瓢虫成虫自残幼虫的研究

在室内对孟氏隐唇瓢虫成虫自残幼虫的行为以及雌虫取食幼虫后的生殖力进行了研究,结果表明,雌成虫在48h内残杀幼虫造成的死亡率为60%,显著高于雄成虫;与其他部位相比幼虫的腹部遭受更多地攻击,幼虫体表覆盖的蜡丝在一定程度上抵御了成虫的攻击。与取食粉蚧的雌虫相比,取食瓢虫幼虫的雌虫产卵率和平均产卵量均显著降低。     

孟氏隐唇瓢虫的触角感受器

本文使用场发射扫描电子显微镜对孟氏隐唇瓢虫Cryptolaemus montrouzieri Mulsant雌雄成虫的触角和触角感受器进行了观察和研究，并对触角形态，感受器形态、类型、数量及分布进行了统计和分析。孟氏隐唇瓢虫雌雄成虫触角均由柄节，梗节及8个鞭小节组成。观察到7种触角感受器：四种锥形感受器（sba），四种刺形感受器（sch），四种毛形感受器（str），一种腔锥形感受器（sco），一种耳形感受器（sau），一种腔形感受器（scl），一种Bhm氏鬃毛（sbm）。毛形感受器和刺形感受器数量最多，其他类型感受器数量都较少。Bhm氏鬃毛只存在于柄节和梗节。第八鞭小节的顶端感受器种类最丰富，被六种感受器稠密覆盖。雌雄触角大小、感受器类型都没有明显差异。根据感受器的分布和以前的相关报道，推测孟氏隐唇瓢虫的毛形感受器可能是信息素接收者，刺形感受器可能是机械性刺激感受器和化学感受器，锥形感受器和腔锥形感受器可能都是植物挥发物接受者，化学和温湿度感受器，耳形感受器或许承担物理或者嗅觉器官的功能，而Bhm氏鬃毛或许能感知触角的位置和活动。     

孟氏隐唇瓢虫对产卵基质的颜色选择行为研究

在室内26±2℃,RH=50±10%,L:D=16h:8h环境下,使用白色、红色、蓝色、紫色、黑色5种棉花作为孟氏隐唇瓢虫Cryptolaemus montrouzieri Mulsant的产卵基质供其产卵,研究该瓢虫对产卵基质的颜色偏好性.结果表明连续10日内该瓢虫在红色棉花上的产卵次数和总产卵量最多,产卵次数与总产...     

温度对红点唇瓢虫实验种群的影响

在７种温度下测定了红点唇瓢虫（Ｃｈｉｌｏｃｏｒｕｓ ｋｕｗａｎａｅ Ｓｉｌｖｅｓｔｒｉ）发育速率,并求得各虫态的发育起点温度和有效积温。其发育速率与温度的关系能很好地用王如松等（１９８２）提出的模型进行拟合。由此模型估计出最低、最高临界温度和最适发育温度,分别为１０．４２－１３．０１－℃、℃３３．５３－３７．０３℃和２４．９９－３０．１２℃。卵期忍耐温度变化的能力最强。４龄幼虫最弱。温度明显地影响     

孟氏隐唇瓢虫生殖系统结构和卵子发生的研究

【目的】孟氏隐唇瓢虫Cryptolaemus montrouzieri Mulsant生殖系统结构和卵细胞发生将为昆虫的系统进化关系及瓢虫分类提供依据,同时可作为瓢虫人工饲料研究开发的参考。【方法】利用组织石蜡切片技术和光学显微镜,观察孟氏隐唇瓢虫生殖系统结构,以及自成虫羽化后不同发育阶段卵巢发育状况和成熟卵巢管卵子发生过程。【结果】孟氏隐唇瓢虫雄性生殖系统包括2对附腺、1对精巢、1对输精管、1对贮精囊、射精管、弯管和阳基。雌性生殖系统包括2片生殖板、生殖腔、受精囊、中输卵管、1对侧输卵管和1对卵巢。单侧卵巢管数量在11~14根之间,卵巢管端部延伸出细长的端丝。卵巢管属于端滋式,分为原卵区和生长区。滋养细胞分散且细胞核几乎充满整个细胞,未见合胞体。卵细胞稀疏地集中在原卵区下端,并且可见营养索向卵巢管顶端延伸。根据卵细胞位置和形态,卵黄积累情况,滤泡细胞形态变化,将卵细胞发生分为前期,中期,中后期和后期。卵细胞发育后期,营养索消失,滤泡细胞排列疏松,细胞间隙增大。【结论】孟氏隐唇瓢虫卵巢管的滋养细胞是端滋式卵巢管滋养细胞中的原始类型,且推测瓢虫科昆虫卵巢管滋养细胞均属于此类。卵细胞早期发育过程中,卵细胞通过营养索从滋养细胞获取营养物质。     

孟氏隐唇瓢虫细胞色素P450基因的克隆与序列分析

采用同源克隆和RACE技术,从孟氏隐唇瓢虫Cryptolaemus montrouzieri Mulsant中克隆到细胞色素P450CYP9Z401基因的c DNA全序列(Gen Bank number:KP164813)。c DNA全长1722 bp,包含3'非编码区域(UTR)为86 bp和5'UTR为49 bp,开放阅读框(ORF)长1587 bp,编码528个氨基酸。预测的分子量为61.27 k D,理论等电点为6.58,无信号肽结构,在2-20氨基酸处存在跨膜螺旋。该基因拥有细胞色素P450特征序列螺旋C、螺旋K和Meander区域,另外还有血红素结合区(1个氨基酸差异)和螺旋I区(2个氨基酸差异)。与其他昆虫的CYP9家族基因比较,与赤拟谷盗Tribolium castaneum同源性最高,为47%,系统发育分析也表明两者的亲缘关系最近。CYP9Z401基因的克隆和比较分析为进一步深入研究孟氏隐唇瓢虫的细胞色素P450基因功能及其进化具有重要意义。     

孟氏隐唇瓢虫qRT-PCR分析中内参基因的筛选

选择合适的内参基因是qRT-PCR研究的关键。本文以孟氏隐唇瓢虫Cryptolaemus montrouzieri Mulsant为研究材料,利用qRT-PCR技术,对孟氏隐唇瓢虫4个候选内参基因Actin、RPS23、GAPDH和β-tubulin的mRNA的表达量进行了分析,并用Ge Norm、Norm Finder和Best Keeper软件分析它们在孟氏隐唇瓢虫不同发育阶段及成虫不同组织中的表达稳定性。结果表明,以成虫不同组织为材料时,综合三种软件分析结果显示4个候选基因表达稳定性平均等级值排名为RPS23(rank=1)β-tubulin(rank=2.3)GAPDH(rank=3)Actin(rank=3.7),以不同发育时期虫体为材料时,综合分析结果显示4个候选内参基因表达稳定性平均等级值排名为RPS23(rank=1.7)Actin(rank=2)GAPDH(rank=2.7)β-tubulin(rank=3.7)。综合分析在瓢虫不同发育阶段及成虫不同组织两种处理下,三种软件的评价效果,4个候选基因表达稳定性等级值的总平均排名为RPS23(rank=1.3)Actin(rank=2.8)=GAPDH(rank=2.8)β-tubulin(rank=3)。RPS23在瓢虫不同发育阶段及成虫不同组织中均显示出较高的表达稳定性及与其它基因之间极大的相关性,可以确定为孟氏隐唇虫不同发育阶段及成虫不同组织基因表达分析中一个稳定表达的基因,可作为单个内参基因或者其它内参基因的协同基因,本实验为开展孟氏隐唇瓢虫功能基因表达分析奠定了方法学基础。     

First study of the biology of Cryptolaemus montrouzieri and its potential to feed on the mealybug Dactylopius opuntiae (Hemiptera: Dactylopiidae) under laboratory conditions in Morocco

Abstract

Dactylopius opuntiae, is known as specific Opuntia cochineal in many countries around the world. This sap-sucking insect was first detected in Morocco in 2014. To address the problem, the feeding potential of different development stages of Cryptolaemus montrouzieri Mulsant, a biological control agent against mealybugs, was investigated on different development stages of D. opuntiae. Fourth instar larvae and adults of C. montrouzieri were the most voracious feeders on different instars of the mealybug. The numbers of mealybug eggs consumed by first, second, third and fourth instar larva and adults of C. montrouzieri were 36.18?±?1.84, 68.08?±?4.17, 280.55?±?5.41, 540.55?±?5.35, 6514.13?±?64.28, respectively. The numbers of mealybug nymphs consumed by the same stages of C. montrouzieri were 35.43?±?1.75, 67.73?±?3.88, 279.85?±?5.58, 539.63?±?5.08 and 6501.7?±?81.94 (first instars) and 34.83?±?1.20, 57.45?±?1.22, 83.80?±?1.92, 213.65?±?3.46 and 6013.23?±?35.46 (second instars), respectively. The corresponding figures for adult female mealybugs were 1.40?±?0.78, 10.65?±?1.83, 18.58?±?1.24, 25.23?±?1.10 and 197.15?±?3.29, respectively. The egg, larval, prepupal, pupal and adult stages occupied 3.36–3.69, 20.21–27.59, 1.31–1.59, 10.62–10.72 and 96.10–102.51?days, respectively when the coccinellid was reared on different stages of D. opuntiae. The results indicate that C. montrouzieri has the potential to be used as a biocontrol agent in Morocco.     

孟氏隐唇瓢虫气味结合蛋白ComOBP1基因的克隆和时空表达

采用RT-PCR和RACE技术,从孟氏隐唇瓢虫Cryptolaemus montrouzieri Mulsant中成功克隆出气味结合蛋白(Com OBP1)基因的全序列(Genbank登陆号:KU170686)。Com OBP1基因全长922 bp,包括5'端长为40 bp的非编码区域(UTR),及3'端长为462 bp的UTR,开放阅读框ORF长为420 bp,编码139个氨基酸,预测的分子量为15.516 k Da,等电点p I为6.57,存在AATAAA加尾信号。N-末端疏水区包含由20个氨基酸构成的信号肽,无跨膜结构,有4个保守的半胱氨酸,属于Minus-C OBP,也是在孟氏隐唇瓢虫中发现的第一个Minus-C OBP。氨基酸序列中有且仅有一个N-糖基化位点为62 NLSA,并存在2个潜在的磷酸化位点。与其它昆虫的Minus-C OBPs进行同源性比较并构建系统发育树,发现与同为鞘翅目Coleoptera昆虫的同源性较高。利用Real-time PCR、RT-PCR技术对Cmon OBP1基因在孟氏隐唇瓢虫不同发育阶段,不同组织,不同营养条件及不同食性下的表达水平进行了测定,结果显示,Cmon OBP1基因在整个发育阶段均有表达,雄性成虫期具有最高表达量,且多在成虫的头部及翅部表达。当营养条件发生变化时,表达丰度不会发生变化,当猎物由天然猎物柑橘粉蚧Planococcus citri Risso变成碗豆修尾蚜Megoura japonica Matsumura时,表达量会明显下降。该结果表明,孟氏隐唇瓢虫的不同发育阶段、不同组织及猎物种类会影响Cmon OBP1基因的表达,从而进一步影响其嗅觉行为。同时,Cmon OBP1基因可能在雄虫相关的信息素感受过程中发挥着重要作用。     

Effects of three mealybug species on the development,survivorship and reproduction of the predatory lady beetle Cryptolaemus montrouzieri Mulsant

The lady beetle Cryptolaemus montrouzieri Mulsant (Coleoptera: Coccinellidae) is an important predator of mealybugs. The development, survivorship, longevity and reproduction of C. montrouzieri feeding on three different mealybug species [Dysmicoccus neobrevipes Beardsley, Ferrisia virgata Cockerell and Planococcus minor (Maskell)] were investigated in the laboratory at 26 ± 1°C, 75-–90% RH and 14:10 (L:D) h photoperiod. Results indicated that, when feeding on different mealybugs, no significant differences were observed between developmental periods and survivorship of C. montrouzieri (from egg to adult), but differences were recorded between the sex ratios, preovipositional periods, adult longevities and reproduction of the differently treated lady beetle populations. The highest sex ratio (0.56), the longest preovipositional period (6.6 days) and adult longevity (84.8 days for females and 93.9 days for males), and the maximum fecundity (659.0 eggs/female) of C. montrouzieri were recorded when feeding on F. virgata. Moreover, C. montrouzieri had a high net reproductive rate (313.66), intrinsic rate of increase (0.0816) and finite rate of increase (1.085) when feeding on F. virgata. Results indicated that the population growth of C. montrouzieri may increase faster when feeding on F. virgata than feeding on either of the other two mealybugs.     

温度对豆荚野螟实验种群的影响

作为豆科蔬菜上的重要害虫 ,豆荚野螟 Maruca testulalis Geyer能够对豆科蔬菜造成严重危害。研究温度对豆荚野螟不同虫期发育的影响 ,可以有利掌握其种群动态 ,以便进行有效控制。研究了室内不同温度条件下豆荚野螟的生长发育情况 ,发现在2 0～ 32℃温度范围内 ,随温度上升发育历期缩短 ;对于不同的生活史阶段 ,豆荚野螟卵、1～ 4龄幼虫、蛹及整个世代受温度影响显著 ;并计算得出完成全世代发育起点温度为 7.84℃ ,有效积温达 4 74 .2 0℃。通过内禀增长率参数的比较 ,认为各参数与温度的关系均达到极显著相关 ,以 2 9～ 32℃为豆荚野螟最适温度     

寄主转换对烟蚜种群特征的影响

试验结果表明：除从油菜上转到桃树上不成功外,烟蚜可在昌,油菜和桃树之间的成功转换,且同寄主之间的转换比异寄主之间的转换生殖前期短,产仔量大。烟蚜寄主转换以ｒｍ值的大小依次为桃树转油菜－烟草转油菜－油菜转烟草－桃树转烟草－烟草转桃树。     

Comparative toxicity of different chemical and biological insecticides against the scale insect Dactylopius opuntiae and their side effects on the predator Cryptolaemus montrouzieri

The contact toxicity of various chemical and biological pesticides for the first and second instar nymphs and adults of the Opuntia cochineal scale insect Dactylopius opuntiae and the predator ladybird Cryptolaemus montrouzieri was determined under Morocco semi field conditions. d-limonene (60?g/l) at 100 and 150?cc/hl, mineral oil (780?g/l) at 2400?cc/hl and malathion (500?g/l) at 300?cc/hl caused the highest mortality (99–100%) among first instar nymphs of D. opuntiae 24?h after treatment. d-limonene (60?g/l) at 150?cc/hl caused greatest mortality (99%) in second instar nymphs. The highest mortality (99%) among adult female D. opuntiae was observed 120?h after treatment with d-limonene (60?g/l) at 150?cc/hl and mineral oil (780?g/l) at 2400?cc/hl. For the predator C. montrouzieri the highest mortality (92–97%) among adults 24?h after treatment was caused by malathion (500?g/l) at 100, 200 and 300?cc/hl and alpha-cypermethrin (100?g/l) at 75, 150 and 225?cc/hl. The most harmful pesticides to C. montrouzieri larvae 24?h after treatment were malathion and alpha-cypermethrin with mortality rates of 89–95%. Mortality in larvae ranged from 87 to 100% 120?h after treatment with chlorpyriphos-methyl (480?g/l) at 75, 150 and 225?cc/hl and spinosad (480?g/l) at 100, 200 and 300?cc/hl. d-limonene (60?g/l) at 50?cc/hl and mineral oil (780?g/l) at 1000?cc/hl had the least impact on C. montrouzieri adults and larvae, causing mortality of 11 and 15%, respectively, 120?h after treatment. d-limonene (60?g/l) and mineral oil (780?g/l) may therefore be viable alternatives to others high-risk chemical pesticides. These two biological insecticides are effective in controlling the Opuntia cochineal scale insect but have little adverse impact on the predator C. montrouzieri.