Wolbachia在螟黄赤眼蜂种群内的分布及其遗传稳定性

本文通过对Wolbachia的wsp及16S rDNA基因的PCR扩增及克隆测序,明确了Wolbachia在华南、华北不同作物田间螟黄赤眼蜂Trichogramma chilonis种群内的分布情况.结果表明,在所有检测的螟黄赤眼蜂个体内都存在着Kue和Pip两组Wolbachia,双重感染率达到了100%.在不同温度下用米蛾Corcyra cephalonica Stainton卵饲养多代后,螟黄赤眼蜂体内的Wolbachia类型及感染率未发生改变,表明Wolbachia能够在螟黄赤眼蜂体内稳定遗传,不受环境温度的直接影响.本研究中螟黄赤眼蜂为两性生殖或孤雌产雄生殖,Wolbachia未能引起赤眼蜂的产雌孤雌生殖.首次报道了Wolbachia在我国螟黄赤眼蜂野生种群内的分布情况及其遗传稳定性.     

松毛虫赤眼蜂和螟黄赤眼蜂羧酸酯酶性质比较研究

该文对松毛虫赤眼蜂Trichogramma dendrolimi和螟黄赤眼蜂T.Chilonis羧酸酯酶的毒理学和生物化学性质进行了初步研究。由时间进程曲线确定了赤眼蜂羧酸酯酶活性测定中的最适反应时间为40min。对两种赤眼蜂单头羧酸酯酶活性的测定表明,松毛虫赤眼蜂羧酸酯酶活性主要分布在0—1OD/(mg·min)之间,而螟黄赤眼蜂羧酸酯酶活性主要分布在1-4OD/(mg·min)之间。比较两种赤眼蜂羧酸酯酶米氏常数(K_m)值,螟黄赤眼蜂羧酸酯酶对底物的亲和力比松毛虫赤眼蜂羧酸酯酶对底物的亲和力高。对氧磷对螟黄赤眼蜂和松毛虫赤眼蜂羧酸酯酶的抑制作用没有明显差异,而对氧磷对松毛虫赤眼蜂羧酸酯酶的抑制作用明显强于磷酸三苯酯(TPP)。该文还将棉铃虫Helicoverpa armigera (Hubner)羧酸酯酶部分性质与两种赤眼蜂作了比较。     

螟黄赤眼蜂的个体发育

螟黄赤眼蜂广泛应用于多种农林害虫的生物防治,常用米蛾卵进行规模化繁育。本文观察了螟黄赤眼蜂在米蛾卵中的个体发育过程。结果发现,在26℃±1℃的条件下,螟黄赤眼蜂从产卵到成虫的个体发育过程历时8 d,其中胚胎期26 h,幼虫期36 h,预蛹期48 h,蛹期84 h,在产卵后的192 h后成虫开始羽化。螟黄赤眼蜂卵为白色透明的棒状,随着胚胎发育时间的增加,其长径和短径逐渐变大,但两者之比逐渐减少;刚进入幼虫期的虫体长径与短径之比较大,虫体上下宽度基本相等,但体积随取食而快速增加;虫体出现梅花斑和米蛾卵卵壳颜色变黑是进入预蛹期的标志;蛹的头、胸和腹部明显分界,梅花斑消失,腹部二条黑带逐渐横贯整个腹背面,复眼随着时间的增加逐渐加深,最后为深红色。经过192 h的发育,螟黄赤眼蜂成虫羽化,咬破米蛾卵壳而出。     

诊断引物应用于我国三种重要赤眼蜂分子鉴定的研究

本研究根据螟黄赤眼蜂rDNA-ITS2序列设计了螟黄赤眼蜂Trichogramma chilonis Ishii特异引物,同时采用文献中发表的松毛虫赤眼蜂Trichogramma dendrolimi Matsumura 和玉米螟赤眼蜂Trichogramma ostriniae Pang et Chen的特异引物以及赤眼蜂属Trichogramma 特异引物对赤眼蜂成虫和寄主卵样品进行了PCR特异扩增分析。结果表明,采用上述特异引物可从单头蜂稳定地扩增出明显的目的DNA条带,并且分子鉴定结果与形态学鉴定结果完全吻合。因此,采用上述3对特异引物可以实现对我国3种重要赤眼蜂种,即松毛虫赤眼蜂、螟黄赤眼蜂和玉米螟赤眼蜂的分子鉴定。     

基于诊断引物的赤眼蜂鉴定和检测技术

通过对松毛虫赤眼蜂Tichogramma dendrolimi Matsumura和玉米螟赤眼蜂T.ostriniae Pang et Chen（膜翅目：赤眼蜂科）核内可转录第二间隔区（简称：ITS2）的克隆、测序,并获取和分析了GenBank中已登录的同源序列,然后设计了松毛虫赤蜂的特异引物以用于该蜂的分子鉴定和检测,经过反复筛选发现：采用鉴定引物通过PCR扩增不仅可以区分鑫头样品,单头样品（雌蜂或雄峰）,而且可鉴定幼期虫和卵,这用传统方法是无法办到的。该鉴定技术比基于形态学鉴定检测技术用来鉴定了从中国大陆不同地域和寄主上采集到的12个样品,结果表明：该方法可用于赤眼蜂田间分子监测和实验室拟寄生行为研究。     

Wolbachia在我国不同地理种群的玉米螟赤眼蜂中的分布

Wolbachia是一类可以引起寄主生殖行为改变的共生菌,广泛存在于节肢动物内。玉米螟赤眼蜂Trichogramma ostriniae Pang et Chen是我国亚洲玉米螟Ostrinia furnacalis(Guene)卵寄生蜂的优势种。本文利用Wolbachia的16SrDNA基因特异性引物进行PCR扩增以及克隆测序,对我国14个地区的玉米螟赤眼蜂种群中Wolbachia的感染情况做了详细的调查。结果发现,被测的1412头野生玉米螟赤眼蜂全部感染有Wolbachia。系统发育分析表明,北京柳沟、山东菏泽、河南郑州、河南驻马店、湖南长沙5个种群存在超感染现象。这是Wolbachia在玉米螟赤眼蜂种群内分布情况的首次报道。     

不同地理飞蝗种群的遗传多样性

用GenBank中飞蝗Locusta migratoriaL.的序列来设计微卫星引物,并对这些引物的有效性进行验证。结果表明,在所设计的3对引物中,只有1对为有效引物,可扩增出微卫星位点。序列分析表明本位点是一个不连续的重复微卫星位点。该多态微卫星位点含有14个等位基因,不同飞蝗地理种群的等位基因数目、大小和频率都存在较大的差异。对该位点各等位基因型进行χ2检验,基因型频率的观察和期望杂合度分别为0.4578和0.8836,该位点不属于Hardy-Weinberg平衡位点(χ2=733.12,P=0.0000)。该微卫星位点表现出高度的多态性说明是分析飞蝗种群遗传多样性的优良分子遗传标记。     

植物不同种属间共用微卫星引物的研究

微卫星标记是近年来发展起来的建立在PCR基础卜的第二代分子标记,具有分布均匀、多态性高、共显性、选择中性等优点,已用于品种鉴定、构建连锁图谱、构建物理图谱、遗传多样性、居群和进化等方面的研究。由于来源于数据库的微卫星序列不能代表整个基因组微卫星的分布情况,而且数据库中的DNA序列仅局限于一些模式植物及经济作物。同时,通过基因组文库筛选获得微卫星序列既耗时又耗资。最近有很多研究表明,从某种植物来的微卫星引物可用于其近缘物种的扩增。本文概括介绍了植物不同种属间共用微卫星引物的研究情况及意义。     

大螟不同地理种群的AFLP遗传分析

为了明确我国大螟Sesamia inferens Walker不同地理种群的遗传多样性水平,利用AFLP方法对我国长江流域和南方稻区大螟的10个地理种群进行了遗传多样性分析,结果表明不同地理种群间具有丰富的遗传多样性,利用27对引物共扩增到1 055条条带,其中837条条带具有遗传多态性,多态性比例为78.75%;种群间分化系数(Gst)为0.7132;Nei’s基因多样性(h)范围在0.2969⁰.4547之间,平均为0.3752;Shannon信息指数介于0.29360.4547之间,平均为0.3752;Shannon信息指数介于0.2936⁰.5043之间,平均为0.3888;10个种群间遗传一致度较高,均大于0.63。遗传距离聚类分析表明,大螟10个地理种群可区分为4类,德阳,龙海,湛江各为一类,其余7个地理种群聚为一类。研究还表明,与其他测试种群的遗传距离普遍较大、相对孤立的地理种群,其聚类结果有一定的随机性,因此,取样点较多、且样点分布较均匀的聚类分析结果才有较高可信度。     

亚洲玉米螟不同地理种群基于COⅠ基因序列初步分析

本研究通过对我国亚洲玉米螟Ostrinia furnacalis(Guenée)6个地理种群的线粒体DNACOⅠ基因进行测序,运用分子进化遗传分析软件MEGA5对序列进行对比和同源性分析,并建立系统发育关系。结果表明,COⅠ基因在6个地理种群中进化具有一定的差异,平均遗传距离为0.013;进化树显示,亚洲玉米螟的三亚种群、广州种群与南昌种群聚为一支,然后与泰安种群聚为一支,再与哈尔滨种群聚为一支;而廊坊种群另聚成一支。遗传分歧度与系统发育进化关系均表明不同地理种群的遗传分化与地理距离之间具有一定的相关性。     

Dispersal of Trichogramma ostriniae in field corn

Trichogramma ostriniae has shown success as a biological control agent for European corn borer (Ostrinia nubilalis) in sweet corn and the species offers potential for suppression of lepidopteran pests of field corn. Field corn is typically planted at higher densities, is taller, and has greater leaf area than sweet corn, presenting a possible restriction on T. ostriniae dispersal and efficacy. Therefore, parasitoid dispersal in field corn from the centre of a 6.25 ha square grid was determined using sticky cards to capture adult T. ostriniae and sentinel eggs of O. nubilalis to monitor parasitism after releases of ~1 million of T. ostriniae each into four fields of corn. Dispersal was rapid and extensive, achieving distances of ~175 m within 4–7 days after release. The pattern of movement fit well with a diffusion model of dispersal, with the greatest level of dispersal occurring from 7 to 10 days post-release. Parasitism of O. nubilalis sentinel egg masses declined linearly with distance from the release foci, and was also greatest 7–10 days post-release. However, measurement of association showed no significant differences between the spatial distributions of sticky trap captures and percentage parasitism of O. nubilalis egg masses. The distances from the release point that encompassed 98% of re-captured T. ostriniae increased over time and were estimated to range from a low of 100 m at 4 days post-release to 365 m at 14 days post-release. The results of this research suggest that T. ostriniae relies initially on random movement to locate host patches, and that a single release locus per hectare would be sufficient in field corn.     

高温对玉米螟赤眼蜂实验种群的影响

以米蛾卵为繁育寄主,组建了玉米螟赤眼蜂在26、29、32和35 ℃下的实验种群生命表,探讨了高温对玉米螟赤眼蜂生长和生殖的影响.结果表明,高温32 ℃～35 ℃对玉米螟赤眼蜂各虫态的生长发育有明显的抑制作用.卵 幼虫的存活率在26 ℃和29 ℃分别为 94.2%和88.6%,32 ℃时下降至80.2%,而35 ℃时仅为53.2%,蛹的存活率也表现出相应的下降趋势.玉米螟赤眼蜂羽化后12 h内的抱卵量、体长及卵粒大小随温度的升高而减少或变小.26 ℃时,玉米螟赤眼蜂羽化后0～12 h的抱卵量为37.9粒,随着温度的升高,抱卵量急剧下降,29 ℃、32 ℃和35 ℃的抱卵量分别为29.6、21.6和13.7粒.在26 ℃及29 ℃时,所有雌蜂均能进行正常交配和产卵,32 ℃时有6.7%的个体不产卵,至35 ℃时,则有60%的个体不能进行正常的生殖活动.     

9个水稻栽培种和1个水稻组培种对稻螟赤眼蜂和赤眼卵蜂的互利作用（英文）

采用培养皿法对营养生长期和开花期的9个水稻栽培种（Pusa Sugandh-2, Pusa Basmati-1, Pusa-2511, Pusa Basmati-370, Pusa-1077, Karnal Local, PRR-78, Jaya, Pusa-1238）和1个组培种（Culture No.34）的正己烷提取物进行了生测,以调查它们对稻螟赤眼蜂Trichogramma japonicum Ashmead和赤眼卵蜂Trichogramma chilonis (Ishii)平均寄生活性指数（PAI）和平均寄生百分率的影响。将不同水稻品种的正己烷提取物进行气相 液相色谱,来测定饱和碳氢化合物。其中,营养生长期的Pusa Sugandh-2叶片提取物激发两种寄生蜂产生最大的反应,平均寄生率最大。而开花期的Pusa Basmati-1使两种寄生蜂的平均寄生率最高。对营养生长阶段的叶片提取物进行的气相 液相色谱分析表明：Pusa Sugandh-2 含有25个碳原子(C25) 和29个碳原子 (C29)的2种化合物。对开花期的叶片提取物进行的气相 液相色谱分析也表明：Basmati-1 含有25个碳原子(C25)、26个碳原子(C26)和29个碳原子 (C29)的3种化合物,可激发寄生蜂产生最大的反应     

Overwintering of Trichogramma ostriniae (Hymenoptera: Trichogrammatidae) within target and non-target host eggs

Trichogramma ostriniae was imported into the USA from China and it continues to be evaluated as a biological control agent against the European corn borer and other lepidopteran pest species. A natural enemy's ability to overwinter is a facet of its biology with important ramifications for biological control and non-target effects. Thus, studies were conducted to examine the ability of the introduced egg parasitoid to survive over winters in central New York State. Eggs of Ostrinia nubilalis, Ephestia kuehniella, Trichoplusia ni, Helicoverpa zea and Utetheisa ornatrix were subjected to parasitism by adult T. ostriniae and then placed out of doors and exposed to winter conditions. For trials initiated in 2003 and 2004, the adult parental wasps were exposed to a diapause-inducing photoperiod and temperature regime in the laboratory; in 2010, parental wasps were conditioned out of doors and prior to the onset of winter conditions. Emergence of their progeny was monitored over time by taking aliquots of parasitised eggs, and holding them under warm conditions until emergence was complete. The level of wasp emergence generally displayed a decline followed by gradual increase until spring. Levels of overwintering ranged from 1% for O. nubilalis to 76% for E. kuehniella, and logistic regression indicated that the odds of overwintering was dependent on the year, host species, time out of doors and varied over exposure time depending on host. The potential to overwinter in New York was further confirmed by positive identification of T. ostriniae from naturally occurring O. nubilalis eggs collected from field sites where augmentative releases had been made in previous years.     

玉米螟赤眼蜂在甜玉米地寄生率和扩散规律的研究

调查研究了玉米螟赤眼蜂Trichogramma ostriniae Pang et Chen在甜玉米地的年消长动态、日活动规律,以及对不同甜玉米品种和不同防治类型地亚洲玉米螟Ostrinia furnacalis (Guenee)卵的寄生率。玉米螟赤眼蜂田间扩散规律为：在苗期,按每点2000头的蜂量相距32m左右释放玉米螟赤眼蜂,即可达到50％以上的寄生率;在穗期,按每点2000头的蜂量相距24m左右释放玉米螟赤眼蜂,可达到80％以上的寄生率。     

温度对麦蛾卵繁育的玉米螟赤眼蜂发育和生殖的影响

玉米螟赤眼蜂Trichogramma ostriniae Pang et Chen是玉米螟的重要卵寄生性天敌,控制适宜的温度有利于室内大规模繁育。本试验以麦蛾卵为寄主,研究了不同温度(18℃、20℃、23℃、25℃、28℃、30℃、33℃、36℃)对玉米螟赤眼蜂的寿命、生殖力、子代羽化率和发育历期的影响。结果表明,20℃-25℃条件下雌蜂寿命最长,达7.9-9.0 d。在18℃-30℃亲代生殖力和子代羽化率无显著差异,亲代产卵量为40.9-61.7粒;羽化率为74.31%-84.22%。玉米螟赤眼蜂从卵到羽化的发育历期随温度的升高而显著缩短,由18℃时的22.6 d缩短至33℃的7.0 d。36℃时,雌蜂的寿命仅为0.56 d,且不能进行正常的生殖活动。因此,室内用麦蛾卵大规模繁育玉米螟赤眼蜂,适宜的温度为28℃-30℃;仅保存蜂种时,可选用温度18℃-20℃。     

Trichogramma ostriniae (Hymenoptera: Trichogrammatidae) response to relative humidity with and without host cues

Trichogramma ostriniae is a parasitoid wasp species that is used as a biological control for several lepidopteran agricultural pests. Our study examined the effects of relative humidity (RH) on the behaviour of female T. ostriniae. In Y-tube RH choice assays, wasps generally chose higher RH over lower RH but the latency time taken to make a choice displayed no clear patterns that could be ascribed to the magnitude of the RH difference between arms of the Y-tube. In trials conducted in glass arenas with fixed humidities, the conditional probabilities of transition from searching to interacting with the leaf disk, and from interacting with leaf disks to antennal egg drumming and oviposition, were not significantly different among the RH levels, nor were there differences in latency times. These findings suggest that the level of RH may influence habitat choice but have little effect on search efficacy.     

挥发性信息化合物对玉米螟赤眼蜂寄主选择行为的影响

在实验室条件下,利用“Y”型嗅觉仪测定了源于寄主亚洲玉米螟Ostrinia furnacalis (Guenée) 鳞片、卵表以及不同生理阶段的雌蛾附腺的不同浓度的提取物对玉米螟赤眼蜂Trichogramma ostriniae Pang et Chen寄主选择行为的影响,并对其引诱作用大小进行了比较。结果表明： 亚洲玉米螟鳞片的正己烷提取物在0.5 mg/mL和 1 mg/mL浓度时对玉米螟赤眼蜂有明显的引诱作用; 寄主卵的正己烷提取物在5块卵/mL和10块卵/mL浓度时对玉米螟赤眼蜂有显著的引诱作用,而在40块卵/mL浓度时对玉米螟赤眼蜂有极显著的驱避作用; 玉米螟赤眼蜂对亚洲玉米螟交配未产卵和产卵后前期的雌蛾附腺提取液有反应,而对处女蛾和产卵后期的附腺提取液没反应。卵表提取物和附腺提取物比鳞片提取液对赤眼蜂引诱作用强,两者对赤眼蜂的引诱作用没有明显差异。     

Wolbachia在玉米螟赤眼蜂内的三重感染

Wolbachia是一类广泛存在于节肢动物体内的共生菌。玉米螟赤眼蜂Trichogramma ostriniae是我国玉米田间的优势赤眼蜂种, 据报道, 赤眼蜂种内有Wolbachia感染。本文利用Wolbachia的16s rDNA和wsp基因引物通过PCR方法对玉米螟赤眼蜂的野生种群进行了调查, 发现以wsp基因为鉴定依据, 检测的所有个体都感染了3种Wolbachia ［wOstGDAa (GenBank accession no. EU157103), wOstGDAb (GenBank accession no. EU157104) 和 wOstGDB (GenBank accession no. EU157105)］。本文首次报道了野生赤眼蜂种群内Wolbachia的三重感染率几乎为100%。根据本研究的结果, 可以推测当不同种赤眼蜂寄生同一寄主时, Wolbachia可能会在不同赤眼蜂种间进行横向传播。