南方部分地区烟粉虱及其寄生蜂内共生菌Cardinium的检测及其系统发育关系

【目的】研究我国南方四个省份6个烟粉虱Bemisia tabaci种群和2个蚜小蜂种群内共生菌Cardinium的感染率及其系统发育关系。【方法】利用PCR技术与DNA测序技术检测来自广州、肇庆、南宁、昆明、厦门烟粉虱种群以及广州2个蚜小蜂种群内Cardinium的感染情况,通过基于mt COⅠDNA序列和16S r RNA序列建立烟粉虱和Cardinium系统进化树,分析烟粉虱、寄生蜂和共生菌Cardinium的协同进化关系。【结果】广州的Asia II1与Asia II7烟粉虱种群、肇庆与南宁的MEAM1烟粉虱种群、昆明与厦门的Mediterranean烟粉虱种群以及广州的丽蚜小蜂Encarsia formosa、双斑蚜小蜂Encarsia bimaculata种群体内都有Cardinium的感染,且以Mediterranean与土著的Asia II1、Asia II7感染率较高(65.2%~92.5%),寄生蜂体内的感染率较低(16.7%~18.6%)。通过进化树对比分析发现,Cardinium与烟粉虱寄主之间无协同进化现象,但是Cardinium在烟粉虱不同地理种群之间,烟粉虱与寄生蜂之间的同源性超过98%。【结论】Cardinium在两种寄主昆虫之间进行相互感染与传播的概率较大,寄生蜂可能是Cardinium在不同烟粉虱种群之间水平传播的桥梁之一。     

烟粉虱及其优势寄生蜂内共生菌的种类及系统发育分析

为了研究入侵我国的2个主要烟粉虱隐种Bemisia tabaci MEAM1和MED及其3种优势寄生蜂(浅黄恩蚜小蜂Encarsia sophia、丽蚜小蜂E.formosa、海氏桨角蚜小蜂Eretmocerus hayati)体内感染内共生菌的种类丰度,并进一步探讨其系统发育关系,本文利用分子生物学手段对昆虫体内细菌的16S rRNA基因序列进行扩增、测序和分析,并采用邻接法(Neighbor-Joining,NJ)和最大似然法(MaximumLikehood,ML)分别构建优势内共生菌的系统发育树。结果表明,烟粉虱2个隐种内共生菌的种类丰度大于其3种优势寄生蜂,3种优势寄生蜂中丽蚜小蜂内共生菌的种类丰度最高;同源性分析发现烟粉虱和寄生蜂所携带的Rickettsia基因同源性达到99%,属于Rickettsia bellii种,进一步的进化树分析也发现所研究物种的Rickettsia和Hamiltonella均可各自聚为同一进化分支。烟粉虱及其优势寄生蜂体内含有种类丰富的内共生菌,其中优势内共生菌Rickettsia和Hamiltonella各自亲缘关系很近,说明内共生菌在烟粉虱和寄生蜂间可能进行水平传播。     

不同食物对烟粉虱三种寄生蜂取食寄生等行为的影响

【目的】就不同食物对烟粉虱Bemisia tabaci(Gennadius)寄生蜂取食、寄生等行为的影响进行分析,以阐明食物或营养来源对蚜小蜂取食、寄生等生物学行为的影响机制。【方法】以古桥桨角蚜小蜂Eretmocerus furuhashii、丽蚜小蜂Encarsia formosa和双斑恩蚜小蜂Encarsia bimaculata为研究对象,分别以蜂蜜水、2~3龄烟粉虱若虫及清水进行饲喂,观察和研究分析3种食物来源对3种蚜小蜂取食与寄生等行为的影响。【结果】3种蚜小蜂均以取食15%蜂蜜水的蚜小蜂的取食、搜索与寄生等行为频次最高、分配时间最长,而取食清水的蚜小蜂取食、搜索与寄生等行为频次最低、分配时间最短;取食不同食物后,3种蚜小蜂各种行为的变化趋势一致。【结论】不同食物或营养来源对蚜小蜂的取食及寄生等行为有着显著的影响,进而可以影响到蚜小蜂对烟粉虱的生物防治效率。     

基于SCAR标记技术的丽蚜小蜂快速识别

丽蚜小蜂Encarsia formosa Gahan作为温室粉虱Trialeurodes vaporariorum Westwood和烟粉虱Bemisia tabaci (Gennadius)等粉虱类害虫的优势寄生蜂而备受关注。针对丽蚜小蜂体型微小, 难以与其他同域近缘种寄生蜂快速、 准确区别的问题, 本研究采用SCAR (sequence characterized amplified region, 特异性扩增区域)标记技术, 筛选出一对丽蚜小蜂特征片段扩增引物(EFZZF/EFZZR), 其扩增片段的大小为287 bp。种特异性检验结果表明, 该对引物只对丽蚜小蜂的基因组DNA具有扩增能力, 对其近缘种属寄生蜂如浅黄恩蚜小蜂Encarsia sophia (Girault & Dodd)、 海氏桨角蚜小蜂Eretmocerus hayati Zolnerowich & Rose、 本地未知种桨角蚜小蜂Eretmocerus sp.、 蒙氏桨角蚜小蜂Eretmocerus mundus Mercet、 刺粉虱黑蜂Amitus hesperidum Silvertri不具有扩增效果, 对丽蚜小蜂的寄主包括不同生物型 (B型、 Q型、 ZHJ 1型和ZHJ 2型)的烟粉虱、 温室粉虱以及我国最常见的黑刺粉虱Aleurocanthus spiniferus (Quaintanca)等亦不具有扩增能力。同时, 该检测技术灵敏度高, 对成虫的最低检出阈值为7.812 ng/μL (相当于1/1 600头成虫)。研究结果对丽蚜小蜂的种类识别、 寄主谱的确定及其有效利用具有重要意义。     

宁波地区烟粉虱及其寄生蜂调查

烟粉虱Bemisia tabaci(Gennadius)是重要的世界性害虫之一,对农业生产造成了严重的危害.利用天敌对烟粉虱进行生物防治已经成为研究重点.本文于2007年6月至2008年11月对宁波地区不同寄主作物上烟粉虱虫口数量及其寄生蜂种类进行了调查.结果查得烟粉虱寄生蜂共7种,它们是恩蚜小蜂属Encarsm Forster的浅黄恩蚜小蜂En.sophia、日本恩蚜小蜂En.japonica、丽蚜小蜂En.formosa、黄盾恩蚜小蜂En.smithi和桨角蚜小蜂属Eretmocerus Haldeman的蒙氏桨角蚜小蜂Er.mundus、狄氏桨角蚜小蜂Er.debachi、盾黑桨角蚜小蜂Er.melanoscutus,其中优势种为浅黄恩蚜小蜂En.sophia、蒙氏桨角蚜小蜂Er.mundus和狄氏桨角蚜小蜂Er.debachi,这3种寄生蜂分别占收集到寄生蜂总数量的35.89%、49.66%和11.01%;此外,田间十字花科蔬菜上的烟粉虱虫13数量最高峰在2007年12月,自然寄生率最高峰在2008年10月,寄生率可达到13.86%;田间黄瓜上的烟粉虱虫口数量最高峰出现在2007年11月,自然寄生率最高峰在2007年8月,寄生率可达到8.22%;田间茄子上的烟粉虱虫口数量高峰出现在2007年7月和9月及2008年10月,自然寄生率高峰出现在2007年7月和9月及2008年9月,分别达到13.25%,10.30%和5.92%.     

烟粉虱两种寄生蜂生物学特性及寄主竞争关系研究

室内比较了寄生烟粉虱Bemisia tabaci的两种寄生蜂——丽蚜小蜂Encarsia formosa和浅黄恩蚜小蜂En. sophia的生物学特性,并研究了以烟粉虱为寄主时两种寄生蜂间的竞争关系。结果表明：两者的后足胫节长度间无显著差异。浅黄恩蚜小蜂的产卵器及卵均长于丽蚜小蜂的,但浅黄恩蚜小蜂的待产卵量显著少于丽蚜小蜂。浅黄恩蚜小蜂检测寄主所花时间短于丽蚜小蜂,但检查圈数显著多于丽蚜小蜂,产卵时间以丽蚜小蜂所花时间为多。以4龄初期的烟粉虱为寄主时,丽蚜小蜂的卵期为48 h,而浅黄恩蚜小蜂24 h即完成卵期的发育。后者的蛹历期也显著短于丽蚜小蜂的蛹历期。丽蚜小蜂由卵到成虫的总发育历期比浅黄恩蚜小蜂的要长72 h左右。一头丽蚜小蜂与一头浅黄恩蚜小蜂组合后的总产卵量为14.0粒,略高于两头丽蚜小蜂组合的总产卵量（10.2粒）,显著高于两头浅黄恩蚜小蜂组合的产卵量（9.5粒）;两种蜂组合的处理中被寄生寄主体内的着卵量为1.73粒,显著高于单独一种蜂组合中被寄生寄主的着卵量,后两者分别为1.29和1.39粒。在寄主竞争情况下,在被两种蜂均寄生的寄主体内丽蚜小蜂和浅黄恩蚜小蜂产入的卵量分别为1.21和1.43粒,显著高于仅被一种蜂寄生的寄主体内的对应蜂种的卵量,后者分别为1.06和1.19粒。结果提示两种蜂均能识别对方的存在,且浅黄恩蚜小蜂表现出更强的竞争能力。     

烟粉虱生物型对浅黄恩蚜小蜂寄主选择及个体发育的影响

为探讨寄生蜂在Q型烟粉虱Bemisia tabaci替代B型烟粉虱的过程中是否起作用, 我们在实验室条件(温度27±1℃, 光周期16L∶8D, 相对湿度RH 70%～80%)下, 观察了浅黄恩蚜小蜂Encarsia sophia寄生B型和Q型烟粉虱若虫的行为, 研究了浅黄恩蚜小蜂对B型和Q型烟粉虱若虫的选择性、 烟粉虱生物型对浅黄恩蚜小蜂取食数量及个体发育的影响。结果发现, 浅黄恩蚜小蜂体外检测时间在B型和Q型烟粉虱若虫间差异不显著, 而寄生Q型烟粉虱若虫时的体内检测和产卵时间(190.2±14.6 s）显著高于寄生B型时所用时间(140.0±7.5 s)。在非选择条件下, 浅黄恩蚜小蜂寄生B型烟粉虱若虫的数量(8.1±0.5头)及总产卵量（9.3±0.6粒）显著高于仅提供Q型烟粉虱的寄生数量(6.3±0.5头)及总产卵量(7.0±0.6粒); 而被寄生若虫单头着卵量在处理间差异不显著。在选择性条件下, 该蜂寄生B型烟粉虱若虫量(3.1±0.4头)、总产卵量(3.8±0.5粒)及被寄生若虫单头着卵量(1.2±0.1粒)都显著高于寄生Q型烟粉虱时的情况(1.8±0.3头、1.8±0.4粒、0.7±0.1粒)。被寄生蜂取食的B型与Q型烟粉虱数量间差异不显著, 但对于同一生物型而言, 交配过的雌蜂能够取食更多的烟粉虱若虫。以B型烟粉虱为寄主时, 浅黄恩蚜小蜂雌蜂卵-黑蛹(7.2±0.1 d)、黑蛹-羽化(5.2±0.1 d)的发育时间与以Q型烟粉虱若虫为寄主时的相应发育时间(7.3±0.1 d, 5.6±0.1 d)间无显著性差异。以B型烟粉虱为寄主时寄生蜂的羽化率（73.55%±1.42%）与以Q型烟粉虱为寄主时的羽化率（68.42%±13.01%）间差异不显著。这些结果表明, 虽然浅黄恩蚜小蜂发育时间、 羽化率在烟粉虱2种生物型间无显著差异, 但该小蜂倾向于B型烟粉虱若虫作为寄主, 而且, 以B型烟粉虱若虫为寄主时, 小蜂的产卵量和寄生若虫数量均增加。但田间浅黄恩蚜小蜂的存在是否有助于Q型烟粉虱成为B型和Q型混合种群的优势种群, 还需进一步研究。     

两种蚜小蜂对烟粉虱寄主的产卵选择性(英文)

双斑恩蚜小蜂和桨角蚜小蜂是华南地区烟粉虱的两种优势种寄生蜂。本文研究了两种寄生蜂对不同龄期烟粉虱寄主的产卵选择特性。结果表明:两种寄生蜂均可寄生烟粉虱的1~4龄若虫。当只有1个龄期的烟粉虱若虫存在时,双斑恩蚜小蜂更多地寄生3龄和4龄若虫,较少寄生1龄和2龄若虫;而桨角蚜小蜂则更多地寄生3龄和2龄若虫,较少寄生1龄和4龄若虫。在4个龄期若虫同时存在时,双斑恩蚜小蜂会明显增加对3龄和4龄若虫的产卵寄生,降低对1龄和2龄若虫的寄生;而桨角蚜小蜂则增加对2龄和3龄若虫的寄生,减少对1龄和4龄若虫的寄生。寄主植物的差异不影响这二种蚜小蜂对各龄期烟粉虱若虫的产卵选择倾向。结果提示,烟粉虱若虫3龄和4龄是双斑恩蚜小蜂最适宜的寄主,而桨角蚜小蜂最适宜的寄主是烟粉虱2龄和3龄若虫。     

两种蚜小蜂对烟粉虱寄主的产卵选择性（英文）

双斑恩蚜小蜂和桨角蚜小蜂是华南地区烟粉虱的两种优势种寄生蜂。本文研究了两种寄生蜂对不同龄期烟粉虱寄主的产卵选择特性。结果表明: 两种寄生蜂均可寄生烟粉虱的1～4龄若虫。当只有1个龄期的烟粉虱若虫存在时,双斑恩蚜小蜂更多地寄生3龄和4龄若虫,较少寄生1龄和2龄若虫;而桨角蚜小蜂则更多地寄生3龄和2龄若虫,较少寄生1龄和4龄若虫。在4个龄期若虫同时存在时,双斑恩蚜小蜂会明显增加对3龄和4龄若虫的产卵寄生,降低对1龄和2龄若虫的寄生;而桨角蚜小蜂则增加对2龄和3龄若虫的寄生,减少对1龄和4龄若虫的寄生。寄主植物的差异不影响这二种蚜小蜂对各龄期烟粉虱若虫的产卵选择倾向。结果提示,烟粉虱若虫3龄和4龄是双斑恩蚜小蜂最适宜的寄主,而桨角蚜小蜂最适宜的寄主是烟粉虱2龄和3龄若虫。     

寄主植物对双斑恩蚜小蜂的发育、存活和繁殖的影响

研究了茄子和扶桑两种不同寄主植物对烟粉虱寄生性天敌双斑恩蚜小蜂Encarsia bimaculata发育、存活和繁殖等特性的影响。在26±0.5℃的条件下,在扶桑上,双斑恩蚜小蜂卵至成虫的发育历期为13.6天,2龄幼虫至成虫的存活率为93.2%;而在茄子上双斑恩蚜小蜂卵至成虫的发育历期和2龄幼虫至成虫的存活率分别为12.1天和91.1％。扶桑上双斑恩蚜小蜂雌性成虫的平均寿命为8.0天,平均单雌产卵量为35.9粒;而在茄子上雌性成虫的平均寿命为6.6天,平均单雌产卵量为27.6粒。双斑恩蚜小蜂在扶桑和茄子植物上的种群内禀增长率分别为0.2081和0.892。两种不同寄主植物对双斑恩蚜小蜂在若虫发育历期和存活率、成虫寿命和平均产卵量以及内禀增长率等方面均存在着显著差异。研究结果表明,寄主植物对双斑恩蚜小蜂的发育、存活和繁殖有较大的影响。     

Interspecific host discrimination and within-host competition between Encarsia formosa and E. pergandiella (Hymenoptera: Aphelinidae), two endoparasitoids of whiteflies (Hemiptera: Aleyrodidae)

Interspecific host discrimination and within-host competition between Encarsia formosa Gahan and Encarsia pergandiella (Howard), two endoparasitoids of whiteflies, were studied under laboratory conditions. Interspecific host discrimination was studied at two time intervals (0 h and 72 h after the first species had oviposited). Parasitized and unparasitized Trialeurodes vaporariorum (Westwood) hosts were accepted for oviposition at the same rate by the two parasitoid species. Host type did not affect the handling time of the two parasitoids. The outcome of within-host competition was investigated after females of the two species parasitized the hosts at various time intervals. In four treatments, E. pergandiella was allowed to oviposit 0, 24, 48 and 72 h after E. formosa while in the other two, E. formosa was allowed to oviposit 0 and 72 h after E. pergandiella. In four of these treatments: E. formosa following E. pergandiella at 0 and 72 h, and E. pergandiella following E. formosa at 0 and 24 h, E. pergandiella prevailed. In the host discrimination experiment (72 h interval), 20% of E. pergandiella eggs were killed by E. formosa females. Interspecific ovicide was also observed in the within-host competition experiment, in which 6% of 72-h-old E. pergandiella eggs were killed by E. formosa females.     

Vitellogenin of the parasitoid wasp, Encarsia formosa (Hymenoptera: Aphelinidae): gene organization and differential use by members of the genus

The vitellogenin (Vg) gene of the parasitoid wasp, Encarsia formosa (Hymenoptera: Aphelinidae), has been cloned and sequenced. The gene codes for a protein consisting of 1814 amino acids in seven exons. The position of the six introns in the E. formosa gene align with those inferred for the Vg gene of the honeybee, Apis mellifera. The position of two introns in the hymenopteran sequences are shared with every full-length insect Vg gene characterized to date. The deduced amino acid sequence of the E. formosa Vg gene most closely resembles that of the ichneumonid parasitoid, Pimpla nipponica (38% identity). The gene product, less the putative signal peptide, contains large quantities of serine (11.3% of total residues) but lacks the extensive polyserine tracts found in the Vgs of insects outside the apocritan Hymenoptera. The gene also codes for the highest level of lysine (9.5%), and lowest levels of phenylalanine (2.6%) and tyrosine (2.3%), observed in any insect Vg characterized to date. The mature gene product retains 12 cysteine residues in positions conserved in other insect Vgs. Ovary homogenates suggest that processed Vg is stored in the egg as an uncleaved molecule of approximately 200 kDa. Vg expression was examined in three additional Encarsia species. The protein was found in female E. sophia and E. luteola, but not in male E. luteola or female E. pergandiella. Despite extensive screening of a phage library prepared from E. pergandiella genomic DNA, a Vg gene was not detected in this species.     

Comparison of searching strategies of five parasitoid species of Bemisia argentifolii Bellows and Perring (Hom., Aleyrodidae)

Amitus bennetti Viggiani and Evans, Encarsia formosa Gahan (two strains), Eretmocerus eremicus Rose and Zolnerowich, Eretmocerus mundus Mercet and Eretmocerus staufferi Rose and Zolnerowich, which are parasitoids of Bemisia argentifolii Bellows and Perring were compared with respect to their searching behaviour as part of a pre-introduction evaluation programme. Within a 5 cm arena, host-finding time was independent of the release distance from the host. Before oviposition, A. bennetti walked fastest, the E. formosa strains walked slowest and the Eretmocerus species intermediate. Leg length was not the most significant factor determining the differences in walking speed. After oviposition, A. bennetti and Er. eremicus had a lower walking speed and higher turn rate which is an indication of area-restricted search. The effect was strongest for A. bennetti . All species showed preference for counter-clockwise turns. Based on the walking speed alone, it is expected that A. bennetti will be the most efficient natural enemy of B. argentifolii , the Eretmocerus species intermediate and the E. formosa strains the least.     

A revision of the Encarsia pergandiella species complex (Hymenoptera: Aphelinidae) shows cryptic diversity in parasitoids of whitefly pests

Encarsia pergandiella Howard, described from North America (USA), and Encarsia tabacivora Viggiani, described from South America (Brazil) (Hymenoptera: Aphelinidae), are two formally recognized taxonomic entities, that have been treated by several authors as synonyms due to lack of strong diagnostic characters. Taxonomy of these species is further complicated because several populations, geographically separated and differing in their biology, have been included under the concept of E. pergandiella. Among these, a population originally collected in Brazil and introduced to North America reproduces by thelytokous parthenogenesis and is infected by the symbiont Cardinium, while a morphologically indistinguishable population, naturally occurring in Texas, is biparental and infected by a related strain of Cardinium that induces cytoplasmic incompatibility. A third population known from California and introduced to the Old World is biparental and uninfected by intracellular symbionts. While adult females of the first two populations have entirely light yellow bodies and pupate face up (light form), those of the third population have largely brown bodies and pupate face down (dark form). Other dark form populations are known from Texas, Florida and New York. Because these parasitoids are economically important biological control agents of cosmopolitan whitefly pests, it is critical to characterize them correctly. In this study, we integrated molecular and morphometric analyses to substantiate observed differences in biological traits, and resolve the complicated taxonomy of this species complex. We sequenced the mitochondrial cytochrome c oxidase subunit I gene and the D2 region of the ribosomal 28S gene for individuals of both light form (from Texas and Brazil) and dark form (from California, Texas, Italy and Canary Islands) originating from laboratory cultures or collected in the field. Phylogenetic analysis unambiguously distinguished three well‐supported groups corresponding to the Texas light form, the Brazil light form and the dark form. Individuals of these three groups, in combination with all available type material (E. pergandiella, its synonym Encarsia versicolor Girault and E. tabacivora) and additional museum specimens of the dark form from New York and Italy, were subjected to multivariate morphometric analyses using Burnaby principal component analysis followed by a linear discriminant analysis, and multivariate ratio analysis. Overall, the analyses showed that: (i) E. pergandiella and E. tabacivora are two distinct species; (ii) the thelytokous Brazil light form corresponds to E. tabacivora; (iii) the biparental Texas light form is a new species formally described here as Encarsia suzannae sp.n. ; (iv) two new biparental species can be referred to the dark form, one described as Encarsia gennaroi sp.n. including the populations sampled in California, Texas, Italy and Canary Islands, and the other corresponding to the population from New York described as Encarsia marthae sp.n. A dichotomous key for both sexes of the species of the E. pergandiella complex is provided for identification. This published work has been registered in ZooBank, http://zoobank.org/urn:lsid:zoobank.org:pub:278475A0‐C2C4‐4400‐A042‐A5716457829D .     

广东省烟粉虱蚜小蜂种类及种群动态调查初报

通过调查采集和鉴定 ,初步获得了广东省内烟粉虱蚜小蜂 6种 ,其中隶属于恩蚜小蜂属EncarsiaFoerster的有 4种 ,隶属于桨角蚜小蜂属ErtmocerusHaldeman有 2种。调查发现 ,蚜小蜂在蔬菜和园林植物上的寄生动态有一定的规律 ,4月中旬至 6月下旬和 9月下旬至 1 1月下旬寄生率较高 ,6月下旬至 8月上旬和 1 1月下旬至翌年 2月下旬寄生率较低。初步确定双斑恩蚜小蜂Encarsiabimaculata和桨角蚜小蜂Eretmocerussp .为广东省内烟粉虱的寄生蜂优势种类。     

Effects of UV-blocking films on the dispersal behavior of Encarsia formosa (Hymenoptera: Aphelinidae)

The parasitoid Encarsia formosa Gahan (Hymenoptera: Aphelinidae) has been used successfully for the control of Trialeurodes vaporariorum (Westwood) (Homoptera: Aleyrodidae). The development of UV-blocking plastic films has added a new component to future integrated pest management systems by disrupting insect pest infestation when UV light is excluded. Because both T. vaporariorum and E. formosa are reported to have similar spectral efficiency, there was a need to identify the impact of UV-blocking films on the dispersal behavior of both the pest and the natural enemy. In field studies, using choice-chamber experiments, E. formosa showed some preference to disperse into compartments where less UV light was blocked. However, further studies indicated that the effect was primarily attributable to the different light diffusion properties of the films tested. Thus, unlike its whitefly host, when the UV-absorbing properties of the films were similar, but the light diffusion properties differed, E. formosa adults preferred to disperse into compartments clad with films that had high light diffusion properties. When the plastic films differed most in their UV-absorbing capacity and had no light-diffusion capability, the initial dispersal of E. formosa between treatments was similar, although a small preference toward the environment with UV light was observed over time. When parasitoid dispersal was measured 3 h after release, more parasitoids were found on plants, suggesting that the parasitoids would search plants for whitefly hosts, even in a UV-blocked light environment. The potential for the integration of UV-blocking films with E. formosa in an advanced whitefly management system is discussed.