不同地区及不同季节的柞蚕寄生蝇RAPD分析

利用RAPD技术对不同地区及不同季节发生危害的柞蚕寄生蝇进行了遗传多态性研究。选取11个随机引物对14份供试材料的基因组DNA进行扩增,共扩增出140条稳定条带,其中128(91.43%)条为多态性条带,表明柞蚕饰腹寄蝇(Blepharipa tibialis)、秋柞蚕寄生蝇(未命名)及栗蚕寄生蝇(未命名)间具有丰富的多态性。各材料间的遗传距离在0.0214~0.5143之间,利用UPGMA法进行聚类分析,14份供试材料可聚为两大类:栗蚕(Dic-tyoplocajaponica)寄生蝇与寄生柞蚕的寄生蝇各聚为一类,其中来自河南省的柞蚕饰腹寄蝇自成一类,来自东北地区的柞蚕饰腹寄蝇与秋柞蚕寄生蝇又各自聚为一类。     

危害野蚕的4种寄生蝇的DNA条形编码鉴定

危害柞蚕(Antheraea pernyi)、栗蚕(Dictyoploca japonica)等野蚕的寄生蝇种类较多,准确鉴定其种类是害虫防治的基础。对采集自不同年份(2007年和2016年)、不同地理区域(辽宁、吉林和河南)寄生柞蚕和栗蚕的50份寄生蝇蛹样品的DNA条形编码进行了研究。来自辽宁6个地区和吉林2个地区的38份柞蚕寄生蝇样品属于柞蚕饰腹寄蝇(Blepharipa tibialis),它们的DNA条形编码序列的碱基一致性达100%,表明辽宁和吉林的柞蚕饰腹寄蝇均来源于一个扩散点,而且扩散历史极短。来自河南的柞蚕寄生蝇样品HN6的DNA条形编码序列与家蚕追寄蝇(Exorista sorbillans)的序列一致性达到96%,属于家蚕追寄蝇;而另外的5份样品HN1~5的序列一致性为100%,推测属于蚕饰腹寄蝇(Blepharipa zebina)。来自辽宁的寄生栗蚕的6份寄生蝇样品DJY1~6的DNA条形编码序列完全一致,但其分类学地位有待于进一步确认。有趣的是,来自河南同一个柞蚕茧内的寄生蝇样品HN3~5和HN6分属于2种寄生蝇。上述结果表明,利用DNA条形编码技术进行危害野蚕的寄生蝇种类鉴定和种群发生监测是可行的。     

灭蚕蝇一号对柞蚕的毒性及防治柞蚕寄蝇的效果试验

通过初步调查发现,河南省寄生危害柞蚕的寄蝇主要有蚕饰腹寄蝇和家蚕追寄蝇2种;同时开展了“灭蚕蝇一号”对柞蚕的毒性及防治柞蚕寄蝇的效果试验。结果表明：在柞蚕大眠起4～8d时,喷施0.107%浓度“灭蚕蝇一号”药液,可有效防治河南省蚕饰腹寄蝇病及家蚕追寄蝇病,防治效果达90%以上;该方法对柞蚕安全无毒害作用。     

柞蚕饰腹寄蝇发生规律及防治策略研究

研究柞蚕饰腹寄蝇病发生规律，有助于指导柞蚕生产上更合理的对该病害进行化学防治。在辽东地区，通过对柞蚕饰腹寄蝇羽化时期调查发现，该虫羽化时间主要集中在5月10日至5月15日之间，占比达到59.7%;通过柞蚕饰腹寄蝇寄生柞蚕时期调查发现，该虫对柞蚕的寄生时期主要集中在5月下旬到6月上中旬，占比达到87.46%。柞蚕饰腹防寄蝇化学防治试验，柞蚕老眠起约50%左右及进营茧场后2次施药的校正防效为91.7%±0.60%;分别在柞蚕三眠起约50%、老眠起约50%以及进营茧场后3次施药的校正防效为93.22%±0.64%;2次施药与3次施药差异不显著(P<0.05),结果表明通过2次施药即可有效防治柞蚕饰腹寄蝇病。     

吉林地区柞蚕饰腹寄蝇发生原因分析及防治对策

近年来柞蚕饰腹寄蝇在吉林地区的发生日趋严重,寄生率逐年上升,个别地区平均寄生率高达49.0%。导致寄蝇发生的主要原因是春蚕放养规模不断扩大,以及柞蚕饰腹寄蝇的自然迁飞。生产中可采用药杀柞蚕体内蝇蛆和妥善处理脱出的蝇蛆的方法控制其危害。     

柞蚕饰腹寄蝇的防治方法

柞蚕饰腹寄蝇也叫柞蚕寄生蝇,蚕民称蛆蛟、蚕蛆,属双翅目寄蝇科。近年柞蚕饰腹寄蝇在辽宁春柞蚕放养中发生较重,多时可寄生70%～90%,严重的影响了春种茧的质量和数量,给农民造成严重的经济损失。所以掌握蚕寄蝇的防治方法尤为重要。1用灭蚕蝇3号浸蚕杀蛆1.1看蚕寄生情况1.1.1在     

利用DNA条形编码探讨云南野柞蚕的分类学地位

2001年在云南曲靖发现的野生柞蚕(云南野柞蚕,A.pernyiwild)拥有一些与放养型柞蚕(A.pernyi)不同的特性。测定了云南野柞蚕线粒体细胞色素酶C亚基I基因5′端的部分片段(658 bp,GenBank:EU532613),并利用该DNA条形编码探讨其分类学地位。基于Kimura-2-Parameter计算的4个放养型柞蚕品种之间的平均遗传距离仅0.003,而云南野柞蚕与放养型柞蚕之间的遗传距离为0.016,小于已确定分类学地位的放养型柞蚕与印度野蚕(A.rolyii)之间的遗传距离(0.028),但与家蚕(B.mori)同其祖先中国野桑蚕(B.mandarinaChina)之间的遗传距离相近(0.015)。NJ树中云南野柞蚕与放养型柞蚕也最先聚在一起,从分子水平证实其仍属于柞蚕种。初步认为,云南野柞蚕可以考虑成为柞蚕种的一个亚种——野柞蚕亚种。     

在多雨年份和多雨地区如何利用灭蚕蝇1号防治柞蚕饰腹寄蝇

多雨年份和多雨地区,春柞蚕用灭蚕蝇1号防治柞蚕饰腹寄蝇。眷柞蚕4龄、5龄采取分别用0．05％与0．08％的灭蚕蝇1号两次喷药．防治柞蚕饰腹寄蝇效果好,成本低。     

柞蚕饰腹寄蝇不同地理种群的杀虫剂靶标酶和解毒酶活性分析

昆虫通过降低靶标酶敏感性及增强对杀虫剂的代谢能力产生抗药性。蝇毒磷是防治柞蚕饰腹寄蝇(Blepharipa tibia-lis Chao)的常用有机磷杀虫剂,为探讨柞蚕饰腹寄蝇不同地理种群对蝇毒磷的抗药性分化程度,分析来自辽宁省瓦房店市、凤城市、庄河市、开原市和西丰县的5个柞蚕饰腹寄蝇地理种群体内的乙酰胆碱酯酶(AChE)、谷胱甘肽S-转移酶(GST)、细胞色素P450(CYP450)和羧酸酯酶(CarE)4种酶的活性及蝇毒磷对这些酶活性的体外抑制率。结果表明:来自瓦房店市和凤城市的柞蚕饰腹寄蝇地理种群的AChE比活力及蝇毒磷对酶活性的体外抑制率均显著高于其它3个地理种群;瓦房店市地理种群的GST比活力显著高于其它4个地理种群,蝇毒磷对酶活性的体外抑制率显著高于除庄河市地理种群外的其它3个地理种群;庄河市地理种群的CYP450比活力显著高于来自瓦房店市、凤城市和西丰县的3个地理种群,但与开原市地理种群相比差异不显著,蝇毒磷对瓦房店市地理种群的CYP450活性的体外抑制率显著高于其它4个地理种群;瓦房店市与凤城市地理种群的CarE比活力均显著高于来自庄河市、开原市和西丰县的3个地理种群;蝇毒磷对瓦房店市和凤城市地理种群的CarE活性的体外抑制率显著高于庄河市地理种群,对其它地理种群间的CarE活性的体外抑制率差异不显著。根据以上检测结果,推测来自庄河市的柞蚕饰腹寄蝇地理种群对蝇毒磷的抗性相对较强,而来自瓦房店市的柞蚕饰腹寄蝇地理种群对蝇毒磷的抗性相对较弱。     

珍稀绢丝昆虫柳蚕的DNA条形编码与系统进化初步分析

利用DNA测序技术获得柳蚕(Actias selene)线粒体细胞色素酶C亚基Ⅰ基因(COI)5′端574 bp的片段,作为用于柳蚕种质资源分子鉴定的DNA条形编码(GenBank:FJ358505)。测定的柳蚕COI基因序列与其它大蚕蛾科绢丝昆虫的COI序列一样,均表现出偏好于碱基T的倾向(AT skew=-0.179)。在所分析的蚕类昆虫中,柳蚕与合目大蚕蛾的遗传距离最小(0.120),而与蓖麻蚕之间的遗传距离最大(0.138)。构建的NJ和UPGMA分子树中,大蚕蛾科的柳蚕属、柞蚕属、蓖麻蚕属、大乌桕蚕属、栗蚕属均各自形成一个支系,并且明显形成两个分支:大蚕蛾族(saturni-ini),包括柳蚕、柞蚕和栗蚕;眉纹大蚕蛾族(attacini),包括蓖麻蚕和大乌桕蚕。这一结果与传统分类一致。     

危害柞蚕的寄生蝇种类记述

寄生蝇是柞蚕的重要天敌昆虫之一。为帮助生产上识别及防治柞蚕寄生蝇,综合已有的研究报道及最新的研究结果,从分类地位、成虫形态特征、地理分布、寄主及发生危害等方面对柞蚕饰腹寄蝇[Blepharipa tibialis(Chao)]、蚕饰腹寄蝇[Blepharipa zebina(Walker)]、家蚕追寄蝇[Exorista sorbillans(Wiedemann)]、坎坦追寄蝇(Exorista cantans Mesnil)、札幌毛瓣寄蝇(Nemoraea sapporensis Kocha)和舞毒蛾克麻蝇[Kramerea schuetzei(Kramer)]共6种危害柞蚕的寄生蝇进行系统概述,其中,舞毒蛾克麻蝇为柞蚕天敌昆虫新记录种。     

辛伐他汀防治柞蚕饰腹寄蝇初报

为筛选防治柞蚕饰腹寄蝇新药剂,采用辛伐他汀片单用及其与"灭蚕蝇1号"混用喷蚕喷叶处理,调查药剂对柞蚕的影响和对柞蚕饰腹寄蝇的防治效果。初步试验结果表明,辛伐他汀单用或混用均未见对柞蚕产生不良影响,辛伐他汀片100mg·L-1与"灭蚕蝇1号"混用较"灭蚕蝇1号"单用防效提高10.1%。     

Molecular identification of polydorid polychaetes (Annelida: Spionidae): is there a quick way to identify pest and alien species?

The early detection and correct identification of polydorid polychaete species is essential as they are often encountered as invasive alien pests in aquaculture facilities or the intertidal where they may modify the ecosystem. Accurate identification is, however, often hampered by high levels of morphological similarity among species. This taxon will therefore benefit from the development of a library of sequences, such as COI barcodes, to aid identification. However, the universal primers for the cytochrome c oxidase I (COI) DNA barcoding marker has failed to consistently amplify this gene for polydorids, greatly hampering the development of such a library. We describe the development of unique PCR primers for the COI gene that work across four genera and nine species of polydorids. We also compared its efficacy with sequence data for mitochondrial cytochrome b and nuclear 18S rRNA, and a concatenated dataset consisting of all three markers. The nuclear 18S rRNA gene showed the least variation both intra- (0.0–1.2%) and interspecifically (0.6–4.3%), and was the most accurate for species identifications among the three markers. Although COI was characterised by higher intraspecific variation compared with Cyt b (0.0–14.5% and 0.0–4.2%, respectively), Cyt b showed considerably higher levels of interspecific variation (16.6–30.2%) compared with COI (2.2–20.7%). Of the two mitochondrial DNA markers, COI was actually less accurate for species identifications, having suggested two species within Boccardia pseudonatrix that was not supported by the other markers. Overall, the concatenated dataset yielded the most consistent intraspecific groupings, suggesting that this is the most accurate means of identifying polydorids using DNA sequence data. Thus, there may not be a quick and easy way to identify these species accurately using only molecular data.     

DNA条形码技术在野生哺乳动物物种鉴定中的可行性

为新疆野生哺乳动物物种鉴定找出最好的方法并建立新疆哺乳动物DNA条形码数据库提供资料,本研究选取新疆野生哺乳动物的肌肉、血液、粪便作为研究材料,运用DNA条形码技术检测野生哺乳动物线粒体DNA COI基因序列。共检测片段长度为642～729 bp的52条COI基因序列,52条COI基因序列通过BLAST网站同源性比较,序列同源性达到98%～100%,相似度达到90%～99%。全部COI基因序列中A、T、C和G的平均含量为26. 3%、30. 3%、26. 0%和17. 4%,A+T的含量(56. 6%)高于C+G含量(43. 4%),有明显的碱基偏倚性。种内遗传距离为0%～2%,种间遗传距离为15%～42%,二者分离度高。邻接法和最小进化法构建的系统发育树,两种方法所得的系统进化树拓扑结构高度一致,分支明显,都分配在单个支上。说明DNA条形码技术可用于新疆野生哺乳动物物种鉴定并弥补其他物种鉴定方法的不足和缺点,在新疆濒危野生动物的管理与保护中有重要意义。     

Evidence of multiple divergent mitochondrial lineages within the southern African diplopod genus Bicoxidens Attems, 1928 (Spirostreptida)

Two recent studies have suggested that divergent mitochondrial lineages may be present within spirostreptid genera such as Bicoxidens Attems, 1928. Bicoxidens, similar to many other endemic soil invertebrates, exhibits low dispersal capabilities and strict microclimate habitat preferences, attributes that often lead to geographic isolation. Given that prolonged geographic isolation often lays the foundation for population genetic differentiation, genetic divergence and possibly speciation, there was good reason to suspect that Bicoxidens may consist of several distinct lineages. On this basis, the mitochondrial cytochrome c oxidase subunit 1 (COI) was used to reconstruct the phylogeny of Bicoxidens and reveal divergent lineages within the genus. Maximum likelihood and Bayesian inference analyses recovered a paraphyletic Bicoxidens phylogram with divergent lineages present in three species – B. friendi, B. flavicollis and B. brincki – suggesting high genetic diversity within the genus. Bayesian genetic cluster analyses suggested the presence of multiple distinct mitochondrial lineages within the genus with four identified in B. flavicollis alone. It was therefore concluded that the divergent lineages observed among Bicoxidens populations may suggest the presence of hidden species.     

Molecular genetic comparison of Onchocerca sp. infecting dogs in Europe with other spirurid nematodes including Onchocerca lienalis

In the past 15 years, subconjunctival onchocercosis has been reported from 63 dogs in south-western United States (Arizona, California, Utah) and Southern and Central Europe (Germany, Greece, Hungary, Portugal, Switzerland). To reveal the taxonomic status of the parasite responsible for these infections, fragments of the mitochondrial cytochrome oxidase subunit I (COI) and NADH dehydrogenase subunit 5 (ND5) genes of three European strains of canine Onchocerca sp. and the 16S ribosomal RNA (16S rRNA) gene of their Wolbachia endosymbionts were sequenced and compared to the homologous sequences of other spirurid nematodes. The evolutionary divergence between COI and ND5 gene sequences of Greek, Hungarian and Portuguese strains of canine Onchocerca sp. were similar in magnitude to that seen within Thelazia callipaeda or Onchocerca lienalis. The evolutionary divergence between the sequences of canine Onchocerca sp. and other Onchocerca spp. including O. lienalis were similar or higher in magnitude to that seen between other Onchocerca spp. The results of the current and earlier phylogenetic analyses indicate that canine Onchocerca sp. separated from other Onchocerca spp. early in the evolution. Based on the similar clinical pictures, the identical morphology of nematodes and the sequence analyses of COI and ND5 genes of the worms and 16S rRNA gene of their wolbachiae, the Onchocerca worms isolated from European dogs appear to belong to the same species. The results support the earlier biological and morphological arguments that a distinct species, most likely O. lupi originally described from the subconjunctival tissues of a Caucasian wolf is responsible for canine ocular onchocercosis in Europe.     

Rapid diagnostic PCR assays for members of the Culicoides obsoletus and Culicoides pulicaris species complexes, implicated vectors of bluetongue virus in Europe

Biting midges of the Culicoides obsoletus Meigen and Culicoides pulicaris L. species complexes (Diptera: Ceratopogonidae) are increasingly implicated as vectors of bluetongue virus in Palearctic regions. However, predicting epidemiological risk and the spread of disease is hampered because whilst vector competence of Culicoides is expressed only in adult females, morphological identification of constituent species is only readily applicable to adult males and some species distinguishing traits have overlapping character states. Furthermore, adult males are typically rare in field collections, making characterisation of Culicoides communities impossible. Here we highlight the utility of mitochondrial cytochrome oxidase subunit I (COI) DNA sequences for taxonomic resolution and species identification of all species within C. obsoletus and C. pulicarus complexes. Culicoides were collected from 18 sites in the UK and Continental Europe, and identified to species level, or species complex level, based on morphological characters. The sample comprised four species from the C. obsoletus complex (n = 88) and five species from the C. pulicaris complex (n = 39). The DNA sequence of the 5' end of the COI gene was obtained from all individuals. Each member species formed a well-supported reciprocally monophyletic clade in a maximum likelihood phylogeny. Levels of DNA sequence divergence were sufficiently high between species to allow the design of species-specific PCR primers that can be used in PCR for identification of members of the C. pulicaris complex or in a multiplex PCR to identify members of the C. obsoletus complex. This approach provides a valuable diagnostic tool for monitoring species composition in mixed field collections of Culicoides.