Wolbachia在Drosophila auraria复合种和Drosophila simul …

用ＰＣＲ方法检测了多种果蝇中共生菌Ｗｏｂａｃｈｉａ的感染，发现，Ｄｒｏｓｏｐｈｉｌａ ａｕｒａｒｉａ复合以及采集于北京地区的Ｄｒｏｓｏｐｈｉｌａ ｓｉｍｕｌａｎｓ为Ｗｏｌｂａｃｈｉａ所感染，ＲＦＬＰ分析证实为单一感染，克隆了编码Ｗｏｌｂａｃｈｉａ外膜蛋白质的ｗｓｐ基因并进行了序列测定。     

Wolbachia 在中国稻田飞虱中的感染和传播

用ＰＣＲ方法检测了采集于不同地域的稻田飞虱共生菌Ｗｏｌｂａｃｈｉａ的感染,发现灰飞虱Ｌａｏｄｅｌｐｈａｘ ｓｔｒｉａｔｅｌｌｕｓ,褐飞虱Ｎｉｌｅｐａｒｖａｔａ ｌｕｇｅｎｓ,白背飞虱Ｓｏｇａｔｅｌｌａ ｆｕｒｃｉｆｅｒａ 为Ｗｏｌｂａｃｈｉａ所感染。克隆了编码Ｗｏｌｂａｃｈｉａ外膜蛋白质的ｗｓｐ基因并进行了序列测定,对ｗｓｐ的ＲＦＬＰ分析证实了这些飞虱为单一Ｗｏｌｂａｃｈｉａ感染,还发现一种能同时     

血吸虫细胞色素C氧化酶亚基Ⅰ基因的克隆与近缘种属关系分析

从生物化学的角度比较血吸虫不同虫株的遗传进化差异.以日本血吸虫湖北株成虫基因组总DNA为模板,设计特异性引物扩增获取血吸虫线粒体细胞色素C氧化酶基因片段,转化大肠杆菌DH5a,筛选阳性克隆,测序,对克隆得到的coI基因作进化分析.结果表明:无论是在核苷酸序列还是在氨基酸序列上,日本血吸虫湖北株与其近地域株(云南、江苏、湖南)均具有较高的同源性,col摹因的相似性分别为98%、98%和97%,但与不同种属血吸虫(曼氏、湄公、埃及、马来)的一致性则较低,分别为76%、84%、76%、85%,由此证实日本血吸虫湖北株与其它近地域株具有较近的亲缘关系.     

基于COI和D-Loop序列豫北虾虎鱼分子系统分析

虾虎鱼在河南省北部广泛分布,但其系统分类和多样性未有报道.从豫北原阳黄寺、安阳小南海、淇县淇河、长垣孙东闸、林州弓上水库等地采集虾虎鱼样本共计23尾,成都岷江样本6尾,分别扩增了线粒体COI基因和D-Loop序列.联合GenBank下载虾虎鱼的COI序列和D-Loop序列,构建了虾虎鱼的系统进化树.结果显示,采自黄寺、小南海、淇县、岷江地区的子陵吻虾虎鱼(Rhinogobius giurinus)聚为一个分支,而采自孙东闸和弓上水库的褐吻虾虎鱼(Rhinogobius brunneus)聚为一个单系群;这两种虾虎鱼遗传分化主要来自群体间.系统发育结果支持子陵吻虾虎鱼的韩国群体与中国群体构成姐妹群关系,中国不同地理种群间则相互交叉.     

大白菜雄性不育系和保持系花药的几种同工酶分析

本文分析了大白菜品种“早皇白”雄性不育系及同型保持系花药三种同工酶谱,不育系均少于保持系.细胞色素氧化酶显示,不育系为3条,保持系为7条;过氧化物酶谱不育系为3条,保侍系为8条;脂酶、不育系为4条,保持系为7条.     

基于线粒体DNA多态性鉴定烟台海鲜市场鱼种

由烟台海鲜市场随机采集具有相对重要商业价值的10种海鱼(小黄鱼、牙片、鲅鱼、面条鱼、鲐鱼、鲳鱼、鲈鱼、偏口、黑鱼、白姑鱼),利用聚合酶链式反应-限制性片段长度多态性(PCR-RFLP)技术对鱼种进行鉴定.首先提取10种海鱼的基因组DNA,利用自行设计的通用引物对其线粒体细胞色素C氧化酶亚基Ⅲ基因(MT-co3)进行PCR扩增,对扩增产物克隆、测序,用NCBI数据库检索,确定了10种海鱼的鱼种.依据测序结果进行限制性内切酶酶切分析,选用NlaⅢ和HinfⅠ2种内切酶进行酶切,获得了各种鱼类特异的酶切片段图谱.研究结果表明,PCR-RFLP能有效地区分以上10种海鱼,在鱼种鉴定上具有简便、准确的优势,可为鱼类食品检测提供科学依据.     

Ice-2和Ice-5基因在家蚕凋亡信号通路中的作用研究

细胞凋亡是细胞自主的有序死亡，caspase家族在细胞凋亡中起着重要的作用，ICE（Interlukin-1β—converting enzyme，白细胞介素-1β转换酶）是Caspase家族的一员，又称Caspase-1。为了了解家蚕ice-2和ice-5基因（GenBank登录号为：DQ360829和DQ360830）在凋亡信号通路中的作用，采用紫外线等刺激家蚕幼虫和家蚕细胞，利用荧光定量PCR的方法测定不同因素刺激下的ice-2和ice-5的表达量的变化。结果显示ice-2和ice-5都与家蚕的细胞信号转导有关，ice-5可能比ice-2起更重要的作用。     

复合PCR技术在混合感染检测中的应用

用传统的方法检测传染性疾病一次只能检测一种病原体,若为混合感染时,检测繁琐、费时且成本高,并且常常造成漏检.为了有效地解决上述问题,本文利用复合PCR分子诊断技术,建立了一套对由沙眼衣原体和解脲支原体造成的混合感染进行检测的方法.结果表明,该方法高效快速,可以开发成诊断试剂盒应用于混合感染的检测中.     

黄河鮈Cytb基因克隆及其在鮈属中的系统发育

采用聚合酶链式反应克隆黄河鮈线粒体细胞色素b基因 (Cyt b),首次报道了该基因的全长序列(1140bp,FJ904648) ,并与鮈属中其他6个物种的Cyt b 基因进行同源性比较,分析了碱基组成和变异情况,以马口鱼属的马口鱼和鯝属的云南鯝作为外群构建了ML和NJ系统发育树.遗传距离和分子系统学分析表明,黄河鮈与犬首鮈具有较低的遗传距离(6.30%),在系统发育树上两者聚在一起,提示黄河鮈与犬首鮈存在较近的亲缘关系.     

亚洲黑熊四川亚种Ursus thibetanus mupinensis线粒体细胞色素C氧化酶亚基Ⅲ基因的克隆与初步分析

根据已报道的部分哺乳动物线粒体细胞色素C氧化酶（Cytochrome C Oxidase）亚基Ⅲ基因（mtCOXⅢ）的相关信息设计引物,运用PCR技术,首次从亚洲黑熊四川亚种（以下简称四川黑熊,Ursus thibetanus mupinensis）的肌肉组织总DNA中成功克隆了mtCOXⅢ基因的编码序列,并对其进行了初步分析.结果表明：四川黑熊mtCOXⅢ基因序列全长784 bp,ORF是783 bp,编码261个氨基酸的蛋白质,该蛋白的等电点为6.47,分子量为29840.6 Da.四川黑熊的mtCOXⅢ与已报道的部分哺乳动物的mtCOXⅢ基因具有很高的同源性.进一步分析发现,四川黑熊线粒体细胞色素C氧化酶亚基Ⅲ功能位点的位置、种类和数量均与已报道的部分哺乳动物具有很高的相似性.     

Infection of Wolbachia may improve the olfactory response of Drosophila

The endosymbiotic bacterium Wolbachia infects various insects and is primarily known for its ability to manipulate host reproduction.Recent investigations reveal that Wolbachia also affects the activity of somatic cells.We here demonstrated by trap method and T-maze that Wolbachia infection had signifi-cant impact on the olfactory response of Drosophila simulans.Wolbachia-infected flies took shorter time to enter the food trap and were more sensitive to odorant in T-maze than those uninfected controls.The t...     

Bidirectional incompatibility between conspecific populations of Drosophila simulans

Cytoplasmic incompatibility (CI) describes the phenomenon whereby eggs fertilized by sperm from insects infected with a rickettsial endosymbiont fail to hatch. Unidirectional CI between conspecific populations of insects is a well documented phenomenon. Bidirectional CI has, however, only been described in mosquito populations, and recently between closely related species of parasitic wasps, where it is of interest as both an unusual form of reproductive isolation and as a potential means of insect population suppression. Here we report on the first known example of bidirectional CI between conspecific populations of Drosophila simulans. Further, we show that defects as early as the first cleavage division are associated with CI. This observation suggests that the cellular basis of CI involves disruption of processes before or during zygote formation and that CI arises from defects in the structure and/or function of the sperm during fertilization.     

山东野生果蝇体内Wolbachia的检测及WSP基因序列分析

Wolbachia是当前分布最广的共生菌类,参与调控寄主的多种生殖活动.通过对山东8个地理种群野生果蝇Wolbachia的WSP基因进行检测,在5个地理种群的黑腹果蝇和3个地理种群拟果蝇中分别分离到长度为632bp和611bp的Wolbachia的WSP基因片段,命名为WSPSD632和WSPSD611,在GeneBank所获注册号分别为EU395833和EU395834.将两序列和CeneBank上公布的WSP基因序列构建系统发育树,表明山东野生黑腹果蝇和拟果蝇所感染的Wolbachia分属于WMel和WRi类群.序列比对结果显示,两核苷酸序列的同源性为86.39%,但碱基替换率和缺失率存在一定差异.     

果蝇姊妹种间亲缘关系的聚类分析

本文以聚丙烯酰胺凝胶电泳（ＰＡＧＥ）技术分析了金色果蝇复合种（Ｄｒｏｓｏｐｈｉｌａａｕｒａｒｉａｓｐｅｃｉｅｓｃｏｍｐｌｅｘ）的５个姊妹种共１７个地理种群及其它种类的５个地理种群的８种同工酶酶谱，根据其中７种酶（Ａｍｙ、Ａｏ、Ｅｓｔ、α－Ｇｐｄｈ、Ｍｄｈ、Ｍｅ、Ｔｙｒ、Ｔｏ）的条带特征按Ｆｏｒｇｕｓｏｎ及Ａｑｕａｄｏ　＆Ａｖｉｓｅ的公式分别计算遗传相似系数及遗传距离，以类平均法（ＵＰＧＭＡ）进行了聚类分析，绘制出８个种类共２２个地理种群的生化系统树，结合姊妹种间无选择杂交可能组合的结果讨论了ａｕｒａｒｉａｓｐｅｃｉｅｓｃｏｒｎｐｌｅｘ各姊妹种间的亲缘关系、种化模式及分布在日本Ｋａｍｅｔｏｋｕ岛上的Ｄ．ｔｒｉａｕｒａｒｉａ发生分化的可能性。     

Somatic stem cell niche tropism in Wolbachia

Wolbachia are intracellular bacteria found in the reproductive tissue of all major groups of arthropods. They are transmitted vertically from the female hosts to their offspring, in a pattern analogous to mitochondria inheritance. But Wolbachia phylogeny does not parallel that of the host, indicating that horizontal infectious transmission must also occur. Insect parasitoids are considered the most likely vectors, but the mechanism for horizontal transfer is largely unknown. Here we show that newly introduced Wolbachia cross several tissues and infect the germline of the adult Drosophila melanogaster female. Through investigation of bacterial migration patterns during the course of infection, we found that Wolbachia reach the germline through the somatic stem cell niche in the D. melanogaster germarium. In addition, our data suggest that Wolbachia are highly abundant in the somatic stem cell niche of long-term infected hosts, implying that this location may also contribute to efficient vertical transmission. This is, to our knowledge, the first report of an intracellular parasite displaying tropism for a stem cell niche.     

Wolbachia variability and host effects on crossing type in Culex mosquitoes

Wolbachia is a common maternally inherited bacterial symbiont able to induce crossing sterilities known as cytoplasmic incompatibility (CI) in insects. Wolbachia-modified sperm are unable to complete fertilization of uninfected ova, but a rescue function allows infected eggs to develop normally. By providing a reproductive advantage to infected females, Wolbachia can rapidly invade uninfected populations, and this could provide a mechanism for driving transgenes through pest populations. CI can also occur between Wolbachia-infected populations and is usually associated with the presence of different Wolbachia strains. In the Culex pipiens mosquito group (including the filariasis vector C. quinquefasciatus) a very unusual degree of complexity of Wolbachia-induced crossing-types has been reported, with partial or complete CI that can be unidirectional or bidirectional, yet no Wolbachia strain variation was found. Here we show variation between incompatible Culex strains in two Wolbachia ankyrin repeat-encoding genes associated with a prophage region, one of which is sex-specifically expressed in some strains, and also a direct effect of the host nuclear genome on CI rescue.     

Molecular Identification of a Wolbachia endosymbiont in Trichogramma dendrolimi

Wolbachia is a common and widespread group of bacteria found in arthropods. These bacteria have evolved various mechanisms for manipulating reproduction of their host. The presence of Wolbachia in a lab strain of the arrhenotokous species Trichogramma dendrolimi was observed by the amplification and sequencing of part of the wsp gene. Aligning the resulting sequences with already published ones, the phylogenetic relationships between Wolbachia found in Trichogramma dendrolimi and in other Trichogramma wasps was established, and the phylogenetic relationships of Wolbachia in Trichogramma were not congruent with their hosts Trichogramma. Some factors contributing to this uncongruence are discussed here.     

Wolbachia在灰飞虱体内的分布

灰飞虱（Laodelphax striatellus)能传播水稻条纹叶枯病,是我国主要的水稻害虫,有些灰飞虱体内含有细胞内共生菌－Wolbachia。Wolbachia是一种细胞质遗传的细胞内共生菌,这类细菌改变了宿主的生殖行为,引起细胞质不相容（CI）,雌性化及孤雌生殖等现象,导致带有Wolbachia的宿主具有生殖优势。采用PCR和Western杂交的方法分析Wolbachia的灰飞虱体内不同组织的分布状况,发现灰飞虱体内Wolbachia除存在于生殖组织外,还广泛分布在头、胸、腹、唾液腺、消化道等非生殖组织中。这种广泛的分布状态说明Wolbachia是在昆虫体内引入、表达与传播外源基因的良好媒介。     

Isolation and characterization of a potential transposable element from Wolbachia

Wolbachia are a group of Rickettsia-like bacteria which parasitize the cells of a wide range of anthropoid. These microorganisms are associated with the reproductive and developmental abnormalities io their hosts. To study the molecular mechanism underlying such phenomena, we analyzed the genomic difference between Wolbachia with different cytoplasmic incompatibility (CI) phenotype using representational difference analysis method. A potential transposable element, which exists in the strong CI-inducing strain wRi, was isolated. This element was designated as Wolbachia insertion sequence element (WISE).