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《African Zoology》2013,48(3):181-185
Recent reports on finding Wolbachia-strain infections in field mosquito species in some West African countries and the potential for developing these as disease vector biocontrol tools have prompted a search for Wolbachia in mosquitoes within the study area. Using a completely randomised design, mosquito traps were set at different locations in a rural and an urbanised community. One hundred and eighty (180) mosquitoes were trapped and pooled on the basis of genus, sex and site of collection, because there have been no earlier reports of Wolbachia isolated from Nigeria. Twenty pools, made up of not more than ten mosquitoes per pool, were homogenised and analysed for Wolbachia-specific DNA. Mosquitoes were trapped within Ede (urbanised community) and Akoda (rural community). Genomic DNA was extracted from trapped mosquito samples and used as a template in a PCR reaction. The Wolbachia sp. specific 16S rRNA gene was amplified, sequence analysis of PCR products was performed and a chromatogram of the sequence was subjected to Basic Local Alignment Search Tool analysis to identify the Wolbachia sp. This sequence was subsequently submitted to GenBank with accession number MK127541. The first evidence of the presence of the endosymbiont, Wolbachia in field-caught mosquitoes is hereby documented. The homology of this strain of Wolbachia bears similarities to those reported recently from other parts of West Africa and forms a single clade with a Wolbachia sp. from Mali, with a strong bootstrap support of 99%. This finding of a Wolbachia strain in mosquitoes at Ede could form the basis for more searches for diverse strains of Wolbachia in Nigeria. 相似文献
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蚜虫内共生菌基因组DNA提取方法的比较和优化 总被引：3，自引：3，他引：0
为探究蚜虫共生菌基因组DNA的提取方案,以桃蚜为试验材料,比较了目前较为常用的4种蚜虫基因组DNA提取方法,从DNA纯度、完整性、PCR扩增效率及稳定性等方面进行了比较和评价,并通过调整蛋白酶K用量和水浴温度及时间对STE法进行了优化。结果表明,4种方法提取的蚜虫共生菌基因组DNA均可用于共生菌的PCR扩增检测;CTAB法和SDS法提取的DNA纯度较高,条带较完整,稳定性相对较高,不易降解,而STE法和PCR缓冲液法操作简便,适于快速提取单头蚜虫共生菌的基因组DNA,但纯度相对较低;可根据试验条件和要求进行选择。STE法优化条件为:用30 μL STE缓冲液将蚜虫匀浆,加入1.5 μL 20 mg/mL蛋白酶K,于56 ℃水浴1.5 h;再加入0.1 μL 10 mg/L RNA酶,于37 ℃培养1 h,95 ℃下处理5 min,5 000 r/min离心3 min,将提取到的DNA于-20 ℃保存或直接用于PCR扩增。优化后的STE法可作为提取蚜虫共生菌基因组DNA经济而快捷有效的方法。 相似文献
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烟粉虱内共生菌groEL基因编码一种与植物病毒传毒相关的GroEL蛋白.以B型烟粉虱为研究对象,利用1对特异引物,对B型烟粉虱体内的groEL基因进行PCR扩增,选取502 bp groEL基因片段构建dsRNA,并将其连接植物表达载体pBIN19,将重组质粒电激转化农杆菌菌株LBA4404.采用农杆菌浸染转化本氏烟草,PCR筛选表明,目的片段已整合至转基因植株的基因组DNA;RT-PCR结果表明,502 bp的groEL基因片段在转基因植株中被转录. 相似文献
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褐飞虱Nilaparvata lugens是我国及东南亚地区水稻上的主要害虫,其体内存在的大量共生菌是褐飞虱生物防治中尚未开发的一类重要微生物资源。褐飞虱内共生菌种类丰富多样,目前研究主要集中于其初级共生真菌——类酵母共生菌（yeast-like symbiotes,YLS）和2类次级共生细菌——沃尔巴克氏体Wolbachia和杀雄菌Arsenophonus。早期关于褐飞虱内共生菌的分离及种类鉴定主要采用体外离体培养、克隆文库构建以及PCR-变性梯度凝胶电泳等传统技术。近年来,宏基因组和高通量测序技术的应用极大拓展了褐飞虱内共生菌种类多样性研究的深度。内共生菌不仅参与褐飞虱的营养代谢,为宿主提供氨基酸、维生素和甾醇等必需营养物质以满足宿主正常的生长发育和繁殖,同时还可以介导宿主的致害性变异、抗药性产生和病原物防御等生理过程。鉴于内共生菌在褐飞虱生长发育及环境适应过程中的重要作用,利用内共生菌对褐飞虱进行生物防控已成为目前研究的一大热点,并展现出良好的应用前景。该文对褐飞虱内共生菌的主要种类及其生物学特性进行介绍,对其分离鉴定技术和多样性分析技术进行总结,并对褐飞虱内共生菌的生物学功能进... 相似文献
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Effect of Cardinium Infection on the Probing Behavior of Bemisia tabaci (Hemiptera: Aleyrodidae) MED
Liu Ying Liu Baiming Li Hongran Ding Tianbo Tao Yunli Chu Dong 《Journal of insect science (Online)》2021,21(3)
Facultative endosymbionts can affect the growth, physiology, and behavior of their arthropod hosts. There are several endosymbionts in the invasive whitefly Bemisia tabaci Mediterranean (MED, Q biotype) that influence host fitness by altering stylet probing behavior. We investigated the probing behavior of B. tabaci MED infected with the facultative endosymbiont Candidatus Cardinium hertigii (Cardinium (Sphingobacteriales: Flexibacteraceae)). We generated genetically similar Cardinium-infected (C*+) and uninfected (C-) clonal sublines and analyzed the probing behavior of newly emerged adult on cotton (Malvales: Malvaceae), Gossypium hirsutum L., using electropenetrography (EPG). The C- subline demonstrated a longer duration of E2 (2.81-fold) and more events of E2 (2.22-fold) than the C*+ subline, indicating a greater level of sustained ingestion of plant phloem. These findings provide insight into the fitness costs (fitness of a particular genotype is lower than the average fitness of the population) of the Cardinium-infected B. tabaci. 相似文献
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为了解水稻褐飞虱中肠内共生菌的种类,用高氏1 号和马铃薯葡萄糖琼脂两种培养基对褐飞虱肠道进
行分离培养,对分离纯化后的菌株用菌类16S rDNA 的通用引物进行PCR 扩增鉴定,测序结果经BLAST 比对分析,
发现其与粉虱科的杀雄菌属内共生菌（Arsenophonus sp.）相似度达99%,说明所分离得到的为杀雄菌属的菌株。将所
得这条序列与7 条来自粉虱科不同种及来自褐飞虱的杀雄菌属内共生菌的16S rDNA 序列用邻接法构建进化关系
树,发现其与来自褐飞虱杀雄菌属内共生菌的16S rDNA（GU124505）距离较近,初步推断为同一种。 相似文献
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《水稻科学》2014,21(1):59-64
The structures of bacterial communities in the brown planthopper (BPH) Nilaparvata lugens (Stål) from different geographic and resistant virulent populations were analyzed by using denatured gradient gel electrophoresis (DGGE). Results showed that the bacterial communities in BPH nymph from the first to the fifth instars varied with nymphal growth and development. The bacterial communities in the first-instar BPH nymph were similar to those in adults. Nine geographic BPH populations were divided into three groups based on the cluster analysis of DGGE fingerprint. The first group was from the Philippines; the second group was from Thailand and Hainan, Yunnan and Zhejiang provinces of China; and the third group was from Vietnam and Guangxi, Hunan and Jiangxi provinces of China. BPH populations adapted to different resistant rice varieties. The BPH populations from Mudgo (with resistant gene Bph1) and ASD7 (with resistant gene bph2) differed with those of the susceptible rice variety TN1. 相似文献
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