Caragata E P, Pais F S, Baton L A, Silva J B L, Sorgine M H F, Moreira L A
Grupo Mosquitos Vetores: Endossimbiontes e Interação Patógeno Vetor, Centro de Pesquisas René Rachou - Fiocruz, Belo Horizonte, Minas Gerais, Brazil.
Grupo de Informática de Biossistemas e Genômica, Centro de Pesquisas René Rachou - Fiocruz, Belo Horizonte, Minas Gerais, Brazil.
BMC Genomics. 2017 Jan 3;18(1):6. doi: 10.1186/s12864-016-3441-4.
Wolbachia is a bacterial endosymbiont that naturally infects a wide range of insect species, and causes drastic changes to host biology. Stable infections of Wolbachia in mosquitoes can inhibit infection with medically important pathogens such as dengue virus and malaria-causing Plasmodium parasites. However, some native Wolbachia strains can enhance infection with certain pathogens, as is the case for the mosquito Aedes fluviatilis, where infection with Plasmodium gallinaceum is enhanced by the native wFlu Wolbachia strain. To better understand the biological interactions between mosquitoes and native Wolbachia infections, and to investigate the process of pathogen enhancement, we used RNA-Seq to generate the transcriptome of Ae. fluviatilis with and without Wolbachia infection.
In total, we generated 22,280,160 Illumina paired-end reads from Wolbachia-infected and uninfected mosquitoes, and used these to make a de novo transcriptome assembly, resulting in 58,013 contigs with a median sequence length of 443 bp and an N50 of 2454 bp. Contigs were annotated through local alignments using BlastX, and associated with both gene ontology and KEGG orthology terms. Through baySeq, we identified 159 contigs that were significantly upregulated due to Wolbachia infection, and 98 that were downregulated. Critically, we saw no changes to Toll or IMD immune gene transcription, but did see evidence that wFlu infection altered the expression of several bacterial recognition genes, and immune-related genes that could influence Plasmodium infection. wFlu infection also had a widespread effect on a number of host physiological processes including protein, carbohydrate and lipid metabolism, and oxidative stress. We then compared our data set with transcriptomic data for other Wolbachia infections in Aedes aegypti, and identified a core set of 15 gene groups associated with Wolbachia infection in mosquitoes.
While the scale of transcriptional changes associated with wFlu infection might be small, the scope is rather large, which confirms that native Wolbachia infections maintain intricate molecular relationships with their mosquito hosts even after lengthy periods of co-evolution. We have also identified several potential means through which wFlu infection might influence Plasmodium infection in Ae. fluviatilis, and these genes should form the basis of future investigation into the enhancement of Plasmodium by Wolbachia.
沃尔巴克氏体是一种细菌内共生体,能自然感染多种昆虫物种,并导致宿主生物学发生剧烈变化。沃尔巴克氏体在蚊子中的稳定感染可抑制登革病毒和疟原虫等重要医学病原体的感染。然而,一些本地的沃尔巴克氏体菌株可增强某些病原体的感染,例如在河流伊蚊中,本地的wFlu沃尔巴克氏体菌株可增强鸡疟原虫的感染。为了更好地理解蚊子与本地沃尔巴克氏体感染之间的生物学相互作用,并研究病原体增强的过程,我们使用RNA测序技术生成了感染和未感染沃尔巴克氏体的河流伊蚊的转录组。
我们总共从感染和未感染沃尔巴克氏体的蚊子中生成了22,280,160条Illumina双端读数,并用于进行从头转录组组装,得到了58,013个重叠群,中位序列长度为443 bp,N50为2454 bp。通过使用BlastX进行局部比对,对重叠群进行注释,并与基因本体论和KEGG直系同源术语相关联。通过baySeq,我们鉴定出159个因沃尔巴克氏体感染而显著上调的重叠群和98个下调的重叠群。至关重要的是,我们未观察到Toll或IMD免疫基因转录的变化,但确实发现有证据表明wFlu感染改变了几种细菌识别基因以及可能影响疟原虫感染的免疫相关基因的表达。wFlu感染还对许多宿主生理过程产生了广泛影响,包括蛋白质、碳水化合物和脂质代谢以及氧化应激。然后,我们将我们的数据集与埃及伊蚊中其他沃尔巴克氏体感染的转录组数据进行比较,并鉴定出一组与蚊子中沃尔巴克氏体感染相关的15个基因群的核心集。
虽然与wFlu感染相关的转录变化规模可能较小,但范围相当大,这证实了即使经过长时间的共同进化,本地沃尔巴克氏体感染与其蚊子宿主仍保持着复杂的分子关系。我们还确定了wFlu感染可能影响河流伊蚊中疟原虫感染的几种潜在途径,这些基因应成为未来研究沃尔巴克氏体增强疟原虫感染的基础。