Diverse Legionella‐Like Bacteria Associated with Testate Amoebae of the Genus Arcella (Arcellinida: Amoebozoa)

Diverse species of Legionella and Legionella‐like amoebal pathogens (LLAPs) have been identified as intracellular bacteria in many amoeboid protists. There are, however, other amoeboid groups such as testate amoeba for which we know little about their potential to host such bacteria. In this study, we assessed the occurrence and diversity of Legionella spp. in cultures and environmental isolates of freshwater arcellinid testate amoebae species, Arcella hemispherica, Arcella intermedia, and Arcella vulgaris, via 16S rRNA gene sequence analyses and fluorescent in situ hybridization (FISH). Analysis of the 16S rRNA gene sequences indicated that A. hemispherica, A. intermedia, and A. vulgaris host Legionella‐like bacteria with 94–98% identity to other Legionella spp. based on NCBI BLAST search. Phylogenetic analysis placed Legionella‐like Arcella‐associated bacteria (LLAB) in three different clusters within a tree containing all other members of Legionella and LLAPs. The intracellular localization of the Legionella within Arcella hosts was confirmed using FISH with a Legionella‐specific probe. This study demonstrates that the host range of Legionella and Legionella‐like bacteria in the Amoebozoa extends beyond members of “naked” amoebae species, with members of the testate amoebae potentially serving an ecological role in the dispersal, protection, and replication of Legionella spp. in natural environments.     

Family‐based genome scan for age at onset of late‐onset Alzheimer's disease in whole exome sequencing data

Alzheimer's disease (AD) is a common and complex neurodegenerative disease. Age at onset (AAO) of AD is an important component phenotype with a genetic basis, and identification of genes in which variation affects AAO would contribute to identification of factors that affect timing of onset. Increase in AAO through prevention or therapeutic measures would have enormous benefits by delaying AD and its associated morbidities. In this paper, we performed a family‐based genome‐wide association study for AAO of late‐onset AD in whole exome sequence data generated in multigenerational families with multiple AD cases. We conducted single marker and gene‐based burden tests for common and rare variants, respectively. We combined association analyses with variance component linkage analysis, and with reference to prior studies, in order to enhance evidence of the identified genes. For variants and genes implicated by the association study, we performed a gene‐set enrichment analysis to identify potential novel pathways associated with AAO of AD. We found statistically significant association with AAO for three genes (WRN, NTN4 and LAMC3) with common associated variants, and for four genes (SLC8A3, SLC19A3, MADD and LRRK2) with multiple rare‐associated variants that have a plausible biological function related to AD. The genes we have identified are in pathways that are strong candidates for involvement in the development of AD pathology and may lead to a better understanding of AD pathogenesis.     

Evaluation of the reference genes for expression analysis using quantitative real‐time polymerase chain reaction in the green peach aphid,Myzus persicae

The green peach aphid, Myzus persicae Sulzer (Hemiptera, Aphididae), is an important cosmopolitan pest. Real time qRT‐PCR has been used for target gene expression analysis on M. persicae. Using real time qRT‐PCR, the expression levels are normalized on the basis of the reliable reference genes. However, to date, the stability of available reference genes has been insufficient. In this study, we evaluated nine candidate reference genes from M. persicae under diverse experimental conditions. The tested candidate genes were comprehensively ranked based on five alternative methods (RefFinder, geNorm, Normfinder, BestKeeper and the comparative ΔC_t method). 18s, Actin and ribosomal protein L27 (L27) were recommended as the most stable reference genes for M. persicae, whereas ribosomal protein L27 (L27) was found to be the least stable reference genes for abiotic studies (photoperiod, temperature and insecticide susceptibility). Our finding not only sheds light on establishing an accurate and reliable normalization of real time qRT‐PCR data in M. persicae but also lays a solid foundation for further studies of M. persicae involving RNA interference and functional gene research.     

水杨酸诱导下蝙蝠蛾拟青霉转录组分析及虫草酸代谢途径关键酶基因挖掘

【目的】虫草酸是虫草中重要的活性成分之一,但其低含量极大地限制了其工业应用。水杨酸(salicylic acid, SA)是一种非生物诱导子,可以显著提高蝙蝠蛾拟青霉中虫草酸的合成,但蝙蝠蛾拟青霉虫草酸代谢途径及其对水杨酸的响应尚不明确。本研究旨在获得蝙蝠蛾拟青霉响应SA处理的转录组学信息,挖掘蝙蝠蛾拟青霉中虫草酸代谢途径关键酶基因。【方法】采用SA诱导培养蝙蝠蛾拟青霉,8 h后选取诱导和未诱导的菌丝进行转录组高通量测序分析。【结果】测序最终获得40.37 Gb的clean data,拼接得到20 317条unigene,平均长度为1 357.13 bp,功能注释共获得13 592条unigene。差异基因分析共筛选出差异基因2 574个,其中有1 135个上调,1 439个下调。KEGG富集分析表明,差异基因主要富集于细胞周期、减数分裂、半乳糖代谢、DNA复制、糖醇脂类生物合成、甘油脂类代谢等KEGG通路中。进一步分析得到与虫草酸代谢相关的基因13条,其中参与虫草酸生物合成的基因glk、gpi、gla、mpi、fbp、mtld在SA处理后表达量上调,而涉及虫草酸消耗的基因mdh在SA...