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Growth medium and incubation temperature alter the Pseudogymnoascus destructans transcriptome: implications in identifying virulence factors

机译:生长培养基和培养温度改变假炎症破碎物转录组:在识别毒力因子方面的意义

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摘要

Pseudogymnoascus destructans is the causal agent of bat white-nose syndrome (WNS), which is devastating some North American bat populations. Previous transcriptome studies provided insight regarding the molecular mechanisms involved in WNS; however, it is unclear how different environmental parameters could influence pathogenicity. This information could be useful in developing management strategies to mitigate the negative impacts of P. destructans on bats. We cultured three P. destructans isolates from Atlantic Canada on two growth media (potato dextrose agar and Sabouraud dextrose agar) that differ in their nitrogen source, and at two separate incubation temperatures (4 C and 15 C) that approximate the temperature range of bat hibernacula during the winter and a temperature within its optimal mycelia) growth range. We conducted RNA sequencing to determine transcript levels in each sample and performed differential gene expression (DGE) analyses to test the influence of growth medium and incubation temperature on gene expression. We also compared our in vitro results with previous RNAsequencing data sets generated from P. destructans growing on the wings of a susceptible host, Myotis lucifugus. Our findings point to a critical role for substrate and incubation temperature in influencing the P. destructans transcriptome. DGE analyses suggested that growth medium plays a larger role than temperature in determining P. destructans gene expression and that although the psychrophilic fungus responds to different nitrogen sources, it may have evolved for continued growth at a broad range of low temperatures. Further, our data suggest that down-regulation of the RNA-interference pathway and increased fatty acid metabolism are involved in the P. destructans-bat interaction. Finally, we speculate that to reduce the activation of host defense responses, P. destructans minimizes changes in the expression of genes encoding secreted proteins during bat colonization.
机译:伪影症破坏者是蝙蝠白鼻综合征(WNS)的因果因子,这是毁灭一些北美蝙蝠种群。以前的转录组研究提供了有关WNS中涉及的分子机制的洞察力;然而,目前尚不清楚不同的环境参数如何影响致病性。这些信息对于制定管理策略,可以减轻P. Destuctans对蝙蝠的负面影响。我们在两种生长培养基(马铃薯右旋糖糖浆和Sabourauge Dextrose琼脂)上培养了三个P. Destructans的分离株,其氮源不同,并且在两个单独的培养温度(4c和15c)中,近似蝙蝠的温度范围冬季血渍和最佳菌丝体内的温度)增长范围。我们进行了RNA测序以确定每个样品中的转录物水平并进行差异基因表达(DGE)分析以测试生长培养基和孵育温度对基因表达的影响。我们还将我们的体外结果与先前的RNASEQUENCING数据集进行了比较,从P. Destructans生长在易感宿主的翅膀上生长,肌霉素。我们的发现指出了影响P. Destructors转录组的底物和孵育温度的关键作用。 DGE分析表明,生长培养基在确定P. destructans基因表达中的温度比温度发挥着更大的作用,并且虽然心理学真菌对不同的氮源作出反应,但它可能已经进化为在广泛的低温下继续生长。此外,我们的数据表明,RNA干扰途径的下调和增加的脂肪酸代谢涉及P. Destructans-Bat相互作用。最后,我们推测,减少宿主防御反应的激活,P. destructans最小化在蝙蝠殖民化期间编码分泌蛋白质的基因表达的变化。

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