RNA seq of H2O, glucose, mannose or galactose-treated Batrachochytrium salamandrivorans zoospores

We report RNAseq data from B. salamandrivorans zoospores treated with different carbohydrates (glucose, mannose or galactose, 50 mM) or H2O as a control, for 1 hour. We found a selective upregulation of putative B. salamandrivorans virulence factors during initial contact with carbohydrates and galactose specifically. Total RNA was isolated from B. salamandrivorans zoospores treated with different carbohydrates. Therefore, zoospores were harvested from 175 cm2 cell culture flasks by replacing the TGhL broth with distilled water, which was filtered using a sterile mesh filter with pore size 10 µm. Six independent pools containing 4 x 10e7 zoospores were obtained (biological replicates). Per pool, the spores were divided in 4 eppendorfs (10e7 zoospores/eppendorf) which were treated for 1 hour at 15°C with H2O (control), 50 mM (α-D-galactose), 50 mM (α-D-glucose) or 50 mM (α-D-mannose). After 1 hour, the zoospores were centrifuged for 5 min at 3000 rpm at 15°C to remove the supernatant, after which RNA was extracted using the RNeasy mini kit (Qiagen). The RNA was treated with Turbo™ DNase, following the manufacturer's instructions. RNA degradation and contamination was monitored on 1% agarose gels. The RNA purity was checked using the NanoPhotometer® spectrophotometer . Finally, the RNA integrity and quantitation were assessed using the RNA Nano 6000 assay kit of the Bioanalyzer 2100 system.

本研究报道了经不同碳水化合物（葡萄糖、甘露糖或半乳糖，终浓度50 mM）或无菌水（对照组）处理1小时的蝾螈壶菌（B. salamandrivorans）游动孢子的RNA测序（RNAseq）数据。研究发现，在与碳水化合物（尤其半乳糖）初始接触阶段，蝾螈壶菌的推定毒力因子呈现选择性上调。本研究的总RNA提取自经不同碳水化合物处理的蝾螈壶菌游动孢子：具体操作如下，研究人员从175 cm²细胞培养瓶中收集游动孢子，将瓶内TGhL培养液替换为蒸馏水，随后通过孔径10 µm的无菌滤网过滤收集孢子；共获得6个独立孢子池（生物学重复），每个孢子池含4×10^7个游动孢子。每个孢子池被均分至4个离心管中（每管含1×10^7个游动孢子），随后于15℃条件下分别用以下试剂处理1小时：无菌水（对照组）、50 mM α-D-半乳糖、50 mM α-D-葡萄糖及50 mM α-D-甘露糖。处理1小时后，将样品于15℃、3000 rpm条件下离心5分钟以去除上清液，随后使用RNeasy迷你试剂盒（Qiagen公司）提取总RNA；按照试剂盒说明书，采用Turbo™ DNase对提取的RNA进行消化以去除基因组DNA污染。通过1%琼脂糖凝胶电泳监测RNA的降解与污染情况，使用NanoPhotometer®分光光度计检测RNA纯度，最后采用2100生物分析仪系统配套的RNA Nano 6000检测试剂盒评估RNA的完整性与浓度。