Landcare Research, Auckland, New Zealand.
Comp Biochem Physiol Part D Genomics Proteomics. 2013 Mar;8(1):24-31. doi: 10.1016/j.cbd.2012.10.005. Epub 2012 Oct 27.
The endemic New Zealand alpine stick insect Micrarchus nov. sp. 2 regularly experiences sub-zero temperatures in the wild. 454-based RNA-Seq was used to generate a de novo transcriptome and differentiate between treatments to investigate the genetic basis of cold tolerance. Non cold-treated individuals were compared to those exposed to 0°C for 1 h followed by a 1 h recovery period at 20°C. We aligned 607,410 Roche 454 reads, generating a transcriptome of 5235 contigs. Differential expression analysis ranked candidate cold responsive genes for qPCR validation by P-value. The top nine up-regulated candidates, together with eight a priori targets identified from previous studies, had their relative expression quantified using qPCR. Three candidate cold responsive genes from the RNA-Seq data were verified as significantly up-regulated, annotated as: prolyl 4-hydroxylase subunit alpha-1 (P4HA1), staphylococcal nuclease domain-containing protein 1 (snd1) and cuticular protein analogous to peritrophins 3-D2 (Cpap3-d2). All three are novel candidate genes, illustrating the varied response to low temperature across insects.
新西兰地方性高山枝角类昆虫 Micrarchus nov. sp. 2 在野外经常经历零度以下的温度。使用基于 454 的 RNA-Seq 生成从头转录组，并对不同处理进行区分，以研究耐寒性的遗传基础。将未经过冷处理的个体与暴露在 0°C 下 1 小时然后在 20°C 下恢复 1 小时的个体进行比较。我们对齐了 607,410 个罗氏 454 读取，生成了一个由 5235 个连续序列组成的转录组。差异表达分析通过 P 值对候选冷响应基因进行 qPCR 验证进行排名。前九个上调候选基因，加上之前研究中确定的八个先验靶标，使用 qPCR 对其相对表达量进行了定量。从 RNA-Seq 数据中验证了三个候选冷响应基因显著上调，注释为：脯氨酰 4-羟化酶亚基α-1 (P4HA1)、葡萄球菌核酸酶结构域蛋白 1 (snd1) 和角质蛋白类似于围食膜蛋白 3-D2 (Cpap3-d2)。这三个都是新的候选基因，说明了昆虫对低温的不同反应。
