目的:TagSeq是一种经济有效的方法,用于需要大量样本的基因表达研究。迄今为止,植物中的TagSeq研究仅限于具有高质量参考基因组的植物。我们在非模型植物中测试了TagSeq参考转录组的适用性,作为圣丽塔实验范围国家生态观测网(NEON)核心站点自然基因表达变异研究的一部分。
方法:TagSeq的组织样本来自四个物种的多个个体(Boutelouaaristidoides和Eragrostislehmanniana[禾本科],铜绿苔藓[Amaranthaceae],和佛罗里达帕金森病[Fabaceae])在三个日期的两个地点(总共56个样品)。每个物种一个样品用于通过标准RNA-seq产生参考转录组。通过参考基因座的回收来评估TagSeq性能,标记比对的特异性,和样本之间的差异。
结果:大部分标签与每个参考对齐并唯一映射。数万个基因座的表达模式是可量化的,揭示了所有物种表达的一致空间差异。
结论:使用从头参考转录组的TagSeq是本研究中定量基因表达的有效方法。标签是高度基因座特异性的,并为四种非模型植物物种产生生物学信息谱。
OBJECTIVE: TagSeq is a cost-effective approach for gene expression studies requiring a large number of samples. To date, TagSeq studies in plants have been limited to those with a high-quality reference genome. We tested the suitability of reference transcriptomes for TagSeq in non-model plants, as part of a
study of natural gene expression variation at the Santa Rita Experimental Range National Ecological Observatory Network (NEON) core site.
METHODS: Tissue for TagSeq was sampled from multiple individuals of four species (Bouteloua aristidoides and Eragrostis lehmanniana [Poaceae], Tidestromia lanuginosa [Amaranthaceae], and Parkinsonia florida [Fabaceae]) at two locations on three dates (56 samples total). One sample per species was used to create a reference transcriptome via standard RNA-seq. TagSeq performance was assessed by recovery of reference loci, specificity of tag alignments, and variation among samples.
RESULTS: A high fraction of tags aligned to each reference and mapped uniquely. Expression patterns were quantifiable for tens of thousands of loci, which revealed consistent spatial differentiation in expression for all species.
CONCLUSIONS: TagSeq using de novo reference transcriptomes was an effective approach to quantifying gene expression in this
study. Tags were highly locus specific and generated biologically informative profiles for four non-model plant species.