Biotechnology Department, University of Verona, Strada Le Grazie 15, I-37134, Verona, Italy.
BMC Genomics. 2013 Jan 18;14:41. doi: 10.1186/1471-2164-14-41.
Plants such as grapevine (Vitis spp.) display significant inter-cultivar genetic and phenotypic variation. The genetic components underlying phenotypic diversity in grapevine must be understood in order to disentangle genetic and environmental factors.
We have shown that cDNA sequencing by RNA-seq is a robust approach for the characterization of varietal diversity between a local grapevine cultivar (Corvina) and the PN40024 reference genome. We detected 15,161 known genes including 9463 with novel splice isoforms, and identified 2321 potentially novel protein-coding genes in non-annotated or unassembled regions of the reference genome. We also discovered 180 apparent private genes in the Corvina genome which were missing from the reference genome.
The de novo assembly approach allowed a substantial amount of the Corvina transcriptome to be reconstructed, improving known gene annotations by robustly defining gene structures, annotating splice isoforms and detecting genes without annotations. The private genes we discovered are likely to be nonessential but could influence certain cultivar-specific characteristics. Therefore, the application of de novo transcriptome assembly should not be restricted to species lacking a reference genome because it can also improve existing reference genome annotations and identify novel, cultivar-specific genes.
葡萄(Vitis spp.)等植物表现出显著的品种间遗传和表型变异。为了厘清遗传和环境因素,必须了解葡萄表型多样性的遗传基础。
我们已经表明,通过 RNA-seq 进行 cDNA 测序是一种用于描述本地葡萄品种(Corvina)与 PN40024 参考基因组之间品种多样性的稳健方法。我们检测到 15161 个已知基因,包括 9463 个具有新剪接异构体的基因,并在参考基因组的非注释或未组装区域鉴定了 2321 个潜在的新蛋白编码基因。我们还在 Corvina 基因组中发现了 180 个明显的特有基因,这些基因在参考基因组中缺失。
从头组装方法允许大量重建 Corvina 的转录组,通过稳健地定义基因结构、注释剪接异构体和检测无注释的基因,从而改进已知基因的注释。我们发现的特有基因可能是非必需的,但可能会影响某些特定品种的特征。因此,从头转录组组装的应用不应仅限于缺乏参考基因组的物种,因为它还可以改进现有的参考基因组注释并鉴定新的、特定于品种的基因。