Vitulo Nicola, Forcato Claudio, Carpinelli Elisa Corteggiani, Telatin Andrea, Campagna Davide, D'Angelo Michela, Zimbello Rosanna, Corso Massimiliano, Vannozzi Alessandro, Bonghi Claudio, Lucchin Margherita, Valle Giorgio
CRIBI Biotechnology Centre, University of Padua, Padua, Italy.
BMC Plant Biol. 2014 Apr 17;14:99. doi: 10.1186/1471-2229-14-99.
Alternative splicing (AS) significantly enhances transcriptome complexity. It is differentially regulated in a wide variety of cell types and plays a role in several cellular processes. Here we describe a detailed survey of alternative splicing in grape based on 124 SOLiD RNAseq analyses from different tissues, stress conditions and genotypes.
We used the RNAseq data to update the existing grape gene prediction with 2,258 new coding genes and 3,336 putative long non-coding RNAs. Several gene structures have been improved and alternative splicing was described for about 30% of the genes. A link between AS and miRNAs was shown in 139 genes where we found that AS affects the miRNA target site. A quantitative analysis of the isoforms indicated that most of the spliced genes have one major isoform and tend to simultaneously co-express a low number of isoforms, typically two, with intron retention being the most frequent alternative splicing event.
As described in Arabidopsis, also grape displays a marked AS tissue-specificity, while stress conditions produce splicing changes to a minor extent. Surprisingly, some distinctive splicing features were also observed between genotypes. This was further supported by the observation that the panel of Serine/Arginine-rich splicing factors show a few, but very marked differences between genotypes. The finding that a part the splicing machinery can change in closely related organisms can lead to some interesting hypotheses for evolutionary adaptation, that could be particularly relevant in the response to sudden and strong selective pressures.
可变剪接(AS)显著增强了转录组的复杂性。它在多种细胞类型中受到差异调节,并在多个细胞过程中发挥作用。在此,我们基于对来自不同组织、胁迫条件和基因型的124次SOLiD RNA测序分析,详细描述了葡萄中的可变剪接情况。
我们利用RNA测序数据更新了现有的葡萄基因预测,新增了2258个编码基因和3336个推定的长链非编码RNA。多个基因结构得到了改进,约30%的基因存在可变剪接现象。在139个基因中显示了可变剪接与微小RNA(miRNA)之间的联系,我们发现可变剪接会影响miRNA的靶位点。对异构体的定量分析表明,大多数剪接基因有一个主要异构体,并且倾向于同时共表达少量异构体,通常为两个,内含子保留是最常见的可变剪接事件。
正如在拟南芥中所描述的那样,葡萄也表现出明显的可变剪接组织特异性,而胁迫条件只会在较小程度上产生剪接变化。令人惊讶的是,在不同基因型之间也观察到了一些独特的剪接特征。富含丝氨酸/精氨酸的剪接因子在不同基因型之间存在一些但非常明显的差异,这一观察结果进一步支持了上述结论。可变剪接机制的一部分在亲缘关系密切的生物体中会发生变化,这一发现可能会引发一些关于进化适应性的有趣假设,这在应对突然而强烈的选择压力时可能尤为重要。