Petunia x hybrida Transcriptome or Gene expression. Petunia x hybrida

We report the application of sequencing-by-synthesis technology for high-throughput profiling of small RNAs involved in Chalcone synthase A (CHS-A) sense cosuppression in petunia. Sense cosuppression is a classical form of eukaryotic post-transcriptional gene silencing. It was first reported in transgenic petunia, where a transgene overexpressing the host Chalcone Synthase-A (CHS-A) gene caused the degradation of the homologous transcripts and the loss of flower pigmentation. Though sense cosuppression is recognized as an RNA silencing mechanism, little evidence has been yet provided demonstrating its association with the generation of individual small interfering RNAs (siRNAs) that are the assumed determinants. In this work, the deep sequencing of small RNAs in cosuppressed transgenic petunia and WT petunia respectively allowed for the identication of siRNAs that vastly predominate in the silenced flower and guide prominent cleavage events in CHS-A mRNA. Overall design: Examination of 2 small RNA populations from WT and cosuppressed petunia petals respectively

本研究报道了边合成边测序（sequencing-by-synthesis）技术在矮牵牛查尔酮合酶A（Chalcone synthase A, CHS-A）正义共抑制相关小RNA高通量分析中的应用。正义共抑制是真核生物转录后基因沉默的经典类型。该现象最早在转基因矮牵牛中被报道：过表达宿主查尔酮合酶A（CHS-A）基因的转基因会导致同源转录本降解以及花朵色素沉着缺失。尽管正义共抑制已被认定为一种RNA沉默机制，但目前鲜有证据表明其与作为假定调控因子的单一小干扰RNA（small interfering RNAs, siRNAs）的生成存在关联。本研究分别对发生共抑制的转基因矮牵牛与野生型（Wild Type, WT）矮牵牛中的小RNA进行深度测序，成功鉴定出在沉默花朵中大量富集、并可介导查尔酮合酶A（CHS-A）mRNA发生显著切割事件的小干扰RNA。整体实验设计：分别分析野生型矮牵牛与共抑制型矮牵牛花瓣中的两类小RNA群体。