羽衣甘蓝红色系叶片形成相关microRNAs的鉴定及分析

Identification and analysis of microRNAs related to the formation of ornamental kale red-color leaf

为了在转录后水平上探索microRNAs(miRNAs)对羽衣甘蓝红色系叶片形成的调控作用,以红和白色系的羽衣甘蓝品种‘紫鸽’‘白鸽’为研究试材,利用高通量测序技术分别对其紫红(Bo-P)和白色(Bo-W)心叶进行了小RNA(sRNA)测序分析。结果表明:1)共鉴定出77个保守miRNAs和33个新miRNAs在Bo-P和Bo-W文库中差异表达;2)基于miRNAs的差异表达和相应靶基因的注释结果,筛选出2个可能与红色系叶片中花青苷生物合成有关的DEMs(miR828和miR858-3);3)荧光定量PCR(qRT-PCR)结果显示与高通量测序结果基本一致,其中2次结果显示miR828在紫红和白色心叶中表达量的差异倍数均很大。基于上述结果,初步确定miR828和miR858-3(特别是miR828)在转录后水平对羽衣甘蓝叶片红色系表型的形成起到关键调控作用,该结果将为今后深入揭示羽衣甘蓝叶色的分子调控机制提供理论基础和新视角。

In order to explore the regulatory role of microRNAs(miRNAs)on red-color leaf formation in ornamental kale at the post-transcriptional level,the red and white leaf series of ornamental kale including the‘Purple Pigeon’and‘White Pigeon’cultivars were selected as research materials in this study.Based on the high-throughput sequencing technology,small RNA(sRNA)sequences of purple red(Bo-P)and white(Bo-W)inner leaves were analyzed.The results showed that:A total of 77 known and 33novel miRNAs were differentially expressed between the Bo-P and Bo-W libraries;Based on the results of differential expression of miRNAs and annotation of the corresponding target genes,two DEMs(miR828 and miR858-3)were also identified,which might be related to anthocyanin biosynthesis of the red leaf;The results of qRT-PCR were basically consistent with the results of high-throughput sequencing.The two results all showed that the expression level of miR828 in purple red and white inner leaves was significantly different.Based on these above results,it is concluded that miR828 and miR858-3(especially miR828)are possibly involved in mediating formation of the red-color leaf phenotype at the post-transcriptional level.The study will provide a theoretical basis and a new perspective for further revealing the molecular regulation mechanism of leaf color in ornamental kale.