Sequencing Analysis Of MiRNAs In Vegetative And Reproductive Ceratopteris Pteridoides

Ceratopteris pteridoides was located within the Ceratopteris genus of the Pteridaceae family,and it was a perennial herb that possessed definitive transition from vegetative growth to reproductive growth.micro RNAs(miRNAs)were a class of small RNA which was non-coding and long as ～ 21 nucleotides(nt),and they mainly controlled expression of genes related to growth development and stress response.miRNA156(miR156)and miR172 were critical to transition of growth phases in angiosperms;however,it was unclear about their roles in ferns.Thus,it was of great significance to study miRNA in C.pteridoides for understanding conservation of the mechanism underlying the growth phase transition within vascular plants.This study collected the complete plants undergoing vegetative growth and reproductive growth for C.pteridoides,constructed sRNA sequencing libraries,and performed second-generation sequencing.This study identified miRNAs,analyzed differential expression,predicted target genes,and conduced quantitative PCR,producing following results:1.It Obtained 21930111 clean reads for the vegetative sample,and 21612191 for the reproductive sample.It identified 42 conserved miRNAs,and they were shown differentially expressed between the vegetative and the reproductive stage,with 25 miRNAs possessing more than 2-fold difference.Nine miRNAs were upregulated in the vegetative phase,whereas the other was upregulated in the reproductive phase.2.miR156 was downregulated in the reproductive stage,and miR172 was upregulated in this stage,whereas their target genes also correspondingly changed.miR171 and miR159 also displayed expected alterations in accumulation level.Thus,the mechanism controlling the transition of growth phases by miRNAs might originate in ancestral euphyllophytes.3.miR160/166/319/394 might be related to the downward incurvature morphology of fertile leaves in C.pteridoides.tasi R-ARF could be produce in C.pteridoides,indicating miR390–TAS3–ARF signaling firstly occurred in ferns.4.175 novel miRNAs were identified,and they all were differentially expression between the vegetative and the reproductive stage,with target prediction and function enrichment suggesting they should be related to multiple activities.5.Fluorescence quantitative PCR validated alterations in expression for target genes of miR156 and miR172.