The role of microRNAs in recovery rates of Arabidopsis thaliana after short term cryo-storage

Abstract

Key message The alteration of expression levels of cold- and oxidative stress-induced genes related-miRNAs play a crucial role in development of successful cryopreservation protocols. Cryopreservation is a safe and cost-effective long-term storage technique which helps to conserve plant germplasm in liquid nitrogen (LN). However, up to now, the molecular regulatory mechanisms that effect survival after cryopreservation have not been satisfactorily elucidated in plants. Thus, the aim of the study concerned to reveal out microRNA (miRNA)-based post-transcriptional regulations during different stages of cryopreservation in 48-h and 72-h in vitro germinated Arabidopsis thaliana since miRNAs not only regulate gene expression in developmental process of plants, but also control transcription in stress conditions by post-transcriptional degradation. Among the screened 10 miRNAs and their selected target genes, differential expression levels were detected in two tested germination time according to exposure of cryopreservation stages, as 6 miRNAs (miR393, miR397, miR398, miR159, miR169 and miR172) showed a mixed pattern in 48-h germinated seedlings while 6 miRNAs (miR397, miR398, miR159, miR169, miR172 and miR408) were significantly down-regulated in OD (Osmoprotection-Dehydration) and LD (Liquid nitrogen-Dilution) samples in comparison to control in 72-h germinated ones. The most striking expression change was obtained in miR393 as it was up-regulated 15-fold in LN treated 48-h seedlings while relatively lower amount induction (twofold) was recorded in cryopreserved 72-h seedlings. The concominant down-regulation of its target TIR1 and HOS1 expression and up-regulation of CBF1 and CBF3 and their downstream genes (COR15 and COR47) were also observed in cryopreserved 48-h germinated seedlings. Thus, the alteration of expression levels of cold-induced genes related-miRNAs plays a crucial role for successful cryopreservation.