Evaluation of gene expression changes of miR156 and miR172 and their targeted genes (AP2 & SPL3; vernalization factors) in two bread wheat (Triticum aestivum L.) cultivars

Floral transition through vernalization has a large influence on cold tolerance and agronomic traits in winter cereals. It is now apparent that in many plants small RNAs play critical roles in determination of the flowering time. There is evidence suggesting that the miR156 and miR172 families play a key role in the flowering transition of plants. In this study, the expression of two temporally regulated miRNA (miR156 and miR172) and their targeted genes (SPL3, AP2) were investigated in the winter bread wheat cv. Norstar and the spring bread wheat cv. Baj in 2017-2018 cropping season. The vernalization was exposed to the cold treatments (4°C) for 2 and 14 days at seedling stage. Time to flowering was estimated using the final leaf number (FLN), which significantly decreased under vernalization treatments, only in winter cultivar 'Norstar'. Moreover, analysis of variance showed that vernalization treatments × cultivarsv interaction effect was significant on FLN. Comparison of gene expression using bootstrapping method showed that the expression of miR172 was significantly down-regulated only in Norstar under vernalization treatments. Similarly, the expression of miR156 was completely different under vernalization treatment in two cultivars. Increased expression of miR156 in Baj cultivar was not significant, but vernalization treatment significantly decreased the expression of this gene in cv. Norstar. Although the induction of AP2 expression during vernalization (14 days) was observed in both cultivars, but, the expression levels of SPL3 were only significantly decreased in cv. Norstar, and this reduction was more in two-day vernalization. Unexpectedly, in this experiment, there was no relationship between up-regulation of both miRNA and down-regulation of their targeted genes in two bread wheat cultivars. These results demonstrated that molecular mechanism of flowering time in bread wheat is complex and still largely unknown.