Bioinformatic and Expression Pattern Analysis of dfr-miR160a and Target Gene DfARF10 in Dryopteris fragrans
To further understand the molecular mechanism underlying miRNA regulation of growth and development of Dryopteris fragrans,we screened the differentially expressed dfr-pri-mir160a through the miRNA database established earlier in the laboratory,and predicted its target gene as DfARF10.The target relationship between dfr-pri-mir160a and DfARF10 was verified by tobacco transient co-transformation,together with double luciferase(LUC)activity.The results showed that the GUS and LUC activity in tobacco leaves co-injected with dfr-pri-mir160a and DfARF10 decreased sig-nificantly.qRT-PCR analysis showed that dfr-miR160a and its target gene DfARF10 were expressed in the gameto-phytes,roots,petioles,leaves and sporangium of D.fragrans,with the highest expression in the leaves and the lowest in the roots.We analyzed the effects of drought,NaCl,high temperature and low temperature stress treatments on dfr-miR160a and its target gene DfARF10 through qRT-PCR.Under drought and high temperature treatment,the relative expression of dfr-miR160a was up-regulated,but under NaCl treatment,the expression of dfr-miR160a was down-regulated.Under low temperature treatment,the expression of dfr-miR160a was down-regulated at 0-1 h and was up-regulated at 3-48 h.The expression of DfARF10 was up-regulated under NaCl,high temperature and low temperature treatments.However,under drought treatment,the expression of DfARF10 decreased,distinct from dfr-miR160a.The above results indicated that target gene of dfr-miR160 was DfARF10,and both of them can respond to abiotic stress treatment.This study provides a new scientific basis for revealing the abiotic stress resistance mechanism of D.fragrans at the molecular level.
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