The microRNA miR399 plays an important role in phosphorus signal transduction pathways in plants.Previously,miR399 was shown to be closely associated with berry ripening in grape(Vitis vinifera).The objective of the p...The microRNA miR399 plays an important role in phosphorus signal transduction pathways in plants.Previously,miR399 was shown to be closely associated with berry ripening in grape(Vitis vinifera).The objective of the present study was to elucidate the evolutionary characteristics of the miR399 gene family in grape and to verify the cleavage effect on the target genes.Grape miR399s were identified by miRNA sequencing and retrieval from the miRBase database.The mature sequences and precursor sequences were subjected to phylogenetic analysis to reconstruct evolutionary trees,as well as secondary structure analysis of the precursor sequence,and prediction of target genes.The cisacting elements in the miR399 promoter were predicted and the cleavage effect of grape miR399b on its target genes was verified.The grape miR399 family comprised nine precursor sequences and nine mature sequences.The precursor sequences formed a typical and stable stem—loop structure.The minimum folding free energy ranged from-55.70 kcal·mol^(-1)to-37.40 kcal·mol^(-1).Multiple sequence alignment revealed that the miR399 family was highly conserved.The grape miR399 family was phylogenetically closely related to peach,apple,and citrus miR399s.Grape miR399s were predicted to target inorganic phosphate transporter 1—3,phospholipase D delta-like,and beta-glucuronosyltransferase.The cleavage effect of grape miR399b on the target genes was verified by means of a dual-luciferase assay and 5’RLM-RACE.Histochemical GUS staining showed that the promoter activity of miR399b was promoted by GA3treatment.展开更多
基金supported by Natural Science Foundation of China(Grant No.U1904113)National Key Research and Development Program of China(Grant No.2018YFD1000105)+3 种基金Program for Innovative Research Team(in Science and Technology)in University of Henan Province(Grant No.21IRTSTHN021)Program for Science&Technology Innovation Talents in Universities of Henan Province(Grant No.21HASTIT035)Scientific and technological breakthroughs in Henan Province(Grant No.222102110083)PhD Research Startup Foundation of Henan University of Science and Technology(Grant Nos.13480067,13480068)。
文摘The microRNA miR399 plays an important role in phosphorus signal transduction pathways in plants.Previously,miR399 was shown to be closely associated with berry ripening in grape(Vitis vinifera).The objective of the present study was to elucidate the evolutionary characteristics of the miR399 gene family in grape and to verify the cleavage effect on the target genes.Grape miR399s were identified by miRNA sequencing and retrieval from the miRBase database.The mature sequences and precursor sequences were subjected to phylogenetic analysis to reconstruct evolutionary trees,as well as secondary structure analysis of the precursor sequence,and prediction of target genes.The cisacting elements in the miR399 promoter were predicted and the cleavage effect of grape miR399b on its target genes was verified.The grape miR399 family comprised nine precursor sequences and nine mature sequences.The precursor sequences formed a typical and stable stem—loop structure.The minimum folding free energy ranged from-55.70 kcal·mol^(-1)to-37.40 kcal·mol^(-1).Multiple sequence alignment revealed that the miR399 family was highly conserved.The grape miR399 family was phylogenetically closely related to peach,apple,and citrus miR399s.Grape miR399s were predicted to target inorganic phosphate transporter 1—3,phospholipase D delta-like,and beta-glucuronosyltransferase.The cleavage effect of grape miR399b on the target genes was verified by means of a dual-luciferase assay and 5’RLM-RACE.Histochemical GUS staining showed that the promoter activity of miR399b was promoted by GA3treatment.
