首页|Identification,characterization,and verification of miR399 target gene in grape

Identification,characterization,and verification of miR399 target gene in grape

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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 charac-teristics 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 cis-acting 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 GA3 treatment.

GrapemiR399Phosphorus metabolismEarly ripening

Maosong Pei、Hainan Liu、Tonglu Wei、Huiying Jin、Yihe Yu、Mengting Ma、Xiaomeng Song、Rundong Dai、Dalong Guo

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Henan Engineering Technology Research Center of Quality Regulation and Controlling of Horticultural Plants,College of Horticulture and Plant Protection,Henan University of Science and Technology,Luoyang,Henan 471023,China

Natural Science Foundation of ChinaNational Key Research and Development Program of ChinaProgram for Innovative Research Team(in Science and Technology)in University of Henan ProvinceProgram for Science & Technology Innovation Talents in Universities of Henan ProvinceScientific and technological breakthroughs in Henan ProvincePhD Research Startup Foundation of Henan University of Science and TechnologyPhD Research Startup Foundation of Henan University of Science and Technology

U19041132018YFD100010521IRTSTHN02121HASTIT0352221021100831348006713480068

2024

10.1016/j.hpj.2023.02.009

园艺学报(英文版)

园艺学报(英文版)

CSTPCD
ISSN：
年,卷(期)：2024.10(1)
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