title: MicroRNA Maturation and MicroRNA Target Gene Expression Regulation Are Severely Disrupted in Soybean dicer-like1 Double Mutants.
abstract: Small nonprotein-coding microRNAs (miRNAs) are present in most eukaryotes and are central effectors of RNA silencing-mediated mechanisms for gene expression regulation. In plants, DICER-LIKE1 (DCL1) is the founding member of a highly conserved family of RNase III-like endonucleases that function as core machinery proteins to process hairpin-like precursor transcripts into mature miRNAs, small regulatory RNAs, 21-22 nucleotides in length. Zinc finger nucleases (ZFNs) were used to generate single and double-mutants of putative soybean DCL1 homologs, DCL1a and DCL1b, to confirm their functional role(s) in the soybean miRNA pathway. Neither DCL1 single mutant, dcl1a or dcl1b plants, exhibited a pronounced morphological or molecular phenotype. However, the dcl1a/dcl1b double mutant expressed a strong morphological phenotype, characterized by reduced seed size and aborted seedling development, in addition to defective miRNA precursor transcript processing efficiency and deregulated miRNA target gene expression. Together, these findings indicate that the two soybean DCL1 paralogs, DCL1a and DCL1b, largely play functionally redundant roles in the miRNA pathway and are essential for normal plant development.
authors.affiliation: Department of Plant Pathology, University of Minnesota, St. Paul, Minnesota, 55108 curti242@umn.edu rstupar@umn.edu.
authors.forename: Shaun J
authors.identifier: http://orcid.org/0000-0002-9528-3335
authors.initials: SJ
authors.lastname: Curtin
authors.affiliation: Department of Agronomy and Plant Genetics, University of Minnesota, St. Paul, Minnesota, 55108.
authors.forename: Jean-Michel
authors.initials: JM
authors.lastname: Michno
authors.affiliation: Department of Agronomy and Plant Genetics, University of Minnesota, St. Paul, Minnesota, 55108.
authors.forename: Benjamin W
authors.initials: BW
authors.lastname: Campbell
authors.affiliation: Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, Minnesota 55455 Center for Genome Engineering, University of Minnesota, Minneapolis, Minnesota 55455.
authors.forename: Javier
authors.initials: J
authors.lastname: Gil-Humanes
authors.affiliation: Department of Plant and Soil Sciences, University of Delaware, Newark, Delaware 19711.
authors.forename: Sandra M
authors.initials: SM
authors.lastname: Mathioni
authors.affiliation: Department of Plant and Soil Sciences, University of Delaware, Newark, Delaware 19711.
authors.forename: Reza
authors.initials: R
authors.lastname: Hammond
authors.affiliation: Department of Agronomy and Plant Genetics, University of Minnesota, St. Paul, Minnesota, 55108.
authors.forename: Juan J
authors.initials: JJ
authors.lastname: Gutierrez-Gonzalez
authors.affiliation: Department of Agronomy and Plant Genetics, University of Minnesota, St. Paul, Minnesota, 55108.
authors.forename: Ryan C
authors.initials: RC
authors.lastname: Donohue
authors.affiliation: Department of Agronomy and Plant Genetics, University of Minnesota, St. Paul, Minnesota, 55108.
authors.forename: Michael B
authors.initials: MB
authors.lastname: Kantar
authors.affiliation: School of Environmental and Life Sciences, The University of Newcastle, Callaghan, New South Wales 2308, Australia.
authors.forename: Andrew L
authors.initials: AL
authors.lastname: Eamens
authors.affiliation: Department of Plant and Soil Sciences, University of Delaware, Newark, Delaware 19711.
authors.forename: Blake C
authors.initials: BC
authors.lastname: Meyers
authors.affiliation: Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, Minnesota 55455 Center for Genome Engineering, University of Minnesota, Minneapolis, Minnesota 55455.
authors.forename: Daniel F
authors.initials: DF
authors.lastname: Voytas
authors.affiliation: Department of Agronomy and Plant Genetics, University of Minnesota, St. Paul, Minnesota, 55108 curti242@umn.edu rstupar@umn.edu.
authors.forename: Robert M
authors.initials: RM
authors.lastname: Stupar
mesh_terms: D000483:Alleles; D001483:Base Sequence; D016000:Cluster Analysis; D020869:Gene Expression Profiling; D018506:Gene Expression Regulation, Plant; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010641:Phenotype; D011485:Protein Binding; D034622:RNA Interference; D020871:RNA Stability; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D043244:Ribonuclease III / Q000235:genetics*; D012260:Ribonucleases / Q000378:metabolism; D013025:Glycine max / Q000235:genetics*; D016335:Zinc Fingers
chemical_list: D035683:MicroRNAs; D012333:RNA, Messenger; D012260:Ribonucleases; D043244:Ribonuclease III
keywords: ZFN; dicer-like; genome engineering; miRNA; soybean