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G3 (Bethesda). 2015 Dec 17;6(2):423-33. doi: 10.1534/g3.115.022137.

MicroRNA Maturation and MicroRNA Target Gene Expression Regulation Are Severely Disrupted in Soybean dicer-like1 Double Mutants.

Author information

1
Department of Plant Pathology, University of Minnesota, St. Paul, Minnesota, 55108 curti242@umn.edu rstupar@umn.edu.
2
Department of Agronomy and Plant Genetics, University of Minnesota, St. Paul, Minnesota, 55108.
3
Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, Minnesota 55455 Center for Genome Engineering, University of Minnesota, Minneapolis, Minnesota 55455.
4
Department of Plant and Soil Sciences, University of Delaware, Newark, Delaware 19711.
5
School of Environmental and Life Sciences, The University of Newcastle, Callaghan, New South Wales 2308, Australia.
6
Department of Agronomy and Plant Genetics, University of Minnesota, St. Paul, Minnesota, 55108 curti242@umn.edu rstupar@umn.edu.

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.

KEYWORDS:

ZFN; dicer-like; genome engineering; miRNA; soybean

PMID:
26681515
PMCID:
PMC4751560
DOI:
10.1534/g3.115.022137
[Indexed for MEDLINE]
Free PMC Article

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