The miR156/ SPL2 module regulates floral organ development and plant fertility by activating AS2 in Arabidopsis thaliana
Citation
The miR156/ SPL2 module regulates floral organ development and plant fertility by activating AS2 in Arabidopsis thaliana
Plain language summary
MicroRNA156 (miR156) regulates plant growth and development by silencing members of the SQUAMOSA PROMOTER BINDING PROTEIN-LIKE (SPL) gene family. Using Next Generation Sequencing we identified SPL2 and ASYMMETRIC LEAVES 2 (AS2) as candidate genes responsible for the small silique phenotype in miR156-overexpression (OE) plants. QRT-PCR analysis revealed that both miR156-OE and dominant negative SPL2 plants had reduced AS2 transcript levels in their siliques relative to the WT control. Furthermore, phenotypic analysis showed similarities between the AS2 loss-of-function mutant (as2) and the miR156-OE and dominant negative SPL2 plants, including reduced sepal, petal, and silique size as well as reduced fertility. To prove that AS2 is a direct target of SPL2, we conducted Chromatin Immunoprecipitation-qPCR (ChIP-qPCR) and demonstrated that SPL2 can bind directly to the 5’UTR of AS2. Together, these results show that the miR156/SPL2 module affects floral organ size and plant fertility by regulating AS2.
Abstract
MicroRNA156 (miR156) regulates plant growth and development by silencing members of the SQUAMOSA PROMOTER BINDING PROTEIN-LIKE (SPL) gene family. Using Next Generation Sequencing we identified SPL2 and ASYMMETRIC LEAVES 2 (AS2) as candidate genes responsible for the small silique phenotype in miR156-overexpression (OE) plants. QRT-PCR analysis revealed that both miR156-OE and dominant negative SPL2 plants had reduced AS2 transcript levels in their siliques relative to the WT control. Furthermore, phenotypic analysis showed similarities between the AS2 loss-of-function mutant (as2) and the miR156-OE and dominant negative SPL2 plants, including reduced sepal, petal, and silique size as well as reduced fertility. To prove that AS2 is a direct target of SPL2, we conducted Chromatin Immunoprecipitation-qPCR (ChIP-qPCR) and demonstrated that SPL2 can bind directly to the 5’UTR of AS2. Together, these results show that the miR156/SPL2 module affects floral organ size and plant fertility by regulating AS2.