title: Genetic Enhancer Analysis Reveals that FLORAL ORGAN NUMBER2 and OsMADS3 Co-operatively Regulate Maintenance and Determinacy of the Flower Meristem in Rice.
abstract: Meristems such as the shoot apical meristem and flower meristem (FM) act as a reservoir of stem cells, which reproduce themselves and supply daughter cells for the differentiation of lateral organs. In Oryza sativa (rice), the FLORAL ORGAN NUMBER2 (FON2) gene, which is similar to Arabidopsis CLAVATA3, is involved in meristem maintenance. In fon2 mutants, the numbers of floral organs are increased due to an enlargement of the FM. To identify new factors regulating meristem maintenance in rice, we performed a genetic screening of mutants that enhanced the fon2 mutation, and found a mutant line (2B-424) in which pistil number was dramatically increased. By using a map-based approach and next-generation sequencing, we found that the line 2B-424 had a complete loss-of-function mutation (a large deletion) in OsMADS3, a class C MADS-box gene that is known to be involved in stamen specification. Disruption of OsMADS3 in the fon2 mutant by CRISPR-Cas9 (clustered regularly interspaced short palindromic repeats-CRISPR-associated protein 9) technology caused a flower phenotype similar to that of 2B-424, confirming that the gene responsible for enhancement of fon2 was OsMADS3. Morphological analysis showed that the fon2 and osmads3 mutations synergistically affected pistil development and FM determinacy. We also found that whorl 3 was duplicated in mature flowers and the FM was enlarged at an early developmental stage in severe osmads3 single mutants. These findings suggest that OsMADS3 is involved not only in FM determinacy in late flower development but also in FM activity in early flower development.
authors.affiliation: Department of Biological Sciences, School of Science, The University of Tokyo, Tokyo, Japan.
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Yasui
authors.affiliation: Department of Biological Sciences, School of Science, The University of Tokyo, Tokyo, Japan.
authors.forename: Wakana
authors.initials: W
authors.lastname: Tanaka
authors.affiliation: Plant Global Education Project, Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, Japan.|Faculty of Life Sciences, Kyoto Sangyo University, Motoyama, Kamigamo, Kita-Ku, Kyoto, Japan.
authors.forename: Tomoaki
authors.initials: T
authors.lastname: Sakamoto
authors.affiliation: Plant Global Education Project, Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, Japan.|Graduate School of Life Sciences, Tohoku University, Sendai, Japan.
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Kurata
authors.affiliation: Department of Biological Sciences, School of Science, The University of Tokyo, Tokyo, Japan.
authors.forename: Hiro-Yuki
authors.initials: HY
authors.lastname: Hirano
mesh_terms: D035264:Flowers / Q000166:cytology* / Q000235:genetics / Q000378:metabolism*; D018506:Gene Expression Regulation, Plant / Q000235:genetics / Q000502:physiology; D026161:MADS Domain Proteins / Q000235:genetics / Q000378:metabolism; D018519:Meristem / Q000166:cytology* / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D012275:Oryza / Q000166:cytology* / Q000235:genetics / Q000378:metabolism*; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000166:cytology / Q000235:genetics / Q000378:metabolism
chemical_list: D026161:MADS Domain Proteins; D010940:Plant Proteins
keywords: Determinacy; FON2; Flower development; MADS-box gene; Meristem; Stem cell