Abstract
OsMADS16, a class B floral organ identity gene, plays a pivotal role in stamen formation in rice. To date, little is known about the interacting partners of OsMADS16 except for several MADS-box proteins. In this study, we constructed a high-quality cDNA library of young panicles (< 5 cm in length) and performed yeast two-hybrid (Y2H) screening using OsMADS16 as bait. Eleven candidate proteins interacting with OsMADS16 were identified by Y2H and validated by BiFC and Co-IP assays. Subcellular localization results further confirmed the possibility of the interactions of OsMADS16 with 10 of the candidate proteins in natural rice cells. Bioinformatics analysis indicated that these partners exerted various molecular, cellular and physiological functions. Some of them were known or likely to be related to reproductive events, such as stamen primordium initiation, differentiation and development (OsMADS2, OsMADS4 and OsCOP9) and pollen development (OsbHLH40 and Os6PGDH). Our results provide an important reference for further research on OsMADS16-mediated regulation mechanism on floral organ development and pollen formation.
Highlights
Based on several studies on the floral organ development of eudicots, including Arabidopsis thaliana, Petunia hybrida, and Antirrhinum majus, the classical “ABC” model was proposed to elucidate the genetic and molecular mechanism of floral meristem determinacy and floral organ identity [1,2,3]
We identified eleven candidate partner proteins of OsMADS16 through Y2H, bimolecular fluorescence complementation (BiFC) and co-immunoprecipitation (Co-IP) assay, and demonstrated by bioinformatics analysis and subcellular localization assay that the interactions of OsMADS16 with its partners might exert distinct biological function mainly in nucleus
Total RNA was isolated from young panicles of MH86 using Trizol Reagent (Vazyme), and cDNA for library was gained from the total RNA using SMARTTM cDNA synthesis technology and amplified by long distance PCR
Summary
Based on several studies on the floral organ development of eudicots, including Arabidopsis thaliana, Petunia hybrida, and Antirrhinum majus, the classical “ABC” model (later modified as the “ABCDE” model) was proposed to elucidate the genetic and molecular mechanism of floral meristem determinacy and floral organ identity [1,2,3]. In this model, the class A/B/C genes function individually or in combination with each other, leading to the formation of sepals, petals, stamens and carpels.
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