Abstract

MADS box genes encode transcription factors that play important regulatory roles at various stages in plant development. Transcripts encoding the MIKC*-type (for MADS DNA-binding domain, Intervening domain, Keratin-like domain, and C-terminal domain) factors, a divergent clade, are enriched in mature pollen. Previous studies have shown that these proteins bind DNA as heterodimers, which form between S- and P-class MIKC* proteins. In this study, Arabidopsis (Arabidopsis thaliana) pollen with little or no MIKC* activity was produced by combining strong loss-of-function alleles of the S-class proteins AGAMOUS-LIKE66 (AGL66) and AGL104. Double mutant plants produce pollen but have severely reduced fertility due to reduced pollen viability, delayed germination, and aberrant pollen tube growth. Microarray analysis of the mutant pollen revealed that the loss of MIKC* regulation has a major impact on pollen gene expression. Pollen competition assays involving various combinations of AGL65, AGL66, AGL104, and AGL94 mutant alleles provided genetic evidence that at least three heterodimers (AGL30-AGL104, AGL65-AGL104, and AGL30-AGL66) form and function in at least a partially redundant fashion in pollen. Analyses of transcript abundance in wild-type and mutant pollen indicated that AGL65-containing complexes are likely to be more abundant than the others and that accumulation of AGL30 and AGL94 transcripts increases in response to reductions in MIKC* activity. These results were combined to create a model to describe MIKC* heterodimer contributions in pollen.