The formation of leaves and other lateral organs in plants depends on the proper specification of adaxial-abaxial (upper-lower) polarity. KANADI1 (KAN1), a member of the GARP family of transcription factors, is a key regulator of abaxial identity, leaf growth, and meristem formation in Arabidopsis thaliana. Here, we demonstrate that the Myb-like domain in KAN1 binds the 6-bp motif GNATA(A/T) and that this motif alone is sufficient to squelch transcription of a linked reporter in vivo. In addition, we report that KAN1 acts as a transcriptional repressor. Among its targets are genes involved in auxin biosynthesis, auxin transport, and auxin response. Furthermore, we find that the adaxializing HD-ZIPIII transcription factor REVOLUTA has opposing effects on multiple components of the auxin pathway. We hypothesize that HD-ZIPIII and KANADI transcription factors pattern auxin accumulation and responsiveness in the embryo. Specifically, we propose the opposing actions of KANADI and HD-ZIPIII factors on cotyledon formation (KANADI represses and HD-ZIPIII promotes cotyledon formation) occur through their opposing actions on genes acting at multiple steps in the auxin pathway.
Expansins are non-hydrolytic cell wall-loosening proteins that are involved in the cell wall modifications that underlie many plant developmental processes. Root hair growth requires the accumulation of cell wall materials and dynamic cell wall modification at the tip region. Although several lines of indirect evidence support the idea that expansin-mediated wall modification occurs during root hair growth, the involvement of these proteins remains to be demonstrated in vivo. In this study, we used RNA interference (RNAi) to examine the biological function of Arabidopsis thaliana EXPANSIN A7 (AtEXPA7), which is expressed specifically in the root hair cell. The root hairspecific AtEXPA7 promoter was used to drive RNAi expression, which targeted two independent regions in the AtEXPA7 transcript. Quantitative reverse transcriptase-PCR analyses were used to examine AtEXPA7 transcript levels. In four independent RNAi transformant lines, RNAi expression reduced AtEXPA7 transcript levels by 25-58% compared to controls. Accordingly, the root hairs of RNAi transformant lines were 25-48% shorter than control plants and exhibited a broader range of lengths than the controls. Our results provide in vivo evidence that expansins are required for root hair tip growth.
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