SUMMARYMYB transcription factors have been implicated in regulation of the development of ovule epidermal cells into the elongated seed fibres of cotton. An R2R3 MYB, GhMYB25-like, identified from its reduced expression in a fibreless mutant of cotton (Xu142 fl), is here shown to play a key role in the very early stages of fibre cell differentiation. A GhMYB25-like promoter-GUS construct was expressed predominantly in the epidermal layers of cotton ovules before anthesis ()3 days post-anthesis, dpa), increasing in expression in 0-dpa ovules, primarily in those epidermal cells expanding into fibres, and then in elongating fibres at +3 dpa, declining thereafter. This was consistent with GhMYB25-like transcript abundance during fibre development. RNA interference suppression of GhMYB25-like resulted in cotton plants with fibreless seeds, but normal trichomes elsewhere, phenocopying the Xu142 fl mutant. Like Xu142 fl these plants had reduced expression of the fibreexpressed MYBs, GhMYB25 and GhMYB109, indicating that GhMYB25-like is upstream from those MYBs. This hierarchy was supported by the absence of any change in transcript level of GhMYB25-like in GhMYB25-and GhMYB109-silenced transgenic lines. Transgenic cotton with an additional copy of the native gene had elevated expression of GhMYB25-like in ovules, but no obvious increase in fibre initials, suggesting that there may be other factors that interact with GhMYB25-like to differentiate epidermal cells into fibre cells.

SUMMARYGossypium hirsutum L. (cotton) fibres are specialized trichomes a few centimetres in length that grow from the seed coat. Few genes directly involved in the differentiation of these epidermal cells have been identified. These include GhMYB25-like and GhMYB25, two related MYB transcription factors that regulate fibre cell initiation and expansion. We have also identified a putative homeodomain leucine zipper (HD-ZIP) transcription factor, GhHD-1, expressed in trichomes and early fibres that might play a role in cotton fibre initiation. Here, we characterize GhHD-1 homoeologues from tetraploid G. hirsutum and show, using reporter constructs and quantitative real-time PCR (qRT-PCR), that they are expressed predominantly in epidermal tissues during early fibre development, and in other tissues bearing epidermal trichomes. Silencing of GhHD-1 reduced trichome formation and delayed the timing of fibre initiation. Constitutive overexpression of GhHD-1 increased the number of fibres initiating on the seed, but did not affect leaf trichomes. Expression of GhHD-1 in cotton silenced for different fibre MYBs suggest that in ovules it acts downstream of GhMYB25-like, but is unaffected in GhMYB25-or GhMYB109-silenced plants. Microarray analysis of silencing and overexpression lines of GhHD-1 indicated that it potentially regulates the levels of ethylene and reactive oxidation species (ROS) through a WRKY transcription factor and calcium-signalling pathway genes to activate downstream genes necessary for cell expansion and elongation.

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