GhTBL3 is required for fiber secondary cell wall (<scp>SCW</scp>) formation via maintaining acetylation of xylan in cotton

TBL family proteins containing the domain of unknown function mainly act as xylan O-acetyltransferases, but the specific molecular mechanism of their functions remains unclear in plants (especially in cotton) so far. In this study, we characterized the TBL family proteins containing the conserved GDS and DxxH motifs in cotton (Gossypium hirsutum). Among them, GhTBL3 is highly expressed in fibers at the stage of secondary cell wall (SCW) formation and mainly functions as O-acetyltransferase to maintain acetylation of xylan in fiber SCW development. Overexpression of GhTBL3 in cotton promoted fiber SCW formation, resulting in increased fiber cell wall thickness. In contrast, suppression of GhTBL3 expression in cotton impaired fiber SCW synthesis, leading to the decreased fiber cell wall thickness, compared with wild type (WT). Furthermore, two fiber SCW-related transcription factors GhMYBL1 and GhKNL1 were found to directly bind to the promoter of GhTBL3 in cotton. GhMYBL1 enhanced the transcription activity of GhTBL3, whereas GhKNL1 inhibited the expression of GhTBL3 in fibers. The acetylation level of xylan was remarkably decreased in fibers of GhMYBL1 RNAi transgenic cotton, but the acetylation level of xylan was significantly increased in fibers of GhKNL1 RNAi cotton, relative to WT. Given together, the above results suggested that GhTBL3 may be under the dual control of GhMYBL1 and GhKNL1 to maintain the suitable acetylation level of xylan required for fiber SCW formation in cotton. Thus, our data provide an effective clue for potentially improving fiber quality by genetic manipulation of GhTBL3 in cotton breeding.