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Background  The  INNER NO OUTER  ( INO ) gene, which encodes a YABBY-type transcription factor, specifies and promotes the growth of the outer integument of the ovule in Arabidopsis.  INO  expression is limited to the abaxial cell layer of the developing outer integument of the ovule and is regulated by multiple regions of the  INO  promoter, including POS9, a positive element that when present in quadruplicate can produce low-level expression in the normal  INO  pattern.    Results  Significant redundancy in activity between different regions of the  INO  promoter is demonstrated. For specific regulatory elements, multimerization or the addition of the cauliflower mosaic virus 35S general enhancer was able to activate expression of reporter gene constructs that were otherwise incapable of expression on their own. A new promoter element, POS6, is defined and is shown to include sufficient positive regulatory information to reproduce the endogenous pattern of expression in ovules, but other promoter regions are necessary to fully suppress expression outside of ovules. The full-length  INO  promoter, but not any of the  INO  promoter deletions tested, is able to act as an enhancer-blocking insulator to prevent the ectopic activation of expression by the 35S enhancer. Sequence conservation between the promoter regions of  Arabidopsis thaliana ,  Brassica oleracea  and  Brassica rapa  aligns closely with the functional definition of the POS6 and POS9 regions, and with a defined  INO  minimal promoter. The  B .  oleracea INO  promoter is sufficient to promote a similar pattern and level of reporter gene expression in Arabidopsis to that observed for the Arabidopsis promoter.    Conclusions  At least two independent regions of the  INO  promoter contain sufficient regulatory information to direct the specific pattern but not the level of  INO  gene expression. These regulatory regions act in a partially redundant manner to promote the expression in a specific pattern in the ovule and suppress expression outside of ovules. Establishment of this pattern requires cooperation and competition between multiple positive and negative regulatory elements.

bmc plant biology

Positive- and negative-acting regulatory elements contribute to the tissue-specific expression of INNER NO OUTER, a YABBY-type transcription factor gene in Arabidopsis