In this study, we carried out an evolutionary, transcriptional, and functional analyses of the trihelix transcription factor family. A total of 319 trihelix members, identified from 11 land plant species, were classified into five clades. The results of phylogeny indicate the binding domains of GT1 and GT2 diverged early in the existence of land plants. Genomic localization revealed that the trihelix family members were highly conserved among cereal species, even though some homeologs generated during the tetraploidy of maize were lost. Three-dimensional structural analyses and an examination of subcellular localization of this family supported the involvement of all five clades in transcriptional regulation. Furthermore, the family members from all clades in sorghum and rice showed a broad and dynamic expression pattern in response to abiotic stresses, indicating regulatory subfunctionalization of their original functions. This finding is further supported by the phenotypes of enhanced tolerance to cold, salt, and drought in transgenic plants overexpressing Sb06g023980 and Sb06g024110. In contrast, few Arobidopsis genes showed inducible expression under abiotic stress conditions, which may indicate a functional shift. Finally, our co-expression analysis points to the involvement of this family in various metabolic processes, implying their further functional divergence.

Jasmonates (jasmonic acid and its relatives) are a group of oxylipin phytohormones that are implicated in the regulation of a range of developmental processes and responses to environmental stimuli in plants. The biosynthesis of JAs occur sequentially in various subcellular compartments including the chloroplasts, peroxisomes and the cytoplasm. The biologically active jasmonoyl-isoleucine (JA-Ile) activates the core JA signaling in the nucleus by binding with its coreceptor, SCF^COI1-JAZ. Five members of a clade of ATP-binding cassette G (ABCG) transporters of Arabidopsis thaliana were identified as the candidates of jasmonate transporters (JATs) in yeast cells. Among these JATs, AtJAT1/AtABCG16, has a dual localization in the plasma membrane and nuclear envelop and mediates the efflux of jasmonic acid (JA) across the plasma membrane and influx of JA-Ile into the nucleus. Genetic, cellular and biochemical analyses have demonstrated that AtJAT1/AtABCG16 is crucial for modulating JA-Ile concentration in the nucleus to orchestrate JA signaling. AtJAT1 could also be involved in modulating the biosynthesis of JA-Ile by regulating the distribution of JA and JA-Ile in the cytoplasm and nucleus, which would contribute to the highly dynamic JA signaling. Furthermore, other JAT members are localized in the plasma membrane and possibly in peroxisomes. Characterization of these JATs will provide further insights into a crucial role of transporter-mediated subcellular distribution in the metabolism and signaling of plant hormones, an emerging theme supported by the identification of increasing number of endomembrane-localized transporters.

Plant epidermis serves important functions in shoot growth, plant defense and lipid metabolism, though mechanisms of related transcriptional regulation are largely unknown. Here, we identified cis-elements specific to shoot epidermis expression by dissecting the promoter of Triticum aestivum lipid transfer protein 1 (TaLTP1). A preliminary promoter deletion analysis revealed that a truncated fragment within 400 bp upstream from the translation start site was sufficient to confer conserved epidermis-specific expression in transgenic Brachypodium distachyon and Arabidopsis thaliana. Further, deletion or mutation of a GC(N4)GGCC motif at position −380 bp caused a loss of expression in pavement cells. With an electrophoretic mobility shift assay (EMSA) and transgenic reporter assay, we found that a light-responsive CcATC motif at position −268 bp was also involved in regulating pavement cell-specific expression that is evolutionary conserved. Moreover, expression specific to leaf trichome cells was found to be independently regulated by a CCaacAt motif at position −303 bp.