The plant cell wall is an important factor for determining cell shape, function and response to the environment. Secondary cell walls, such as those found in xylem, are composed of cellulose, hemicelluloses and lignin and account for the bulk of plant biomass. The coordination between transcriptional regulation of synthesis for each polymer is complex and vital to cell function. A regulatory hierarchy of developmental switches has been proposed, although the full complement of regulators remains unknown. Here we present a protein–DNA network between Arabidopsis thaliana transcription factors and secondary cell wall metabolic genes with gene expression regulated by a series of feed-forward loops. This model allowed us to develop and validate new hypotheses about secondary wall gene regulation under abiotic stress. Distinct stresses are able to perturb targeted genes to potentially promote functional adaptation. These interactions will serve as a foundation for understanding the regulation of a complex, integral plant component.

Plants sense neighbor proximity as a decrease in the ratio of red to far-red light, which triggers a series of developmental responses. In Arabidopsis, phytochrome B (PHYB) is the major sensor of shade, but PHYB excitation has not been linked directly to a growth response. We show that the basic helix–loop–helix (bHLH) transcription factor PIF7 (phytochrome-interacting factor 7), an interactor of PHYB, accumulates in its dephosphorylated form in shade, allowing it to bind auxin biosynthetic genes and increase their expression. New auxin synthesized through a PIF7-regulated pathway is required for shade-induced growth, linking directly the perception of a light quality signal to a rapid growth response.

Correct daily phasing of transcription confers an adaptive advantage to almost all organisms, including higher plants. In this study, we describe a hypothesis-driven network discovery pipeline that identifies biologically relevant patterns in genome-scale data. To demonstrate its utility, we analyzed a comprehensive matrix of time courses interrogating the nuclear transcriptome of Arabidopsis thaliana plants grown under different thermocycles, photocycles, and circadian conditions. We show that 89% of Arabidopsis transcripts cycle in at least one condition and that most genes have peak expression at a particular time of day, which shifts depending on the environment. Thermocycles alone can drive at least half of all transcripts critical for synchronizing internal processes such as cell cycle and protein synthesis. We identified at least three distinct transcription modules controlling phase-specific expression, including a new midnight specific module, PBX/TBX/SBX. We validated the network discovery pipeline, as well as the midnight specific module, by demonstrating that the PBX element was sufficient to drive diurnal and circadian condition-dependent expression. Moreover, we show that the three transcription modules are conserved across Arabidopsis, poplar, and rice. These results confirm the complex interplay between thermocycles, photocycles, and the circadian clock on the daily transcription program, and provide a comprehensive view of the conserved genomic targets for a transcriptional network key to successful adaptation. As the earth rotates, environmental conditions oscillate between illuminated warm days and dark cool nights. Plants have adapted to these changes by timing physiological processes to specific times of the day or night. Light and temperature signaling and the circadian clock regulate this adaptive response. To determine the contributions of each of these factors on gene regulation, we analyzed microarray time course experiments interrogating light, temperature, and circadian conditions. We discovered that almost all Arabidopsis genes cycle in at least one condition. From a signaling perspective, this suggests that light, temperature, and circadian clock play an important role in modulating many physiological pathways. To clarify the contribution of transcriptional regulation on this process, we mined the promoters of cycling genes to identify DNA elements associated with expression at specific times of day. This confirmed the importance of several DNA motifs such as the G-box and the evening element in the regulation of gene expression by light and the circadian clock, but also facilitated the discovery of new elements linked to a novel midnight regulatory module. Identification of orthologous promoter elements in rice and poplar revealed a conserved transcriptional regulatory network that allows global adaptation to the ever-changing daily environment.

Transcriptional feedback loops constitute the molecular circuitry of the plant circadian clock. In Arabidopsis , a core loop is established between CCA1 and TOC1. Although CCA1 directly represses TOC1 , the TOC1 protein has no DNA binding domains, which suggests that it cannot directly regulate CCA1 . We established a functional genomic strategy that led to the identification of CHE, a TCP transcription factor that binds specifically to the CCA1 promoter. CHE is a clock component partially redundant with LHY in the repression of CCA1 . The expression of CHE is regulated by CCA1, thus adding a CCA1/CHE feedback loop to the Arabidopsis circadian network. Because CHE and TOC1 interact, and CHE binds to the CCA1 promoter, a molecular linkage between TOC1 and CCA1 gene regulation is established.

▪ Abstract Plants harbor four families of kinases that have been implicated in Ca²⁺ signaling (CDPKs, CRKs, CCaMKs, and SnRK3s). Although each family appears to respond to Ca²⁺ via different mechanisms, they all utilize Ca²⁺ sensors that bind Ca²⁺ through multiple EF-hands. The CDPK (Ca²⁺-dependent protein kinase) family is represented by the most genes, with 12 subfamilies comprised of 34 isoforms in Arabidopsis and 27 in rice. Some of the calcium-regulated kinases also show potential for regulation by lipid signals and kinase cascades. Thus, Ca²⁺-regulated kinases provide potential nodes of cross-talk for multiple signaling pathways that integrate Ca²⁺ signals into all aspects of plant growth and development.

Summary Plants have diverse strategies to cope with stress, including early flowering to “escape” abiotic stress and late flowering to mitigate biotic stress. Plants are usually exposed to multiple stresses simultaneously, but little is known about the impact of multiple co-occurring stresses on plant evolution. We tested for adaptation to both aridity and interspecific competition of the model plant Brachypodium spp., collected along the aridity gradient in Israel. We recorded flowering time and estimated fitness in a controlled watering experiment, with treatments mimicking Mediterranean and arid precipitation, and in two common gardens located in the extremes of the gradient (i.e., desert and mesic Mediterranean). At the latter we also manipulated interspecific competition to examine the combined effect of competition and aridity. Plants from arid environments always flowered earlier, but we found no selection on flowering time in the watering experiment. In the common gardens, however, the direction of selection on flowering time differed between sites and competition treatments. We conclude that interactions between aridity and competition drive local adaptation of Brachypodium in the Eastern Mediterranean basin. Variation in flowering time is an important adaptive mechanism to aridity and multiple selection agents can have interactive effects on the evolution of this trait.