In natural environments, plants compete with neighbouring plants for resources such as light, water and nutrients. To detect neighbours, plants have evolved mechanisms that are poorly understood at the molecular-genetic level. This study examined the impact of competition on the growth and reproductive success of Arabidopsis thaliana grown in crowded settings together with conspecifics or with the grass Lolium perenne. Intraspecific and interspecific competition resulted in strongly reduced shoot branching and silique production. The reduction in shoot branching correlated with greatly altered gene expression in lateral buds, in particular of hormone- and defence-related genes, while it was independent of the hub transcription factor BRANCHED1. Mutants defective in strigolactone signalling retained a response to competition. Similarly, competitors unable to synthesize strigolactones caused a normal inhibitory effect, indicating that strigolactones are not required for a response of Arabidopsis to competition. Fertilization did not overcome the inhibitory effect of competition, showing that plants under competition did not experience a lack of mineral nutrients. When the roots of focal and competitor plants were separated by water-impermeable below-ground partitions, plants did not respond to competition. We suggest that below-ground communication, together with a sensing of soil volume, participates in the response to competition.

Abstract The COP1/SPA ubiquitin ligase is a key repressor of photomorphogenesis that is inactivated by photoreceptors to initiate light signalling. The four SPA proteins (SPA1-SPA4) confer functional specificity to COP1 during plant growth, yet the underlying molecular mechanisms remain unclear. Here, we used a domain-swap approach in transgenic seedlings to address the functional divergence of SPA2 and SPA3. We show that the respective N-terminal kinase domain determines the contrasting protein stabilities of SPA2 and SPA3 in light-grown seedlings. The instability of SPA2 correlates with a specific ability of the SPA2 N-terminal domain to bind phytochrome A in the light, suggesting that phytochrome A promotes the CUL4 DET1/COP1 -dependent degradation of SPA2 but not of SPA3. We uncover that the coiled-coiled and WD-repeat domains of SPA2 and SPA3 substantially differ in their activity in repression of photomorphogenesis, with those of SPA2 being more active repressors than those of SPA3. Thus, SPA2 combines a potent repressor activity with light-induced instability. We conclude that the evolution of SPA2 instability in response to light counterbalances its inherent strong repressor activity, thereby allowing seedling etiolation in darkness followed by rapid reduction in COP1 activity through SPA2 degradation upon light-exposure as seedlings emerge from soil to initiate photosynthetic growth. Highlight The repressor of light signaling SPA2 combines a phytochrome A-interacting instability domain with a potent repressor domain to allow greatly contrasting activities of COP1 in skoto- and photomorphogenesis.

During seedling etiolation in darkness, the biosynthesis of protochlorophyllide (Pchlide) and the development of etioplasts must be strictly controlled to prevent photooxidative damage upon light exposure. The transcription factors GLK1 and GLK2 are central regulators of chlorophyll biosynthesis and chloroplast biogenesis. Here, we show that GLK1 and GLK2 interact with ABSCISIC ACID INSENSITIVE 4 (ABI4). We reveal that GLKs and ABI4 have antagonistic functions in cotyledon greening of light-exposed etiolated seedlings: compared to the wild type, abi4 mutants, similar to a transgenic line overexpressing GLK2, accumulated more Pchlide in dark-grown seedlings, while glk1 glk2 mutants contained less Pchlide. The high Pchlide levels in etiolated abi4 mutants and GLK2 overexpressors were inefficiently photoreduced upon light exposure, leading to a significant accumulation of 1O2 in the cotyledons after the dark-to-light transition. This corresponded to low cotyledon greening rates and low seedling survival. Additionally, we identified eight PhANGs involved in Pchlide biosynthesis and etioplast development, whose transcript accumulation patterns may contribute to the photobleaching of etiolated abi4 mutants and GLK2 overexpressors. Importantly, the high Pchlide content, low cotyledon greening rate, high 1O2 level and high PhANG induction in abi4 mutant seedlings were fully dependent on GLK1 and GLK2, indicating that ABI4 acts upstream of GLKs. Since ABI4 does not regulate GLK transcript level, and ABI4 physically interacts with GLK proteins, these data suggest that ABI4 inhibits GLK1 and GLK2 activities in etiolated seedlings to prevent high Pchlide accumulation which would lead to high 1O2 levels and seedling death upon exposure to light.

COP1 is a conserved ubiquitin ligase found in plants and animals. In plants, COP1 acts together with SPA proteins to suppress light signaling in darkness by promoting the degradation of transcription factors involved in photomorphogenesis. Substrates of this ubiquitin ligase share a Valine-Proline (VP) motif that interacts with the WD-repeat domain of COP1 in plants and humans. Here, we have identified the transcription factor TCP3 as a noncanonical substrate of COP1/SPA that lacks a VP motif. The TCP domain of TCP3 directly interacts with the WD-repeat domains of COP1 and SPA1. TCP3 requires the VP-binding cleft of COP1 for protein–protein interaction. We further show that the TCP3 protein is degraded in darkness and preferentially in short day through a COP1-dependent manner, while TCP3 is stabilized by red, far-red, blue light, and long day conditions. COP1/SPA-mediated degradation of TCP3 inhibits anthocyanin accumulation by reducing the expression of anthocyanin biosynthesis genes. COP1/SPA-mediated degradation of TCP3 is also important in regulating flowering time. Taken together, our results have identified a noncanonical substrate of the COP1/SPA ubiquitin ligase, thereby also uncovering TCPs as a transcription factor family that is targeted by COP1/SPA. Since the COP1/SPA-interacting TCP domain is conserved among TCPs, it is possible that other members of the TCP family—having divergent functions including cell fate determination and hormone signaling—are targets of COP1/SPA as well.

COP1 is a conserved ubiquitin ligase found in plants and animals. In plants, COP1 acts together with SPA proteins to suppress light signaling in darkness by promoting the degradation of transcription factors involved in photomorphogenesis. Substrates of this ubiquitin ligase share a Valine-Proline (VP) motif that interacts with the WD-repeat domain of COP1 in plants and humans. Here, we have identified the transcription factor TCP3 as a noncanonical substrate of COP1/SPA that lacks a VP motif. The TCP domain of TCP3 directly interacts with the WD-repeat domains of COP1 and SPA1. TCP3 requires the VP-binding cleft of COP1 for protein-protein interaction. We further show that the TCP3 protein is degraded in darkness and preferentially in short day through a COP1-dependent manner, while TCP3 is stabilized by red, far-red, blue light, and long day conditions. COP1/SPA-mediated degradation of TCP3 inhibits anthocyanin accumulation by reducing the expression of anthocyanin biosynthesis genes. COP1/SPA-mediated degradation of TCP3 is also important in regulating flowering time. Taken together, our results have identified a noncanonical substrate of the COP1/SPA ubiquitin ligase, thereby also uncovering TCPs as a transcription factor family that is targeted by COP1/SPA. Since the COP1/SPA-interacting TCP domain is conserved among TCPs, it is possible that other members of the TCP family-having divergent functions including cell fate determination and hormone signaling-are targets of COP1/SPA as well.

In blue light, cryptochrome photoreceptors inhibit the key repressor of light signaling, the COP1/SPA ubiquitin ligase, to promote photomorphogenic responses. This inhibition relies on the direct interaction between COP1 and cryptochromes. Here, we analyzed the molecular mechanism of CRY1-mediated inhibition of COP1. We show that the VP motif in the C-terminal domain of CRY1 is essential for the COP1-CRY1 interaction in Arabidopsis. Phenotypic analysis of transgenic Arabidopsis plants harboring a mutation in the VP motif reveals that the VP motif of CRY1 is required for blue light-induced responses, such as seedling de-etiolation and anthocyanin biosynthesis. Via its VP motif, CRY1 inhibits the interaction between COP1 and the COP1 substrate transcription factors PAP2 and HY5. Replacing the VP motif of CRY1 with that of the human COP1 interactor TRIB1 produces a functional photoreceptor in transgenic plants. Since HY5, PAP2 and CRY1 interact with COP1 through their respective VP motifs, our results demonstrate that CRY1 inhibits the activity of COP1 by competitively displacing substrates from COP1. Taken together with previous results showing VP-dependent substrate displacement by photoactivated CRY2 and UVR8 photoreceptors, our results highlight the conservation of this mechanism across multiple photoreceptors.

The zygnematophytes are the closest relatives of land plants and comprise several lineages that adapted to a life on land. Species of the genus Serritaenia form colorful, mucilaginous capsules, which surround the cells and block harmful solar radiation, one of the major terrestrial stressors. In eukaryotic algae, this 'sunscreen mucilage' represents a unique photoprotective strategy, whose induction and chemical background are unknown. We generated a de novo transcriptome of Serritaenia testaceovaginata and studied its gene regulation under moderate UV radiation (UVR) that triggers sunscreen mucilage under experimental conditions. UVR induced the repair of DNA and the photosynthetic apparatus as well as the synthesis of aromatic specialized metabolites. Specifically, we observed pronounced expressional changes in the production of aromatic amino acids, phenylpropanoid biosynthesis genes, potential cross-membrane transporters of phenolics, and extracellular, oxidative enzymes. Interestingly, the most up-regulated enzyme was a secreted class III peroxidase, whose embryophyte homologs are involved in apoplastic lignin formation. Overall, our findings reveal a conserved, plant-like UVR perception system (UVR8 and downstream factors) in zygnematophyte algae and point to a polyphenolic origin of the sunscreen pigment of Serritaenia, whose synthesis might be extracellular and oxidative, resembling that of plant lignins.

Exposure of plants to ultraviolet-B (UV-B) radiation initiates transcriptional responses that modify metabolism, physiology and development to enhance viability in sunlight. Many of these regulatory responses to UV-B radiation are mediated by the photoreceptor UV RESISTANCE LOCUS 8 (UVR8). Following photoreception, UVR8 interacts directly with multiple proteins to regulate gene expression, but the mechanisms that control differential protein binding to initiate distinct responses are unknown. Here we show that UVR8 is phosphorylated at several sites and that UV-B stimulates phosphorylation at Serine 402. Site-directed mutagenesis to mimic Serine 402 phosphorylation promotes binding of UVR8 to REPRESSOR OF UV-B PHOTOMORPHOGENESIS (RUP) proteins, which negatively regulate UVR8 action. Complementation of the uvr8 mutant with phosphonull or phosphomimetic variants suggests that phosphorylation of Serine 402 modifies UVR8 activity and promotes flavonoid biosynthesis, a key UV-B-stimulated response that enhances plant protection and crop nutritional quality. This research provides a basis to understand how UVR8 interacts differentially with effector proteins to regulate plant responses to UV-B radiation.

The Arabidopsis COP1/SPA ubiquitin ligase suppresses photomorphogenesis in darkness. In the light, photoreceptors inactivate COP1/SPA to allow a light response. While SPA genes are specific to the green lineage, COP1 also exists in humans. This raises the question of when in evolution plant COP1 acquired the need for SPA accessory proteins. We addressed this question by generating Physcomitrium Ppcop1 mutants and comparing their visible and molecular phenotypes with those of Physcomitrium Ppspa mutants. The phenotype of Ppcop1 nonuple mutants resembles that of Ppspa mutants. Most importantly, both mutants produce green chloroplasts in complete darkness. They also exhibit dwarfed gametophores, disturbed branching of protonemata and absent gravitropism. RNA-sequencing analysis indicates that both mutants undergo weak constitutive light signaling in darkness. PpCOP1 and PpSPA proteins form a complex and they interact via their WD repeat domains with the VP motif of the cryptochrome CCE domain in a blue light-dependent manner. This resembles the interaction of Arabidopsis SPA proteins with Arabidopsis CRY1, and is different from that with Arabidopsis CRY2. Taken together, the data indicate that PpCOP1 and PpSPA act together to regulate growth and development of Physcomitrium. However, in contrast to their Arabidopsis orthologs, PpCOP1 and PpSPA proteins execute only partial suppression of light signaling in darkness. Hence, additional repressors may exist that contribute to the repression of a light response in dark-exposed Physcomitrium.

Plant terrestrialization brought forth the land plants (embryophytes). Embryophytes account for most of the biomass on land and evolved from streptophyte algae in a singular event. Recent advances have unravelled the first full genomes of the closest algal relatives of land plants; among the first such species was Mesotaenium endlicherianum . Here we used fine-combed RNA sequencing in tandem with a photophysiological assessment on Mesotaenium exposed to a continuous range of temperature and light cues. Our data establish a grid of 42 different conditions, resulting in 128 transcriptomes and ~1.5 Tbp (~9.9 billion reads) of data to study the combinatory effects of stress response using clustering along gradients. Mesotaenium shares with land plants major hubs in genetic networks underpinning stress response and acclimation. Our data suggest that lipid droplet formation and plastid and cell wall-derived signals have denominated molecular programmes since more than 600 million years of streptophyte evolution—before plants made their first steps on land.

Cryptochromes (CRYs) are blue light photoreceptors that regulate growth, development, and metabolism in plants. In Arabidopsis thaliana (Arabidopsis), CRY1 and CRY2 possess partially redundant and overlapping functions. Upon exposure to blue light, the monomeric inactive CRYs undergo phosphorylation and oligomerization, which are crucial to CRY function. Both the N- and C-terminal domains of CRYs participate in light-induced interaction with multiple signaling proteins. These include the COP1/SPA E3 ubiquitin ligase, several transcription factors, hormone signaling intermediates and proteins involved in chromatin-remodeling and RNA N6 adenosine methylation. In this review, we discuss the mechanisms of Arabidopsis CRY signaling in photomorphogenesis and the recent breakthroughs in Arabidopsis CRY research.

Light initiates chloroplast biogenesis in Arabidopsis by eliminating PHYTOCHROME-INTERACTING transcription FACTORs (PIFs), which in turn de-represses nuclear photosynthesis genes, and synchronously, generates a nucleus-to-plastid (anterograde) signal that activates the plastid-encoded bacterial-type RNA polymerase (PEP) to transcribe plastid photosynthesis genes. However, the identity of the anterograde signal remains frustratingly elusive. The main challenge has been the difficulty to distinguish regulators from the plethora of necessary components for plastid transcription and other essential chloroplast functions, such as photosynthesis. Here, we show that the genome-wide induction of nuclear photosynthesis genes is insufficient to activate the PEP. PEP inhibition is imposed redundantly by multiple PIFs and requires PIF3's activator activity. Among the nuclear-encoded components of the PEP holoenzyme, we identify four light-inducible, PIF-repressed sigma factors as anterograde signals. Together, our results elucidate that light-dependent inhibition of PIFs activates plastid photosynthesis genes via sigma factors as anterograde signals in parallel with the induction of nuclear photosynthesis genes.

Yeast-based interaction assays to determine protein-protein and protein-nucleic acid interactions commonly rely on the reconstitution of chimeric transcription factors that activate the expression of target reporter genes. The enzyme β-galactosidase (β-gal), coded by the LacZ gene of Escherichia coli, is a widely used reporter in yeast systems, and its expression is commonly assessed by evaluating its activity. X-gal (5-bromo-4-chloro-3-indolyl-β-d-galactopyranoside) is an inexpensive and sensitive substrate of β-gal, whose hydrolysis results in an intensely blue colored and easily detectable end product, 5,5'-dibromo-4,4'-dichloro-indigo. The insoluble nature of this end product, however, makes X-gal-based assays unsuitable for direct spectrophotometric absorbance quantification. As such, the use of X-gal is mostly restricted to solid-support approaches, such as colony lift or agar plate assays, which often only provide a qualitative readout. In this article, we describe a quantitative solid-phase X-gal assay to measure protein-protein interaction strength in yeast cells using a simple and low-cost experimental setup. We have optimized multiple aspects of the assay, namely sample preparation, reaction time, and quantification method, for speed and consistency. By integrating the use of a freely available ImageJ-based plugin, we have further standardized the assay for reliability and reproducibility. This improved quantitative X-gal assay can be performed in a standard molecular biology lab without the need for any specialized equipment other than an inexpensive and widely accessible smartphone camera. To exemplify the protocol, we provide detailed step-by-step instructions to perform a quantitative X-gal assay to assess the interaction between two Arabidopsis thaliana proteins, SUPPRESSOR OF PHYA-105 1 (SPA1) and PRODUCTION OF ANTHOCYANIN PIGMENT 2 (PAP2). To demonstrate the sensitivity of our assay in detecting weaker interactions, we also compare the results with a liquid-phase assay that uses ONPG (ortho-nitrophenyl-β-galactopyranoside) as a substrate for β-gal. The quantitative X-gal assay described here can easily be adapted for high-throughout interaction studies and protein domain mapping, even in yeast strains with low levels of LacZ expression. © 2022 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Preparation of competent yeast cells and transformation Alternate Protocol 1: In-house preparation of yeast competent cells for use in lithium acetate (LiAc)-mediated yeast transformation Support Protocol: Long-term storage and revival of frozen yeast strain stocks Basic Protocol 2: Measuring β-galactosidase activity via the quantitative X-gal assay Alternate Protocol 2: Quantification of interaction strength using liquid ONPG assay.

CONSTITUTIVE PHOTOMORPHOGENIC 1 functions as an E3 ubiquitin ligase in plants and animals. Discovered originally in Arabidopsis thaliana, COP1 acts in a complex with SPA proteins as a central repressor of light-mediated responses in plants. By ubiquitinating and promoting the degradation of several substrates, COP1/SPA regulates many aspects of plant growth, development and metabolism. In contrast to plants, human COP1 acts as a crucial regulator of tumorigenesis. In this review, we discuss the recent important findings in COP1/SPA research including a brief comparison between COP1 activity in plants and humans.

Phytochromes are photoreceptors regulating growth and development in plants. Using the model plant Arabidopsis, we identified a novel signalling pathway downstream of the far-red light-sensing phytochrome, phyA, that depends on the highly conserved CCR4-NOT complex. CCR4-NOT is integral to RNA metabolism in yeast and animals, but its function in plants is largely unknown. NOT9B, an Arabidopsis homologue of human CNOT9, is a component of the CCR4-NOT complex, and acts as negative regulator of phyA-specific light signalling when bound to NOT1, the scaffold protein of the complex. Light-activated phyA interacts with and displaces NOT9B from NOT1, suggesting a potential mechanism for light signalling through CCR4-NOT. ARGONAUTE 1 and proteins involved in splicing associate with NOT9B and we show that NOT9B is required for specific phyA-dependent alternative splicing events. Furthermore, association with nuclear localised ARGONAUTE 1 raises the possibility that NOT9B and CCR4-NOT are involved in phyA-modulated gene expression.

The biosynthesis of defensive secondary metabolites, such as glucosinolates (GSLs), is a costly process, which requires nutrients, ATP, and reduction equivalents, and, therefore, needs well-orchestrated machinery while coordinating defense and growth. We discovered that the key repressor of light signaling, the CONSTITUTIVE PHOTOMORPHOGENIC 1/SUPPRESSOR OF PHYTOCHROME A-105 (COP1/SPA) complex, is a crucial component of GSL biosynthesis regulation. Various mutants in this COP1/SPA complex exhibited a strongly reduced level of GSL and a low expression of jasmonate (JA)-dependent genes. Furthermore, cop1, which is known to accumulate DELLA proteins in the dark, shows reduced gibberellin (GA) and JA signaling, thereby phenocopying other DELLA-accumulating mutants. This phenotype can be complemented by a dominant gain-of-function allele of MYC3 and by crossing with a mutant having low DELLA protein levels. Hence, SPA1 interacts with DELLA proteins in a yeast two-hybrid screen, whereas high levels of DELLA inhibit MYC function and suppress JA signaling. DELLA accumulation leads to reduced synthesis of GSL and inhibited growth. Thus, the COP1/SPA-mediated degradation of DELLA not only affects growth but also regulates the biosynthesis of GSLs.

Arabidopsis (Arabidopsis thaliana) CONSTITUTIVELY PHOTOMORPHOGENIC1 (COP1) and members of the SUPPRESSOR OF PHYTOCHROMEA-105 (SPA) protein family form an E3 ubiquitin ligase that suppresses light signaling in darkness by polyubiquitinating positive regulators of the light response. COP1/SPA is inactivated by light to allow photomorphogenesis to proceed. Mechanisms of inactivation include light-induced degradation of SPA1 and, in particular, SPA2, corresponding to a particularly efficient inactivation of COP1/SPA2 by light. Here, we show that SPA3 and SPA4 proteins are stable in the light, indicating that light-induced destabilization is specific to SPA1 and SPA2, possibly related to the predominant function of SPA1 and SPA2 in dark-grown etiolating seedlings. SPA2 degradation involves cullin and the COP10-DEETIOLATED-DAMAGED-DNA BINDING PROTEIN (DDB1) CDD complex, besides COP1. Consistent with this finding, light-induced SPA2 degradation required the DDB1-interacting Trp-Asp (WD)-repeat domain of SPA2. Deletion of the N-terminus of SPA2 containing the kinase domain led to strong stabilization of SPA2 in darkness and fully abolished light-induced degradation of SPA2. This prevented seedling de-etiolation even in very strong far-red and blue light and reduced de-etiolation in red light, indicating destabilization of SPA2 through its N-terminal domain is essential for light response. SPA2 is exclusively destabilized by phytochrome A in far-red and blue light. However, deletion of the N-terminal domain of SPA2 did not abolish SPA2-phytochrome A interaction in yeast nor in vivo. Our domain mapping suggests there are two SPA2-phytochrome A interacting domains, the N-terminal domain and the WD-repeat domain. Conferring a light-induced SPA2-phyA interaction only via the WD-repeat domain may thus not lead to COP1/SPA2 inactivation.

The Arabidopsis COP1/SPA complex acts as a cullin4-based E3 ubiquitin ligase to suppress photomorphogenesis in darkness. It is a tetrameric complex of two COP1 and two SPA proteins. Both COP1 and SPA are essential for the activity of this complex, and they both contain a C-terminal WD-repeat domain responsible for substrate recruitment and binding of DDB1. Here, we used a WD domain swap-approach to address the cooperativity of COP1 and SPA proteins. We found that expression of a chimeric COP1 carrying the WD-repeat domain of SPA1 mostly complemented the cop1-4-mutant phenotype in darkness, indicating that the WD repeat of SPA1 can replace the WD repeat of COP1. In the light, SPA1-WD partially substituted for COP1-WD. In contrast, expression of a chimeric SPA1 protein carrying the WD repeat of COP1 did not rescue the spa-mutant phenotype. Together, our findings demonstrate that a SPA1-type WD repeat is essential for COP1/SPA activity, while a COP1-type WD is in part dispensible. Moreover, a complex with four SPA1-WDs is more active than a complex with only two SPA1-WDs. A homology model of SPA1-WD based on the crystal structure of COP1-WD uncovered two insertions and several amino acid substitutions at the predicted substrate-binding pocket of SPA1-WD.

Phytochromes are red/far-red light receptors in plants involved in the regulation of growth and development. Phytochromes can sense the light environment and contribute to measuring day length; thereby, they allow plants to respond and adapt to changes in the ambient environment. Two well-characterized signalling pathways act downstream of phytochromes and link light perception to the regulation of gene expression. The CONSTITUTIVELY PHOTOMORPHOGENIC 1/SUPPRESSOR OF PHYA-105 (COP1/SPA) E3 ubiquitin ligase complex and the PHYTOCHROME INTERACTING FACTORs (PIFs) are key components of these pathways and repress light responses in the dark. In light-grown seedlings, phytochromes inhibit COP1/SPA and PIF activity and thereby promote light signalling. In a yeast-two-hybrid screen for proteins binding to light-activated phytochromes, we identified COLD-REGULATED GENE 27 (COR27). COR27 and its homologue COR28 bind to phyA and phyB, the two primary phytochromes in seed plants. COR27 and COR28 have been described previously with regard to a function in the regulation of freezing tolerance, flowering and the circadian clock. Here, we show that COR27 and COR28 repress early seedling development in blue, far-red and in particular red light. COR27 and COR28 contain a conserved Val-Pro (VP)-peptide motif, which mediates binding to the COP1/SPA complex. COR27 and COR28 are targeted for degradation by COP1/SPA and mutant versions with a VP to AA amino acid substitution in the VP-peptide motif are stabilized. Overall, our data suggest that COR27 and COR28 accumulate in light but act as negative regulators of light signalling during early seedling development, thereby preventing an exaggerated response to light.

Photomorphogenesis is repressed in the dark mainly by an E3 ubiquitin ligase complex comprising CONSTITUTIVE PHOTOMORPHOGENIC 1 (COP1) and four homologous proteins called SUPPRESSOR OF PHYA-105 (SPA1-SPA4) in Arabidopsis. This complex induces the ubiquitination and subsequent degradation of positively acting transcription factors (e.g., HY5, HFR1, PAP1 and others) in the dark to repress photomorphogenesis. Genomic evidence showed a large number of genes regulated by COP1 in the dark, of which many are direct targets of HY5. However, the genomic basis for the constitute photomorphogenic phenotype of spaQ remains unknown. Here, we show that >7200 genes are differentially expressed in the spaQ background compared to wild-type in the dark. Comparison of the RNA Sequencing (RNA-Seq) data between cop1 and spaQ revealed a large overlapping set of genes regulated by the COP1-SPA complex. In addition, many of the genes coordinately regulated by the COP1-SPA complex are also regulated by HY5 directly and indirectly. Taken together, our data reveal that SPA proteins repress photomorphogenesis by controlling gene expression in concert with COP1, likely through regulating the abundance of downstream transcription factors in light signaling pathways. Moreover, SPA proteins may function both in a COP1-dependent and –independent manner in regulating many biological processes and developmental pathways in Arabidopsis. Summary statement Comparison of transcriptome analyses between cop1 and spaQ mutants reveal overlapping pathways regulated by COP1 and SPAs.