Control Of Hypocotyl Elongation Research Articles

Myb-like transcription factors have epistatic effects on circadian clock function but additive effects on plant growth.

The functions of closely related Myb-like repressor and Myb-like activator proteins within the plant circadian oscillator have been well-studied as separate groups, but the genetic interactions between them are less clear. We hypothesized that these repressors and activators would interact additively to regulate both circadian and growth phenotypes. We used CRISPR-Cas9 to generate new mutant alleles and performed physiological and molecular characterization of plant mutants for five of these core Myb-like clock factors compared with a repressor mutant and an activator mutant. We first examined circadian clock function in plants likely null for both the repressor proteins, CIRCADIAN CLOCK ASSOCIATED 1 (CCA1) and LATE ELONGATED HYPOCOTYL (LHY), and the activator proteins, REVEILLE 4 (RVE4), REVEILLE (RVE6), and REVEILLE (RVE8). The rve468 triple mutant has a long period and flowers late, while cca1 lhy rve468 quintuple mutants, similarly to cca1 lhy mutants, have poor circadian rhythms and flower early. This suggests that CCA1 and LHY are epistatic to RVE4, RVE6, and RVE8 for circadian clock and flowering time function. We next examined hypocotyl elongation and rosette leaf size in these mutants. The cca1 lhy rve468 mutants have growth phenotypes intermediate between cca1 lhy and rve468 mutants, suggesting that CCA1, LHY, RVE4, RVE6, and RVE8 interact additively to regulate growth. Together, our data suggest that these five Myb-like factors interact differently in regulation of the circadian clock versus growth. More generally, the near-norm al seedling phenotypes observed in the largely arrhythmic quintuple mutant demonstrate that circadian-regulated output processes, like control of hypocotyl elongation, do not always depend upon rhythmic oscillator function.

Coordination of matrix attachment and ATP-dependent chromatin remodeling regulate auxin biosynthesis and Arabidopsis hypocotyl elongation.

Hypocotyl elongation is extensively controlled by hormone signaling networks. In particular, auxin metabolism and signaling play key roles in light-dependent hypocotyl growth. The nuclear matrix facilitates organization of DNA within the nucleus, and dynamic interactions between nuclear matrix and DNA are related to gene regulation. Conserved scaffold/matrix attachment regions (S/MARs) are anchored to the nuclear matrix by the AT-HOOK MOTIF CONTAINING NUCLEAR LOCALIZED (AHL) proteins in Arabidopsis. Here, we found that ESCAROLA (ESC)/AHL27 and SUPPRESSOR OF PHYTOCHROME B-4 #3 (SOB3)/AHL29 redundantly regulate auxin biosynthesis in the control of hypocotyl elongation. The light-inducible AHL proteins bind directly to an S/MAR region of the YUCCA 9 (YUC9) promoter and suppress its expression to inhibit hypocotyl growth in light-grown seedlings. In addition, they recruit the SWI2/SNF2-RELATED 1 (SWR1) complex and promote exchange of H2A with the histone variant H2A.Z at the YUC9 locus to further elaborately control auxin biosynthesis. Consistent with these results, the long hypocotyl phenotypes of light-grown genetic mutants of the AHLs and H2A.Z-exchanging components were suppressed by potent chemical inhibitors of auxin transport and YUC enzymes. These results suggest that the coordination of matrix attachment and chromatin modification underlies auxin biosynthesis in light-dependent hypocotyl growth.

Supraoptimal ethylene acts antagonistically with exogenous gibberellins during Solanum lycopersicum (Solanaceae) hypocotyl growth.

In many plant species, ethylene and gibberellins interact to regulate plant growth and development. In some cases, these hormones can act in a synergistic way whereas in others they can be antagonistic. To date, the control of hypocotyl elongation by ethylene and gibberellins has been poorly explored in tomato. In this paper, we report that, application of exogenous ethylene to tomato seedlings or high endogenous ethylene production, as in the epinastic mutant, strongly prevent the effect of gibberellic acid (GA3) application. Moreover, constitutive activation of gibberellin signal in a DELLA deficient mutant is not able to counteract the inhibitory effect of ethylene on hypocotyl elongation, suggesting that ethylene acts independently from DELLA-mediated gibberellin response. Interestingly, when ethylene perception is blocked, the GA3 promotive effect on hypocotyl length is less effective, indicating that the presence of a basal level of ethylene could synergistically enhance hypocotyl growth. Taken together, these observations may suggest that, in tomato, supraoptimal concentrations of ethylene are able to antagonize gibberellin effect but normal levels seem to promote gibberellin-induced hypocotyl elongation.

SCAB3 Is Required for Reorganization of Actin Filaments during Light Quality Changes

The STOMATAL CLOSURE-RELATED ACTIN BINDING PROTEIN (SCAB) family is plant-specific, and its members all contain a novel actin binding domain. Here, we report that SCAB3, a homolog of SCAB1, binds, stabilizes and bundles actin filaments. The SCAB3 promoter contains a cis-element which could be bound by the FHY3/FAR1 transcription factors. Consistently, the expression of SCAB3 is induced when plants were transferred from white light to far red light (T-Far Red) conditions. The scab3 mutants show defects in the control of hypocotyl elongation under T-Far Red condition, which may result from an impaired reorganization of actin filaments. Together, our results suggest that SCAB3 plays an important role in plant growth under changes of light conditions possibly by regulating actin filament dynamics.

The Arabidopsis E3 ubiquitin ligase HOS1 contributes to auxin biosynthesis in the control of hypocotyl elongation

Photomorphogenesis is a key developmental process involving numerous developmental and physiological changes in response to light, including inhibition of hypocotyl elongation. Light-dependent hypocotyl development is intensively regulated by all plant hormones. In agreement with this, a number of hormone metabolic and signaling components are involved in the light regulation of plant morphogenesis. Here, we report that the RING finger-containing E3 ubiquitin ligase HIGH EXPRESSION OF OSMOTICALLY RESPONSIVE GENES1 (HOS1) negatively regulates auxin biosynthesis in the control of hypocotyl elongation in Arabidopsis. The HOS1-deficient mutants exhibited a long hypocotyl in light conditions but a short hypocotyl in the dark, as observed in auxin-overproducing mutants. We found that auxin biosynthetic genes, such as YUCCAs (YUCs), TRYPTOPHAN AMINOTRANSFERASE OF ARABIDOPSIS 1, and CYP79B2, were up-regulated in the hos1–3 mutant, and endogenous auxin levels were accordingly increased. Furthermore, the long hypocotyl phenotype of light-grown hos1–3 seedlings was rescued by the application of auxin transport inhibitors, 1-naphthylphthalamic acid and 2,3,5-triiodobenzoic acid. Taken together, our data demonstrate that HOS1 modulates auxin biosynthesis and probably links light and auxin signaling in the control of hypocotyl elongation.

CCA1 and ELF3 Interact in the Control of Hypocotyl Length and Flowering Time in Arabidopsis

The circadian clock is an endogenous oscillator with a period of approximately 24 h that allows organisms to anticipate, and respond to, changes in the environment. In Arabidopsis (Arabidopsis thaliana), the circadian clock regulates a wide variety of physiological processes, including hypocotyl elongation and flowering time. CIRCADIAN CLOCK ASSOCIATED1 (CCA1) is a central clock component, and CCA1 overexpression causes circadian dysfunction, elongated hypocotyls, and late flowering. EARLY FLOWERING3 (ELF3) modulates light input to the clock and is also postulated to be part of the clock mechanism. elf3 mutations cause light-dependent arrhythmicity, elongated hypocotyls, and early flowering. Although both genes affect similar processes, their relationship is not clear. Here, we show that CCA1 represses ELF3 by associating with its promoter, completing a CCA1-ELF3 negative feedback loop that places ELF3 within the oscillator. We also show that ELF3 acts downstream of CCA1, mediating the repression of PHYTOCHROME-INTERACTING FACTOR4 (PIF4) and PIF5 in the control of hypocotyl elongation. In the regulation of flowering, our findings show that ELF3 and CCA1 either cooperate or act in parallel through the CONSTANS/FLOWERING LOCUS T pathway. In addition, we show that CCA1 represses GIGANTEA and SUPPRESSOR OF CONSTANS1 by direct interaction with their promoters, revealing additional connections between the circadian clock and the flowering pathways.

Dual role of TOC1 in the control of circadian and photomorphogenic responses in Arabidopsis.

To examine the role of the TOC1 (TIMING OF CAB EXPRESSION1) gene in the Arabidopsis circadian system, we generated a series of transgenic plants expressing a gradation in TOC1 levels. Silencing of the TOC1 gene causes arrhythmia in constant darkness and in various intensities of red light, whereas in blue light, the clock runs faster in silenced plants than in wild-type plants. Increments in TOC1 gene dosage delayed the pace of the clock, whereas TOC1 overexpression abolished rhythmicity in all light conditions tested. Our results show that TOC1 RNA interference and toc1-2 mutant plants displayed an important reduction in sensitivity to red and far-red light in the control of hypocotyl elongation, whereas increments in TOC1 gene dosage clearly enhanced light sensitivity. Furthermore, the red light-mediated induction of CCA1/LHY expression was decreased in TOC1 RNA interference and toc1-2 mutant plants, indicating a role for TOC1 in the phytochrome regulation of circadian gene expression. We conclude that TOC1 is an important component of the circadian clock in Arabidopsis with a crucial function in the integration of light signals to control circadian and morphogenic responses.

Control of hypocotyl elongation in Arabidopsis thaliana by photoreceptor interaction.

In order to test the interaction of different phytochromes and blue-light receptors, etiolated seedlings of wild-type Arabidopsis thaliana (L.) Heynh., a phytochrome (phy) B-overexpressor line (ABO), and the photoreceptor mutants phyA-201, phyB-5, hy4-2.23n, fha-1, phyA-201/phyB-5, and phyA-201/hy4-2.23n were exposed to red and far-red light pulses after various preirradiations. The responsiveness to the inductive red pulses is primarily mediated by phyB which is rather stable in its far-red-absorbing form as demonstrated by a very slow loss of reversibility. Without preirradiation the red pulses had an impact on hypocotyl elongation only in PHYA mutants but not in the wild type. This indicates a suppression of phyB function by the presence of phyA. Preirradiation with either far-red or blue light resulted in an inhibition of hypocotyl elongation by red pulses in the wild type. Responsiveness amplification by far-red light is mediated by phyA and disappears slowly in the dark. The extent of responsiveness amplification by blue light was identical in the wild type and in the absence of phyA, or the cryptochromes cryl (hy4-2.23n) or cry2 (fha-1). Therefore, we conclude that stimulation of phyB by blue light preirradiation is either mediated by an additional still-unidentified blue-light-absorbing pigment or that phyA, cry1 and cry2 substitute for each other completely. Both blue and red preirradiation established responsiveness to red pulses in phyA-201/phyB-5 double mutants. These results demonstrate that inhibition of hypocotyl elongation by red pulses is not only mediated by phyB but also by a phytochrome(s) other than phyA and phyB.

Photomorphogenic responses to UV radiation: Involvement of phytochrome and UV photoreceptors in the control of hypocotyl elongation in Lycopersicon esculentum

Photomorphogenic responses to UV radiation: Involvement of phytochrome and UV photoreceptors in the control of hypocotyl elongation in Lycopersicon esculentum

Photomorphogenic responses to UV radiation: Involvement of phytochrome and UV photoreceptors in the control of hypocotyl elongation in Lycopersicon esculentum

The photo-inhibition of Lycopersicon esculentum Mill, hypocotyl growth induced by UV radiation may be mediated by both phytochrome and UV-absorbing receptors. The inhibition of growth induced by continuous irradiation with high fluence rate UV radiation is similar in the au mutant, which is severely deficient in spectrophoto metrically and immunochemically detectable phytochrome, and in the isogenic wild type. Parallel irradiation with 692 nm light, which is equivalent to UV radiation for the phytochrome system in our experimental conditions, induced at high photon fluence rates a significant increase in hypocotyl growth in the au mutant. The same light treatments inhibited the hypocotyl growth of the wild type. The responses of water-grown seedlings and chlorophyll-free seedlings (streptomycin and norflurazon treated seedlings) were compared. Water-grown and chlorophyll-free seedlings responded similarly to UV radiation. The presence of chlorophyll correlates with a significant increase in hypocotyl growth of au mutants irradiated with 692 nm light. These results support the conclusion that UV-induced inhibition of growth in the au mutant is independent of phytochrome.

Photophysiology and Phytochrome Content of Long-Hypocotyl Mutant and Wild-Type Cucumber Seedlings

Photomorphogenetic responses have been studied in a cucumber (Cucumis sativus L.) mutant (lh), which has long hypocotyls in white light (WL). While etiolated seedlings of this mutant have a similar phytochrome content and control of hypocotyl elongation as wild type, deetiolation is retarded and WL-grown seedlings show reduced phytochrome control. Spectrophotometric measurements exhibit that WL-grown tissues of the lh mutant (flower petals and Norflurazon-bleached leaves) contain 35 to 50% of the phytochrome level in the wild type. We propose that this is a consequence of a lack of light-stable phytochrome, in agreement with our hypothesis proposed on the basis of physiological experiments. The lh mutant lacks an end-of-day far-red light response of hypocotyl elongation. This enables the end-of-day far-red light response, clearly shown by the wild type, to be ascribed to the phytochrome, deficient in the lh mutant. Growth experiments in continuous blue light (BL) and continuous BL + red light (RL) show that when RL is added to BL, hypocotyl growth remains inhibited in the wild type, whereas the lh mutant exhibits significant growth promotion compared to BL alone. It is proposed that the hypocotyls fail to grow long in low fluence rate BL because photosynthesis is insufficient to sustain growth.