Dark Growth Research Articles

The production and characteristics of glycogen synthesized by various strains of the thermoacidophilic red microalgae Galdieria grown heterotrophically

AbstractRed microalgae from the Cyanidiophyceae, particularly Galdieria sulphuraria and Cyanidioschyzon merolae, are primitive photosynthetic thermoacidophiles that thrive in acidic hot springs and geysers. Unlike most Cyanidiophyceae, Galdieria strains are metabolically flexible as they can switch from photoautotrophic growth in the light to heterotrophic growth in complete darkness. Galdieria sulphuraria is especially noteworthy for its accumulation of various commercially valuable, functional compounds such as glycogen and phycocyanin. Glycogen, a branched fractal-like polysaccharide composed of several thousands of anhydroglucopyranose units, can be added to cosmetic products and sports drinks as a moisturizer or slow-digestible carbohydrate. While the production and structural characteristics of the glycogen of G. sulphuraria 108.79, isolated from Yellowstone National Park, have been previously described, our investigation aimed to explore glycogen production and properties across various Galdieria strains from different locations. Our findings reveal that all examined strains produce substantial amounts of highly branched glycogen when grown heterotrophically on glycerol in the dark. Notably, the structural characteristics of Galdieria glycogen distinguish it from both eukaryotic and prokaryotic glycogen, exhibiting a significantly higher degree of branching, substantially shorter side chains, and a considerable extent of indigestibility. These findings support the hypothesis that this highly branched, small glycogen is a long-term energy store, enabling survival during extended periods of complete darkness.

Proteomic analysis of unicellular cyanobacterium Crocosphaera subtropica ATCC 51142 under extended light or dark growth.

The daily light-dark cycle is a recurrent and predictable environmental phenomenon to which many organisms, including cyanobacteria, have evolved to adapt. Understanding how cyanobacteria alter their metabolic attributes in response to subjective light or dark growth may provide key features for developing strains with improved photosynthetic efficiency and applications in enhanced carbon sequestration and renewable energy. Here, we undertook a label-free proteomic approach to investigate the effect of extended light (LL) or extended dark (DD) conditions on the unicellular cyanobacterium Crocosphaera subtropica ATCC 51142. We quantified 2287 proteins, of which 603 proteins were significantly different between the two growth conditions. These proteins represent several biological processes, including photosynthetic electron transport, carbon fixation, stress responses, translation, and protein degradation. One significant observation is the regulation of over two dozen proteases, including ATP dependent Clp-proteases (endopeptidases) and metalloproteases, the majority of which were upregulated in LL compared to DD. This suggests that proteases play a crucial role in the regulation and maintenance of photosynthesis, especially the PSI and PSII components. The higher protease activity in LL indicates a need for more frequent degradation and repair of certain photosynthetic components, highlighting the dynamic nature of protein turnover and quality control mechanisms in response to prolonged light exposure. The results enhance our understanding of how Crocosphaera subtropica ATCC51142 adjusts its molecular machinery in response to extended light or dark growth conditions.

Automated Imaging to Evaluate the Exogenous Gibberellin (Ga3) Impact on Seedlings from Salt-Stressed Lettuce Seeds.

Salinity stress is a common challenge in plant growth, impacting seed quality, germination, and general plant health. Sodium chloride (NaCl) ions disrupt membranes, causing ion leakage and reducing seed viability. Gibberellic acid (GA3) treatments have been found to promote germination and mitigate salinity stress on germination and plant growth. 'Bauer' and 'Muir' lettuce (Lactuca sativa) seeds were soaked in distilled water (control), 100 mM NaCl, 100 mM NaCl + 50 mg/L GA3, and 100 mM NaCl + 150 mg/L GA3 in Petri dishes and kept in a dark growth chamber at 25 °C for 24 h. After germination, seedlings were monitored using embedded cameras, capturing red, green, and blue (RGB) images from seeding to final harvest. Despite consistent germination rates, 'Bauer' seeds treated with NaCl showed reduced germination. Surprisingly, the 'Muir' cultivar's final dry weight differed across treatments, with the NaCl and high GA3 concentration combination yielding the poorest results (p < 0.05). This study highlights the efficacy of GA3 applications in improving germination rates. However, at elevated concentrations, it induced excessive hypocotyl elongation and pale seedlings, posing challenges for two-dimensional imaging. Nonetheless, a sigmoidal regression model using projected canopy size accurately predicted dry weight across growth stages and cultivars, emphasizing its reliability despite treatment variations (R2 = 0.96, RMSE = 0.11, p < 0.001).

The Use of Imaging to Quantify the Impact of Seed Aging on Lettuce Seed Germination and Seedling Vigor.

The decline in seed quality over time due to natural aging or mishandling requires assessing seed vigor for resilience in adverse conditions. Accelerated aging (AA) methods simulate seed deterioration by subjecting seeds to high temperatures and humidity. Saturated salt accelerated aging (SSAA) is an AA method adopted for small seeds like lettuce (Lactuca sativa). In this study, we subjected seeds of two lettuce cultivars ('Muir' and 'Bauer') to SSAA by sealing them in a box containing 40 g/100 mL of a sodium chloride (NaCl) solution in a dark growth chamber at 41 °C for 24, 48, and 72 h with a control. We monitored their vigor using embedded computer cameras, tracking the projected canopy size (PCS) daily from sowing to harvest. The cultivar 'Muir' exhibited consistent PCS values across the treatments, while 'Bauer' showed PCS variations, with notable declines after prolonged aging. The germination rates dropped significantly after 48 and 72 h of SSAA. A nonlinear regression model revealed a strong relationship between PCS and shoot dry weight across harvests and cultivars (R2 = 0.93, RMSE = 0.15, p < 0.001). The research found that the projected canopy size and shoot dry weight increased over time with significant differences in treatments for the cultivar 'Bauer' but not for 'Muir,' with the canopy size being a strong predictor of dry weight and no significant impact from the SSAA treatments. This study highlights cultivar-specific responses to aging and demonstrates the efficacy of our imaging tool in predicting lettuce dry weight despite treatment variations. Understanding how aging affects different lettuce varieties is crucial for seed management and crop sustainability.

Cotton sprouts as potential vegetable source: Nutrient compositions and alterations in metabolomics and transcriptomics between light and dark growth conditions

Cottonseed is a by-product of cotton industry, but its broad application in human diets has not been fully elucidated. This study aims to analyse the nutritional composition of cotton sprouts and investigate metabolic and transcriptomic differences under light and darkness. The findings revealed that the glanded ZM41 yellow cotton sprouts were rich in protein (2.8 %), vitamin E (0.443 mg/100 g), and minerals, with < 0.01 % free gossypol. Moreover, protein, aspartic acid (1,026 mg/100 g), and serine (136 mg/100 g) reached their highest levels in nine-day-old sprouts, while no significant differences were observed in the other nutrients on the 7th, 9th, or 11th day. The average quantitative values of 64 cottonseed varieties found gossypol (27.9 mg/kg), protein (3.6 %), potassium (1,487.3 mg/kg), calcium (506.2 mg/kg) and magnesium (330.2 mg/kg). Integrated metabolome and transcriptome analyses revealed that 317 out of 1,880 metabolites were differentially accumulated between light and dark conditions, wherein the downregulation of structural genes encoding CHI, F3H, FLS, DFR, F3′H, ANS, LAR and ANR resulted in decreased flavonoid levels in dark-cultivated sprouts, while the upregulation of HMGR and 2-ODD-1 enhanced the biosynthesis of gossypol. This study provides insights into the nutritional and metabolic aspects of cotton sprouts, offering a reference for potential commercial applications.

A system for the study of roots 3D kinematics in hydroponic culture: a study on the oscillatory features of root tip

BackgroundThe root of a plant is a fundamental organ for the multisensory perception of the environment. Investigating root growth dynamics as a mean of their interaction with the environment is of key importance for improving knowledge in plant behaviour, plant biology and agriculture. To date, it is difficult to study roots movements from a dynamic perspective given that available technologies for root imaging focus mostly on static characterizations, lacking temporal and three-dimensional (3D) spatial information. This paper describes a new system based on time-lapse for the 3D reconstruction and analysis of roots growing in hydroponics.ResultsThe system is based on infrared stereo-cameras acquiring time-lapse images of the roots for 3D reconstruction. The acquisition protocol guarantees the root growth in complete dark while the upper part of the plant grows in normal light conditions. The system extracts the 3D trajectory of the root tip and a set of descriptive features in both the temporal and frequency domains. The system has been used on Zea mays L. (B73) during the first week of growth and shows good inter-reliability between operators with an Intra Class Correlation Coefficient (ICC) > 0.9 for all features extracted. It also showed measurement accuracy with a median difference of < 1 mm between computed and manually measured root length.ConclusionsThe system and the protocol presented in this study enable accurate 3D analysis of primary root growth in hydroponics. It can serve as a valuable tool for analysing real-time root responses to environmental stimuli thus improving knowledge on the processes contributing to roots physiological and phenotypic plasticity.

Arabidopsis hypocotyl growth in darkness requires the phosphorylation of a microtubule-associated protein.

Rapid hypocotyl elongation allows buried seedlings to emerge, where light triggers de-etiolation and inhibits hypocotyl growth mainly by photoreceptors. Phosphorylation/dephosphorylation events regulate many aspects of plant development. Only recently we have begun to uncover the earliest phospho-signaling responders to light. Here, we reported a large-scale phosphoproteomic analysis and identified 20 proteins that changed their phosphorylation pattern following a 20 min light pulse compared to darkness. Microtubule-associated proteins were highly overrepresented in this group. Among them, we studied CIP7 (COP1-INTERACTING-PROTEIN 7), which presented microtubule (MT) localization in contrast to the previous description. An isoform of CIP7 phosphorylated at Serine915 was detected in etiolated seedlings but was undetectable after a light pulse in the presence of photoreceptors, while CIP7 transcript expression decays with long light exposure. The short hypocotyl phenotype and rearrangement of MTs in etiolated cip7 mutants are complemented by CIP7-YFP and the phospho-mimetic CIP7S915D-YFP, but not the phospho-null CIP7S915A-YFP suggesting that the phosphorylated S915CIP7 isoform promotes hypocotyl elongation through MT reorganization in darkness. Our evidence on Serine915 of CIP7 unveils phospho-regulation of MT-based processes during skotomorphogenic hypocotyl growth.

Identification and characterization of the low molecular mass ferredoxins involved in central metabolism in Heliomicrobium modesticaldum.

The homodimeric Type I reaction center (RC) from Heliomicrobium modesticaldum lacks the PsaC subunit found in Photosystem I and instead uses the interpolypeptide [4Fe-4S] cluster FX as the terminal electron acceptor. Our goal was to identify which of the small mobile dicluster ferredoxins encoded by the H. modesticaldum genome are capable of accepting electrons from the heliobacterial RC (HbRC) and pyruvate:ferredoxin oxidoreductase (PFOR), a key metabolic enzyme. Analysis of the genome revealed seven candidates: HM1_1462 (PshB1), HM1_1461 (PshB2), HM1_2505 (Fdx3), HM1_0869 (FdxB), HM1_1043, HM1_0357, and HM1_2767. Heterologous expression in Escherichia coli and studies using time-resolved optical spectroscopy revealed that only PshB1, PshB2, and Fdx3 are capable of accepting electrons from the HbRC and PFOR. Modeling studies using AlphaFold show that only PshB1, PshB2, and Fdx3 should be capable of docking on PFOR at a positively charged patch that overlays a surface-proximal [4Fe-4S] cluster. Proteomic analysis of wild-type and gene deletion strains ΔpshB1, ΔpshB2, ΔpshB1pshB2, and Δfdx3 grown under nitrogen-replete conditions revealed that Fdx3 is undetectable in the wild-type, ΔpshB1, and Δfdx3 strains, but it is present in the ΔpshB2 and ΔpshB1pshB2 strains, implying that Fdx3 may substitute for PshB2. When grown under nitrogen-deplete conditions, Fdx3 is present in the wild-type and all deletion strains except for Δfdx3. None of the knockout strains demonstrated significant impairment during chemotrophic dark growth on pyruvate, photoheterotrophic light growth on pyruvate, or phototrophic growth on acetate+CO2, indicating a high degree of redundancy among these three electron transfer proteins. Loss of both PshB1 and PshB2, but not FdxB, resulted in poor growth under N2-fixing conditions.

Tracking the Ti4+ substitution in phlogopite by spectroscopic imaging: A tool for unravelling the growth of micas at HP-HT conditions

Phlogopite solid-solutions have a wide pressure–temperature (P-T) stability field and are ubiquitous in a wide variety of geological settings, from deep lithosphere magmatic environments to upper crust metamorphic domains. Phlogopite composition represents therefore a valuable physical–chemical archive and may provide important information regarding its crystallization and the petrogenesis of the host-rock. In this paper we examine the phlogopite phenocrysts from the well-known Fort Regent mica-bearing lamprophyre minette from St. Helier (Island of Jersey, UK). Phlogopite phenocrystals from lamprophyres generally show normal-step and continuous compositional zoning, however those from the Fort Regent minette show a peculiar texture characterized by dark brown high-Ti (average TiO2 ≈ 8.5 wt.%) cores enveloped by euhedral low- to mid-amplitude zonation due to oscillatory contents in Ti, Fe and Mg. Thermo-barometry modelling based on biotite-only composition yields relatively high P-T estimates (T ≈ 970 ± 54 °C at P ≈ 0.73 ± 0.13 GPa) for cores whereas lower values (T ≈ 790 ± 54 °C at P ≈ 0.29 ± 0.13 GPa) are obtained for the outer rims. Comparable temperatures (T ≈ 1075 ± 54 °C) but extremely high and anomalous pressure values (P ≈ 1.82 ± 0.13 GPa) are obtained for the yellowish inner rims. The combination of electron micro probe (EMP) analysis and single-crystal infra-red (FTIR) imaging in the OH-stretching region shows that the exceptional and oscillatory Ti contents are due to the Ti-vacancy substitution, typical of crystallization and growth processes of HP/HT environments. Raman imaging provides additional insight for this process, confirming the dominant dioctahedral nature for the Ti-Fe-rich cores and outer rims. Interpretation of thermobaric estimates obtained from the phlogopite composition-only model, based on the fine-scale compositional evolution, shows that pressure–temperature values from low-Ti high-Mg domains should be carefully evaluated because the substitution mechanisms during the dark mica growth are not univocally related to pressure–temperature variation of the crystallizing environment. Our results demonstrate how a multidisciplinary approach based on the combination of chemical investigations and vibrational spectroscopies could represent a valuable tool to evaluate pressure–temperature estimates from biotite composition-only thermo-barometry models and therefore to correctly unravel HP/HT petrogenetic processes at a very fine scale.

Three cases of pseudoepitheliomatous hyperplasia mimicking squamous cell carcinoma of the foot: A case series

Abstract Pseudoepitheliomatous hyperplasia (PEH) is a benign condition characterized by hyperplasia and proliferation of the epidermis and adnexal epithelium which can mimic squamous cell carcinoma (SCC). In our first case, a 55-year-old female with a prior history of split skin graft presented with dark verrucous growth over the graft site with a nonhealing ulcer of 3 cm × 2 cm seen over the ventral aspect of the left foot. Our second case, a 59-year-old male with a prior history of split skin grafting presented with dark verrucous growth and pain over the right forefoot for 3 months. The third case, a 58-year-old man with diabetic foot ulcer with verrucous growth over the toes. In all the cases, the biopsy was done and diagnosed to be PEH. We report this case series of PEH for the proper distinction from the SCC of the skin.

GROWTH PERFORMANCE OF CLARIAS GARIEPINUS (BURCHELL, 1822) REARED IN GLASS AQUARIA UNDER DIFFERENT LIGHT CONDITIONS

Growth performance of Clarias gariepinus (Burchell, 1822) reared in glass aquaria under different light conditions was studied from hatching to 75 days post hatching (dph). The objective was to generate baseline data/information on artificial seed production of catfish under different rearing conditions. Eggs of C. gariepinus were hatched at three photoperiods in complete darkness (DD), full illumination (LL) and control 12 h dark and 12 h light (normal) growth conditions. African catfish showed higher survival rates in the dark conditions than in the light conditions. The larvae also showed significantly increased cannibalistic behavior under light condition. Highest survival rate (92.8%) was observed for C. gariepinus that grew under darker condition. The higher activity of C. gariepinus under dark conditions resulted in reduced rates of cannibalism and higher rates of survival than light conditions. The sequence of development and ossification was faster in dark group by a day than in the light and by two days in the control group, whereas light group faster by a day compared to the control group. Some of the organ development and appearance completed within an hour gap and it was difficult to look sequential ontogeny on dark growth conditions. Dark growth conditions may be recommended as a best, effective technique for intensification of production of C. gariepinus, as it improves catfish survival rates.

Immunophilin FKB20-2 participates in oligomerization of Photosystem I in Chlamydomonas.

PSI is a sophisticated photosynthesis protein complex that fuels the light reaction of photosynthesis in algae and vascular plants. While the structure and function of PSI have been studied extensively, the dynamic regulation on PSI oligomerization and high light response is less understood. In this work, we characterized a high light-responsive immunophilin gene FKB20-2 (FK506-binding protein 20-2) required for PSI oligomerization and high light tolerance in Chlamydomonas (Chlamydomonas reinhardtii). Biochemical assays and 77-K fluorescence measurement showed that loss of FKB20-2 led to the reduced accumulation of PSI core subunits and abnormal oligomerization of PSI complexes and, particularly, reduced PSI intermediate complexes in fkb20-2. It is noteworthy that the abnormal PSI oligomerization was observed in fkb20-2 even under dark and dim light growth conditions. Coimmunoprecipitation, MS, and yeast 2-hybrid assay revealed that FKB20-2 directly interacted with the low molecular weight PSI subunit PsaG, which might be involved in the dynamic regulation of PSI-light-harvesting complex I supercomplexes. Moreover, abnormal PSI oligomerization caused accelerated photodamage to PSII in fkb20-2 under high light stress. Together, we demonstrated that immunophilin FKB20-2 affects PSI oligomerization probably by interacting with PsaG and plays pivotal roles during Chlamydomonas tolerance to high light.

Unlocking microalgal host-exploring dark-growing microalgae transformation for sustainable high-value phytochemical production.

Microalgae have emerged as a promising, next-generation sustainable resource with versatile applications, particularly as expression platforms and green cell factories. They possess the ability to overcome the limitations of terrestrial plants, such non-arable land, water scarcity, time-intensive growth, and seasonal changes. However, the heterologous expression of interested genes in microalgae under heterotrophic cultivation (dark mode) remains a niche area within the field of engineering technologies. In this study, the green microalga, Chlorella sorokiniana AARL G015 was chosen as a potential candidate due to its remarkable capacity for rapid growth in complete darkness, its ability to utilize diverse carbon sources, and its potential for wastewater treatment in a circular bioeconomy model. The aims of this study were to advance microalgal genetic engineering via dark cultivation, thereby positioning the strain as promising dark-host for expressing heterologous genes to produce high-value phytochemicals and ingredients for food and feed. To facilitate comprehensive screening based on resistance, eleven common antibiotics were tested under heterotrophic condition. As the most effective selectable markers for this strain, G418, hygromycin, and streptomycin exhibited growth inhibition rates of 98%, 93%, and 92%, respectively, ensuring robust long-term transgenic growth. Successful transformation was achieved through microalgal cell cocultivation with Agrobacterium under complete darkness verified through the expression of green fluorescence protein and β-glucuronidase. In summary, this study pioneers an alternative dark-host microalgal platform, using, Chlorella, under dark mode, presenting an easy protocol for heterologous gene transformation for microalgal host, devoid of the need for expensive equipment and light for industrial production. Furthermore, the developed genetic transformation methodology presents a sustainable way for production of high-value nutrients, dietary supplements, nutraceuticals, proteins and pharmaceuticals using heterotrophic microalgae as an innovative host system.

Influence of Cytokinins, Dark Incubation and Air-Lift Bioreactor Culture on Axillary Shoot Proliferation of Al-Taif Rose (Rosa damascena trigintipetala (Diek) R. Keller)

Rose is a widely favored floriculture crop that is commercially propagated through the application of tissue culture techniques. Here, we report an effective method for axillary shoot proliferation in Al-Taif rose, an important cultivar for rose oil industry. Stem nodes were excised from an adult donor Al-Taif rose shrub and cultured for 4 weeks on Murashige and Skoog’s (MS) medium supplemented with 6-benzylaminopurine (BAP) or gibberellic acid (GA3) at 0 and 3 mg·L−1 to induce the sprouting of axillary shoots. Al-Taif rose shoots were cultured in vitro for 6 weeks on MS medium fortified with different concentrations of cytokinins, light/dark incubation and different culture types (gelled and liquid/bioreactor culture). The culture conditions that were applied had a noteworthy impact on the responses of Al-Taif rose shoot proliferation. The supplementation of the medium with 6-benzylaminopurine (BAP) resulted in an augmented rate of shoot proliferation in comparison to other cytokinins. Additionally, dark incubation limited foliage growth, leaf yellowing and abscission and favored shoot proliferation compared with light incubation. Liquid culture using bioreactors provided higher axillary shoot proliferation and growth as compared with gelled culture. A continuous immersion system with a net provided the highest axillary shoots (four shoots per explant) and shoot length (16.5 cm), whereas an immersion system without a net provided the highest fresh weight of axillary shoots (499 mg per explant). These findings will improve commercial propagation and contribute to the rose oil industry of Al-Taif rose.

Role of plastids and mitochondria in the early development of seedlings in dark growth conditions.

Establishment of the seedlings is a crucial stage of the plant life cycle. The success of this process is essential for the growth of the mature plant. In Nature, when seeds germinate under the soil, seedlings follow a dark-specific program called skotomorphogenesis, which is characterized by small, non-green cotyledons, long hypocotyl, and an apical hook-protecting meristematic cells. These developmental structures are required for the seedlings to emerge quickly and safely through the soil and gain autotrophy before the complete depletion of seed resources. Due to the lack of photosynthesis during this period, the seed nutrient stocks are the primary energy source for seedling development. The energy is provided by the bioenergetic organelles, mitochondria, and etioplast (plastid in the dark), to the cell in the form of ATP through mitochondrial respiration and etio-respiration processes, respectively. Recent studies suggest that the limitation of the plastidial or mitochondrial gene expression induces a drastic reprogramming of the seedling morphology in the dark. Here, we discuss the dark signaling mechanisms involved during a regular skotomorphogenesis and how the dysfunction of the bioenergetic organelles is perceived by the nucleus leading to developmental changes. We also describe the probable involvement of several plastid retrograde pathways and the interconnection between plastid and mitochondria during seedling development. Understanding the integration mechanisms of organellar signals in the developmental program of seedlings can be utilized in the future for better emergence of crops through the soil.

Light-Induced Changes in Secondary Metabolite Production of Trichoderma atroviride.

Many studies aim at maximizing fungal secondary metabolite production but the influence of light during cultivation has often been neglected. Here, we combined an untargeted isotope-assisted liquid chromatography-high-resolution mass spectrometry-based metabolomics approach with standardized cultivation of Trichoderma atroviride under three defined light regimes (darkness (PD), reduced light (RL) exposure, and 12/12 h light/dark cycle (LD)) to systematically determine the effect of light on secondary metabolite production. Comparative analyses revealed a similar metabolite profile upon cultivation in PD and RL, whereas LD treatment had an inhibiting effect on both the number and abundance of metabolites. Additionally, the spatial distribution of the detected metabolites for PD and RL was analyzed. From the more than 500 detected metabolites, only 25 were exclusively produced upon fungal growth in darkness and 85 were significantly more abundant in darkness. The majority were detected under both cultivation conditions and annotation revealed a cluster of substances whose production followed the pattern observed for the well-known T. atroviride metabolite 6-pentyl-alpha-pyrone. We conclude that cultivation of T. atroviride under RL can be used to maximize secondary metabolite production.

Two types of GLR channels cooperate differently in light and dark growth of Arabidopsis seedlings

BackgroundGLutamate Receptor-like (GLR) channels are multimeric, ionotropic, ligand-gated plant transmembrane receptors. They are homologous to mammalian glutamate receptors, iGLuRs, which are critical to neuronal function. GLRs have been reported several times to play a role in photomorphogenesis. However, to date, no study has looked at the mechanism of their involvement in this process. Here we focused on examining the impact of GLRs on the regulation of early seedling growth in blue light, red light, and in the dark.ResultsWild type and six photoreceptor mutant seedlings were grown on media supplemented with known iGLuR/GLR channel antagonists: MK-801, which non-competitively blocks NMDA channels in mammalian cells, and CNQX, known for competitive blocking of AMPA channels in mammalian cells. The lengths of hypocotyls and roots were measured in seedlings of phyA, phyB, phot1, phot2, cry1, and cry2 mutants after 7 days of in vitro culture. Changes in growth parameters, both in light and in darkness upon application of chemical antagonists, show that both types of GLR channels, NMDA-like and AMPA-like, are involved in the regulation of seedling growth irrespective of light conditions. Analysis of seedling growth of photoreceptor mutants indicates that the channels are influenced by signaling from phot1, phot2, and cry1. To extend our analysis, we also evaluated the elicitation of a calcium wave, which is likely to be partially driven by GLRs, in Arabidopsis seedlings. The changes in cellobiose-induced calcium waves observed after applying GLR inhibitors suggest that both types of channels likely cooperate in shaping Arabidopsis seedling growth and development.ConclusionsOur work provides the first experimental evidence that two types of GLR channels function in plants: NMDA-like and AMPA-like. We also demonstrate that the channels are involved in seedling growth and development, at least partially through modulation of calcium signaling, but they are unlikely to play a major role in photomorphogenesis.

RGS4 impacts carbohydrate and siderophore metabolism in Trichoderma reesei

BackgroundAdaptation to complex, rapidly changing environments is crucial for evolutionary success of fungi. The heterotrimeric G-protein pathway belongs to the most important signaling cascades applied for this task. In Trichoderma reesei, enzyme production, growth and secondary metabolism are among the physiological traits influenced by the G-protein pathway in a light dependent manner.ResultsHere, we investigated the function of the SNX/H-type regulator of G-protein signaling (RGS) protein RGS4 of T. reesei. We show that RGS4 is involved in regulation of cellulase production, growth, asexual development and oxidative stress response in darkness as well as in osmotic stress response in the presence of sodium chloride, particularly in light. Transcriptome analysis revealed regulation of several ribosomal genes, six genes mutated in RutC30 as well as several genes encoding transcription factors and transporters. Importantly, RGS4 positively regulates the siderophore cluster responsible for fusarinine C biosynthesis in light. The respective deletion mutant shows altered growth on nutrient sources related to siderophore production such as ornithine or proline in a BIOLOG phenotype microarray assay. Additionally, growth on storage carbohydrates as well as several intermediates of the D-galactose and D-arabinose catabolic pathway is decreased, predominantly in light.ConclusionsWe conclude that RGS4 mainly operates in light and targets plant cell wall degradation, siderophore production and storage compound metabolism in T. reesei.

WAVE-DAMPENED2-LIKE4 modulates the hyper-elongation of light-grown hypocotyl cells.

Light, temperature, water, and nutrient availability influence how plants grow to maximize access to resources. Axial growth, the linear extension of tissues by coordinated axial cell expansion, plays a central role in these adaptive morphological responses. Using Arabidopsis (Arabidopsis thaliana) hypocotyl cells to explore axial growth control mechanisms, we investigated WAVE-DAMPENED2-LIKE4 (WDL4), an auxin-induced, microtubule-associated protein and member of the larger WDL gene family shown to modulate hypocotyl growth under changing environmental conditions. Loss-of-function wdl4 seedlings exhibited a hyper-elongation phenotype under light conditions, continuing to elongate when wild type Col-0 hypocotyls arrested and reaching 150-200% of wild type length before shoot emergence. wdl4 seedling hypocotyls showed dramatic hyper-elongation (500%) in response to temperature elevation, indicating an important role in morphological adaptation to environmental cues. WDL4 associated with microtubules under both light and dark growth conditions, and no evidence was found for altered microtubule array patterning in loss-of-function wdl4 mutants under various conditions. Examination of hormone responses showed altered sensitivity to ethylene and evidence for changes in the spatial distribution of the auxin-dependent DR5:GFP reporter. Our data provide evidence that WDL4 regulates hypocotyl cell elongation without substantial changes to microtubule array patterning, suggesting an unconventional role in axial growth control.

Chemical and genetic carotenoid deficiency delays growth in dark-grown Euglena gracilis.

Light-independent functions of carotenoids in photosynthetic organisms are poorly understood. Here we investigated the growth properties of microalga, Euglena gracilis, under altered light and temperature using norflurazon-treated carotenoid-deficient cells and genetically modified strains, including non-photosynthetic SM-ZK and colorless cl4. Norflurazon treatment decreased carotenoid and chlorophyll contents, causing cell bleaching. SM-ZK strain had lower carotenoid content than WT strain, and it was below the detectable level in the cl4 strain. Norflurazon treatment decreased phytoene synthase EgCrtB levels, although EgcrtB was transcriptionally induced. Carotenoid deficiency in norflurazon-treated cells and the cl4 strain caused similar extent of delayed growth under light and dark conditions at 25 °C, indicating that carotenoids promote growth in darkness. Both WT and SM-ZK strains exhibited similar growth rates. Dark conditions at 20 °C enhanced growth delay of norflurazon-treated cells and the cl4 strain. These results indicate that carotenoids impart environmental stress tolerance to E. gracilis in light-dependent and -independent manners.