Continuous Light Conditions Research Articles

Advanced puberty triggered by bi-weekly changes in reproductive factors during the photolabile period in a male teleost fish, Dicentrarchus labrax L.

This study evaluated the impact of continuous light (LL) within the photolabile period on advanced puberty in juvenile male European sea bass. The exposure to an LL regime for 1 month, from August 15 to September 15 (LLa/s), was compared to a constant simulated natural photoperiod (NP) and constant continuous light conditions year-round (LLy). Somatic growth, hormone plasma levels, rates of testicular maturation and spermiation, as well as the mRNA levels of some reproductive genes were analyzed. Our results demonstrated that both LLa/s and LLy treatments, which include LL exposure during the photolabile period, were highly effective in inhibiting the gametogenesis process that affects testicular development, and clearly reduced the early sexual maturation of males. Exposure to an LL photoperiod affected body weight and length of juvenile fish during early gametogenesis and throughout the first year of life. Interestingly, LL induced bi-weekly changes in some reproductive factors affecting Gnrh1 and Gnrh2 content in the brain, and also reduced pituitary fshβ expression and plasmatic levels of 11-KT, E2, Fsh throughout early gametogenesis. We suggest that low levels of E2 in early September in the LL groups, which would be concomitant with the reduced number of spermatogonial mitoses in these groups, might indicate a putative role for estrogens in spermatogonial proliferation during the early gonadal development of this species. Furthermore, a significant decrease in amh expression was observed, coinciding with low plasma levels of 11-KT under LL regimes, which is consistent with the idea that this growth factor may be crucial for the progress of spermatogenesis in male sea bass.

Seed and germination traits of the summer weed Trianthema portulacastrum L.

Trianthema portulacastrum L. is one of the problematic terrestrial summer weeds. It competes with various agricultural crops. The present investigation aimed to investigate the traits, which enable the seeds to disperse, germinate, grow and adapt to habitat conditions. The plant produces large number (1931 seeds) of small seeds (1.08 mg seed-1). Scanning electron microscopy of the seed testa revealed that the anticlinal walls of the epidermal cells were penta-hexagonal, and highly raised, while the periclinal walls were sunken. This pattern leads to the porous appearance of the testa. The effects of temperature on seed germination were monitored under continuous light and continuous dark conditions. Under light conditions, the percentages of seed germination were significantly higher than at the dark conditions. Higher temperatures of 35, 40 and 45 °C promoted the seed germination. In the temperature range from 30 to 45 °C, the plumule lengths of the seedlings germinated in the light conditions were smaller than those grown under dark conditions. Under all conditions, the length of radicle was higher than the plumule. The traits of seeds, germination, and seedlings grown under experimental conditions are discussed in terms of the seed dispersal and potential adaptations of seedlings to the environment.

Metabolic engineering of cyanobacteria for photoautotrophic production of heparosan, a pharmaceutical precursor of heparin

Heparosan is an unsulfated polysaccharide potentially important for its wide range of cosmetic and pharmaceutical applications, particularly as the precursor for the extensively used anticoagulant, heparin. Generally sourced from animals, commercially available heparin may encounter various immunological and contamination risks. Thus, safe and sustainable microbial platforms could serve as an alternative heparin source. Synechococcus, due to their fast photoautotrophic growth, strong sugar phosphate metabolisms and generally regarded as safe (GRAS) nature, may serve as photo-biorefineries for manufacturing heparosan. In this study, we have synthesized an integrative plasmid pUPm48 for cloning galU and PmHS2 genes in Synechococcus elongatus PCC 7942. The engineered recombinants (pgp7942) exhibited significant production of heparosan under different culture conditions, where the products were present in both supernatant and cell biomass. The maximum yield of 0.7 ± 0.2 μg/g-DCW (dry cell weight) and a titer of 2.8 ± 0.3 μg/L was achieved by pgp7942 under shake flask and continuous light conditions. Large scale plastic-bag cultures with natural diurnal light exhibited heparosan production of 0.5 μg/g-DCW with a titer of 0.44 μg/L. The analysis also found PCC 7942 encodes a promiscuous uridyltransferase for UDP-glucose synthesis and naturally produces multiple glycosaminoglycans including chondroitin sulfate (CS). This study demonstrates for the first-time cyanobacteria as a promising photoautotrophic refinery for producing a high-value polysaccharide commonly from animals.

Herbivory elicits changes in green leaf volatile production via jasmonate signaling and the circadian clock.

The timing of plant volatile emissions is important for a robust indirect defense response. Green leaf volatiles (GLVs) are emitted by plants upon damage but can be suppressed by herbivore-associated elicitors, and the abundance and composition of GLVs vary depending on the timing of herbivore attack. We show that the GLV biosynthetic enzyme HYDROPEROXIDE LYASE (HPL) is transcriptionally regulated by the circadian clock in Nicotiana attenuata. In accordance with transcript abundance of NaHPL, GLV aldehyde pools in intact leaves peaked at night and at subjective night under diurnal and continuous light conditions, respectively. Moreover, although the basal abundance of NaHPL transcripts is upregulated by jasmonate (JA) signaling, JA does not regulate the reduction of NaHPL transcript abundance in damaged leaves by simulated herbivore treatment. Unexpectedly, the plant circadian clock was strongly altered when Manduca sexta larvae fed on N.attenuata, and this was also independent of JA signaling. Lastly, the temporal dynamics of NaHPL transcripts and total GLV emissions were strongly altered by M.sexta larval feeding. Our data suggest that the temporal dynamics of emitted GLV blends result from a combination of damage, JA signaling, herbivore-associated elicitors, and the plant circadian clock.

Durability Testing of Photoelectrochemical Hydrogen Production under Day/Night Light Cycled Conditions

AbstractThis work investigates long‐term photoelectrochemical hydrogen evolution (82 days) in 1 M HClO4 using a TiO2:H protected crystalline Si‐based photocathode with metal‐oxide‐semiconductor (MOS) junctions. It is shown that day/night cycling leads to relatively rapid performance degradation while the photocurrent under the continuous light condition is relatively stable. We observed that the performance loss is mainly due to contamination of the catalytically active surface with carbonaceous material. By ultraviolet (UV) light exposure, we also observed that the activity can be restored, most likely due to photocatalytic degradation of organic compounds by the UV light excited TiO2 protection layer.

The plastid-nucleus located DNA/RNA binding protein WHIRLY1 regulates microRNA-levels during stress in barley (Hordeum vulgare L.)

ABSTRACTIn this article a novel mechanism of retrograde signaling by chloroplasts during stress is described. This mechanism involves the DNA/RNA binding protein WHIRLY1 as a regulator of microRNA levels. By virtue of its dual localization in chloroplasts and the nucleus of the same cell, WHIRLY1 was proposed as an excellent candidate coordinator of chloroplast function and nuclear gene expression. Comparison of wild-type and transgenic plants with an RNAi-mediated knockdown of WHIRLY1 showed, that the transgenic plants were unable to cope with continuous high light conditions. They were impaired in production of several microRNAs mediating post-transcriptional responses during stress. The results support a central role of WHIRLY1 in retrograde signaling and also underpin a so far underestimated role of microRNAs in this process.

Study of the beneficial effects of green light on lettuce grown under short-term continuous red and blue light-emitting diodes.

Red and blue light are the most important light spectra for driving photosynthesis to produce adequate crop yield. It is also believed that green light may contribute to adaptations to growth. However, the effects of green light, which can trigger specific and necessary responses of plant growth, have been underestimated in the past. In this study, lettuce (Lactuca sativa L.) was exposed to different continuous light (CL) conditions for 48 h by a combination of red and blue light-emitting diodes (LEDs) supplemented with or without green LEDs, in an environmental-controlled growth chamber. Green light supplementation enhanced photosynthetic capacity by increasing net photosynthetic rates, maximal photochemical efficiency, electron transport for carbon fixation (JPSII ) and chlorophyll content in plants under the CL treatment. Green light decreased malondialdehyde and H2 O2 accumulation by increasing the activities of superoxide dismutase (EC 1.15.1.1) and ascorbate peroxidase (EC 1.11.1.11) after 24 h of CL. Supplemental green light significantly increased the expression of photosynthetic genes LHCb and PsbA from 6 to 12 h, and these gene expressions were maintained at higher levels than those under other light conditions between 12 and 24 h. However, a notable downregulation of both LHCb and PsbA was observed during 24 to 48 h. These results indicate that the effects of green light on lettuce plant growth, via enhancing activity of particular components of antioxidative enzyme system and promoting of LHCb and PsbA expression to maintain higher photosynthetic capacity, alleviated a number of the negative effects caused by CL.

Effect of temperature, light on germination and morphological characteristics of Bipolaris sorokiniana

Wheat is one of the most nutritious, proteinaceous and basic stable food crop is attacked by several factors. Among the biotic factors, the most notorious disease of wheat is spot blotch caused by Bipolaris sorokiniana. The behaviour of the pathogen with different combinations of weather regimes were studied in-vitro, in the dark at different temperatures. Maximum conidial germination (92.04%) was observed at 30 °C followed by 91.69% at 25 °C temperature after 4 h of incubation. Lowest germination was recorded at 10 °C. Maximum length of unipolar germ tube (26.28 µm) was recorded at 15 °C which was statistically superior than other treatments. Whereas, maximum width of unipolar germinated germ tube (4.49 µm) was observed at 25 °C followed by 15 °C. The length and width of germ tubes of the two poles of bipolar germinated B. sorokiniana spores were 19.74, 22.61 and 3.92, 4.11 µm, respectively at 15 °C followed by 20 °C temperature. Significantly lowest time required for secondary conidiophore formation (25 h) and conidia formation (39 h) was recorded at 25 °C. Highest time required for formation of secondary conidiophore (35 h) at 20 °C, conidia at 30 °C (80 h) but at 10 and 35 °C temperature secondary conidiophore and conidia formation was not observed within 80 h of incubation. Higher mycelial growth and no sporulation were observed under continuous light condition.

External validations of a non-obtrusive practical method to measure personal lighting conditions in offices

Health symptoms may be influenced, supported, or even controlled via a lighting control system which includes personal lighting conditions and personal factors (health characteristics). In order to be effective, this lighting control system requires both continuous information on the lighting and health conditions at the individual level. A new practical method to determine these continuous personal lighting conditions has been developed: location-bound estimations (LBE). This method was validated in the field in two case studies; comparisons were made between the LBE and location-bound measurements (LBM) in case study 1 and between the LBE and person-bound measurements (PBM) in case study 2. Overall, the relative deviation between the LBE and LBM was less than 15%, whereas the relative deviation between the LBE and PBM was 32.9% in the best-case situation. The relative deviation depends on inaccuracies in both methods (i.e., LBE and PBM) and needs further research. Adding more input parameters to the predictive model (LBE) will improve the accuracy of the LBE. The proposed first approach of the LBE is not without limitations; however, it is expected that this practical method will be a pragmatic approach of inserting personal lighting conditions into lighting control systems.

Circadian clocks of both plants and pollinators influence flower seeking behavior of the pollinator hawkmoth Manduca sexta

Most plant-pollinator interactions occur during specific periods during the day. To facilitate these interactions, many flowers are known to display their attractive qualities, such as scent emission and petal opening, in a daily rhythmic fashion. However, less is known about how the internal timing mechanisms (the circadian clocks) of plants and animals influence their daily interactions. We examine the role of the circadian clock in modulating the interaction between Petunia and one of its pollinators, the hawkmoth Manduca sexta. We find that desynchronization of the Petunia circadian clock affects moth visitation preference for Petunia flowers. Similarly, moths with circadian time aligned to plants show stronger flower-foraging activities than moths that lack this alignment. Moth locomotor activity is circadian clock-regulated, although it is also strongly repressed by light. Moths show a time-dependent burst increase in flight activity during subjective night. In addition, moth antennal responsiveness to the floral scent compounds exhibits a 24-hour rhythm in both continuous light and dark conditions. This study highlights the importance of the circadian clocks in both plants and animals as a crucial factor in initiating specialized plant-pollinator relationships.

Draft genomes and phenotypic characterization of Tisochrysis lutea strains. Toward the production of domesticated strains with high added value

Tisochrysis lutea is a microalga species currently used in aquaculture as a feed for shellfish, oysters and shrimps. It also has many other potential industrial applications, such as the production of neutral lipids for biofuels or the production of ω-3 fatty acids for nutraceuticals (human food complements). To efficiently exploit the potential of this microalga, however, higher lipid productivities are needed. To this end, improvement programs need to be developed and optimized. The diversity of strains available in microalgae has not yet been exploited in such improvement programs.In this study, the intra-strain diversity was observed and exploited to increase neutral lipid productivity. New clonal strains with higher neutral lipid productivity were successfully selected. The best clonal strain selected accumulated 520% more triacylglycerols, with a similar growth rate to the wild-type strain in continuous light and nitrogen starvation conditions. In a photoperiod culture condition, this clonal stain also accumulated 84% more storage lipids and 30% less carbohydrates, compared to the wild-type strain. This clonal strain thus had a higher productivity which is of great interest for feed or biofuel applications.This study also focused on identifying the genomic mechanisms responsible for the improvements in these clonal strains. With this objective, the genome of Tisochrysis lutea was sequenced for the first time. It is the third genome of a Haptophyte microalga sequenced so far. Different genetic polymorphisms were identified between the sequenced genomes of the wild-type strain and clonal strains. Activity of transposable elements seems to have been involved in the genome reshuffling obtained through the improvement program. The contribution of transposable elements to the adaptive capacity of microalgae remains to be demonstrated.

The light cycle controls the hatching rhythm in Bombyx mori via negative feedback loop of the circadian oscillator.

Hatching behavior is a key target in silkworm (Bombyx mori) rearing, especially for the control of Lepidoptera pests. According to previous research, hatching rhythms appear to be controlled by a clock mechanism that restricts or "gates" hatching to a particular time. However, the underlying mechanism remains elusive. Under 12-h light:12-h dark photoperiod (LD) conditions, the transcriptional levels of the chitinase5 (Cht5) and hatching enzyme-like (Hel) genes, as well as the enzymatic activities of their gene products, oscillated in time with ambient light cycles, as did the transcriptional levels of the cryptochrome 1, cryptochrome 2, period (per), and timeless genes, which are key components of the negative feedback loop of the circadian rhythm. These changes were related to the expression profile of the ecdysteroid receptor gene and the hatching behavior of B. mori eggs. However, under continuous light or dark conditions, the hatching behavior, the expression levels of Cht5 and Hel, as well as the enzymatic activities of their gene products, were not synchronized unlike under LD conditions. In addition, immunohistochemistry experiments showed that light promoted the translocation of PER from the cytoplasm to the nucleus. In conclusion, LD cycles regulate the hatching rhythm of B. mori via negative feedback loop of the circadian oscillator.

The Tomato DOF Daily Fluctuations 1, TDDF1 acts as flowering accelerator and protector against various stresses

Adaptation to environmental changes is an important fitness trait for crop development. Photoperiod is an essential factor in seasonal control of flowering time. Sensing of day-length requires an interaction between the Photoperiod and the endogenous rhythms that is controlled by plant circadian clock. Thus, circadian clock is a critical regulator and internal molecular time-keeping mechanism, controlling key agricultural traits in crop plants such as the ability to adjust their growth and physiology to anticipate diurnal environmental changes. Here, we describe the gene Tomato Dof Daily Fluctuations 1 (TDDF1), which is involved in circadian regulation and stress resistance. Large daily oscillations in TDDF1 expression were retained after transferring to continuous dark (DD) or light (LL) conditions. Interestingly, overexpressing TDDF1 induce early flowering in tomato through up-regulation of the flowering-time control genes, moreover, by protein-protein interaction with the floral inducer SFT gene. Notably, overexpressing TDDF1 in tomato was associated with chlorophyll overaccumulation by up-regulating the related biosynthetic genes. TDDF1 expression results in improved drought, salt, various hormones stress tolerance alongwith resistance to late blight caused by Phytophthora infestans. This study can be a distinctive strategy to improve other economically important crops.

Impact of constant versus fluctuating temperatures on the development and life history parameters of Tetranychus urticae (Acari: Tetranychidae).

The impact of daily temperature fluctuations on arthropod life history parameters is inadequately studied compared with the ample amount of research that has been conducted on the effects of constant temperatures. Fluctuating temperatures are likely to be more realistic, as they are ecologically more similar to what these arthropods experience in nature. Here, we compared the impact of 11 constant temperatures that ranged from 10 to 35°C with fluctuating temperatures with the same corresponding mean temperature and an amplitude of 10°C between high (12h) and low (12h) temperatures on the development and life history parameters of Tetranychus urticae under continuous light conditions. No eggs hatched at constant 10°C, whereas 81.5% of eggs successfully completed development at fluctuating 10°C (15/5°C). Egg-to-female adult development was faster under fluctuating temperatures from 12.5 to 27.5°C than under constant temperatures, whereas the opposite trend was observed at >30°C. The lower thermal thresholds (T) were 11.63 and 8.63°C, and thermal constants (K) were 127.81 and 150.69 degree-days for egg-to-female adults at constant and fluctuating temperatures, respectively. The numbers of oviposition days were significantly higher at fluctuating 15°C than at the corresponding constant temperature, whereas the opposite trend was observed from 20 to 30°C. The intrinsic rate of increase (r) was higher at fluctuating than at constant 15°C. The net reproductive rate (R 0) was also higher at fluctuating than at constant 15 and 35°C, but showed an opposite trend at 20 and 25°C. We conclude that fluctuating temperatures should be considered to accurately predict spider mite population dynamics in nature.

English

The artificial lighting conditions which promoted growth of the gametophytes and sporophytes of brown alga Undaria pinnatifida were examined. The seaweed was subjected to continuous or intermittent white, blue, or red light. There were notable, but not significant, differences in gametophyte and sporophyte growth between continuous and intermittent (104 Hz) white light conditions. Gametophyte growth was promoted most notably by white, followed by blue light. Sporophyte growth length was promoted most notably under intermittent white light, while body length and blade area were promoted notably under continuous white light. Sporophytes under blue or red light withered considerably. The results showed that white light is more beneficial for growth of both U. pinnatifida gametophytes and sporophytes compared with blue or red light. Male and female gametophyte grew more robustly under white light regardless of whether the pattern was intermittent or continuous light. However, the results further indicated that overall continuous white light promoted growth to a greater degree than did intermittent white light. Finally, white light promoted U. pinnatifida sporophyte growth to a greater degree than blue or red light. Key words: Light wavelength, intermittent light, Undaria pinnatifida, seaweed culture, sporophyte growth.

Pulsed Light-Emitting Diodes for a Higher Phytochemical Level in Microgreens.

A novel research of pulsed light-emitting diode (LED) lighting versus continuous lighting was conducted by analyzing phytochemical levels in microgreens. Red pak choi (Brassica rapa var. chinensis), mustard (Brassica juncea L.), and tatsoi (Brassica rapa var. rosularis) were grown indoors under HPS lamps supplemented with monochromatic (455, 470, 505, 590, and 627 nm) LEDs [total photosynthetic photon flux density (PPFD) of 200 ± 10 μmol m-2 s-1, for 16 h day-1]. For pulsed light treatments, the frequencies at 2, 32, 256, and 1024 Hz with a duty cycle of 50% monochromatic LEDs were applied. The results were compared to those under the continuous light (0 Hz) condition in terms of total phenolic content, anthocyanins, and antiradical activity (DPPH). The summarized data suggested that pulsed light affected accumulation of secondary metabolites both positive and negative in microgreens. The significant differences in the response of phytochemicals between pulsed light at several frequencies and continuous light were determined. The most positive effects of 2, 256, and 1024 Hz for total phenolic compounds in mustard under all wavelength LEDs were achieved. The LED frequencies at 2 and 32 Hz were the most suitable for accumulation of anthocyanins in red pak choi and tatsoi. The highest antiradical activity under the treatments of 32, 256, and 1024 Hz in mustard and under the 2 Hz frequency in red pak choi and tatsoi was determined.

Real-time monitoring of PtaHMGB activity in poplar transactivation assays

BackgroundPrecise control of gene expression is essential to synchronize plant development with the environment. In perennial plants, transcriptional regulation remains poorly understood, mainly due to the long time required to perform functional studies. Transcriptional reporters based on luciferase have been useful to study circadian and diurnal regulation of gene expression, both by transcription factors and chromatin remodelers. The high mobility group proteins are considered transcriptional chaperones that also modify the chromatin architecture. They have been found in several species, presenting in some cases a circadian expression of their mRNA or protein.ResultsTransactivation experiments have been shown as a powerful and fast method to obtain information about the potential role of transcription factors upon a certain reporter. We designed and validated a luciferase transcriptional reporter using the 5′ sequence upstream ATG of Populus tremula × alba LHY2 gene. We showed the robustness of this reporter line under long day and continuous light conditions. Moreover, we confirmed that pPtaLHY2::LUC activity reproduces the accumulation of PtaLHY2 mRNA. We performed transactivation studies by transient expression, using the reporter line as a genetic background, unraveling a new function of a high mobility group protein in poplar, which can activate the PtaLHY2 promoter in a gate-dependent manner. We also showed PtaHMGB2/3 needs darkness to produce that activation and exhibits an active degradation after dawn, mediated by the 26S proteasome.ConclusionsWe generated a stable luciferase reporter poplar line based on the circadian clock gene PtaLHY2, which can be used to investigate transcriptional regulation and signal transduction pathway. Using this reporter line as a genetic background, we established a methodology to rapidly assess potential regulators of diurnal and circadian rhythms. This tool allowed us to demonstrate that PtaHMGB2/3 promotes the transcriptional activation of our reporter in a gate-dependent manner. Moreover, we added new information about the PtaHMGB2/3 protein regulation along the day. This methodology can be easily adapted to other transcription factors and reporters.

Transcriptome Analysis of Genes Associated with the Artemisinin Biosynthesis by Jasmonic Acid Treatment under the Light in Artemisia annua.

Artemisinin is a sesquiterpene lactone endoperoxide extracted from a traditional Chinese medicinal plant Artemisia annua. Artemisinin-based combination therapies (ACTs) are recommended as the best treatment of malaria by the World Health Organization (WHO). Both the phytohormone jasmonic acid (JA) and light promote artemisinin biosynthesis in A. annua. Interestingly, we found that the increase of artemisinin biosynthesis by JA was dependent on light. However, the relationship between the two signal pathways mediated by JA and light remains unclear. Here, we collected the A. annua seedlings of 24 h continuous light (Light), 24 h dark treatment (Dark), 4 h MeJA treatment under the continuous light conditions (Light-MeJA-4h) and 4 h MeJA treatment under the dark conditions (Dark-MeJA-4h) and performed the transcriptome sequencing using Illumina HiSeq 4000 System. A total of 266.7 million clean data were produced and assembled into 185,653 unigenes, with an average length of 537 bp. Among them, 59,490 unigenes were annotated and classified based on the public information. Differential expression analyses were performed between Light and Dark, Light and Light-MeJA-4h, Dark and Dark-MeJA-4h, Light-MeJA-4h, and Dark-MeJA-4h, respectively. Furthermore, transcription factor (TF) analysis revealed that 1588 TFs were identified and divided into 55 TF families, with 284 TFs down-regulated in the Dark relative to Light and 96 TFs up-regulated in the Light-MeJA-4h relative to Light. 8 TFs were selected as candidates for regulating the artemisinin biosynthesis and one of them was validated to be involved in artemisinin transcriptional regulation by Dual-Luciferase (Dual-LUC) assay. The transcriptome data shown in our study offered a comprehensive transcriptional expression pattern influenced by the MeJA and light in A. annua seedling, which will serve as a valuable resource for further studies on transcriptional regulation mechanisms underlying artemisinin biosynthesis.

Expression profiles of genes for mitochondrial respiratory energy-dissipating systems and antioxidant enzymes in wheat leaves during de-etiolation

Mitochondrial respiratory components participate in the maintenance of chloroplast functional activity. This study investigates the effects 48h de-etiolation of spring wheat seedlings (Triticum aestivum L., var. Irgina) on the expression of genes that encode energy-dissipating respiratory components and antioxidant enzymes under continuous light conditions. The expression of AOX1a following the prolonged darkness exhibited a pattern indicating a prominent dependence on light. The expression of other respiratory genes, including NDA2, NDB2, and UCP1b, increased during de-etiolation and dark-to-light transition; however, changes in the expression of these genes occurred later than those in AOX1a expression. A high expression of NDA1 was detected after 12h of de-etiolation. The suppression of AOX1a, NDA2, NDB2, and UCP1b was observed 24h after de-etiolation when the photosynthetic apparatus and its defence systems against excess light were completely developed. The expression patterns of the respiratory genes and several genes encoding antioxidant enzymes (MnSOD, Cu-ZnSOD, t-APX, GR, and GRX) were quite similar. Our data indicate that the induction of nuclear genes encoding respiratory and antioxidant enzymes allow the plants to control reactive oxygen species (ROS) production and avoid oxidative stress during de-etiolation.

Chloroplast overexpression of rice caffeic acid O‐methyltransferase increases melatonin production in chloroplasts via the 5‐methoxytryptamine pathway in transgenic rice plants

Recent analyses of the enzymatic features of various melatonin biosynthetic genes from bacteria, animals, and plants have led to the hypothesis that melatonin could be synthesized via the 5-methoxytryptamine (5-MT) pathway. 5-MT is known to be synthesized in vitro from serotonin by the enzymatic action of O-methyltransferases, including N-acetylserotonin methyltransferase (ASMT) and caffeic acid O-methyltransferase (COMT), leading to melatonin synthesis by the subsequent enzymatic reaction with serotonin N-acetyltransferase (SNAT). Here, we show that 5-MT was produced and served as a precursor for melatonin synthesis in plants. When rice seedlings were challenged with senescence treatment, 5-MT levels and melatonin production were increased in transgenic rice seedlings overexpressing the rice COMT in chloroplasts, while no such increases were observed in wild-type or transgenic seedlings overexpressing the rice COMT in the cytosol, suggesting a 5-MT transport limitation from the cytosol to chloroplasts. In contrast, cadmium treatment led to results different from those in senescence. The enhanced melatonin production was not observed in the chloroplast COMT lines relative over the cytosol COMT lines although 5-MT levels were equally induced in all genotypes upon cadmium treatment. The transgenic seedlings with enhanced melatonin in their chloroplasts exhibited improved seedling growth vs the wild type under continuous light conditions. This is the first report describing enhanced melatonin production in chloroplasts via the 5-MT pathway with the ectopic overexpression of COMT in chloroplasts in plants.