Shift in the Light Quality of Night Interruption Affects Flowering and Morphogenesis of Petunia hybrida.

Petunia hybrida Hort. "Easy Wave Pink", a qualitative long-day plant (LDP), was investigated to study the effects of the night interruption light (NIL) provided by light-emitting diodes (LEDs) quality shifting on the morphogenesis, blooming, and transcription of photoreceptor genes. Plants were grown in a closed-type plant factory employing white (W) LEDs at an intensity of 180 µmol·m-2·s-1 PPFD provided for short day (SD, 10 h light, 14 h dark), long day (LD, 16 h light, 8 h dark), or SD with 4 h night interruption (NI) with LEDs at an intensity of 10 µmol·m-2·s-1 PPFD. The NIL quality was shifted from one light spectrum to another after the first 2 h of NI. Light treatments consisting of all possible pairings of W, far-red (Fr), red (R), and blue (B) light were tested. The SD and LD were referenced as the control, while 12 NI treatments involved altering LED NIL qualities, as follows: from R to B (NI-RB), from B to R (NI-BR), from Fr to R (NI-FrR), from R to Fr (NI-RFr), from Fr to B (NI-FrB), from B to Fr (NI-BFr), from B to W (NI-BW), from W to B (NI-WB), from W to Fr (NI-WFr), from Fr to W (NI-FrW), from W to R (NI-WR), and from R to W (NI-RW). The NI-RFr resulted in the longest shoots, while the NI-WR and NI-RW resulted in the shortest shoots. NI-WR, NI-RW, NI-BW, NI-WB, NI-RFr, NI-RB, NI-BR, and LD all exhibited flowering. High-level expressions of photoreceptor genes were confirmed in the NI-RFr, NI-FrR, NI-BFr, NI-RW, and NI-WR treatments. Morphogenesis and blooming were both impacted by the photoperiod. The first NIL had no effects on the flowering or the morphogenesis, but the second NIL had a profound impact on both.

Plant Growth-Promoting Microorganism Pseudarthrobacter sp. NIBRBAC000502770 Enhances the Growth and Flavonoid Content of Geum aleppicum.

Plant growth-promoting rhizobacteria are known to enhance the growth and antioxidant activity of several plants. However, the effects of such rhizobacteria on Geum aleppicum, a plant with pharmacological potential in Korea are unknown. In this study, we investigated the effects of Pseudarthrobacter sp. NIBRBAC000502770 treatment (100 mL/pot, every two weeks for 55 days), in the form of culture medium, 100-fold diluted culture, culture supernatant, and pelleted cells resuspended in water, on the growth, antibacterial activity and flavonoid content of G. aleppicum. The NIBRBAC000502770 strain showed high indole-3-acetic acid (IAA) content of 461.81 µg∙mL-1. The dry weight of the roots was significantly higher in the supernatant, diluted culture, and pellet-treated plants compared to that in the control plants. Additionally, the plant height, root length, leaf length, leaf width, chlorophyll content, biomass, and dry weight of the shoot were highest in the pellet-treated plants. Further, methanol extracts of pellet-treated plants showed significantly high flavonoid content compared to that in the control plants (28 mg∙g-1 vs. 7.5 mg∙g-1) and exhibited strong antibacterial activity against Gram-positive and negative bacteria. These results demonstrate the beneficial effects of Pseudarthrobacter sp. NIBRBAC000502770 on the growth and flavonoid content of G. aleppicum.

Red Light Enhances the Antioxidant Properties and Growth of Rubus hongnoensis.

The purpose of this study was to determine the effect of light quality on R.hongnoensis growth, physiology, and antioxidant properties. Five light conditions were employed, including white (control), red (R), blue (B), combined LED of R, green (G), and B at 7:1:2 (RGB), as well as combined LED of R, G, B, and far-red (Fr) at 7:1:2:1 (RGBFr). R light had the greatest growth-promoting effect based on plant height, leaf length, leaf width, stem diameter, and leaf area. However, leaf width and root length exhibited the greatest growth under RGB. The fresh and dry weight of shoots and roots were highest under R and RGB light. Photosynthesis was highest under RGB and lowest under B. Transpiration was highest in RGBFr. Stomatal conductance and photosynthetic water use efficiency were greatest under RGBFr. Total phenol content and radical scavenging activity were highest under R, while total flavonoid content was highest under RGB. Superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX) activities were upregulated under W, whereas guaiacol peroxidase (GPX) activity was highest under RGB. The present results suggest that, among the tested light treatments, R light was most conductive for vegetative growth and antioxidant capacity in R. hongnoensis.

Optimization of Fermentation Medium for Indole Acetic Acid Production by Pseudarthrobacter sp. NIBRBAC000502770.

Indole acetic acid (IAA) has been an important compound for plant growth and is widely known to be produced by plant growth-promoting rhizobacteria (PGPR). The isolate producing the maximum amount of IAA from the Korea shooting range soil was identified as Pseudarthrobacter sp. NIBRBAC000502770, using 16S rRNA gene sequencing. IAA production was determined in Luria-Bertani (LB) broth and optimized using different temperatures, agitation rates, L-tryptophan concentrations, carbon and nitrogen sources, and inorganic salts. The strain NIBRBAC000502770 showed better production of IAA at temperature 30 °C (29.47 mg·L-1) and at an agitation rate of 200 rpm (32.65 mg·L-1). Maltose (0.5%) was found to be the best carbon source for the strain (yielding 36.48 mg·L-1 IAA). IAA yield was 19.17 mg·L-1 and 24.73 mg·L-1 at 1% yeast extract and 1% tryptone as nitrogen sources, respectively. qRT-PCR showed the transcript levels of amiE and aldH genes, which had been predicted to encode indole-3-acetamide hydrolase and indole-3-acetaldehyde dehydrogenase, to be significantly upregulated in response to tryptophan. This study has examined that NIBRBAC000502770 has significant effects as a biological agent such as plant growth promotion, and development of optimal medium could significantly reduce the cost of mass production of microorganisms.

Effect of Iron Source and Medium pH on Growth and Development of Sorbus commixta In Vitro.

Sorbus commixta is a valuable hardwood plant with a high economical value for its medicinal and ornamental qualities. The aim of this work was to investigate the effects of the iron (Fe) source and medium pH on the growth and development of S. commixta in vitro. The Fe sources used, including non-chelated iron sulfate (FeSO4), iron ethylenediaminetetraacetic acid (Fe-EDTA), and iron diethylenetriaminepentaacetic acid (Fe-DTPA), were supplemented to the Multipurpose medium with a final Fe concentration of 2.78 mg·L-1. The medium without any supplementary Fe was used as the control. The pH of the agar-solidified medium was adjusted to either 4.70, 5.70, or 6.70. The experiment was conducted in a culture room for six weeks with 25 °C day and night temperatures, and a 16-h photoperiod with a light intensity of 50 mmol·m-2·s-1 photosynthetic photon flux density (PPFD). Both the Fe source and pH affected the growth and development of the micropropagated plants in vitro. The leaves were greener in the pH 4.70 and 5.70 treatments. The tissue Fe content decreased with the increase of the medium pH. The leaf chlorophyll content was similar between plants treated with FeSO4 and those with Fe-EDTA. The numbers of the shoots and roots of plantlets treated with FeSO4 were 2.5 and 2 times greater than those of the control, respectively. The fresh and dry weights of the shoot and the root were the greatest for plants treated with Fe-EDTA combined with pH 5.70. The calcium, magnesium, and manganese contents in the plantlets increased in the pH 5.70 treatments regardless of the Fe source. Supplementary Fe decreased the activity of ferric chelate reductase. Overall, although the plantlets absorbed more Fe at pH 4.70, Fe-EDTA combined with pH 5.70 was found to be the best for the growth and development of S. commixta in vitro.

How Supplementary or Night-Interrupting Low-Intensity Blue Light Affects the Flower Induction in Chrysanthemum, A Qualitative Short-Day Plant.

This research examined the effects of the supplementary or night-interrupting (NI) blue (B) light supplied at a low intensity on the flowering, gene expression, and morphogenesis of chrysanthemum, a qualitative short-day plant. White (W) light-emitting diodes (LEDs) were used to provide light with a photosynthetic photon flux density (PPFD) of 180 µmol·m-2·s-1 during the photoperiod to grow the plants in a plant factory. The control group was constructed with plants that were exposed to a 10-h short day (SD10) treatment without any blue light. The B light in this research was used for 4 h to either (1) extend the photoperiod for plants at the end of a 9-h short day (SD) treatment as the sole light source (SD9 + 4B), (2) provide night interruption (NI) to plants in the 13-h long-day (LD) treatment (LD13 + NI - 4B), (3) provide NI to plants in the 10-h SD treatment (SD10 + NI - 4B), or (4) supplement the W LEDs at the end of a 13-h LD treatment (LD13 + 4B). Blue LEDs were used to provide the supplementary/NI light at 10 µmol·m-2·s-1 PPFD. The LD13 + NI - 4B treatment resulted in the greatest plant height, followed by LD13 + 4B. Plants in all treatments flowered. It is noteworthy that despite the fact that chrysanthemum is a qualitative SD plant, chrysanthemum plants flowered when grown in the LD13 + 4B and LD13 + NI - 4B treatments. Plants grown in the LD13 + 4B had the greatest number of flowers. Plants grown in the LD13 + 4B treatment had the highest expression levels of the cryptochrome 1, phytochrome A, and phytochrome B genes. The results of this study indicate that a 4-h supplementation of B light during the photoperiod increases flower bud formation and promotes flowering, and presents a possibility as an alternative method to using blackout curtains in LD seasons to practically induce flowering. The B light application methods to induce flowering in SD plants requires further research.

Physiological and Proteomic Insights Into Red and Blue Light-Mediated Enhancement of in vitro Growth in Scrophularia kakudensis-A Potential Medicinal Plant.

The current study has determined the effect of red and blue lights on the enhancement of growth, antioxidant property, phytochemical contents, and expression of proteins in Scrophularia kakudensis. In vitro-grown shoot tip explants of S. kakudensis were cultured on the plant growth regulator-free Murashige and Skoog (MS) medium and cultured under the conventional cool white fluorescent lamp (control), blue light-emitting diodes (LED) light, or red LED light. After 4 weeks, growth, stomatal ultrastructure, total phenols and flavonoids, activities of antioxidant enzymes, and protein expressions were determined. Interestingly, blue or red LED treatment increased the shoot length, shoot diameter, root length, and biomass on comparison with the control. In addition, the LED treatments enhanced the contents of phytochemicals in the extracts. The red LED treatment significantly elicited the accumulation of flavonoids in comparison with the control. In accordance with the secondary metabolites, the LED treatments modulated the activities of antioxidant enzymes. Moreover, the proteomic insights using two-dimensional gel electrophoresis system revealed the proteins involved in transcription and translation, carbohydrate mechanism, post-translational modification, and stress responses. Taken together, the incorporation of blue or red LED during in vitro propagation of S. kakudensis can be a beneficial way to increase the plant quality and medicinal values of S. kakudensis.

Flowering and Morphogenesis of Kalanchoe in Response to Quality and Intensity of Night Interruption Light.

The effects of the quality and intensity of night interruption light (NIL) on the flowering and morphogenesis of kalanchoe (Kalanchoe blossfeldiana) 'Lipstick' and 'Spain' were investigated. Plants were raised in a closed-type plant factory under 250 µmol·m-2·s-1 PPFD white light emitting diodes (LEDs) with additional light treatments. These treatments were designated long day (LD, 16 h light, 8 h dark), short day (SD, 8 h light, 16 h dark), and SD with a 4 h night interruption (NI). The NIL was constructed from 10 µmol·m-2·s-1 or 20 µmol·m-2·s-1 PPFD blue (NI-B), red (NI-R), white (NI-W), or blue and white (NI-BW) LEDs. In 'Spain', the SPAD value, area and thickness of leaves and plant height increased in the NI treatment as compared to the SD treatment. In 'Lipstick', most morphogenetic characteristics in the NI treatment showed no significant difference to those in the SD treatment. For both cultivars, plants in SD were significantly shorter than those in other treatments. The flowering of Kalanchoe 'Lipstick' was not affected by the NIL quality, while Kalanchoe 'Spain' flowered when grown in SD and 10 µmol·m-2·s-1 PPFD NI-B. These results suggest that the NIL quality and intensity affect the morphogenesis and flowering of kalanchoe, and that different cultivars are affected differently. There is a need to further assess the effects of the NIL quality and intensity on the morphogenesis and flowering of short-day plants for practical NIL applications.

Proteomic Analysis of Aphid-Resistant and -Sensitive Rose (Rosa Hybrida) Cultivars at Two Developmental Stages.

The rose is one the most commercially grown and costly ornamental plants because of its aesthetic beauty and aroma. A large number of pests attack its buds, flowers, leaves, and stem at every growing stage due to its high sugar content. The most common pest on roses are aphids which are considered to be the major cause for product loss. Aphid infestations lead to major changes in rose plants, such as large and irregular holes in petals, intact leaves and devouring tissues. It is hypothesized that different cut rose cultivars would have different levels of sensitivity or resistance to aphids, since different levels of infestation are observed in commercially cut rose production greenhouses. The present work compared four cut rose cultivars which were bred in Korea and were either resistant or sensitive to aphid infestation at different flower developmental stages. An integrative study was conducted using comprehensive proteome analyses. Proteins related to ubiquitin metabolism and the stress response were differentially expressed due to aphid infestation. The regulations and possible functions of identified proteins are presented in detail. The differential expressions of the identified proteins were validated by immunoblotting and blue native page. In addition, total sugar and carbohydrate content were also observed.

Proteomic Analysis Reveals the Dynamic Role of Silicon in Alleviation of Hyperhydricity in Carnation Grown In Vitro.

The present study depicted the role of silicon in limiting the hyperhydricity in shoot cultures of carnation through proteomic analysis. Four-week-old healthy shoot cultures of carnation "Purple Beauty" were sub-cultured on Murashige and Skoog medium followed with four treatments, viz. control (-Si/-Hyperhydricity), hyperhydric with no silicon treatment (-Si/+Hyperhydricity), hyperhydric with silicon treatment (+Si/+Hyperhydricity), and only silicon treated with no hyperhydricity (+Si/-Hyperhydricity). Comparing to control morphological features of hyperhydric carnations showed significantly fragile, bushy and lustrous leaf nature, while Si supply restored these effects. Proteomic investigation revealed that approximately seventy protein spots were differentially expressed under Si and/or hyperhydric treatments and were either up- or downregulated in abundance depending on their functions. Most of the identified protein spots were related to stress responses, photosynthesis, and signal transduction. Proteomic results were further confirmed through immunoblots by selecting specific proteins such as superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT), PsaA, and PsbA. Moreover, protein-protein interaction was also performed on differentially expressed protein spots using specific bioinformatic tools. In addition, stress markers were analyzed by histochemical localization of hydrogen peroxide (H2O2) and singlet oxygen (O21-). In addition, the ultrastructure of chloroplasts in hyperhydric leaves significantly resulted in inefficiency of thylakoid lamella with the loss of grana but were recovered in silicon supplemented leaves. The proteomic study together with physiological analysis indicated that Si has a substantial role in upholding the hyperhydricity in in vitro grown carnation shoot cultures.

Chemical Elicitor-Induced Modulation of Antioxidant Metabolism and Enhancement of Secondary Metabolite Accumulation in Cell Suspension Cultures of Scrophularia kakudensis Franch.

Scrophularia kakudensis is an important medicinal plant with pharmaceutically valuable secondary metabolites. To develop a sustainable source of naturaceuticals with vital therapeutic importance, a cell suspension culture was established in S. kakudensis for the first time. Friable calli were induced from the leaf explants cultured on a Murashige and Skoog (MS) medium containing 3.0 mg·L(-1) 6-benzyladenine (BA) in a combination with 2 mg·L(-1) 2,4-dichlorophenoxy acetic acid (2,4-D). From the callus cultures, a cell suspension culture was initiated and the cellular differentiation was investigated. In addition, the effect of biotic elicitors such as methyl jasmonate (MeJa), salicylic acid (SA), and sodium nitroprusside (SNP) on the accumulation of secondary metabolites and antioxidant properties was demonstrated. Among the elicitors, the MeJa elicited the accumulation of total phenols, flavonoids, and acacetin, a flavonoid compound with multiple pharmaceutical values. Similarly, the higher concentrations of the MeJa significantly modulated the activities of antioxidant enzymes and enhanced the scavenging potentials of free radicals of cell suspension extracts. Overall, the outcomes of this study can be utilized for the large scale production of pharmaceutically important secondary metabolites from S. kakudensis through cell suspension cultures.

In Vitro Propagation, Phytochemical Analysis, and Evaluation of Free Radical Scavenging Property of Scrophularia kakudensis Franch Tissue Extracts.

The current study deals with in vitro propagation, antioxidant property estimation, and assessment of acacetin content in Scrophularia kakudensis Franch. Adventitious shoot induction was achieved from the nodal explant with the highest number of adventitious shoots per explant (17.4) on Murashige and Skoog's (MS) medium fortified with 2.0 mg·L(-1) 6-benzyladenine (BA) and 0.5 mg L(-1) indole-3-acetic acid (IAA). Maximum number of roots per plant (16.5) was noted in half strength MS medium supplemented with 0.5 mg·L(-1) IAA. The regenerated plants displayed successful survival ratio (95%) in the greenhouse. The highest content of acacetin, a pharmaceutically important flavonoid, was observed in the shoot extracts (in vitro: 32.83 µg·g(-1) FW; in vivo: 30.05 µg·g(-1) FW) followed by root extracts. Total phenol and flavonoid contents along with free radical scavenging assays revealed the occurrence of larger amount of antioxidants in shoot extract in comparison with callus and root extracts of S. kakudensis. Thus, the outcome of the present study can be highly beneficial for the germplasm conservation and commercial cultivation of S. kakudensis for therapeutic purposes.

Morphogenesis, Flowering, and Gene Expression of Dendranthema grandiflorum in Response to Shift in Light Quality of Night Interruption.

The impact of shifts in the spectral quality of light on morphogenesis, flowering, and photoperiodic gene expression during exposure to light quality of night interruption (NI) was investigated in Dendranthema grandiflorum. The circadian rhythms of plants grown in a closed walk-in growth chamber were interrupted at night for a total of 4 h, using light-emitting diodes with an intensity of 10 µmol·m⁻²·s⁻¹ PPF. The light quality of the NI was shifted from one wavelength to another after the first 2 h. Light treatments consisting of all possible pairings of blue (B), red (R), far-red (Fr), and white (W) light were tested. Plants in the NI treatment groups exposed to Fr light grew larger than plants in other treatment groups. Of plants in NI treatment groups, those in the NI-WB treatment grew the least. In addition, the impact of shifts in the light quality of NI on leaf expansion was greater in treatment groups exposed to a combination of either B and R or R and W light, regardless of their order of supply. Flowering was observed in the NI-RB, NI-FrR, NI-BFr, NI-FrB, NI-WB, NI-FrW, NI-WFr, NI-WR, and SD (short-day) treatments, and was especially promoted in the NI-BFr and NI-FrB treatments. In a combined shift treatment of B and R or B and W light, the NI concluded with B light (NI-RB and NI-WB) treatment induced flowering. The transcriptional factors phyA, cry1 and FTL (FLOWERING LOCUS T) were positively affected, while phyB and AFT were negatively affected. In conclusion, morphogenesis, flowering, and transcriptional factors were all significantly affected either positively or negatively by shifts in the light quality of NI. The light quality of the first 2 h of NI affected neither morphogenesis nor flowering, while the light quality of the last 2 h of NI significantly affected both morphogenesis and flowering.

Proteomic study related to vascular connections in watermelon scions grafted onto bottle-gourd rootstock under different light intensities.

Although grafting is broadly used in the production of crops, no information is available about the proteins involved in vascular connections between rootstock and scion. Similarly, proteome changes under the light intensities widely used for grafted seedlings are of practical use. The objective of this study was to determine the proteome of vascular connections using watermelon (Citrullus vulgaris Schrad.) 'Sambok Honey' and 'Speed' as the scion and bottle gourd (Lagenaria siceraria Stanld.) 'RS Dongjanggun' as the rootstock grown under different light intensities (25, 50, 75 and 100 µmol m-2 s-1). Our proteomic analysis revealed 24 and 27 differentially expressed proteins in 'Sambok Honey' and 'Speed', respectively, under different light intensities. The identified proteins were largely involved in ion binding, amino acid metabolism, transcriptional regulation and defense response. The enhancement of ion-binding, transcriptional regulation, amino acid metabolism, and defense response proteins suggests a strengthening of the connection between the rootstock and scion under high light intensity. Indeed, the accumulation of key enzymes in the biological processes described above appears to play an important role in the vascular connections of grafted seedlings. Moreover, it appears that 100 µmol m-2 s-1 results in better protein expression responses in grafted seedlings.

Physiological and proteomic analysis in chloroplasts of Solanum lycopersicum L. under silicon efficiency and salinity stress.

Tomato plants often grow in saline environments in Mediterranean countries where salt accumulation in the soil is a major abiotic stress that limits its productivity. However, silicon (Si) supplementation has been reported to improve tolerance against several forms of abiotic stress. The primary aim of our study was to investigate, using comparative physiological and proteomic approaches, salinity stress in chloroplasts of tomato under silicon supplementation. Tomato seedlings (Solanum lycopersicum L.) were grown in nutrient media in the presence or absence of NaCl and supplemented with silicon for 5 days. Salinity stress caused oxidative damage, followed by a decrease in silicon concentrations in the leaves of the tomato plants. However, supplementation with silicon had an overall protective effect against this stress. The major physiological parameters measured in our studies including total chlorophyll and carotenoid content were largely decreased under salinity stress, but were recovered in the presence of silicon. Insufficient levels of net-photosynthesis, transpiration and stomatal conductance were also largely improved by silicon supplementation. Proteomics analysis of chloroplasts analyzed by 2D-BN-PAGE (second-dimensional blue native polyacrylamide-gel electrophoresis) revealed a high sensitivity of multiprotein complex proteins (MCPs) such as photosystems I (PSI) and II (PSII) to the presence of saline. A significant reduction in cytochrome b6/f and the ATP-synthase complex was also alleviated by silicon during salinity stress, while the complex forms of light harvesting complex trimers and monomers (LHCs) were rapidly up-regulated. Our results suggest that silicon plays an important role in moderating damage to chloroplasts and their metabolism in saline environments. We therefore hypothesize that tomato plants have a greater capacity for tolerating saline stress through the improvement of photosynthetic metabolism and chloroplast proteome expression after silicon supplementation.