The red/blue light ratios from light-emitting diodes affect growth and flower quality of Hippeastrum hybridum 'Red Lion'.

Light quality strongly impacts the growth and flower quality of ornamental plants. The optimum light quality for the growth and flowering of Hippeastrum remains to be validated. In the present study, we investigated the effect of the red/blue light ratio of LEDs on the growth and flowering quality of H. hybrid 'Red Lion'. Two LEDs with red/blue light ratio of 1:9 (R10B90) and 9:1 (R90B10) were designed. LEDs of white light were the control. In the earlier vegetative and reproductive growth phase, R90B10 increased the biomass of the bulbs, leaves, and flowers. Compared with the control and R10B90 group, R90B10 LEDs delayed flowering by 2.30 d and 3.26 d, respectively. Based on chlorophyll contents, photosynthetic capacity, chlorophyll fluorescence parameters, and carbohydrate contents, the photosynthesis rate was higher in the R10B90 group. Optimal red and blue light intensity promoted the accumulation of carbohydrates and early flowering and prolonged the flowering period of H. hybrid. Microscopic analysis showed that stomatal density was high, and the number of chloroplasts was large in the R10B90 treatment group, which enhanced photosynthesis. Particularly, R10B90 promoted the expression of seven key genes related to chlorophyll synthesis. R10B90 also promoted early overexpression of the HpCOL gene that promotes early flowering. Thus, higher blue light and 10% red light intensities promote early and extended flowering, while higher red light and 10% blue light promote vegetative plant growth but delay flowering.

Chemical Profile and Skin-Beneficial Activities of the Petal Extracts of Paeonia tenuifolia L. from Serbia.

Without being aware of its chemical makeup, many ancient societies have used Steppe peony in their traditional medicine. Given that modern phytopreparation intended for use on human skin requires, above all, knowledge of its chemical composition, the goal of this study was to make a screening of the composition of aqueous and methanolic extracts of the petals of P. tenuifolia L. and to examine them for various skin-beneficial properties. The extracts were prepared by maceration, ultrasound-assisted, and microwave-assisted extraction procedures. The chemical profiling was conducted by the use of UHPLC-LTQ-OrbiTrap MS and UHPLC/MS, and spectrophotometric methods for the determination of total polyphenol and total flavonoid contents. The biological activities entailed antioxidant ABTS, DPPH, CUPRAC (Cupric Ion Reducing Antioxidant Capacity), and FRAP (Ferric Reducing Antioxidant Power) assays, antimicrobial (antibacterial and antifungal) and antibiofilm activities, cytotoxicity, wound healing potential, as well as the adhesion and invasion of Staphylococcus lugdunensis. The results showed that the petals are rich in phenolic acids and flavonoids, which are commonly associated with numerous biological activities. The aqueous extracts were more efficient in the majority of the bioactivity assays then the methanolic ones, whereas the optimal extraction method varied between the assays. This study is the first step towards the safe use of the aqueous extracts of P. tenuifolia petals for therapeutic skin treatments.

Karyotype Analysis, Genomic and Fluorescence In Situ Hybridization (GISH and FISH) Reveal the Ploidy and Parental Origin of Chromosomes in Paeonia Itoh Hybrids.

Itoh hybrids are intersectional hybrids in Paeonia L. with sect. Moutan and sect. Paeonia as paternal and maternal parents, respectively. Therefore, these hybrids have herbaceous stems with improved ornamental value introduced by the paternal parent. Although both of their parents are diploids, Itoh hybrids are triploids. Moreover, the parental origin of their chromosomes has not been extensively studied. This study systematically analyzed the genome size, ploidy, and karyotype of Itoh hybrids and compared them with their parental taxa. Although the monoploid genome size of Itoh hybrids was different, it was not significantly different from that of the parents. However, the size of varieties in the two parental taxa was significantly different from the wild species, probably due to genome rearrangements caused by artificial selection. Further karyotype analysis, correlation analysis, and hierarchical clustering could not identify the parental origin of chromosomes in Itoh hybrids. Verification through genomic and fluorescence in situ hybridization (GISH and FISH) suggested that for the three sets of chromosomes in Itoh hybrids, two were from the paternal parent, and one was from the maternal parent. One of the first two sets was from wild species, and the other from a cultivated variety. GISH could not label the chromosomes of cultivated peonies from the sect. Moutan, probably due to the huge and complex genomes compared with the wild species. Meanwhile, 5S rDNA-based FISH was first applied in Paeonia, which may be used for ploidy assessment. This work may give insights into the utilization of Itoh hybrid resources.

A Comparative Transcriptome Analysis Reveals the Molecular Mechanisms That Underlie Somatic Embryogenesis in Peaonia ostii 'Fengdan'.

Low propagation rate is the primary problem that limits industry development of tree peony. In this study, a highly efficient regeneration system for tree peony using somatic embryogenesis (SE) was established. The transcriptomes of zygotic embryo explants (S0), non-embryonic callus (S1), embryonic callus (S2), somatic embryos (S3), and regenerated shoots (S4) were analyzed to determine the regulatory mechanisms that underlie SE in tree peony. The differentially expressed genes (DEGs) were identified in the pairwise comparisons of S1-vs-S2 and S1-vs-S3, respectively. The enriched DEGs were primarily involved in hormone signal transduction, stress response and the nucleus (epigenetic modifications). The results indicated that cell division, particularly asymmetric cell division, was enhanced in S3. Moreover, the genes implicated in cell fate determination played central roles in S3. Hormone signal pathways work in concert with epigenetic modifications and stress responses to regulate SE. SERK, WOX9, BBM, FUS3, CUC, and WUS were characterized as the molecular markers for tree peony SE. To our knowledge, this is the first study of the SE of tree peony using transcriptome sequencing. These results will improve our understanding of the molecular mechanisms that underly SE in tree peony and will benefit the propagation and genetic engineering of this plant.

Nutrient Supply Is Essential for Shifting Tree Peony Reflowering Ahead in Autumn and Sugar Signaling Is Involved.

The flowering time of tree peony is short and concentrated in spring, which limits the development of its industry. We previously achieved tree peony reflowering in autumn. Here, we further shifted its reflowering time ahead through proper gibberellin (GA) treatment plus nutrient supply. GA treatment alone initiated bud differentiation, but it aborted later, whereas GA plus nutrient (G + N) treatment completed the opening process 38 days before the control group. Through microstructural observation of bud differentiation and starch grains, we concluded that GA plays a triggering role in flowering induction, whereas the nutriment supply ensured the continuous developing for final opening, and both are necessary. We further determined the expression of five floral induction pathway genes and found that PsSOC1 and PsLFY probably played key integral roles in flowering induction and nutrient supply, respectively. Considering the GA signaling, PsGA2ox may be mainly involved in GA regulation, whereas PsGAI may regulate further flower formation after nutrient application. Furthermore, G + N treatment, but not GA alone, inhibited the expression of PsTPS1, a key restricting enzyme in sugar signaling, at the early stage, indicating that sugar signaling is also involved in this process; in addition, GA treatment induced high expression of PsSnRK1, a major nutrient insufficiency indicator, and the induction of PsHXK1, a rate-limiting enzyme for synthesis of sugar signaling substances, further confirmed the nutrient shortage. In short, besides GA application, exogenous nutrient supply is essential to shift tree peony reflowering ahead in autumn under current forcing culture technologies.

Acidic and Alkaline Conditions Affect the Growth of Tree Peony Plants via Altering Photosynthetic Characteristics, Limiting Nutrient Assimilation, and Impairing ROS Balance.

Exposure to acidic and alkaline conditions were found to cause the excess accumulation of reactive oxygen species in tree peony, thereby causing damage and inhibiting plant growth and development. The activities of antioxidant enzymes were also found to be significantly up-regulated, especially under alkaline conditions; this explained why tree peony is better adapted to alkaline than to acidic conditions. Through pairwise comparisons, 144 differentially expressed genes (DEGs) associated with plant growth, photosynthesis, and stress were identified. The DEGs related to stress were up-regulated, whereas the remaining DEGs were almost all down-regulated after acid and alkaline treatments. The nutrient assimilation was greatly inhibited. Chlorophyll synthesis genes were suppressed, and chlorophyll content was reduced. The development and structures of stomata and chloroplasts and the transcription of related genes were also influenced. Among photosynthesis-related DEGs, electron transport chains were the most sensitive. The suppressed expression of photosynthesis genes and the reduced light-harvesting capacity, together with the impairment of chloroplasts and stomata, finally led to a sharp decrease in the net photosynthetic rate. Carbohydrate accumulation and plant biomass were also reduced. The present study provides a theoretical basis for the response mechanisms of tree peony to adverse pH conditions and enriches knowledge of plant adaptation to alkaline conditions.

Seed Proteomic Profiles of Three Paeonia Varieties and Evaluation of Peony Seed Protein as a Food Product.

Peony (Paeonia) has high ornamental, edible, and medicinal values. In order to distinguish seeds varieties, describe the proteomic profiles correlated with stress tolerance, and evaluate peony seed protein (PSP) as a functional food product, we characterized the seed protein profiles of these three species and their glucosidase inhibition activities. Results showed that the intensity of protein bands in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and specific protein ID (especially for specifically expressed proteins (SEPs)) was effective to distinguish these peony seed varieties. Proteomic analysis of the three species showed that P. ostii "Fengdan" has heat and pathogen tolerance-related proteins, while P. rockii has higher content of proteins related to cold resistance, which were all highly consistent with their adaptation of heat or cold habitat. Moreover, stress-related proteins were also accumulated in P. lactiflora Pall "Hangshao" seeds, showing its potential for stress resistance. Further protein analysis showed that the primary composition of PSP was albumin and globulin. And the solubility of PSP was good. Furthermore, PSP also showed high glucosidase inhibition activity, indicating that PSP might have some potential function for the remission of hyperglycemia. And P. ostii "Fengdan" seeds may be a better source for protein production than seeds of the other two species in terms of protein solubility and the content of total protein, albumin, and globulin. In addition, an optimal protocol of microwave-assisted alkali extraction was developed to produce PSP. In conclusion, the evaluated stress-related proteins in three peony seed species by proteomic analysis quite agreed with their adaptation of heat or cold stress; proteomics could also be a very useful tool for distinguishing species in the production; and peony seeds may be a good source for protein production.

Molecular characterization and expression analysis of the SQUAMOSA PROMOTER BINDING PROTEIN-LIKE gene family in Paeonia suffruticosa.

KEY MESSAGE: A total of 16 PsSPL genes were identified in tree peony. PsSPLs potentially regulated flowering time, lateral bud and seed development, and the juvenile-to-adult phase transition. SQUAMOSA PROMOTER BINDING PROTEIN-LIKE (SPL) transcription factors are important for plant growth and development. Here, we report the identification of 16 full-length PsSPLs in tree peony (Peaonia suffruticosa Andr.) and 9 PsSPLs that have miR156 target sites. Phylogenetic analysis of the relationship of SPLs in P. suffruticosa and Arabidopsis suggested that they can be classified into six groups, and PsSPLs were highly correlated with Arabidopsis SPLs counterparts in the same group. Cis-element of promoter region analysis suggested that PsSPL genes play roles in physiological processes and developmental events. Expression analysis indicated that most PsSPL genes exhibited high expression levels in the tissues and organs examined here. The increasing expression levels of PsSPL1, PsSPL2, PsSPL8, PsSPL9, PsSPL12, and PsSPL16, and decreasing expression levels of PsSPL1A and PsSPL1B in buds over time suggested that they were probably regulated by the juvenile-to-adult phase transition. In addition, the expression profiles of PsSPL genes in different developmental buds and seeds suggested that PsSPL2, PsSPL3, PsSPL9, PsSPL10, PsSPL13, and PsSPL13A were important genes for regulating the flowering time of the tree peony; PsSPL2 and PsSPL8 might play a role in suppressing lateral bud development, and PsSPL2, PsSPL13, and PsSPL14 positively controlled grain size and number, and pod branching. These results provide a foundation for future functional analysis of PsSPL genes in tree peony growth and development.

Defoliation, not gibberellin, induces tree peony autumn reflowering regulated by carbon allocation and metabolism in buds and leaves.

Short and concentrated natural fluorescence hinders tree peony (Paeonia suffruticosa) annual production, and defoliation and gibberellin (GA) application is used to induce its reflowering in autumn. Here, the individual roles of defoliation and GA treatment were determined by monitoring morphological and soluble sugar changes in buds and leaves, and by investigating carbon allocation- and metabolism-related gene expression. Both defoliation and GA treatment induced early bud development, but induction was faster using the GA treatment. Only defoliation, not GA treatment, induced the final reflowering, although their combination accelerated it. Furthermore, defoliation decreased the sucrose content in buds much faster than the GA treatment. This sucrose reduction may play a key role in tree peony reflowering, and the higher carbon metabolism activity in young leaves after defoliation may further help the reflowering process. Defoliation enhanced the expression of sucrose transporters PsSUT4 and PsSWEET12 in buds, and their expression in young leaves was greater than after GA treatment. This indicated that PsSUT4 and PsSWEET12 may help transport carbon into buds after defoliation. In addition, the invertases, PsCIN2 and PsCWIN1 in young leaves were more highly expressed after defoliation, indicating that they may contribute to reflowering after defoliation by accelerating sucrose hydrolysis in young leaves. In addition, the expression levels of PsVIN1 and PsVIN2 in leaves, and PsVIN2 in buds were more highly induced by GA treatment than by defoliation, indicating that PsVINs may mainly respond to GA treatment. These results may help improve the tree peony forcing culture technology and related industrial production.

Synergistic Effect of NaCl Pretreatment and PVP on Browning Suppression and Callus Induction from Petal Explants of Paeonia Lactiflora Pall. 'Festival Maxima'.

Browning is prevalent in tissue cultures of Paeonia lactiflora Pall. (herbaceous peony), and severely affects and restricts the growth and differentiation of the explants. In this study, dipping excised explants in a sodium chloride (NaCl) solution as a pretreatment, adding polyvinyl pyrrolidone (PVP) to the culture medium, storing planted explants at 4 °C for 24 h, and transferring planted explants to a new medium after 24 h were considered as browning-suppression methods in tissue cultures of herbaceous peony 'Festival Maxima'. The treated petal explants were cultured in a culture room with a 16-hour photoperiod, 25 °C temperature, and 80% relative humidity in darkness for 4 to 8 weeks. The results demonstrated that dipping excised explants in a 0.5 g·L-1 NaCl solution, adding 0.5 g·L-1 PVP to the medium, storing planted explants at 4 °C for 24 h, and transferring planted explants to the same fresh medium after 24 h could effectively inhibit browning. Adding PVP to the medium led to the greatest browning suppression percentage of 95%. Storing planted explants at 4 °C for 24 h reduced the effectiveness of other treatments in suppressing browning. After 8 weeks, dipping excised explants in a NaCl solution resulted in the highest callus induction percentage of 75%, while storing explants at 4 °C for 24 h suppressed callus formation. It was observed in all treatments that decreases in browning was accompanied with higher levels of phenols and lower activities of phenylalanine ammonia-lyase (PAL) and polyphenoloxidase (PPO). Overall, the results suggest that dipping in a NaCl solution was effective in alleviating the browning issues of herbaceous peony tissue cultures, and had positive synergistic effects with PVP on browning suppression and callus induction.

Transcriptome sequencing and identification of key callus browning-related genes from petiole callus of tree peony (Paeonia suffruticosa cv. Kao) cultured on media with three browning inhibitors.

Tree peony (Paeonia suffruticosa Andrews) has ornamental, oil, and medicinal values, and demand in the markets for uniform tree peony seedlings is increasing. Micropropagation could quickly propagate uniform seedlings. However, the heavy browning phenomenon hinders large-scale development of uniform tree peony seedlings. In this paper, we measured the total phenolic compounds content, and sequenced the transcriptomes of tree peony 'Kao' petiole calluses cultured on media with three browning antagonist treatments and fresh petioles to identify the key genes involved in callus browning. Polyvinylpyrrolidone (PVP) treatment can reduce production of phenolic compounds and promote callus regeneration. A total of 218,957 unigenes were obtained from fresh petiole and three kinds of browning petiole calluses by transcriptome sequencing. The average sequence length of unigenes was 446 bp with an N50 of 493 bp. Functional annotation analysis revealed that 43,428, 45,357, 31,194, 30,019, and 21,357 unigenes were annotated using the NCBI-NR database, Swiss-Prot, KOG, GO, and KEGG, respectively. In total, 33 differentially expressed genes (DEGs) were identified as potentially associated with callus browning. Among these DEGs, 12 genes were predicted to participate in phenolic compounds biosynthesis, three genes were predicted to be involved in phenolic compounds oxidation, and six genes were predicted to participate in callus regeneration. Moreover, six transcription factors were observed to be differentially expressed in the fresh petiole and three treated petioles in tree peony. This study comprehensively identifies browning-related gene resources and will possibly help in deciphering the molecular mechanisms of callus browning of tree peony in the future.

Dry storage improves the vase quality of cut peony by increasing water uptake efficiency through aquaporins regulation.

Proper storage prolongs peony market supply. Here, we determined the changes in fresh weight and expression of four aquaporin genes under dry storage (DS) and wet storage (WS). It has showed that after harvesting, the fresh weight change was accompanied with flower opening. After both short- and long-term of storage, the water uptake efficiency in DS group was greater during the first few vase days, providing a direct material basis of DS improved vase quality. The gene expression results showed that PlPIP1;3 and PlTIP2;1 were mainly expressed in petals, whereas PlNIP1;2-like and PlSIP2;1 were mainly expressed in the green tissues. In addition, the expression of PlTIP2;1 in the petals was consistent with the flower opening process, indicating that it may play a major role in facilitating water uptake. During cold storage, the expression of PlPIP1;3 and PlTIP2;1 was higher or more rapidly induced in the DS group, and thus we deduced that they play important roles in improving the vase quality of DS. Furthermore, the expression of PlNIP1;2-like in the early stage of the DS group was more stable than in WS, which may also be partially responsible for the vase quality improvement. In contrast, PlSIP2;1 may not be involved, since no significant change was observed between the DS and WS group. In short, the expression of PlPIP1;3 and PlTIP2;1 in the DS group during storage may improve water uptake efficiency during the vase period and then improving the vase quality of cut peony.

Night Temperature Affects the Growth, Metabolism, and Photosynthetic Gene Expression in Astragalus membranaceus and Codonopsis lanceolata Plug Seedlings.

Astragalus membranaceus and Codonopsis lanceolata are two important medical herbs used in traditional Oriental medicine for preventing cancer, obesity, and inflammation. Night temperature is an important factor that influences the plug seedling quality. However, little research has focused on how the night temperature affects the growth and development of plug seedlings of these two medicinal species. In this study, uniform plug seedlings were cultivated in three environmentally controlled chambers for four weeks under three sets of day/night temperatures (25/10 °C, 25/15 °C, or 25/20 °C), the same relative humidity (75%), photoperiod (12 h), and light intensity (150 µmol·m-2·s-1 PPFD) provided by white LEDs. The results showed that night temperature had a marked influence on the growth and development of both species. The night temperature of 15 °C notably enhanced the quality of plug seedlings evidenced by the increased shoot, root, and leaf dry weights, stem diameter, and Dickson's quality index. Moreover, a night temperature of 15 °C also stimulated and increased contents of primary and secondary metabolites, including soluble sugar, starch, total phenols and flavonoids. Furthermore, the 15 °C night temperature increased the chlorophyll content and stomatal conductance and decreased the hydrogen peroxide content. Analysis of the gene expression showed that granule-bound starch synthase (GBSS), ribulose bisphosphate carboxylase large chain (RBCL), and ferredoxin (FDX) were up-regulated when the night temperature was 15 °C. Taken together, the results suggested that 15 °C is the optimal night temperature for the growth and development of plug seedlings of A. membranaceus and C. lanceolata.

A Two-Stage Culture Method for Zygotic Embryos Effectively Overcomes Constraints Imposed by Hypocotyl and Epicotyl Seed Dormancy in Paeonia ostii 'Fengdan'.

The effect of the exogenous hormone and light quality on breaking hypocotyl and epicotyl dormancy was studied. The results showed that the greatest percentage of hypocotyl dormancy breaking was observed with the Murashige and Skoog (MS) medium supplemented with or without 1.0 mg·L-1 gibberellin 3 (GA3), while ABA and endosperm greatly inhibited hypocotyl dormancy breaking. This suggests that hypocotyl dormancy of the Paeonia ostii 'Fengdan' embryo could be easily overcome by removing constraints of the surrounding endosperm, and ABA may be one of the constraint factors contained in the endosperm. The percentage of epicotyl dormancy breaking was also greatly affected by the concentration of 6-benzylaminopurine (BA) and GA3. Compared to BA by itself, adding GA3 to the medium containing BA highly enhanced epicotyl dormancy breaking, with the greatest percentage of epicotyl dormancy breaking in MS medium supplemented with both 0.5 mg·L-1 BA and 0.5-1.0 mg·L-1 GA3. The percentage of hypocotyl and epicotyl dormancy breaking was also affected by light and its quality. Red light-emitting diodes (LEDs) had the same effect as a dark condition on the hypocotyl dormancy breaking, while blue LEDs and a combination of red and blue LEDs had a negative effect on the hypocotyl dormancy breaking. Unexpectedly, blue LEDs greatly enhanced, whereas red LEDs inhibited, epicotyl dormancy breaking. Conclusively, a two-stage culture method was recommended for breaking the hypocotyl and epicotyl dormancy: hypocotyl dormancy was broken first using the MS medium without any plant growth regulators in the dark (25 °C), and epicotyl dormancy was subsequently broken with the MS medium supplemented with both 1.0 mg·L-1 GA3 and 0.5 mg·L-1 BA under blue light.

Supplementary Light Source Affects Growth, Metabolism, and Physiology of Adenophora triphylla (Thunb.) A.DC. Seedlings.

Adenophora triphylla (Thunb.) A.DC., a well-known herbaceous medicinal species, has been reported to protect against human obesity, cancer, and inflammation. Supplementary lighting is a practical strategy to improve crop quality, especially at a propagation stage. However, there has been no study available on the optimal supplementary light source for the commercial production of A. triphylla seedlings. In this study, plug seedlings were cultivated in a greenhouse for four weeks under an average daily light intensity of 490 µmol·m-2·s-1 PPFD coming from the sun and a supplemental lighting (16 h per day) at 120 µmol·m-2·s-1 PPFD provided by high pressure sodium (HPS), metal halide (MH), far-red (FR) light, white LED (red: green: blue = 2:4:3, LED-w), or mixed (red: green: blue = 4:1:4) LED (LED-mix). The results showed that LED-mix, with a higher percentage of red and blue light, substantially promoted seedling growth compared to other treatments by increasing stem diameter, biomass, specific leaf weight, and root to shoot ratio. The LED-mix also promoted accumulation of soluble sugar, starch, and chlorophyll in the tissue and increased contents of total phenols and flavonoids. Moreover, stomata density and pore area per leaf area under the LED-mix were remarkably greater than those under other treatments. Furthermore, the Western blot analysis revealed that the expression of photosynthetic protein, D1, was notably enhanced by the LED-mix as compared with other light sources. In addition, the LED-mix alleviated the oxidative damage of seedlings by improving enzymatic and nonenzymatic antioxidant systems. Collectively, these results suggest that the LED-mix was the optimal supplementary light source for the production of highest quality A. triphylla seedlings.

Light elicited growth, antioxidant enzymes activities and production of medicinal compounds in callus culture of Cnidium officinale Makino.

Cnidium officinale Makino is an important medicinal plant of oriental clinics and is considered as the main source of phthalides, polyphenols, and flavonoids. However, there is no available report regarding the effect of different light colors on the secondary metabolites composition of C. officinale. In this study different light (dark, white, blue, red and red: blue) conditions were arranged to raise callus on MS medium containing 0.5 mg·L-1 of each 2,4-D and BAP. Callus grown in dark condition showed maximum (2.0 g) fresh weight with lower total phenolic and flavonoids contents. Also, in dark condition callus faced higher catalase (CAT) and guaiacol peroxidase (GPX) activities to avoid free radicals. Mix (red: blue) light condition favored the synthesis of phenolics and flavonoids in callus at the cost of higher ascorbate peroxidase (APX) and superoxide dismutase (SOD) enzymes expression. However, DPPH free radical scavenging activity was less variable among the samples from the different light conditions. Interestingly, the HPLC profile showed higher (28.3 µg·g-1 DW) phthalide accumulation in dark grown-cultures. Compared to other light conditions, 3-butyledinephthalide accumulation was higher (0.43 µg·g-1 DW) in white light-grown callus. These findings suggest that light conditions play an important role in the regulation of in vitro callus growth and synthesis of important medicinal compounds of C. officinale.

CircRASSF2 promotes laryngeal squamous cell carcinoma progression by regulating the miR-302b-3p/IGF-1R axis.

Background: Circular RNAs (circRNAs) are a class of non-coding RNAs (ncRNAs) broadly expressed in cells of various species. However, the molecular mechanisms that link circRNAs with laryngeal squamous cell carcinoma (LSCC) are not well understood. In the present study, we attempted to provide novel basis for targeted therapy for LSCC from the aspect of circRNA-microRNA (miRNA)-mRNA interaction.Methods: We investigated the expression of circRNAs in three paired LSCC tissues and adjacent non-tumor tissues by microarray analysis. Differentially expressed circRNAs were identified between LSCC tissues and non-cancerous matched tissues, including 527 up-regulated circRNAs and 414 down-regulated circRNAs. We focused on hsa_circ_0059354, which is located on chromosome 20 and derived from RASSF2, and thus we named it circRASSF2.Results: circRASSF2 was found to be significantly up-regulated in LSCC tissues and LSCC cell lines compared with paired adjacent non-tumorous tissues and normal cells. Moreover, knockdown of circRASSF2 significantly inhibited cell proliferation and migration in vitro, which was blocked by miR-302b-3p inhibitor. Bioinformatics analysis predicted that there is a circRASSF2/miR-302b-3p/ insulin-like growth factor 1 receptor (IGF-1R) axis in LSCC progression. Dual-luciferase reporter system validated the direct interaction of circRASSF2, miR-302b-3p, and IGF-1R. Western blot verified that inhibition of circRASSF2 decreased IGF-1R expression. Furthermore, silencing circRASSF2 suppressed LSCC growth in vivo Importantly, we demonstrated that circRASSF2 was up-regulated in serum exosomes from LSCC patients. Altogether, silencing circRASSF2 suppresses progression of LSCC by interacting with miR-302b-3p and decreasing inhibiting IGF-1R expression.Conclusion: In conclusion, these data suggest that circRASSF2 is a central component linking circRNAs to progression of LSCC via an miR-302b-3p/IGF-1R axis.

Supplementary Light Source Affects the Growth and Development of Codonopsis lanceolata Seedlings.

Codonopsis lanceolata is widely used in traditional medicine and diets. However, there is no optimal protocol for the commercial production of C. lanceolata seedlings. This study was carried out to find the optimum supplementary light source for the production of C. lanceolata seedlings. Seedlings were grown for four weeks in a glasshouse with an average daily light intensity of 490 µmol·m-2·s-1 photosynthetic photon flux density (PPFD) coming from the sun and a 16-h daily supplementary lighting at 120 µmol·m-2·s-1 PPFD from either high-pressure sodium (HPS), metal halide (MH), far-red (FR), white LED (LED-w), or mixed (white: red: blue = 1:2:1) LEDs (LED-mix). The results showed that the greatest total biomass, stem diameter, ratio of shoot weight to shoot length, root biomass, and ratio of root weight to shoot weight were found in seedlings grown under supplementary LED-mix. Meanwhile, the stomatal properties and soluble sugar contents were improved for seedlings in LED-mix. The contents of starch, total phenols, and flavonoids were the greatest for seedlings in LED-w and LED-mix. The expression of photosynthetic proteins and genes in seedlings was also enhanced by LED-mix. Overall, these results suggest that LED-mix is advantageous to the photosynthetic potential and the accumulation of biomass, carbohydrates and secondary metabolites in C. lanceolata.

Effect of explant type and plant growth regulators on callus induction, growth and secondary metabolites production in Cnidium officinale Makino.

This study reports the effect of explant type and plant growth regulators (PGRs) on callus induction in Cnidium officinale. Compared to stem, root explant showed maximum percent callus formation of 75% on Murashige and Skoog (MS) medium containing 2.3 µM 2,4-dichlorophenoxyacetic acid (2,4-D) and 2.2 µM benzyladenine (BA). At 30th day of callus culture on the said medium, callus fresh weight was sevenfold higher than other tested PGRs treatments. It was noted that MS medium supplemented with 27.1 µM 2,4-D showed the highest 0.30 mg g-1 DW of total phenols, while total flavonoids content reached to a maximum of 0.05 mg g-1 DW on the MS medium supplemented with 4.5 µM 2,4-D and 2.2 µM BA. Conversely, maximum (83.9%) DPPH free radical scavenging activity was observed in calli grown on the MS medium supplemented with 2.3 µM 2,4-D and 2.2 µM BA. The high-performance liquid chromatography (HPLC) analysis revealed higher phthalide content in callus than intact roots of in vitro plants. While 3-butylidenephthalide content in callus was comparable to the intact shoots and roots of in vitro grown C. officinale. The concentrations of 2,4-D played a significant role in the production of phthalide and 3-butylidenephthalide. Additional measures are recommended to further enhance their production in vitro.