The effect of different light treatments on morphogenesis, phenolic compound accumulation and antioxidant potential of Dracocephalum forrestii transformed shoots cultured in vitro.

The aim of this study was to evaluate the effects of wavelengths of light emitted from LEDs on cultured in vitro transformed shoots of Dracocephalum forrestii. The shoots were grown on MS agar medium with 0.5 mg/l BPA (N-benzyl-9-(tetrahydropyranyl)-adenine) and 0.2 mg/l IAA (indole-3-acetic acid) under four light environments: blue, red, red/blue (7:3) and white (control). After four weeks of culture, shoot multiplication rate, biomass and morphology were evaluated, as well as bioactive phenolic content, antioxidant capacities and antioxidant enzyme activities. The hydromethanolic extracts from shoots were analyzed using UHPLC method, and antioxidant potential was evaluated using radical scavenging (1,1-diphenyl-2-picrohydrazyl and superoxide anion), and ferric reducing antioxidant power (FRAP), and enzymatic methods, i.e. sodium dismutase (SOD), catalase (CAT) and peroxidase (POD) activity. It was found that the blue and red/blue light had the strongest effect on morphogenesis and shoot propagation; in these conditions, more than five new shoots were obtained per explant. The blue light cultures demonstrated the highest fresh (0.41 g/tube FW) and dry weights (0.045 g/tube DW), the highest levels of polyphenols (99.7 mg/g DW), i.e. almost three times greater than under white light (35.4 mg/g DW), as well as the highest antioxidant potential. Therefore, LED culture appears to be a beneficial strategy for enhancing the production of the medicinal value of transformed D. forrestii shoot culture.

Effect of Shading on Development, Yield and Quality of Bastard Balm Herb (Melittis melissophyllum L.).

The aim of the study was to assess the effects of Melittis melissophyllum shading on its development and accumulation of phenolics. Their content (verbascoside, apiin, luteolin-7-O-glucoside, coumarin, 3,4-dihydroxycoumarin, o-coumaric acid 2-O-glucoside as well as o-coumaric, p-coumaric, chlorogenic, caffeic, ferulic and cichoric acid) was determined in the herb using HPLC-DAD. The results showed that the content of abovementioned flavonoids and phenolic acids was highest in plants grown under full sunlight. On the other hand, a higher content of coumarin was observed in shaded plants, especially after the seed-setting stage. A similar tendency was noted for the amount of chlorophyll a and b. The content of hydrogen peroxide and malondialdehyde, the activity of polyphenol oxidase and catalase and the antioxidant capacity of plant extracts (measured using DPPH, ABTS and FRAP assays) were found to be the highest in the plants grown in full sunlight. However, the plants grown in moderate (30%) shade were found to thrive best.

Physiological and molecular responses to drought stress in teak (Tectona grandis L.f.).

Drought stress is an increasingly common and worrying phenomenon because it causes a loss of production in both agriculture and forestry. Teak is a tropical tree which needs alternating rainy and dry seasons to produce high-quality wood. However, a robust understanding about the physiological characteristics and genes related to drought stress in this species is lacking. Consequently, after applying moderate and severe drought stress to teak seedlings, an infrared gas analyzer (IRGA) was used to measure different parameters in the leaves. Additionally, using the root transcriptome allowed finding and analyzing the expression of several drought-related genes. As a result, in both water deficit treatments a reduction in photosynthesis, transpiration, stomatal conductance and leaf relative water content was found. As well, an increase in free proline levels and intrinsic water use efficiency was found when compared to the control treatment. Furthermore, 977 transcripts from the root contigs showed functional annotation related to drought stress, and of these, TgTPS1, TgDREB1, TgAREB1 and TgPIP1 were selected. The expression analysis of those genes along with TgHSP1, TgHSP2, TgHSP3 and TgBI (other stress-related genes) showed that with moderate treatment, TgTPS1, TgDREB1, TgAREB1, TgPIP1, TgHSP3 and TgBI genes had higher expression than the control treatment, but with severe treatment only TgTPS1 and TgDREB1 showed higher expression than the control treatment. At the end, a schematic model for the physiological and molecular strategies under drought stress in teak from this study is provided. In conclusion, these physiological and biochemical adjustments in leaves and genetic changes in roots under severe and prolonged water shortage situations can be a limiting factor for teak plantlets' growth. Further studies of those genes under different biotic and abiotic stress treatments are needed.

Evaluation of gamma-irradiated aromatic herbs: Chemometric study of samples submitted to extended storage periods.

The preserving capacity of gamma radiation (10 kGy) on the chemical, nutritional and antioxidant components of Aloysia citrodora Paláu, Melissa officinalis L., Melittis melissophyllum L. and Mentha piperita L., stored for 12 and 18 months, was evaluated. Despite the maintenance of the main characteristics during the first 12 months of storage, the additional 6 months induced several significant changes in individual compounds. In general, the analyzed species reacted dissimilarly throughout time, but it was possible to verify that the fatty acids, tocopherols and antioxidant capacity presented the most significant changes after 18 months of storage, inclusively in samples submitted to gamma radiation. In fact, the applied treatment (10 kGy) did not seem to be effective to prevent the decrease of free sugars, organic acids and tocopherols, especially considering the 18 months period. On the other hand, the evolution in color parameters indicated a greener color (yet slightly more yellow) among irradiated samples. Likewise, gamma radiation had a positive effect on oleic acid, ß-carotene bleaching inhibition (in infusions), DPPH scavenging activity and reducing power (in methanolic extracts). Nevertheless, it might be generally concluded that gamma radiation is less suitable than electron-beam to maintain the characteristics of dried herbs during extended storage periods.

Molecular cloning and expression profiling of a chalcone synthase gene from Lamiophlomis rotata.

Lamiophlomis rotata is a renowned Chinese medicinal plant. Chalcone synthase (CHS) is important in flavonoid and isoflavonoid biosynthesis, catalysing the formation of naringenin chalcone in plants. A full-length cDNA encoding the CHS gene was cloned from L. rotata based on the highly conserved CHS gene sequences of Labiatae plants. A blast search showed its homology (named LrCHS) with other CHS genes of Labiate plants. The full-length genomic DNA of LrCHS was 2026 bp with one intron of 651 bp, two exons of 178 bp and 998 bp, flanked by a 73 bp 5'-UTR and a 126 bp 3'-UTR. The cDNA sequence of the LrCHS gene had an 1176 bp open reading frame encoding a 391 amino acid protein of 42,798 Da. The CHS protein predicted from L. rotata showed 79-86% identity with CHS of other plant species. We conducted a phylogenetic analysis of nine families containing 48 plants and L. rotata based on the full amino acid sequences of CHS proteins. Consequently, LrCHS was located in the Labiatae branch. Additionally, we examined LrCHS gene expression patterns in different tissues by quantitative real-time PCR with specific primers. The expression analysis showed preferential expression of LrCHS in flowers and leaves during the flowering stage. Total flavonoid content and CHS gene expression exhibited similar patterns during L. rotata organ development. In agreement with its function as an elicitor-responsive gene, LrCHS expression was coordinated by methyl jasmonate and UV light, and induced between 6 and 18 h. These results provide a molecular basis for additional functional studies of LrCHS in L. rotata.

Adaptive changes in photosynthetic performance and secondary metabolites during white dead nettle micropropagation.

The white dead nettle, Lamium album L., is an herb that has been successfully cultivated under in vitro conditions. The L. album micropropagation system offers a combination of factors (light intensity, temperature, carbon dioxide (CO2) level, humidity) that are limiting for plant growth and bioactive capacity. To get a better understanding of the mechanism of plant acclimation towards environmental changes, we performed a comparative investigation on primary and secondary metabolism in fully expanded L. album leaves during the consecutive growth in in situ, in vitro, and ex vitro conditions. Although the genetic identity was not affected, structural and physiological deviations were observed, and the level of bioactive compounds was modified. During in vitro cultivation, the L. album leaves became thinner with unaffected overall leaf organization, but with a reduced number of palisade mesophyll layers. Structural deviation of the thylakoid membrane system was detected. In addition, the photosystem 2 (PS2) electron transport was retarded, and the plants were more vulnerable to light damage as indicated by the decreased photoprotection ability estimated by fluorescence parameters. The related CO2 assimilation and transpiration rates were subsequently reduced, as were the content of essential oils and phenolics. Transfer of the plants ex vitro did not increase the number of palisade numbers, but the chloroplast structure and PS2 functionality were recovered. Strikingly, the rates of CO2 assimilation and transpiration were increased compared to in situ control plants. While the phenolics content reached normal levels during ex vitro growth, the essential oils remained low. Overall, our study broadens the understanding about the nature of plant responses towards environmental conditions.

Chemical composition and antioxidant properties of γ-irradiated Iranian Zataria multiflora extracts.

CONTEXT: Irradiation is the process of exposing food such as herbal plant to ionizing radiation to destroy microorganisms. Zataria multiflora Boiss (Lamiaceae), known as Avishan-e-Shirazi in Persian, is a thyme-like plant that grows naturally in central and southern parts of Iran and is used in traditional folk medicine. OBJECTIVE: In this study, the effects of γ-radiation on chemical composition and antioxidant properties of Z. multiflora were investigated. MATERIALS AND METHODS: The plants were first irradiated with Co60 source (0, 10, and 25 kGy) and then subjected to Clevenger extraction to obtain essential oils. The composition of the oil was analyzed by a gas chromatography and compared with samples pretreated under different conditions. In parallel, the hydroalcoholic extract was prepared and used for measuring flavonoid content. Thereafter, the free-radical scavenging and antioxidant properties of essential oils and hydroalcoholic extract were examined. RESULTS: Despite the minor change in the individual oil constituents, the total percentage of the main components remained unaffected before and after irradiation (~95%). In addition, the total flavonoid content of hydroalcoholic extract was also unchanged due to irradiation (~32 mg QE/g extract). The high radical scavenging activity of the oil (~67%) and hydroalcoholic extract (~71%), in addition, the antioxidant properties of the oil (~91%) and hydroalcoholic extract (~95%), were unaffected after irradiation. DISCUSSION AND CONCLUSIONS: These findings may suggest the sustainability of Z. multiflora extract properties pretreated with γ-radiation. With a view to its antioxidant applications, resistance of Z. multiflora and its properties against radiation effects are promising findings.

[Determination of ten metal elements in Ziziphora clinopodioides Lam by microwave digestion-FAAS].

Comprehensive utilization of traditional Uighur medicine has been increasingly emphasized, and the relationship between metal elements and traditional Uighur medicine has attracted great attention, so it is quite important to determine the contents of traditional Uighur medicine. The Ziziphora clinopodioides Lam powder was digested with HNO3 by microwave digestion before determination. Ten metal elements in Ziziphora clinopodioides Lam were determined by FAAS. The work conditions, accuracy and precision of the method were studied. The linear correlations of standard curves are good (r = 0.999 0-0.999 8). The recovery (n = 6) is 95%-108%, and the RSD(n = 6) is 0.45%-1.53%. The results showed that there were comparatively rich metal elements, among which are comparatively high calcium, magnesium and potassium in Ziziphora clinopodioides Lam. The method offers traits of low detection limit, high sensitivity, speediness and exactness, and wasapplied to the determination of metal elements in samples with satisfactory results. It provided useful data for discussing the relationship between the content of the metal elements in Ziziphora clinopodioides Lam and clinical application of the Uighur medicine.

Effects of gamma-irradiation on the free radical and antioxidant contents in nine aromatic herbs and spices.

Nine spice and aromatic herb samples (i.e., basil, bird pepper, black pepper, cinnamon, nutmeg, oregano, parsley, rosemary, and sage) were gamma-irradiated at a dose of 10 kGy according to commercial practices. The effects of the disinfection treatment on the content of organic radicals and some nutrients (namely, vitamin C and carotenoids) in the samples were investigated by chromatographic and spectroscopic techniques. Irradiation resulted in a general increase of quinone radical content in all of the investigated samples, as revealed by electron paramagnetic resonance spectroscopy. The fate of these radicals after storage for 3 months was also investigated. The cellulose radical was clearly observed in a few samples. Significant losses of total ascorbate were found for black pepper, cinnamon, nutmeg, oregano, and sage, whereas a significant decrease of carotenoids content was observed for cinnamon, oregano, parsley, rosemary, bird pepper, and sage.

Long-distance signaling within Coleus x hybridus leaves; mediated by changes in intra-leaf CO2?

Rapid long-distance signaling in plants can occur via several mechanisms, including symplastic electric coupling and pressure waves. We show here in variegated Coleus leaves a rapid propagation of electrical signals that appears to be caused by changes in intra-leaf CO2 concentrations. Green leaf cells, when illuminated, undergo a rapid depolarization of their membrane potential (Vm) and an increase in their apoplastic pH (pHa) by a process that requires photosynthesis. This is followed by a slower hyperpolarization of Vm and apoplastic acidification, which do not require photosynthesis. White (chlorophyll-lacking) leaf cells, when in isolated white leaf segments, show only the slow response, but when in mixed (i.e. green and white) segments, the rapid Vm depolarization and increase in pHa propagate over more than 10 mm from the green to the white cells. Similarly, these responses propagate 12-20 mm from illuminated to unilluminated green cells. The fact that the propagation of these responses is eliminated when the leaf air spaces are infiltrated with solution indicates that the signal moves in the apoplast rather than the symplast. A depolarization of the mesophyll cells is induced in the dark by a decrease in apoplastic CO2 but not by an increase in pHa. These results support the hypothesis that the propagating signal for the depolarization of the white mesophyll cells is a photosynthetically induced decrease in the CO2 level of the air spaces throughout the leaf.

Detection of radiation-induced hydrocarbons and 2-alkylcyclobutanones in irradiated perilla seeds.

The method consists of the extraction of fat from perilla seeds, separation of hydrocarbons and 2-alkylcyclobutanones with florisil column chromatography, and identification of hydrocarbons and 2-alkylcyclobutanones by gas chromatography-mass spectroscopy (GC-MS). Concentrations of hydrocarbons and 2-alkylcyclobutanones increased with the irradiation dose. The major hydrocarbons in the irradiated perilla seeds were 8-heptadecene and 1,7-hexadecadiene from oleic acid and 6,9-heptadecadiene and 1,7,10-hexadecatriene from linoleic acid. One of the 2-alkylcyclobutanones, 2-(5'-tetradecenyl)cyclobutanone, was found in the highest concentration in the irradiated perilla seeds. Radiation-induced hydrocarbons in the perilla seeds were detected at doses of 0.5 kGy and higher, and radiation-induced 2-alkylcyclobutanones were detected at doses of 1 kGy and higher. These compounds were not detected in nonirradiated perilla seeds.

A light-inducible Myb-like gene that is specifically expressed in red Perilla frutescens and presumably acts as a determining factor of the anthocyanin forma.

The Myb-p1 gene was isolated by screening for differentially expressed Myb-related genes in red (anthocyanin-producing) and green (anthocyanin nonproducing) forms of Perilla frutescens. Expression of Myb-p1 is increased 10-fold in the red relative to the green form of P. futescens, and the gene is induced by light. MYB-P1 has only one DNA-binding region, which corresponds to repeat III in the general structure of MYB proteins. In the yeast two-hybrid system, it was shown that MYB-P1 interacted with MYC-RP, a MYC-related transcriptional regulatory protein involved in the control of anthocyanin biosynthesis in P. frutescens. In yeast, MYB-P1 was able to bind to a dihydroflavonol reductase (DFR) gene promoter isolated from red P. frutescens. These data suggest that Myb-p1 may be involved in the regulation of anthocyanin biosynthesis and could therefore be responsible for determining anthocyanin formation in red P. frutescens.

[Effects of HE particles on medicinal plant Agastache rugosus (Fisch. et Mey.) O. Ktze].

The biological effect of HE particles on the seeds of Agastache rugosus was probed in experiments on board a retrievable satellite. The result shows that the germination rate of the seeds pierced by HE particles radiation appears rather low. The seeds hit by HE particles (piercing radiation or not) start to germinate two days earlier than those in the control group, and the first leaf emerges four to five days earlier than that in the control group. The resultant seedlings grow markedly faster. Variations take place in the nuclear types of chromosome. The yield of essential oils becomes slightly higher. No marked changes have been observed in the major chemical components of these oils.