Beyond vegetables: effects of indoor LED light on specialized metabolite biosynthesis in medicinal and aromatic plants, edible flowers, and microgreens.

Specialized metabolites from plants are important for human health due to their antioxidant properties. Light is one of the main factors modulating the biosynthesis of specialized metabolites, determining the cascade response activated by photoreceptors and the consequent modulation of expressed genes and biosynthetic pathways. Recent developments in light emitting diode (LED) technology have enabled improvements in artificial light applications for horticulture. In particular, the possibility to select specific spectral light compositions, intensities and photoperiods has been associated with altered metabolite content in a variety of crops. This review aims to analyze the effects of indoor LED lighting recipes and management on the specialized metabolite content in different groups of crop plants (namely medicinal and aromatic plants, microgreens and edible flowers), focusing on the literature from the last 5 years. The literature collection produced a total of 40 papers, which were analyzed according to the effects of artificial LED lighting on the content of anthocyanins, carotenoids, phenols, tocopherols, glycosides, and terpenes, and ranked on a scale of 1 to 3. Most studies applied a combination of red and blue light (22%) or monochromatic blue (23%), with a 16 h day-1 photoperiod (78%) and an intensity greater than 200 µmol m-2  s-1 (77%). These treatment features were often the most efficient in enhancing specialized metabolite content, although large variations in performance were observed, according to the species considered and the compound analyzed. The review aims to provide valuable indications for the definition of the most promising spectral components toward the achievement of nutrient-rich indoor-grown products. © 2021 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

[Analysis of shading on DNA methylation by MSAP in Pinellia ternata].

Pinellia ternata is a medicinal herb of Araceae, and its tubers are used as medicines. It is a common Chinese herbal medicine in China and has a large market demand. When exposing to strong light intensity and high temperature during the growth process, P. ternata withers in a phenomenon known as "sprout tumble", which largely limits tuber production. Shade can effectively delay sprout tumble formation and increase its yield, however the relevant regulation mechanism is unclear. DNA methylation, as a self-modifying response to environmental changes, is often involved in the regulation of plant growth and development. In this study, P. ternata grown under natural light and 90% shading were selected as the control group and the experimental group for genomic DNA methylation analysis by using methylate sensitive amplification polymorphism(MSAP). The results showed that a total of 617 loci were detected with 20 pairs of primers, of which 311 were in the natural light group and 306 in the shading group. The methylation sites in the light and shading groups accounted for 58.2% and 71.57%, respectively, and the methylation ratios in the methylation sites were 27.65% and 29.41%, respectively, indicating that shading significantly induced the genome DNA methylation of P. ternata. Compared to the natural light group, shading promoted 32.51% of the genes methylation, while inducing 16.25% gene demethylation. This study reveals the DNA methylation variation of P. ternata under shading conditions, which lays a preliminary theoretical foundation for further analysis of the mechanism of shading regulation of P. ternata growth from epigenetic level.

Phosphoproteomics Reveals the Biosynthesis of Secondary Metabolites in Catharanthus roseus under Ultraviolet-B Radiation.

Ultraviolet (UV)-B radiation acts as an elicitor to enhance the production of secondary metabolites in medicinal plants. To investigate the mechanisms, which lead to secondary metabolites in Catharanthus roseus under UVB radiation, a phosphoproteomic technique was used. ATP content increased in the leaves of C. roseus under UVB radiation. Phosphoproteins related to calcium such as calmodulin, calcium-dependent kinase, and heat shock proteins increased. Phosphoproteins related to protein synthesis/modification/degradation and signaling intensively changed. Metabolomic analysis indicated that the metabolites classified with pentoses, aromatic amino acids, and phenylpropanoids accumulated under UVB radiation. Phosphoproteomic and immunoblot analyses indicated that proteins related to glycolysis and the reactive-oxygen species scavenging system were changed under UVB radiation. These results suggest that UVB radiation activates the calcium-related pathway and reactive-oxygen species scavenging system in C. roseus. These changes lead to the upregulation of proteins, which are responsible for the redox reactions in secondary metabolism and are important for the accumulation of secondary metabolites in C. roseus under UVB radiation.

Use of greenhouse-covering films with tailored UV-B transmission dose for growing 'medicines' through plants: rocket salad case.

BACKGROUND: The effects of ultraviolet B (UV-B) radiation on plants are well known and have recently attracted a great deal of attention due to the production of large quantities of secondary metabolites, which are very beneficial for human health. Recent studies have demonstrated the possibility of exploiting UV-B radiation to induce metabolic changes in fruit, vegetables, and herbs. The role of UV-B rays in inducing secondary plant metabolites is enhanced by new plastic films, which, as a result of their optical properties, permit the necessary dosage of UV-B to be transmitted into the greenhouse to stimulate such metabolites without altering the harvest. RESULTS: The main goal of the present paper is to demonstrate that, by using a greenhouse plastic film with appropriate transmittance of UV-B for rocket salad cultivation, it is possible to increase the nutraceutical elements in comparison with the same species grown in absence of such radiation. Tests compared nutritional elements extracted from rocket salad grown under greenhouses covered with several plastic films differing in UV-B transmittance. We found that rocket salad grown under plastic with 27% UV-B transmittance exhibited very high luteolin and quercetin content in comparison with rocket salad cultivated under film blocking UV-B radiation. CONCLUSIONS: Our experimental results confirm the possibility of exploiting UV-B radiation in the correct amounts by appropriate greenhouse plastic covers, to produce natural 'medicines' using the plants and to satisfy increasing consumer demand for natural health-promoting food products. © 2019 Society of Chemical Industry.

[Progress of effects of light on plant flavonoids].

As an important secondary metabolites of medicinal plant, flavonoids plays a very important role on itself including light protection and antioxidant. Light is one of the important environmental factors which impacts the secondary metabolites of plant and has a significant impact on biological synthesis of flavonoids. This paper reviews the recent progress of the effects of light on flavonoids on the plants focusing from light intensity and light quality, and summarizes briefly functions of the phenylalanine ammonia lyase (PAL) in plant secondary metabolism and the upstream key enzyme chalcone synthase (CHS) in flavonoid biosynthetic pathway.

[Effects of Different Shading Treatment and Planting Patterns on Several Main Characters and Yield of Elephantopus scaber].

OBJECTIVE: To study the growth and yield of Elephantopus scaber under different light conditions. METHODS: Several main characters and yield performances were studied under six shading treatment as well as two planting patterns. RESULTS: The plant height, leaf number, root length and root-shoot ratio were increased under moderate shading. With the increase of shading ratio, the process of Elephantopus scaber vegetative growth to reproductive growth were shortened, seed yield, dry biomass and root yield decreased as well. Among different shading treatments, dry seed-yield showed 8. 46 ~31. 10 kg/667 m2 dry biomass showed 327. 28 ~ 800. 95 kg/ 667 m2 and dry root yield showed 30. 65 ~ 70. 72 kg/667 m2. CONCLUSION: Elephantopus scaber is a light-demanding but shade-tolerant plant. The patterns of hole seeding were suggested in planting, and not more than 60% shade density may be good under plantations.

Phenolic profile of dark-grown and photoperiod-exposed Hypericum perforatum L. Hairy root cultures.

Hypericum perforatum L. is a medicinal plant considered as an important natural source of secondary metabolites with a wide range of pharmacological attributes. Hairy roots (HR) were induced from root segments of in vitro grown seedlings from H. perforatum after cocultivation with Agrobacterium rhizogenes A4. Investigations have been made to study the production of phenolic compounds in dark-grown (HR1) and photoperiod-exposed (HR2) cultures. The chromatographic analysis of phenolic acids, flavonols, flavan-3-ols, and xanthones revealed marked differences between HR1 and HR2 cultures. The production of quinic acid, kaempferol, and seven identified xanthones was increased in HR2. Moreover, HR2 showed a capability for de novo biosynthesis of two phenolic acids (3-p-coumaroylquinic acid and 3-feruloylquinic acid), three flavonol glycosides (kaempferol hexoside, hyperoside, and quercetin acetylglycoside), and five xanthones (tetrahydroxy-one-methoxyxanthone, 1,3,5-trihydroxy-6-methoxyxanthone, 1,3,5,6-tetrahydroxy-2-prenylxanthone, paxanthone, and banaxanthone E). On the other side, HR1 cultures were better producers of flavan-3-ols (catechin, epicatechin, and proanthocyanidin dimers) than HR2. This is the first comparative study on phenolic profile of H. perforatum HR cultures grown under dark and photoperiod conditions.

Assessment of litter degradation in medicinal plants subjected to ultraviolet-B radiation.

Litter decomposition is an important component of global carbon budget. Elevated influx of ultraviolet-B radiation (UV-B) as a consequence of depletion of stratospheric ozone (O3) layer may affect litter decomposition directly or/modifying the plant tissue quality. Chemical composition of plant can affect litter decomposition. In the present study, three important medicinal plant species i.e. Acorus calamus, Ocimum sanctum and Cymbopogon citratus were exposed to two levels of supplemental UV-B (sUV and sUV,) during the growth period and examined the changes in leaf quality and degradation of leaf litters. The sUV, treatment (+3.6 kJ m(-2) d(-1)) increased the rate of decomposition by 45% and 31% respectively; in leaf litters from O. sanctum and C. citratus, while no significant effect was noticed in A. calamus leaf litter. Higher accumulation of sclerenchymatous tissue around vascular bundles and increased concentrations of total phenols by 39 mg g(-1) probably lowered the decomposition rate; finding k value: 0.0049 g g(-1) d(-1) in leaf litters of A. calamus. The C/N ratio was increased by 14% at sUV2 in C. citratus, whereas in O. sanctum it decreased by 13.6% after treatment. Results of the present experiment illustrates that firstly UV-B can modify the decomposition rate of leaf litter of test plant species, secondly it can alter the tissue chemistry particularly leaf phenolics, N and P concentrations strongly and thus affecting the decay rate and thirdly UV-B effects on decay rate and leaf chemistry is species specific.

Variation of medicinal components in a unique geographical accession of horny goat weed Epimedium sagittatum Maxim. (Berberidaceae).

Herbal Epimedium species have been widely in Traditional Chinese Medicine for sexual enhancement, immunity improvement, anticancer and anti-aging treatment, with flavonoids and polysaccharides being the major active components. However, exhaustive depletion of wild sources warrants germplasm evaluation and quality resource exploration. A preliminarily analysis had previously indicated that a specific local geographic accession of Epimedium sagittatum found in Luotian (LT) county of Hubei Province (China) had a much higher content of total flavonoids and polysaccharides. In this study, we further investigated the medicinal component variation in the LT type under different light intensities and in different regions by the common-garden experiment. The results indicated a light intensity range of 40-160 µmol/m²/s was the most suitable for the synthesis and accumulation of total flavonoids, while polysaccharide accumulation was negatively correlated with the light intensity. Icariin was the component displaying the highest content among flavonoids, and the content of major flavonoid bioactive components was relatively stable in the third year after cultivation. There was significant correlation between the major flavonol glycoside constituents and the geographic location, and Central China followed by Northern China were the highly suitable regions for cultivation of LT type E. sagittatum. The results revealed that there was a functional balance between flavonoids and polysaccharides at different developmental stages, and the best harvesting stage should consider the primary contents of interest. This study provides important information on the exploration of quality resources, further breeding approaches and cultivation practices of E. sagittatum, and thus the important insights to enhance our understanding of quality control of traditional medicinal plants.

[Advances in influence of UV-B radiation on medicinal plant secondary metabolism].

Stratospheric ozone depletion results in an increased level of solar UV-B radiation (UV-B, 280-320 nm) reaching the earth surface. By the effect of UV-B radiation, various medicinal active ingredients changed because of the change of gene expression, enzyme activity and secondary metabolism, clinical effect is also changed. The research status of UV-B radiation and the accumulation of plant secondary metabolites in the past 10 years were summarized in this paper to supply reference for cultivation and exploitation of the medicinal plants.

[Cultivation of protocorms of Dendrobium candidum in air-lift bioreactors].

OBJECTIVE: To explore the factors affecting the growth of protocorms of Dendrobium candidum and substance synthesis in a reactor, in order to provide a new method for mass production of raw materials of D. candidum. METHOD: Protocorms in vitro were used as experimental materials to study the effect of inoculum volume, light intensity and air volume on the growth of protocorms of D. candidum and the accumulation of polysaccharide and dendrobine in a 3 L-air lift balloon type bioreactor. RESULT: After 30 days of cultivation in a bioreactor, protocorms became dark green and grew well at the inoculum volume of 10 g x L(-1). The polysaccharide content in protocorms showed no difference at various inoculum volumes; whereas the dendrobine content showed differences (with the highest treatment at the inoculum volume of 10 g x L(-1)), particularly the productions of polysaccharide and alkaloid reached the maximum at the inoculum volume of 10 g x L(-1). The condition of 1 600 lx of light intensity was the most favorable for the growth of protocorms. Though light played a role of improving the accumulation of polysaccharide in protocorms of D. candidum, it could inhibit the accumulation of dendrobine. Polysaccharide content and production were better under light conditions of 1 600 and 2 400 lx than dark conditions. Despite the maximum dendrobine content in dark conditions, the dendrobine production showed the maximum in the light condition of 1 600 lx due to poor growth of protocorms. Protocorms grew well and became dark green at the air volume of 0.2 vvm (air volume culture volume per minute) , which was better than at 0.1 and 0.3, with maximum polysaccharide and dendrobine contents and productions. CONCLUSION: In a 3 L-air lift balloon type bioreactor with a working volume of 2 L, the conditions of 10 inoculum volume, 1 600 lx light intensity and 0.2 air volume were favorable for the growth of protocorms and the production of dendrobine. This demonstrates that the cultivation of D. candidum and substance synthesis in a reactor is an effectie approach for mass production of polysaccharide and dendrobine.

[Response of isozyme and stress indexes of Coptis chinensis to UV-B radiation].

OBJECTIVE: To study the physiological mechanism of anti-stress of Coptic chinensis and provide theoretical basis for its cultivation and promoting its quality. METHODS: Different degrees of the range of time and intensity of UV-B radiation were set in the experiment. Used the technique of polyacrylamide gelatin vertical board electrophoresis (PAGE) to analyse the isozyme and related stress index. RESULTS: The isoenzymic bands of SOD1 (Rf = 0.125), SOD2 (Rf = 0.312), CAT1 (Rf = 0.428), POD3 (Rf = 0.290), POD4 (Rf = 0.636) were induced by UV-B radiation after 3 hours, with the increase of the time of UV-B radiation, those isoenzymic bands was going to vanish or became unclear. Moreover, isoenzymic bands of CAT1 (Rf = 0.428), POD3 (Rf = 0.290) disappeared in advance under heavy intensity of UV-B radiation. Furthermore, the contents of MDA, soluble sugar, proline were higher dramatically than those of control group under UV-B radiation. However, excluding the increases of proline in UL group, the content of MDA, soluble sugar, proline of other groups commenced to decrease slowly and isoenzymic bands of soluble protein increase after 7 hours of UV-B radiation. CONCLUSION: The increase of the expression of antioxidase isozyme, accumulation of soluble sugar, soluble protein and other antioxidase matter is induced by the short-time UV-B radiation, which can protect Coptis chinensis from being harmed by UV-B radiation. However, regulation system of Coptis chinensis are broken, metabolism is disordered, the bands of antioxidase isozyme vanish or weaken, the bands of soluble protein are increased and widened, these phenomenon is caused by 7 hours of UV-B radiation.

Identification of phenolic compounds in isolated vacuoles of the medicinal plant Catharanthus roseus and their interaction with vacuolar class III peroxidase: an H2O2 affair?

Class III peroxidases (Prxs) are plant enzymes capable of using H(2)O(2) to oxidize a range of plant secondary metabolites, notably phenolic compounds. These enzymes are localized in the cell wall or in the vacuole, which is a target for secondary metabolite accumulation, but very little is known about the function of vacuolar Prxs. Here, the physiological role of the main leaf vacuolar Prx of the medicinal plant Catharanthus roseus, CrPrx1, was further investigated namely by studying its capacity to oxidize co-localized phenolic substrates at the expense of H(2)O(2). LC-PAD-MS analysis of the phenols from isolated leaf vacuoles detected the presence of three caffeoylquinic acids and four flavonoids in this organelle. These phenols or similar compounds were shown to be good CrPrx1 substrates, and the CrPrx1-mediated oxidation of 5-O-caffeoylquinic acid was shown to form a co-operative regenerating cycle with ascorbic acid. Interestingly, more than 90% of total leaf Prx activity was localized in the vacuoles, associated to discrete spots of the tonoplast. Prx activity inside the vacuoles was estimated to be 1809 nkat ml(-1), which, together with the determined concentrations for the putative vacuolar phenolic substrates, indicate a very high H(2)O(2) scavenging capacity, up to 9 mM s(-1). Accordingly, high light conditions, known to increase H(2)O(2) production, induced both phenols and Prx levels. Therefore, it is proposed that the vacuolar couple Prx/secondary metabolites represent an important sink/buffer of H(2)O(2) in green plant cells.

[Effect of different light of LED light quality on growth and antioxidant enzyme activities of Ganoderma lucidum].

OBJECTIVE: To study the effect of light quality on growth, antioxidant enzyme activities of Ganoderma lucidum mycelium. METHOD: G. lucidum mycelium was cultured under different light qualities by light emitting diodes (LED). The growth G. lucidum mycelium was observed and antioxidant enzyme activities was determined in different growth periods. RESULT: Under the red LED, the blue LED and dark condition (CK), the mycelium grew faster than that under other light qualities. The white LED resulted in a largest increase in the amount of the mycelium and always kept the activities of CAT high level. Major fluctuations of POD activities emerged under the green LED, while enhanced severely in the late phase. Under the yellow LED, the activities of SOD appeared high level. However, SOD activities on dark (CK) raised obviously in late period. At the late stage, the content of mycelium polysaccharides was significant higher than that under the blue LED. CONCLUSION: The light quality could influence the growth and metabolism of G. lucidum mycelium.

Ultraviolet-B mediated biochemical and metabolic responses of a medicinal plant Adhatoda vasica Nees. at different growth stages.

In the present study, the effects of elevated UV-B (eUVB; ambient ± 7.2 kJ m-2 day-1) were evaluated on the biochemical and metabolic profile of Adhatoda vasica Nees. (an indigenous medicinal plant) at different growth stages. The results showed reduction in superoxide radical production rate, whereas increase in the content of hydrogen peroxide which was also substantiated by the histochemical localization. Malondialdehyde content, which is a measure of oxidative stress, did not show significant changes at any of the growth stages however photosynthetic rate and chlorophyll content showed reduction at all growth stages under eUV-B exposure. Increased activities of the enzymatic and non-enzymatic antioxidants were noticed except ascorbic acid, which was reduced under eUV-B exposure. The metabolic profile of A. vasica revealed 43 major compounds (assigned under different classes) at different growth stages. Triterpenes, phytosterols, unsaturated fatty acids, diterpenes, tocopherols, and alkaloids showed increment, whereas reduction in saturated fatty acids and sesquiterpenes were observed under eUV-B treatment. Vasicinone and vasicoline, the two important alkaloids of A. vasica, showed significant induction under eUV-B exposure as compared to control. Treatment of eUV-B leads to the synthesis of some new compounds, such as oridonin oxide (diterpene) and α-Bisabolol oxide-B (sesquiterpene), which possess potent anti-inflammatory and anticancerous activities. The study displayed that differential crosstalk between antioxidants and secondary metabolites at different growth stages, were responsible for providing protection to A. vasica against eUV-B induced oxidative stress and enhancing its medicinal properties.

Development of radiation indicators to distinguish between irradiated and non-irradiated herbal medicines using HPLC and GC-MS.

The effects of high dose γ-irradiation on six herbal medicines were investigated using gas chromatography-mass spectrometry (GC/MS) and high-performance liquid chromatography (HPLC). Herbal medicines were irradiated at 0-50 kGy with (60)Co irradiator. HPLC was used to quantify changes of major components including glycyrrhizin, cinnamic acid, poncirin, hesperidin, berberine, and amygdalin in licorice, cinnamon bark, poncirin immature fruit, citrus unshiu peel, coptis rhizome, and apricot kernel. No significant differences were found between gamma-irradiated and non-irradiated samples with regard to the amounts of glycyrrhizin, berberine, and amygdalin. However, the contents of cinnamic acid, poncirin, and hesperidin were increased after irradiation. Volatile compounds were analyzed by GC/MS. The relative proportion of ketone in licorice was diminished after irradiation. The relative amount of hydrocarbons in irradiated cinnamon bark and apricot kernel was higher than that in non-irradiated samples. Therefore, ketone in licorice and hydrocarbons in cinnamon bark and apricot kernel can be considered radiolytic markers. Three unsaturated hydrocarbons, i.e., 1,7,10-hexadecatriene, 6,9-heptadecadiene, and 8-heptadecene, were detected only in apricot kernels irradiated at 25 and 50 kGy. These three hydrocarbons could be used as radiolytic markers to distinguish between irradiated (>25 kGy) and non-irradiated apricot kernels.

Ecological filtering by a dominant herb selects for shade tolerance in the tree seedling community of coastal dune forest.

The regeneration niche is commonly partitioned along a gradient from shade-tolerant to shade-intolerant species to explain plant community assembly in forests. We examined the shade tolerance of tree seedlings in a subtropical coastal forest to determine whether the ecological filtering effect of a dominant, synchronously monocarpic herb (Isoglossa woodii) selects for species at either end of the light response continuum during the herb's vegetative and reproductive phases. Photosynthetic characteristics of seedlings of 20 common tree species and the herb were measured. Seedlings were grown in the greenhouse at 12-14% irradiance, and their light compensation points measured using an open-flow gas exchange system. The light compensation points for the tree species were low, falling within a narrow range from 2.1 ± 0.8 µmol m(-2) s(-1) in Celtis africana to 6.4 ± 0.7 µmol m(-2) s(-1) in Allophylus natalensis, indicating general shade tolerance, consistent with a high and narrow range of apparent quantum yield among species (0.078 ± 0.002 mol CO(2) mol(-1) photon). Rates of dark respiration were significantly lower in a generalist pioneer species (Acacia karroo) than in a forest pioneer (C. africana), or in late successional phase forest species. We argue that the general shade tolerance, and phenotypic clustering of shade tolerance, in many tree species from several families in this system, is a result of ecological filtering by the prevailing low light levels beneath the I. woodii understorey, which excludes most light-demanding species from the seedling community.

Evaluation of 6°Co-gamma radiosterilization on Chinese medicines with HPLC/FTIR.

The aim of this study was to evaluate the effect of radiosterilization on 30 Chinese medicines using γ-rays from the isotope 6°Co. Two groups of Chinese medicines, non-treated and dry samples, were treated using a 6°Co irradiation source at the doses 0, 3, 6 and 9 kGy. After storage for 3 months, characterizations of chemical compounds and functional groups were performed by high-performance liquid chromatography (HPLC) and Fourier transform infrared spectroscopy. The results of radiosterlization showed that nearly all of the medicines were decontaminated under the dose of 9 kGy. In most samples, chemical compounds and functional groups were not altered by the irradiation treatment. However, minor changes were found in the molecular structures of 14 medicines under the reported 'safety dose' (10 kGy). The drying process before irradiation could decrease the chemical changes caused by γ-rays to 50%. The HPLC analysis of nine medicines revealed minor changes at a dose of 3 kGy. The findings in this study provide important information that may suggest the need for a re-evaluation of the reported safety dose. Therefore, further investigation may be warranted to insure the safety of γ-radiosterlization of Chinese medicines.

Response of ultraviolet-B induced antioxidant defense system in a medicinal plant, Acorus calamus.

Ultraviolet-B (UV-B) radiation generates an oxidative stress in plant cells due to excessive generation of reactive oxygen species (ROS). ROS can denature enzymes and damage important cellular components. In the present study, an important medicinal plant Acorus calamus (Sweet flag) was subjected to two doses of supplemental UV-B radiation (sUV-B): sUV1 (+ 1.8 kJ m(-2) d(-1)) and sUV2 (+3.6 kJ m(-2) d(-1)) to evaluate the relative response of antioxidant defense potential. Stimulation of activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and glutathione reductase (GR) was observed at initial growth period while the activities of CAT and SOD decreased at later age of sampling. sUV-B induced lipid peroxidation (LPO) was observed showing alteration of membrane properties. No definite trend of change was observed for ascorbic acid (AsA), while increments in thiol, proline, phenol and protein contents were observed due to sUV-B. Results suggested that sUV-B radiation may stimulate the enzymatic and non-enzymatic defense system of Acorus plants, showing its better adaptation at lower dose of sUV-B.

Effects of gamma radiation treatment on three different medicinal plants: Microbial limit test, total phenolic content, in vitro cytotoxicity effect and antioxidant assay.

BACKGROUND AND AIM: The aim of this study is to evaluate the effects of gamma radiation treatment on three medicinal plants, namely Euodia malayana, Gnetum gnemon and Khaya senegalensis at two different forms; methanol leaf extracts and dried leaves respectively. EXPERIMENTAL PROCEDURE: The microbial limit test (MLT) studies indicated the suitable dosage of minimum and maximum gamma irradiation for leaf extracts as well as dried leaves of all the tested medicinal plants. Quantitative analysis of total phenolic content (TPC) analysis is based on calorimetric measurements determined using the Folin-Ciocalteu reagent with gallic acid (GA) used as the reference. In vitro cytotoxicity assay by using fibroblast (L929) cell lines was performed on each plant to determine the toxicity effect which sodium dodecyl sulfate (SDS) as the positive control. DPPH (2,2-diphenyl-1-picryl-hydrazyl) assay was conducted by using vitamin C and GA as the positive controls to determine the antioxidant property of each plant. RESULTS AND CONCLUSION: The MLT analysis indicated that the suitable dosage gamma irradiation for leaf extracts was 6-12 kGy and dried leaves were 9-13 kGy. The amount of GA concentration in each plant increased significantly from 30-51 mg GAE g-1 before treatment to 57-103 mg GAE g-1 after treatment with gamma radiation. This showed no significant effect of in vitro cytotoxicity activity before and after treatment with gamma irradiation in this study. Effective concentration (EC50) values of Khaya senegalensis plant reduced significantly (P ≤ 0.005) from 44.510 µg/ml before treatment to 24.691 µg/ml after treatment with gamma radiation, which indicate an increase of free radical scavenging activity.