Investigating the simultaneous effect of chitosan and arbuscular mycorrhizal fungi on growth, phenolic compounds, PAL enzyme activity and lipid peroxidation in Salvia nemorosa L.

Considering the importance of Salvia nemorosa L. in the pharmaceutical and food industries, and also beneficial approaches of arbuscular mycorrhizal fungi (AMF) symbiosis and the use of bioelicitors such as chitosan to improve secondary metabolites, the aim of this study was to evaluate the performance of chitosan on the symbiosis of AMF and the effect of both on the biochemical and phytochemical performance of this plant and finally introduced the best treatment. Two factors were considered for the factorial experiment: AMF with four levels (non-inoculated plants, Funneliformis mosseae, Rhizophagus intraradices and the combination of both), and chitosan with six levels (0, 50, 100, 200, 400 mg L-1 and 1% acetic acid). Four months after treatments, the aerial part and root length, the levels of lipid peroxidation, H2O2, phenylalanine ammonia lyase (PAL) activity, total phenol and flavonoid contents and the main secondary metabolites (rosmarinic acid and quercetin) in the leaves and roots were determined. The flowering stage was observed in R. intraradices treatments and the highest percentage of colonization (78.87%) was observed in the treatment of F. mosseae × 400 mg L-1 chitosan. Furthermore, simultaneous application of chitosan and AMF were more effective than their separate application to induce phenolic compounds accumulation, PAL activity and reduce oxidative compounds. The cluster and principal component analysis based on the measured variables indicated that the treatments could be classified into three clusters. It seems that different treatments in different tissues have different effects. However, in an overview, it can be concluded that 400 mg L-1 chitosan and F. mosseae × R. intraradices showed better results in single and simultaneous applications. The results of this research can be considered in the optimization of this medicinal plant under normal conditions and experiments related to abiotic stresses in the future.

Range of factors in the reduction of hyperhydricity associated with in vitro shoots of Salvia santolinifolia Bioss / Gama de fatores na redução da hiperidricidade associada a brotos in vitro de Salvia santolinifolia Bioss

Hyperhydricity is a serious physiological disorder and affects In vitro propagation of many plants and as well of Salvia santolinifolia. The donor material to initiate the in vitro culture was the callus taken from the in vitro shoots produced on Murashig and Skoogs (MS) medium at 4.0 mg/l BA. This callus formed numerous hyperhydric shoots on culturing upon the medium of the same composition. The aim was to systematically evaluate the effect of cytokinins (Benzyladnine (BA) and N6-(-2-isopentenyl) adenine (2iP), culture vessels magnitude, medium solidification, source of nitrogen and calcium chloride for the alleviation of hyperhydricity. In the tissue cultures of S. santolinifolia BA and 2iP induced severe hyperhydricity, when other factors i.e. culture vessels magnitude and a suitable concentration of agar, ammonium nitrate (NH4NO3), potassium nitrate (KNO3) & calcium chloride (CaCl2.2H2O) were not optimized. After 30 days' culture, we observed 83.82% hyperhydric shoots at increased level (1.5 mg/l 2iP) and 81.59% at decreased levels (1.0 mg/l 2iP). On the other hand, hyperhydricity percentage at decreased (0.4%) and at increased (0.8%) levels of agar were 72.37% and 39.08%, respectively. MS medium modification with NH4NO3 (412 mg/l), KNO3 (475 mg/l) and CaCl2.2H2O (880 mg/l) was found the best medium to reduced hyperhydricity (23.6%).

A hiperidricidade é um distúrbio fisiológico sério e afeta a propagação in vitro de muitas plantas e também da Salvia santolinifolia. O material doador para iniciar a cultura in vitro foi o calo retirado dos brotos in vitro produzidos em meio Murashig e Skoogs (MS) a 4,0 mg / l BA. Esse calo formou numerosos rebentos hiperídricos em cultura no meio da mesma composição. O objetivo foi avaliar sistematicamente o efeito das citocininas (Benziladnina (BA) e N6 - (- 2-isopentenil) adenina (2iP), magnitude dos vasos de cultura, solidificação do meio, fonte de nitrogênio e cloreto de cálcio para o alívio da hiperidricidade. culturas de tecidos de S. santolinifolia BA e 2iP induziram hiperidricidade severa, quando outros fatores, como magnitude dos vasos de cultura e uma concentração adequada de ágar, nitrato de amônio (NH4NO3), nitrato de potássio (KNO3) e cloreto de cálcio (CaCl2.2H2O), não foram otimizados. Após 30 dias de cultura, observamos 83,82% de brotos hiperídricos em níveis aumentados (1,5 mg / l 2iP) e 81,59% em níveis reduzidos (1,0 mg / l 2iP). Por outro lado, a porcentagem de hiperidricidade diminuiu (0,4%) e em níveis aumentados (0,8%) de ágar foram 72,37% e 39,08%, respectivamente. A modificação do meio MS com NH4NO3 (412 mg / l), KNO3 (475 mg / l) e CaCl2.2H2O (880 mg / l) foi encontrada melhor hiperidricidade média a reduzida (23,6%).

Osmoprotection in Salvia hispanica L seeds under water stress attenuators / Osmoproteção em sementes de Salvia hispanica L sob atenuadores do estresse hídrico

Salvia hispanica cultivation is recent in Brazil and occurs in the off-season, when there is lower water availability in the soil. Water deficit is one of the abiotic factors that most limit germination for compromising the sequence of metabolic events that culminate with seedling emergence. Several attenuating substances have been used to mitigate the effects resulting from this stress and give higher tolerance to the species. Thus, the objective of this study was to evaluate the action of different agents as water stress attenuators in the germination and accumulation of organic compounds in S. hispanica seedlings. The treatments consisted of pre-soaking the seeds for 4 hours in salicylic acid (1 mM.L-¹), gibberellic acid (0.4 mM.L-¹), distilled water and control treatment (without soaking). The seeds were germinated at osmotic potentials of 0.0, -0.1, -0.2, -0.3 and -0.4 MPa, using PEG 6000 as an osmotic agent. The variables germination percentage, germination speed index, shoot and primary root lengths, total dry mass, proline, total soluble sugars and total free amino acids were analyzed. Salicylic acid and gibberellic acid led to the best results among the attenuators tested, increasing germination, length, dry mass and biochemical components of S. hispanica seedlings under water deficit. Therefore, salicylic and gibberellic acids are efficient in mitigating water stress in S. hispanica seeds up to the potential of -0.4 MPa.

O cultivo da Salvia hispanica é recente no Brasil e se dá no período de entressafra, quando há menor disponibilidade hídrica no solo. O déficit hídrico é um dos fatores abióticos que mais limitam a germinação por comprometer a sequência de eventos metabólicos que culminam com a emergência da plântula. Diversas substâncias atenuadoras têm sido empregadas com a finalidade de mitigar os efeitos resultantes desse estresse e conferir maior tolerância às espécies. Desse modo, objetivou-se avaliar a ação de diferentes agentes como atenuadores do estresse hídrico na germinação e acúmulo de compostos orgânicos em plântulas de S. hispanica. Os tratamentos consistiram na pré-embebição das sementes durante 4 horas em ácido salicílico (1 mM.L-¹), ácido giberélico (0,4 mM.L-¹), água destilada e o tratamento controle (sem embebição). As sementes foram germinadas sob os potenciais osmóticos 0,0, -0,1, -0,2, -0,3 e -0,4 MPa, utilizando PEG 6000 como agente osmótico. Analisaram-se as variáveis porcentagem de germinação, índice de velocidade de germinação, comprimento da parte aérea e da raiz primária, massa seca total, prolina, açúcares solúveis totais e aminoácidos livres totais. O ácido salicílico e o ácido giberélico apresentaram os melhores resultados, dentre os atenuadores testados, incrementando a germinação, o comprimento, a massa seca e os componentes bioquímicos de plântulas de S. hispanica sob déficit hídrico. Logo, os ácidos salicílico e giberélico são eficientes na mitigação do estresse hídrico em sementes de S. hispanica até o potencial -0,4 MPa.

Rapid Hormetic Responses of Photosystem II Photochemistry of Clary Sage to Cadmium Exposure.

Five-day exposure of clary sage (Salvia sclarea L.) to 100 µM cadmium (Cd) in hydroponics was sufficient to increase Cd concentrations significantly in roots and aboveground parts and affect negatively whole plant levels of calcium (Ca) and magnesium (Mg), since Cd competes for Ca channels, while reduced Mg concentrations are associated with increased Cd tolerance. Total zinc (Zn), copper (Cu), and iron (Fe) uptake increased but their translocation to the aboveground parts decreased. Despite the substantial levels of Cd in leaves, without any observed defects on chloroplast ultrastructure, an enhanced photosystem II (PSII) efficiency was observed, with a higher fraction of absorbed light energy to be directed to photochemistry (ΦPSΙΙ). The concomitant increase in the photoprotective mechanism of non-photochemical quenching of photosynthesis (NPQ) resulted in an important decrease in the dissipated non-regulated energy (ΦNO), modifying the homeostasis of reactive oxygen species (ROS), through a decreased singlet oxygen (1O2) formation. A basal ROS level was detected in control plant leaves for optimal growth, while a low increased level of ROS under 5 days Cd exposure seemed to be beneficial for triggering defense responses, and a high level of ROS out of the boundaries (8 days Cd exposure), was harmful to plants. Thus, when clary sage was exposed to Cd for a short period, tolerance mechanisms were triggered. However, exposure to a combination of Cd and high light or to Cd alone (8 days) resulted in an inhibition of PSII functionality, indicating Cd toxicity. Thus, the rapid activation of PSII functionality at short time exposure and the inhibition at longer duration suggests a hormetic response and describes these effects in terms of "adaptive response" and "toxicity", respectively.

Application of 1H-NMR combined with qRT-PCR technology in the exploration of rosmarinic acid biosynthesis in hair roots of Salvia miltiorrhiza Bunge and Salvia castanea f. tomentosa Stib.

MAIN CONCLUSION: Methyl jasmonate promotes the synthesis of rosmarinic acid in Salvia miltiorrhiza Bunge and Salvia castanea f. tomentosa Stib, and it promotes the latter more strongly. Salvia miltiorrhiza Bunge (SMB) is a traditional Chinese medicinal material, its water-soluble phenolic acid component rosmarinic acid has very important medicinal value. Salvia castanea f. tomentosa Stib (SCT) mainly distributed in Nyingchi, Tibet. Its pharmacological effects are similar to SMB, but its rosmarinic acid is significantly higher than the former. Methyl jasmonate (MJ) as an inducer can induce the synthesis of phenolic acids in SMB and SCT. However, the role of MJ on rosmarinic acid in SMB is controversial. Therefore, this study used SMB and SCT hair root as an experimental material and MJ as a variable. On one hand, exploring the controversial reports in SMB; on the other hand, comparing the differences in the mechanism of action of MJ on the phenolic acids in SMB and SCT. The content of related metabolites and the expression of key genes in the synthesis pathway of rosmarinic acid was analyzed by 1H-NMR combined with qRT-PCR technology. Our research has reached the following conclusions: first of all, MJ promotes the accumulation of rosmarinic acid and related phenolic acids in the metabolic pathways of SMB and SCT. After MJ treatment, the content of related components and gene expression are increased. Second, compared to SMB, SCT has a stronger response to MJ. It is speculated that the different responses of secondary metabolism-related genes to MJ may lead to different metabolic responses of salvianolic acid between the two.

Increased phenolic acid and tanshinone production and transcriptional responses of biosynthetic genes in hairy root cultures of Salvia przewalskii Maxim. treated with methyl jasmonate and salicylic acid.

The purpose of this study is to reveal the impact of the plant hormone salicylic acid (SA) and methyl jasmonate (MeJA) on the growth, effective components accumulation, and related gene expression of the hairy root of Salvia przewalskii Maxim. Various concentrations of SA (0, 25, 50, 100, 200 µM) or MeJA (0, 50, 100, 200, 400, 600 µM) were added to the culture medium of Salvia przewalskii Maxim. Low concentrations of SA promoted the growth of hairy root, while a high concentration inhibited it. 0 to 400 µM MeJA promoted the growth of hairy root, but 600 µM MeJA starts to inhibit its growth. 50 µM SA and 400 µM MeJA significantly enhanced the production of caffeic acid, rosmarinic acid, salvianolic acid B, cryptotanshinone, and tanshinone IIA. In general, 50 µM SA can be used to accumulate of tanshinone in hairy roots of S. przewalskii with 6 days. 400 µM MeJA can be used to accumulate of phenolic acids in hairy roots of S. przewalskii with 3 days. The selected genes in the tanshinone and phenolic acid biosynthetic pathway were upregulated with elicitation. To obtain a higher yield and content of secondary metabolites, it is advisable to use 50 µM SA or 400 µM MeJA as the optimal doses to cultivate the hairy root of S. przewalskii. This study provides, for the first time, an efficient tanshinone and phenolic acid production method for S. przewalskii.

Multiple stressors in multiple species: Effects of different RDX soil concentrations and differential water-resourcing on RDX fate, plant health, and plant survival.

Response to simultaneous stressors is an important facet of plant ecology and land management. In a greenhouse trial, we studied how eight plant species responded to single and combined effects of three soil concentrations of the phytotoxic munitions constituent RDX and two levels of water-resourcing. In an outdoor trial, we studied the effects of high RDX soil concentration and two levels of water-resourcing in three plant species. Multiple endpoints related to RDX fate, plant health, and plant survival were evaluated in both trials. Starting RDX concentration was the most frequent factor influencing all endpoints. Water-resourcing also had significant impacts, but in fewer cases. For most endpoints, significant interaction effects between RDX concentration and water-resourcing were observed for some species and treatments. Main and interaction effects were typically variable (significant in one treatment, but not in another; associated with increasing endpoint values for one treatment and/or with decreasing endpoint values in another). This complexity has implications for understanding how RDX and water-availability combine to impact plants, as well as for applications like phytoremediation. As an additional product of these greenhouse and outdoor trials, three plants native or naturalized within the southeastern United States were identified as promising species for further study as in situ phytoremediation resources. Plumbago auriculata exhibited relatively strong and markedly consistent among-treatment mean proportional reductions in soil RDX concentrations (112% and 2.5% of the means of corresponding values observed within other species). Likewise, across all treatments, Salvia coccinea exhibited distinctively low variance in mean leaf chlorophyll content index levels (6.5% of the means of corresponding values observed within other species). Both species also exhibited mean wilting and chlorosis levels that were 66% and 35%, and 67% and 84%, of corresponding values observed in all other plants, respectively. Ruellia caroliniensis exhibited at least 43% higher mean survival across all treatments than any other test species in outdoor trials, despite exhibiting similar RDX uptake and bioconcentration levels.

Impacts of foliar exposure to multi-walled carbon nanotubes on physiological and molecular traits of Salvia verticillata L., as a medicinal plant.

Owing to the growing applications of the multi-walled carbon nanotubes (MWCNTs) in the communications and energy industries, they have attracted increasing attention for their effects on the environment and plants. Therefore, we investigated the impact of foliar exposure to MWCNTs on the oxidative stress responses in the Salvia verticillata as a medicinal plant. Furthermore, we evaluated the possible correlations between gene expression and activity of the key enzymes in the phenolic acids biosynthesis pathways and their accumulation in the treated leaves. The leaves of two-month-old plants were sprayed with different concentrations (0-1000 mg L-1) of MWCNTs. Raman's data and Transmission Electron Microscopy images have confirmed the absorption of MWCNTs via epidermal cells layer into the parenchymal cells of the exposed leaves. The results showed that exposure to MWCNTs led to a decrease in the photosynthetic pigments and increases in the oxidative stress indices (enzymatic and non-enzymatic antioxidants) in the leaves with a dose-dependent manner. The content of rosmarinic acid as a main phenolic acid was increased in the MWCNTs-exposed leaves to 50 and 1000 mg L-1, nearly four times relative to the control. Unlike with other examined enzymes, a positive correlation was deduced between the activity and gene expression patterns of the rosmarinic acid synthase with the rosmarinic acid accumulation in the treatments. Overall, MWCNTs at the low concentrations could promote the production of the pharmaceutical metabolites by the changes in the ROS generation. However, at the higher concentrations, MWCNTs were toxic and induced the oxidative damages in S. verticillata.

The Stimulatory Effect of Purine-Type Cytokinins on Proliferation and Polyphenolic Compound Accumulation in Shoot Culture of Salvia Viridis.

The present study demonstrates hormonal control of Salvia viridis growth and development using four different purine-type cytokinins at different concentrations. The addition of cytokinins significantly increased biomass of cultures, proliferation rate, and, interestingly, secondary metabolite production. The best response in terms of multiplication ratio was recorded on Murashige and Skoog medium supplemented with 0.5 mg/L BPA (N-benzylotetrahydropyranyl adenine), while the greatest biomass accumulation was achieved when supplemented with 1 mg/L m-T (meta-topoline). Quantitative UPLC-DAD analysis of the hydromethanolic extract from S. viridis culture revealed the presence of 12 polyphenols: seven phenolic acids and five phenylethanoids. The highest total content of polyphenolic compounds was found in shoots cultivated on medium with 2 mg/L BPA (18.66 mg/g DW): almost twice that of control shoots. The medium was also the most optimal for the biosynthesis of rosmarinic acid, the predominant phenolic acid. However, the greater phenylethanoid accumulation was stimulated by 1 mg/L m-T: the metabolite content was above three times higher than that found in shoots grown on the control medium (8.03 mg/g DW vs. 2.37 mg/g DW). Hence, it was demonstrated that phytohormones are capable of influencing not only vital physiological processes, but therapeutic potential of plants as well. Therefore, the cytokinin-based sage cultures may be also considered as the alternative sources of bioactive compounds.

Effects of antimony on enzymatic and non-enzymatic antioxidants in a metallicolous and a non-metallicolous population of Salvia spinosa L.

The present study aimed at elucidating the role of antioxidants and stress metabolites in antimony (Sb) tolerance in a metallicolous (M), Sb[V]-hypertolerant population, and a non-metallicolous (NM) population of Salvia spinosa, particularly with regard to the question of whether they could be involved in constitutive Sb tolerance or, specifically, in Sb[V] hypertolerance in the M population. Plants were exposed in hydroponics to 0, 8, 24, 74, 221 µM Sb (Ш or V). Superoxide dismutase (SOD), ascorbate peroxidase (APX) and catalase (CAT) activities, and the concentrations of phenolics, flavonoids, and proline in leaves were measured after 20 d. As potential stress/tolerance markers, the concentrations of chlorophyll a and b, anthocyanins, and those of total soluble and reducing sugars were also measured. Chlorophyll a concentration reflected the difference, both in Sb[III] and Sb[V] tolerance, between N and NM, and the higher toxicity of Sb[III], compared to Sb[V]. APX and proline accumulation were more induced in M than in NM, and more by Sb[V] than by Sb[III], which is theoretically compatible with a role in Sb[V] hypertolerance. CAT was more induced in M than in NM, but more by Sb[III] than Sb[V], suggesting that is not functional in Sb[V] hypertolerance. The other enzymes and compounds did not exhibit significant Sb redox status*population interactions, suggesting that they don't play a role in, specifically, Sb[V] hypertolerance in M, but at most in the constitutive Sb[III] or Sb[V] tolerance of the species.

Thermodynamic and kinetic insights into plant-mediated detoxification of lead, cadmium, and chromium from aqueous solutions by chemically modified Salvia moorcroftiana leaves.

Thermodynamic and kinetic aspects for the biosorptive removal of Pb, Cd, and Cr metals from water using Chemically Modified Leaves of Salvia moorcroftiana (CMSML) were determined. Different parameters including pH, temperature, metal's initial concentration, biomass dosage, and contact time were optimized. Optimum biosorptions of Pb, Cd, and Cr were attained at pH values of 6.0, 7.0, and 3.0 respectively. Batch experiments showed maximum removal of both Pb and Cd at 40 °C and that of Cr at 30 °C. Biosorption capability of CMSML was observed to decrease with raising temperature. Optimal equilibrium times for Pb, Cd, and Cr uptake were 120, 60, and 120 min respectively. Based on the values of regression correlation coefficients (R2), the current data is explained better by applying Langmuir isotherms than the Freundlich model. Maximum biosorbent capabilities (qmax) for Pb, Cd, and Cr were approximately 270.27, 100.00, and 93.45 mg/g respectively. Thermodynamically, removal of all the three metal ions was shown to be exothermic and spontaneous.

Effect of sowing date and water availability on growth of plants of chia (Salvia hispanica L) established in Chile.

From 2010 to 2014 two trials were performed to assess the effect of sowing date (SD1, SD2) and irrigation treatments (IT1, IT2) on the growth of chia in central Chile, measuring leaf area (LA) and dry matter (DM) as primary parameters and relative growth rate (RGR), net assimilation rate (NAR), leaf weight ratio (LWR), crop growth rate (CGR) and specific leaf weight (SLW) as secondary parameters. Both LA and DM reached maximum values between 640 and 1150 accumulated degree days (ADD). However, LA and DM were 25% greater for sowing dates than for available water. Flowering date was also not affected by sowing date or water availability; plants flowered at 1140 and 942 ADD in SD1 and SD2 respectively, and at 499 ADD in the water availability trial. Sowing date had a significant effect on RGR 0.15 g g-1 d-1 for SD1 and 0.12 g g-1 d-1 for SD2 at 410 ADD. Greater water availability increased RGR by 60% compared to stressed plants, however NAR was similar between sowing dates with a tendency to greater values in SD2 plants; maximum values were recorded at 514 ADD in IT1 and IT2, with a tendency toward higher values in IT1. Thus, the primary growth variables LA, DM and flowering are genetically determined. However, the derived growth variables RGR, LWR, NAR, CGR and SLW were affected by sowing date and water availability, with significant differences at p≤ 0.05. The results showed that the sowing date and water availability influence significantly the growth parameters. The physiological component (NAR) show a strong influence on the growth rate of the chia (RGR), above the morphological component (SLW and LAR).

Changes in Photosynthetic Pigments, Total Phenolic Content, and Antioxidant Activity of Salvia coccinea Buc'hoz Ex Etl. Induced by Exogenous Salicylic Acid and Soil Salinity.

Salvia coccinea (Lamiaceae) is a promising source of potential antioxidants, and its extracts can be used in pharmaceutical industry, as well as in food products and cosmetics. Salicylic acid (SA) affects many physiological and metabolic processes in vascular plants under salinity stress. The aim of this study was to investigate the response of S. coccinea to either SA, or sodium chloride (NaCl), or a combination of both. The plants were sprayed with a solution of 0.5 or 1.0 mM SA and watered with 0, 100, 200, or 300 mM NaCl. Exogenous application of SA increased the number of branches, fresh herbal weight, and total chlorophyll content vs control plants. Salinity-exposed plants showed reduced growth, content of photosynthetic pigments total polyphenols, and antioxidant activity. However, foliar application of SA relieved the adverse effects of 100 mM NaCl, as demonstrated by increased number of branches, greater fresh herbal weight, higher content of total chlorophyll, total carotenoids, and total polyphenols, as well as antioxidant potential, detected using ferric-reducing ability of plasma (FRAP) and 2.2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), compared with untreated plants.

THE EFFECTS OF COLCHICINEINDUCED AUTOTETRAPLOIDY ON SELECTED CHARACTERISTICS OF NURUOZAK (SALVIA LERIIFOLIA).

Nuruozak (Salvia leriifolia Benth), is a perennial herbaceous plant that is endemic to Iran and has recently been introduced as a medicinal plant. Artificial polyploidy is an efficient method to increase the production of secondary metabolites and can result in a whole spectrum of genetic, molecular and physiological modifications. In order to produce an autotetraploid population of nuruozak, various concentrations of colchicine (0.00, 0.05, 0.10, 0.20 or 0.50 % w/v) were applied to the seeds and shoot apical meristems of young seedlings at the fourth leaf-stage. Microscopic studies, flow cytometry analysis and chromosome counting were conducted to select tetraploid nuruozak plants. Furthermore, the effects of ploidy level on the essential oil content and composition and biomass production of nuruozak plants, as well as selected structural and physiological characteristics were studied. Based on the number of the obtained tetraploids, treatment of shoot apical meristems was more efficient than seed treatment. Structural and phytochemical characteristics, chlorophyll content and photosynthetic rate were affected by the increase in ploidy level. In addition to the higher potential in biomass production, tetraploid plants produced eight new compounds which were absent in diploids.

Increasing the synthesis of bioactive abietane diterpenes in Salvia sclarea hairy roots by elicited transcriptional reprogramming.

KEY MESSAGE: Transcriptional activation of genes belonging to the plastidial MEP-derived isoprenoid pathway by elicitation with methyl jasmonate and coronatine enhanced the content of bioactive abietane diterpenes in Salvia sclarea hairy roots. We have shown that aethiopinone, an abietane diterpene synthesized in Salvia sclarea roots is cytotoxic and induces apoptosis in human melanoma cells. To develop a production platform for this compound and other abietane diterpenes, hairy root technology was combined with the elicitation of methyl jasmonate (MeJA) or the phytotoxin coronatine (Cor). Both MeJA and Cor induced a significant accumulation of aethiopinone, but prolonged exposure to MeJA irremediably caused inhibition of hairy root growth, which was unaffected by Cor treatment. Considering together the fold increase in aethiopinone content and the final hairy root biomass, the best combination was a Cor treatment for 28 days, which allowed to obtain up to 105.34 ± 2.30 mg L-1 of this compound to be obtained, corresponding to a 24-fold increase above the basal content in untreated hairy roots. MeJA or Cor elicitation also enhanced the synthesis of other bioactive abietane-quinone diterpenes. The elicitor-dependent steering effect was due to a coordinated transcriptional activation of several biosynthetic genes belonging to the plastidial MEP-derived isoprenoid pathway. High correlations between aethiopinone content and MeJA or Cor-elicited level of gene transcripts were found for DXS2 (r 2 = 0.99), DXR (r 2 = 0.99), and GGPPS (r 2 = 0.98), encoding enzymes acting upstream of GGPP, the common precursor of diterpenes and other plastidial-derived terpenes, as well as CPPS (r 2 = 0.99), encoding the enzyme involved in the first cyclization steps leading to copalyl-diphosphate, the precursor of abietane-like diterpenes. These results point to these genes as possible targets of metabolic engineering approaches to establish a more efficient production platform for such promising anti-proliferative plant-derived compounds.

Establishment of Salvia castanea Diels f. tomentosa Stib. hairy root cultures and the promotion of tanshinone accumulation and gene expression with Ag⁺, methyl jasmonate, and yeast extract elicitation.

Salvia castanea Diels f. tomentosa Stib. is an endemic medicinal plant distributed in China, and the notably high content of tanshinone IIA in the root is proven effective for the therapy of heart diseases. Hairy root induction of this Salvia species was inoculated with Agrobacterium rhizogenes strain ATCC 15834. Transformed hairy root was cultured in 6,7-V liquid medium for growth kinetics assessment and elicitation. An S curve was present in the hairy root cultures based on the fresh and dry weights with an interval of 3 days. An optimum concentration of the applied elicitors (15 µM Ag(+), 200 µM methyl jasmonate, and 200 mg l(-1) yeast extract elicitor) benefitted both the growth status and tanshinone accumulation in the hairy root cultures. Tanshinone IIA contents were mostly stimulated 1.8-fold and 1.99-fold compared with the control by Ag(+) and methyl jasmonate elicitation, respectively. Yeast extract dramatically enhanced dry mass accumulation, while it promoted cryptotanshinone content of 2.84 ± 0.33 mg g(-1) dry weight at most in the hairy root cultures. Selected elicitors diversely influenced tanshinone accumulation in the time courses of hairy root cultures within 7 days. Furthermore, transcripts of selected genes in the tanshinone biosynthetic pathway were remarkably upregulated with elicitation. Yeast extract elicitor heightened 13.9-fold of isopentenyl diphosphate isomerase expression level at 12 h, while it increased 16.7-fold of geranylgeranyl diphosphate synthase transcript at 24 h compared with that of the control, which was more effective than Ag(+) and methyl jasmonate. This study provided a convenient hairy root culture system of S. castanea Diels f. tomentosa Stib. for tanshinone production for the first time.

Edible film production from chia seed mucilage: Effect of glycerol concentration on its physicochemical and mechanical properties.

This study investigated the physicochemical and mechanical properties of a novel edible film based on chia mucilage (CM) hydrocolloid. CM (1% w/v) films were prepared by incorporation of three concentrations of glycerol (25%, 50%, and 75% w/w, based on CM weight). As glycerol concentration increased, water vapor permeability (WVP), elongation at break (EB), and water solubility of CM films increased while their tensile strength (TS), and Young's modulus (YM) decreased significantly (p<0.05). CM films containing a high concentration of glycerol were slightly reddish and yellowish in color but still had a transparent appearance. CM films exhibited excellent absorption of ultraviolet light, and good thermal stability. The scanning electron micrographs showed that all CM films had a uniform appearance. This study demonstrated that the chia mucilage hydrocolloid has important properties and potential as an edible film, or coating.

Application of Heavy Metal Rich Tannery Sludge on Sustainable Growth, Yield and Metal Accumulation by Clarysage (Salvia sclarea L.).

A field experiment was conducted to evaluate the effective utilization of tannery sludge for cultivation of clarysage (Salvia sclarea) at CIMAP research farm, Lucknow, India during the year 2012-2013. Six doses (0, 20, 40, 60, 80, 100 tha(-1)) of processed tannery sludge were tested in randomised block design with four replications. Results revealed that maximum shoot, root, dry matter and oil yield were obtained with application of 80 tha(-1)of tannery sludge and these were 94, 113 and 61% higher respectively, over control. Accumulation of heavy metals (Cr, Ni, Fe, Pb) were relatively high in shoot portion of the plant than root. Among heavy metals, magnitude of chromium accumulation was higher than nickel, iron and lead in shoot as well as in root. Linalool, linalyl acetate and sclareol content in oil increased by 13,8 and 27% respectively over control, with tannery sludge application at 80 tha(-1). Heavy metals such as chromium, cadmium and lead content reduced in postharvest soil when compared to initial status. Results indicated that clarysage (Salvia sclarea) can be grown in soil amended with 80 tha(-1)sludge and this can be a suitable accumulator of heavy metals for phytoremediation of metal polluted soils.

Composition of the Essential Oil of Salvia ballotiflora (Lamiaceae) and Its Insecticidal Activity.

Essential oils can be used as an alternative to using synthetic insecticides for pest management. Therefore, the insectistatic and insecticidal activities of the essential oil of aerial parts of Salvia ballotiflora (Lamiaceae) were tested against the fall armyworm Spodoptera frugiperda (Lepidoptera: Noctuidae). The results demonstrated insecticidal and insectistatical activities against this insect pest with concentrations at 80 µg·mL(-1) resulting in 20% larval viability and 10% pupal viability. The larval viability fifty (LV50) corresponded to a concentration of 128.8 µg·mL(-1). This oil also increased the duration of the larval phase by 5.5 days and reduced the pupal weight by 29.2% withrespect to the control. The GC-MS analysis of the essential oil of S. ballotiflora showed its main components to be caryophyllene oxide (15.97%), and ß-caryophyllene (12.74%), which showed insecticidal and insectistatical activities against S. frugiperda. The insecticidal activity of ß-caryophyllene began at 80 µg·mL(-1), giving a larval viability of 25% and viability pupal of 20%. The insectistatic activity also started at 80 µg·mL(-1) reducing the pupal weight by 22.1% with respect to control. Caryophyllene oxide showed insecticidal activity at 80 µg·mL(-1) giving a larval viability of 35% and viability pupal of 20%.The insectistatic activity started at 400 µg·mL(-1) and increased the larval phase by 8.8% days with respect to control. The LV50 values for these compounds were 153.1 and 146.5 µg·mL(-1), respectively.

[Effects of EDTA on lead accumulation in Tagetes patula and Salvia spendens].

Pot experiments were conducted to investigate the effects of EDTA on the Pb availability in rhizospheric soil and the Pb accumulation in Tagetes patula and Salvia spendens. The results showed that 7 days after applying EDTA (3.0 mmol x kg(-1)) into soil, the concentration of ammonium acetate (1 mol x L(-1)) -extracted Pb in rhizospheric soil increased significantly (P < 0.01), and the Pb content in T. patula and S. spendens leaves reached to 2415.88 and 1083.68 mg x kg(-1), being 61.36 and 5.37 times of the control, respectively. The Pb content also had a marked increase in T. patula and S. spendens stems, but had a slight decrease in their roots.