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.

Cytokinin-Based Tissue Cultures for Stable Medicinal Plant Production: Regeneration and Phytochemical Profiling of Salvia bulleyana Shoots.

Salvia bulleyana is a rare Chinese medicinal plant that due to the presence of polyphenols lowers the risk of some chronic diseases especially those related to the cardiovascular system. The present study examines the organogenic competence of various combinations and concentrations of plant growth regulators to develop an efficient protocol for in vitro regeneration of S. bulleyana via leaf explants, maintaining the high production of active constituents. The purpose of the study was also to assess the possibilities of using a cytokinin-based regeneration to effectively produce therapeutic compounds. The adventitious shoot formation was observed through direct organogenesis on media with purine derivatives (meta-topolin, mT and benzylaminopurine, BAP), and through indirect organogenesis on media with urea derivatives (tidiazuron, TDZ and forchlorfenuron, CPPU). The highest regeneration frequency (95%) with 5.2 shoots per explant was obtained on leaves cultured on Murashige and Skoog (MS) medium containing 0.1 mg/L naphthalene-1-acetic acid (NAA) and 2 mg/L BAP. Following inter simple sequence repeat (ISSR) marker-based profiling, the obtained organogenic shoot lines revealed a similar banding pattern to the mother line, with total variability of 4.2-13.7%, indicating high level of genetic stability. The similar genetic profile of the studied lines translated into similar growth parameters. Moreover, HPLC analysis revealed no qualitative differences in the profile of bioactive metabolites; also, the total polyphenol content was similar for different lines, with the exception of the shoots obtained in the presence of CPPU that produced higher level of bioactive compounds. This is the first report of an effective and rapid in vitro organogenesis protocol for S. bulleyana, which can be efficiently employed for obtaining stable cultures rich in bioactive metabolites.

Metabolomic analysis and antioxidant activity of wild type and mutant chia (Salvia hispanica L.) stem and flower grown under different irrigation regimes.

BACKGROUND: Chia (Salvia hispanica L.) is a functional food from Central America. Interest in it is growing rapidly due to the many health benefits from the seed. However, when chia is grown at high latitudes, seed yield may be low whereas a high stem biomass and immature inflorescences are produced. Little is known about the chemical composition and the properties of stems and flowers. In this work, the metabolite profile, the antioxidant activity, and the total polyphenol content of stems and inflorescences were evaluated in a factorial experiment with different chia populations (commercial black chia and long-day flowering mutants G3, G8, and G17) and irrigation (100% and 50% of evapotranspiration). RESULTS: The results show the influence of irrigation and seed source on the antioxidant activity and total polyphenol content of chia flower and stem. Inflorescences exhibit higher antioxidant activity, suggesting their potential use as natural antioxidant. The mutants G3 and G8, at 50% irrigation, contained the highest amounts of compounds with nutraceutical value, especially within the flower. The mutant G17 showed lower antioxidant activity and polyphenol content compared to other seed sources but exhibited high omega 3 content in flowers but low in stems. This indicates that chia varieties should be chosen according to the objective of cultivation. CONCLUSION: These findings, indicating a close relation of metabolite content with irrigation and seed source, may provide the basis for the use of chia flower and stem for their nutraceutical value in the food, feed, and supplement industries. © 2021 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Improvement of Chia Seeds with Antioxidant Activity, GABA, Essential Amino Acids, and Dietary Fiber by Controlled Germination Bioprocess.

Chia (Salvia hispanica L.) plant is native from southern Mexico and northern Guatemala. Their seeds are a rich source of bioactive compounds which protect consumers against chronic diseases. Germination improves functionality of the seeds due to the increase in the bioactive compounds and associated antioxidant activity. The purpose of this study was to obtain functional flour from germinated chia seeds under optimized conditions with increased antioxidant activity, phenolic compounds, GABA, essential amino acids, and dietary fiber with respect to un-germinated chia seeds. The effect of germination temperature and time (GT = 20-35 °C, Gt = 10-300 h) on protein, lipid, and total phenolic contents (PC, LC, TPC, respectively), and antioxidant activity (AoxA) was analyzed by response surface methodology as optimization tool. Chia seeds were germinated inside plastic trays with absorbent paper moisturized with 50 mL of 100 ppm sodium hypochlorite dissolution. The sprouts were dried (50 °C/8 h) and ground to obtain germinated chia flours (GCF). The prediction models developed for PC, LC, TPC, and AoxA showed high coefficients of determination, demonstrating their adequacy to explain the variations in experimental data. The highest values of PC, LC, TPC, and AoxA were obtained at two different optimal conditions (GT = 21 °C/Gt = 157 h; GT = 33 °C/Gt = 126 h). Optimized germinated chia flours (OGCF) had higher PC, TPC, AoxA, GABA, essential amino acids, calculated protein efficiency ratio (C-PER), and total dietary fiber (TDF) than un-germinated chia seed flour. The OGCF could be utilized as a natural source of proteins, dietary fiber, GABA, and antioxidants in the development of new functional beverages and foods.

Effect of Sunlight Radiation on the Growth and Chemical Constituents of Salvia plebeia R.Br.

This study investigated the chemical composition changes of Salvia plebeia R.Br. cultivated under different light sources, including florescent light and sunlight. The plants were exposed to fluorescent light for four months and sunlight and then examined for the next 5-7 months. Plants were harvested monthly during the seven months, and we examined whether the difference in light source affected the phenolic and flavonoid contents and antioxidant activity. A simple and reliable HPLC method using a PAH C18 column was applied for the quantitative analysis of two triterpenoids from the S. plebeia groups. Oleanolic acid (OA) and ursolic acid (UA) showed good linearity (R² > 0.9999) within the test ranges (0.005-0.05 mg/mL), and the average percentage recoveries of the OA and UA were 95.1-104.8% and 97.2-107.1%, respectively. The intra- and inter-day relative standard deviations (RSDs) were less than 2.0%. After exposure to sunlight, the phenolic contents, including rosmarinic acid, showed a reduced tendency, whereas the flavonoid contents, including homoplantaginin and luteolin 7-glucoside, were increased. The content of the triterpenoids also showed an increased tendency under sunlight irradiation, but the variance was not larger than those of the phenolic and flavonoid contents. Among experimental groups, the group harvested at six months, having been exposed to sunlight for two months, showed the most potent antioxidant activity. Therefore, these results showed that the chemical composition and antioxidant activities of S. plebeia R.Br. was affected from environmental culture conditions, such as light source. Our studies will be useful for the development of functional materials using S. plebeia R.Br.

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.

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.

Chemical description and essential oil yield variability of different accessions of Salvia lavandulifolia.

The amount and chemical composition of essential oils are crucial for the modulation of the flavor, scent and therapeutic properties of aromatic and medicinal plants. The aim of this study was to evaluate the effects of phenology and weather conditions on the essential oil yield obtained from the aerial parts of Salvia lavandulifolia Vahl. Besides, we tried to carry out an approach to the chemical composition at the time of full bloom. Essential oil production of several accessions was monitored throughout the whole phenological cycle, both, at the original location growing wild and at the experimental plot as cultivated plants. Local pedoclimatic conditions seem to be crucial for the plant essential oil production. Our results showed high conditioning rates from both yearly climatic conditions and developmental stage of the plants. Maximum yield production was reported at the full seed maturation stage (average 1.74%) and after a slight dry period (average 2.16%). Phytochemical differences were maintained when plants were forced to grow under common pedoclimatic conditions. Thereby, essential oil analysis showed some populations formed by clearly distinct individuals while others had more homogenous plants. Compounds such alpha-pinene, beta-pinene + myrcene, limonene, 1,8-cineol, camphor and beta-caryophyllene were the main compounds of the essential oils of S. lavandulifolia.

Antioxidant and enzyme inhibitory activities of Plebeian herba (Salvia plebeia R. Br.) under different cultivation conditions.

An adaptation of cultural management to the specific cultural system, as well as crop demand, can further result in the improvement of the quality of horticultural products. Therefore, this study focused on the antioxidant and enzyme inhibitory activities of Plebeian herba (Salvia plebeia R. Br.) grown in hydroponics in comparison with those of the plant grown in soil. The antioxidant activities of Plebeian herba extract were measured as 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging abilities as well as the reducing power by decreasing nitric oxide (NO) and superoxide dismutase activity (SOD) in vitro. Interestingly, by comparison with hydroponics and traditional cultivation, Plebeian herba cultivated in nutrition-based soil improved inhibitory effect on free radicals of DPPH, ABTS, and NO and increased the contents of phenolics such as caffeic acid (1), luteolin-7-glucoside (2), homoplantaginin (3), hispidulin (4), and eupatorin. Free radical scavenging and SOD activity, as well as α-glucosidase inhibitory effect, were higher in Plebeian herba grown in nutrition-based soil than in plants grown in hydroponics and traditional condition.

Dietary supplementation with Salvia sahendica attenuates acetylcholinesterase activity and increases mitochondrial transcription factor A and antioxidant proteins in the hippocampus of amyloid beta-injected rats.

OBJECTIVES: We evaluated the acetylcholinesterase (AChE) inhibitory and potential antioxidant effects of Salvia sahendica extract to investigate whether these molecules are involved in learning and memory improvement in rats injected with fibrillar amyloid beta (Aß) peptide in the CA1 region of their hippocampus. METHODS: Rats were fed with S. sahendica (100 mg/kg/day) for 1 week before Aß injection. Western blot analysis and enzymatic assays were carried out 7 days after injections. KEY FINDINGS: Our results indicated that S. sahendica extract decreased AChE activity. Besides, S. sahendica prevented reduction in the level of nuclear respiratory factor-1 and mitochondrial transcription factor A (TFAM). Our data indicated the lack of sensitivity in citrate synthase and reduction in the activity of malate dehydrogenase in the presence of Aß that was prevented with S. sahendica consumption. Pretreatment with S. sahendica extract impeded reduction of glutathione redox ratio, catalase and superoxide dismutase activity, while decreased the malondialdehyde level. CONCLUSIONS: Based on the importance of AChE activity inhibition and increased TFAM level in the prevention of cognitive impairment, the use of S. sahendica could open a new protective issue in therapeutic fields of neurodegenerative disease.

[The quality of Salvia miltforthiza from Beijing suburbs with different growing modes].

OBJECTIVE: Study the yield and quality of Salvia miltforthiza in the poplars and apple trees intercropping system. METHODS: Assay the yield, heavy metals, organochlorine pesticides and active components of Salvia miltforthiza in different intercropping systems are different. RESULTS: The contents of Cd exceeds the allowed figure seriously though other heavy metals such as Pb, As, Hg, Cu fell in the standard range. The contents of organochlorine pesticides of different Salvia miltforthiza fell in the standard range. The active components of biennial Salvia miltforthiza intercropping with apple trees, including Tanshinone II A and Salvianolic acid B complied with the quality standards of Pharmacopoeia. CONCLUSION: Soil fertility, heavy metals of intercropping system and planting years should be considered in order to ensure the quality and stability of Chinese medicine.

Propagación y descripción morfo-anatómica de salvia chilena (Lepechinia chamaedryoides) / Propagation and morpho-anatomical description of Chilean salvia (Lepechinia chamaedryoides)

The aim of this work was to identify an effective method to improve the rate and percentage of germination and the rooting of seeds and cuttings of Lepechinia chamaedryoides (Balb.) Epling. Morphological changes experienced by the seed during the germination process also were followed by microscopy. This research was conducted during 2009 and 2010 in the Faculty of Agronomy of the University of Concepción. Four germination tests were assayed: a) sulfuric acid scarification and further stratification, b) leaching of inhibitors with different water temperatures plus different concentrations of gibberellic acid, c) thermal scarification plus stratification and d) gibberellic acid application at different concentrations. The highest percentage of germination was obtained with thermal scarification and stratification. Vegetative propagation test consisted of different concentrations of indole butyric acid (IBA) applied to the stem base. Significant differences between treatments were not found. The morphological changes of the seed during the germination process were observed by scanning electron microscope (SEM). These findings allowed identifying characteristics of highly lignified pericarp.

Con el propósito de identificar un método efectivo para obtener un alto porcentaje de germinación y enraizamiento en semillas y estacas de Lepechinia chamaedryoides (Balb.) Epling y observar los cambios morfológicos que experimenta la semilla durante el proceso de germinación, se realizó esta investigación durante el año 2009 y 2010 en la Facultad de Agronomía de la Universidad de Concepción. Se efectuaron cuatro ensayos de germinación: a) escarificación con ácido sulfúrico y estratificación, b) lixiviación de inhibidores con agua a diferentes temperaturas más concentraciones de ácido giberélico, c) escarificación térmica más estratificación y d) aplicación de ácido giberélico, en diferentes concentraciones. El mayor porcentaje de germinación se obtuvo con el tratamiento de escarificación térmica y estratificación. Con respecto a la propagación vegetativa, se realizaron ensayos con diferentes concentraciones de ácido indolbutírico (IBA) aplicado a la base de las estacas, no observándose diferencias significativas entre tratamientos. Los cambios morfológicos de la semilla durante el proceso de germinación se observaron mediante Microscopio Electrónico de Barrido (MEB) lo que permitió identificar características anatómicas y estructurales de un pericarpio altamente lignificado.

Chemical composition and phytotoxic effects of essential oils of Salvia hierosolymitana Boiss. and Salvia multicaulis Vahl. var. simplicifolia Boiss. growing wild in Lebanon.

The chemical composition of the essential oils of S. hierosolymitana Boiss. and S. multicaulis Vahl. var. simplicifolia Boiss. collected in Lebanon was studied by means of GC and GC-MS analysis. In all 115 compounds were identified: 82 for S hierosolymitana and 72 for S. multicaulis var. simplicifolia. The presence of carbonylic compounds (17%) characterizes the oil from S. hierosolymitana,while S. multicaulis var. simplicifolia oil is rich of monoterpenes (34.5%) and sesquiterpenes (46.9%). The effects of the essential oils on germination and initial radical elongation of Raphanus sativus L. (radish) and Lepidium sativum L. (garden cress) were studied, indicating in a different activity against radical elongation of the species tested.

The seed's protein and oil content, fatty acid composition, and growing cycle length of a single genotype of chia (Salvia hispanica L.) as affected by environmental factors.

As a botanical source, variability in chia seed composition could be expected between growing locations, and between years within a location, due to genotype and environment effects as well genetic x environment's interactions. The objective of the present study was to determine the location effect on the growing cycle length, and seed's protein content, lipid content, and fatty acid profiles, of a single chia genotype. Seeds of chia genotype Tzotzol grown on eight sites in five different ecosystems were tested. One site was in Argentina, in the Semi-Arid Chaco ecosystem (T(5)); one was in Bolivia, in the Sub-Humid Chaco ecosystem (T(4)); and six in Ecuador, one in the Coastal Desert (T(3)), two on the Tropical Rain Forest (T(2)), and three in the Inter-Andean Dry Valley ecosystem (T(1)). Seeds from plants grown in T(4) and in T(3) contained significantly (P <0.05) more protein percentage than did seeds from the other three ecosystems. No significant (P <0.05) differences in protein content were found between T(3) and T(4), and between T(1), T(2), and T(5). Seeds from T(1) and T(5) ecosystems, with 33.5 and 32.2%, respectively, were the numerically highest oil content producers, but their results were only significantly (P <0.05) higher when compared with the T(2) seeds. Significant (P <0.05) differences in palmitic, stearic, oleic, linoleic and alpha-linolenic fatty acids between oils from seeds grown in different ecosystems were detected, however. Oil of seeds grown in the T(3) ecosystem had the palmitic, stearic and oleic fatty acids' highest contents. Palmitic and oleic fatty acid levels were significantly (P <0.05) higher when were compared to that of seeds grown in the T(1) ecosystem, and stearic when was compared to that of seeds grown in the T(5) ecosystem; omega-6 linoleic fatty acid content was significantly (P <0.05) lower in oils of seeds produced in T(1), and T(2) than in those produced in T(3), T(4), and T(5) ecosystems; omega-3 alpha-linolenic fatty acid content was significantly (P <0.05) higher in seeds produced in T(1), than in those produced in T(3), T(4), and T(5), but not in those produced in T(2).

[Studies on the chemical constituents of Salvia miltiorrhiz of Lijiang].

OBJECTIVE: To study chemical constituents of Salvia miltiorrhiz of Lijiang. METHODS: The constituents were separated and purified by column chromatography with silica gel. Their structures were identified on the basis of physical and spectral data. RESULTS: Eleven compounds were isolated and identified as: (1) ferlalic acid; (2) O-hydroxybenzoic acid; (3) protocatechualdehyde; (4) beta-3,4-dihydroxyphenyl-lactic acid; (5) ursolic acid; (6) 6,7-dimethoxy-5,4'-dihydroxy-flavonol-3-O-glucoside; (7) carnosol; (8) tanshinone II(A); (9) tanshinone I; (10) 5,6-dehydrosugiol; (11) crypotanshinone.

[Study on induction of polyploidy in Salvia bowleyana by colchicine treatment].

OBJECTIVE: To explore the technique of induction of polyploidy in Salvia bowleyana by colchicine treatment. METHOD: The three kinds of explant of bud, leaf and calli were induced by colchicine treatment. RESULT: The induction effects were better when the calli was treated by colchicines (15 mg x L(-1)) and the leaf was pre-cultured for one week. The doubling rate was 33.33%, while the majority were wholy doubled plants, and the leaves were thicker and broader, the color was darker, the root was thicker and the stoma size was obviously bigger than the diploid plants. The number of chromosome were 8 to 64. Isoenzyme analysis showed that the enzyme activities between the polyploid and the diploid plants were quite different. CONCLUSION: Induction of polyploidy by colchicine treatment is efficacious. The part of the doubled plants were identified as homologmous tetraploids.

[Review of germplasm resources studies on Salvia miltiorrhiza].

Research development of germplasm resources of Salvia miltiorrhiza was reviewed in the aspects of species resource, geographic distribution and varieties, biological characters and culture varieties, and cell and tissue culture. It was suggested that more attention be paid to waters-soluble constituents when evaluating the quality of the medicine, and much more basic studies be done in the course of establishing standard plant base for high quality herb and in the aspect of new variety selection.