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.

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.

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.

Molecular Cloning and Characterization of FAD6 Gene from Chia (Salvia hispanica L.).

Plastidial Δ12 fatty acid desaturase (FAD6) is a key enzyme for linoleic acid (LA) and α-linolenic acid (ALA) biosynthesis. Chia (Salvia hispanica L.) is a revived omega-3 plant source that is richest in ALA level. In this study, based on the RACE method, one full-length cDNA sequence encoding FAD6, named ShFAD6, was isolated from chia. There exist three alternative transcription start sites and five alternative poly(A) tailing sites in ShFAD6. The 5'UTR of ShFAD6 contains a purine-stretch of 44 bp. ShFAD6 has an ORF of 1335 bp encoding a 444 aa protein of 51.33 kDa. ShFAD6 contains a conserved Delta12-FADS-like domain together with three strong trans-membrane helices and three histidine motifs. There also exists a chloroplast transmit peptide in ShFAD6 N-terminal. Phylogenetic analyses validated its identity of dicot FAD6 protein and suggested some critical evolutionary features of plant FAD6 genes. Heterologous yeast expression confirmed the catalytic activity of ShFAD6. The qRT-PCR assay showed that ShFAD6 is mainly expressed in leaves, stems, flowers, buds and early-stage seeds, and also responded to various stresses and hormone treatments. Under Sclerotinia infection, qRT-PCR and fluorescence imaging illustrated the possible correlation of ShFAD6 expression and photosynthesis. This study provides insight for further function study of ShFAD6 in oil quality improvement in staple oilseed crops as well as stress response and adaptation in plants.

Changes in the essential oil content and composition of Salvia limbata C.A. Mey at different growth stages and altitudes.

Salvia limbata is of great importance to the pharmaceutical industry owing to its various biological effects. Therefore, it is important to investigate the main factors that affect its essential oil composition. Although some investigations have been performed with regard to the phytochemistry of S. limbata, this study investigates, for the first time, the effect of growth stage and altitude on the content and chemical composition of essential oil extracted from S. limbata. For this purpose, the essential oil was extracted from 45 air-dried samples by hydrodistillation and analyzed by GC-MS and GC-flame methods. The highest content of essential oil was obtained from aerial parts in the vegetative stage at an altitude of 1500 m (0.86% v/w). Our findings show that the vegetative stage at 1500 m is the optimal harvest time to extract the highest content of oil while the highest content of monoterpenes (including α-pinene and ß-pinene) could be obtained in the same phenological stage at 2000 m. By contrast, the content of sesquiterpenes increased to the highest values in the ripening stage at 1500 and 2500 m. The results of this study help to find the optimal conditions to obtain the highest content of S. limbata essential oil, but additional studies are warranted.

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.

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.

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.

Comparative transcriptomic analysis provides insights into the development of a Salvia splendens Ker-Gawler mutant, SX919M.

Salvia splendens is a perennial, ornamental herbaceous flower that is widely cultivated as a bedding plant in gardens. The development of novel S. splendens cultivars and investigating the relevant molecular mechanisms are of great significance. In this study, RNA-sequencing and real-time PCR methods were used to analyze the possible molecular mechanism of S. splendens mutant, SX919M. From the wild-type S. splendens 919CK, we firstly selected a natural mutant, SX919M, which displayed multiple branches, clustered spheroids, and radial symmetrical inflorescence with higher numbers of calyces, ovules, stamens, and perianth tubes. Further, the RNA-seq was used to identify the differentially expressed genes (DEGs) in the mutant which included a total of 3568 upregulated and 3290 downregulated unigenes. We further observed that the indole alkaloid biosynthesis pathway showed the highest DEG enrichment, which was supported by a significant increase in the IAA content in mutant SX919M. In addition, we validated three DEGs, namely, CL2200.Contig2_All encoding methyl IAA esterase, CL12462.Contig1_All and CL12462.Contig2_All, which encoded strictosidine synthase, upregulated in mutant SX919M. We selected a novel S. splendens germplasm SX919M with a high ornamental value and determined that the upregulation of IAA biogenesis may be associated with its development.

Altitude, habitat type and herbivore damage interact in their effects on plant population dynamics.

Insects represent one of the most abundant groups of herbivores, and many of them have significant impacts on the dynamics of plant populations. As insects are very sensitive to changes in climatic conditions, we hypothesize that their effects on plant population dynamics will depend on climatic conditions. Knowledge of the variation in herbivore effects on plant population dynamics is, however, still rather sparse. We studied population dynamics and herbivore damage at the individual plant level of Salvia nubicola along a wide altitudinal gradient representing a range of climatic conditions. Using integral projection models, we estimated the effect of changes in herbivore pressure on plant populations in different climates and habitat types. Since we recorded large differences in the extent of herbivore damage along the altitudinal gradient, we expected that the performance of plants from different altitudes would be affected to different degrees by herbivores. Indeed, we found that populations from low altitudes were better able to withstand increased herbivore damage, while populations from high altitudes were suppressed by herbivores. However, the pattern described above was evident only in populations from open habitats. In forest habitats, the differences in population dynamics between low and high altitudes were largely diminished. The effects of herbivores on plants from different altitudes were thus largely habitat specific. Our results indicate potential problems for plant populations from high altitudes in open habitats because of increased herbivore damage. However, forest habitats may provide refuges for the plants at these high altitudes.

The effect of sowing time on the growth of chia (Salvia hispanica L.): What do nonlinear mixed models tell us about it?

Chia (Salvia hispanica L.) is an annual short-day plant whose growth has not been studied extensively in low-altitudes and at temperatures outside of its optimal range. The objective of this study was to describe the growth dynamics of a chia crop from an experimental plantation in south-east Mexico, on three different sowing dates. The chia grew at temperatures (18-37°C) and an altitude (9 m a.s.l.) outside of the recommended conditions (20-30°C, 500-1000 m a.s.l.). Three individual-plant responses were measured weekly, before seed harvest: height, number of leaves and number of inflorescences. Three theoretical nonlinear growth models were fitted to the data, a different model for each response. Mixed-effect model parameters were estimated by maximum likelihood, and the goodness of fit for each model was evaluated using two criteria: Modeling Efficiency and Root Mean Square Error. Chia seed yield was also measured in each treatment. Estimated parameters for plant height confirmed that medium sowing time (MST) and late sowing time (LST) plants had smaller heights than the early sowing time (EST) plants. Moreover, at the end of their life cycle, EST plants had a greater number of leaves and inflorescences, and higher seed yield. All of these differences were associated to the extended time of vegetative growth of EST plants favored by optimal photoperiod and temperature. Growth dynamics of chia during its ontogenic phases was explored, in more detail, with relative growth parameters derived from fitted models: a decrease in photoperiod influences the beginning of the reproductive phase, with the consequent reduction in speed of vegetative growth. In addition, nonlinear mixed-effects models can be useful in understanding the relation between growth parameters, plant maturity, and the suitable time for chia seed harvest. Our results suggest chia crops are adaptable to non-conventional environmental conditions.

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.

Influencia de reguladores de crecimiento sobre el establecimiento de cultivos in vitro de Salvia hispanica L. ("chía") y sobre su contenido de ácidos grasos / Influence of growth regulators on the establishment of in vitro cultures of Salvia hispanica L. ("chia") and its content of fatty acids

Salvia hispanica L. (Labiateae), comúnmente conocida como "chía", es una especie herbácea anual cuyo cultivo está ampliamente extendido por América del Sur. Es utilizada como cultivo industrial y para la ela-boración de alimentos funcionales por su contenido en proteínas, antioxidantes, fibras y lípidos esenciales. El aceite de sus semillas contiene la mayor proporción (68 %) de omega-3 que cualquier fuente vegetal conocida. El objetivo de este trabajo fue establecer cultivos in vitro de "chía" y analizar la influencia de diferentes reguladores de crecimiento sobre la inducción de callos, sobre el crecimiento de estos y sobre su contenido de ácidos grasos. Se iniciaron cultivos de callos in vitro a partir de explantos de tallos sin nudos y de hojas de plántulas axénicas de 20 días de edad, utilizando 3 tratamientos de reguladores de crecimiento diferentes. El medio Murashige & Skoog modificado (MSRT) con el agregado de ácido 2,4-diclorofenoxia-cético (2,4-D) a una concentración de 2,25 µM y un fotoperíodo de 16 horas fueron las condiciones óptimas para la inducción de callos. Para el mantenimiento de los callos el tratamiento más adecuado resultó ser bencilaminopurina (BAP) a una concentración de 1 µM. La cinética de crecimiento se caracterizó por un período de latencia hasta el día 20 de cultivo, seguido de un período de crecimiento exponencial entre los días 20 y 54. El tratamiento con 2,4-D (2,25 µM) mostró la más alta velocidad específica de crecimiento (0,22 ± 0,01 /día), el tiempo de duplicación más bajo (31,51 ± 1,00 día) y la mayor biomasa máxima (1,46 ± 0,01 g PF). El contenido de ácidos grasos en los callos de "chía" fue 0,83 % en promedio de todos los tratamientos después de 6 meses en cultivo y no mostró variaciones significativas (0,2 % - 0,3 %) entre los tratamientos aplicados (p < 0,05). (AU)

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.

Post-Fire Recovery in Coastal Sage Scrub: Seed Rain and Community Trajectory.

Disturbance is a primary mechanism structuring ecological communities. However, human activity has the potential to alter the frequency and intensity of natural disturbance regimes, with subsequent effects on ecosystem processes. In Southern California, human development has led to increased fire frequency close to urban areas that can form a positive feedback with invasive plant spread. Understanding how abiotic and biotic factors structure post-fire plant communities is a critical component of post-fire management and restoration. In this study we considered a variety of mechanisms affecting post-fire vegetation recovery in Riversidean sage scrub. Comparing recently burned plots to unburned plots, we found that burning significantly reduced species richness and percent cover of exotic vegetation the first two years following a 100-hectare wildfire. Seed rain was higher in burned plots, with more native forb seeds, while unburned plots had more exotic grass seeds. Moreover, there were significant correlations between seed rain composition and plant cover composition the year prior and the year after. Collectively, this case study suggests that fire can alter community composition, but there was not compelling evidence of a vegetation-type conversion. Instead, the changes in the community composition were temporary and convergence in community composition was apparent within two years post-fire.

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.