Stevia rebaudiana

Silver nanoparticles were synthesized using leaf extracts of three selected plants belonging to different family of brassicaceae(Brassica nepus), meliacea (Azadhiractaindica) and asteracea (Stevia rebaudiana). Each of the three plants housed a large number of phytochemicals that are capable of reducing ionic silver to atomic silver. Nano silver particles were produced by mixing 90ml of 1mM aqueous solution of silver nitrate with 10ml of the extract in each case. Characterization was done by physical observation of colour change, UV-Vis spectroscopy and transmission electron mi- croscopy (TEM). There were variations in colour changes which were attributed to the participating phytochemical presence in each of the plants. Size variations were also found with the average of 12.10nm, 20.30nm and 18.80 using the extracts from brassica, neem and stevia respectively.

En el presente estudio se evaluó la capacidad de adsorción del carbón activado y el efecto del hidróxido de calcio grado alimentico (cal grado alimenticio) para la remoción de color presente en los extracto de Stevia. Se preparó un extracto para la cuantificación de clorofila A, B y C por el método espectrofotométrico de Hansmann. Posteriormente se midió la capacidad de adsorción del carbón activado y de remoción de la cal sobre la clorofila A, B y C. Los procesos con carbón activado mostraron que con 0.05 g en 100 ml en un tiempo de residencia de 10 min se adsorbió el 43.30% de clorofilas. El proceso de adsorción sobre carbón activado se ajustó al modelo de Langmuir, con una capacidad máxima de adsorción de clorofilas en monocapas de qmax = 962 mgg–1 y un valor de la constante de Langmuir de b = 159.413 gml–1. Para la precipitación con cal se usaron 10 g por cada 100 ml en un tiempo de residencia de 10 min, retirándose el 70.30% de clorofilas.

Resultados para la evaluación del producto final correspondiente al total de los consumidores. 13.2. Resultado para la evaluación del producto final correspondiente a consumidores con edad menor ó igual a 39 años. 13.3. Resultados para la evaluación del producto final correspondiente a consumidores con edad mayor ó igual a 40 años. 13.4. Resultado para la evaluación del producto final correspondiente a consumidores de sexo femenino. 13.5. Resultado para la evaluación del producto final correspondiente a consumidores de sexo masculino.

Resultados para la evaluación del producto final correspondiente al total de los consumidores. 13.2. Resultado para la evaluación del producto final correspondiente a consumidores con edad menor ó igual a 39 años. 13.3. Resultados para la evaluación del producto final correspondiente a consumidores con edad mayor ó igual a 40 años. 13.4. Resultado para la evaluación del producto final correspondiente a consumidores de sexo femenino. 13.5. Resultado para la evaluación del producto final correspondiente a consumidores de sexo masculino.

This study aimed to determine the effects of selected plant growth-promoting silver nanoparticles (AgNPs) on certain physiological traits, plant growth, enzyme activity, and plant nutrient content of lettuce grown under different irrigation regimes. In this research, four irrigation regimes were tested as experiments in a randomized complete block design layout: I 100 (full irrigation), I 80 (20% deficit), I 60 (40% deficit), and I 40 (60% deficit). Additionally, four nanoparticle doses (AgNP 0 , 0 ppm; AgNP 20 , 20 ppm; AgNP 40 , 40 ppm; and AgNP 80 , 80 ppm) were applied to the crop. The results demonstrated that the head diameter of lettuce decreased by 28% and 30% under the I 60 and I 40 water stress conditions, respectively. However, AgNPs enhanced the head diameter of lettuce by 10.9% compared to the control (non-application of AgNPs). The application of AgNPs increased the nutrient content of lettuce, including potassium (K), sodium (Na), iron (Fe), copper (Cu), and manganese (Mn), by 38%, 19%, 23%, 367%, and 20%, respectively. Plant height was 98.2% higher in the full irrigation treatment (I 100 ) than in the 40% irrigation treatment (I 40 ). Dry matter content was 10.2% higher in the AgNP 80 treatment compared to the AgNP 0 treatment. In addition, the highest catalase (CAT) enzyme content was obtained in the AgNP 40 treatment. The present research suggests that applying AgNPs represents a sustainable and eco-friendly strategy for improving lettuce's plant growth and nutrient uptake under water stress conditions. Therefore, AgNPs can be used in drought stress tolerance studies.

The essential oil extracted by hydrodistillation from the flowering twigs of Stevia rebaudiana Bertoni (Asteraceae) was fractioned by chromatography. Fortythree constituents were characterized with the help of GC, GC-MS and other spectroscopic techniques. The essential oil was found to be a complex mixture of mono-and sesqui-terpenes. The cytotoxicity of the essential oil and its fractions was evaluated by sulforhodamine B (SRB) based assay against two cancer cell types viz. C-6 (rat glioma cells) and CHOK1 (Chinese hamster ovary cells). The essential oil and its fractions showed promising cytotoxicity against both cell lines. The highest activity (95.6±0.6%) was show by the essential oil on the C-6 cell line at a concentration of 400 μg/mL, which was comparable with that of the standard drug vinblastin.

The impact of particle size on bioactive compounds recovery was investigated for two valuable plant matters. This project was to assess the effect of grinding on bioactives extraction of New Zealand Manuka leaves of the Leptospermum scoparium and Stevia rebaudiana plants. Non-grounded and grounded Manuka with particle sizes ranging from 68 μm to 1400 μm were processed using n-hexane extraction for 0–60 min. Moreover, the extraction of two sweeteners from Stevia powder (d ≤ 200 µm) was carried out using hot water extraction and Ultrasound as a non-conventional method. As particle size was reduced, the extraction of bioactives increased, which is expected. However, it was observed that for the small particle size, which was obtained by sever grinding, most of the extraction was achieved as soon as the powder was exposed to the solvent (at zero time) with no further extraction at an extended period. This indicates that short-time exposure appears to be sufficient to recover most of the...

Steviol glycoside sweeteners are extracted and purified from the Stevia rebaudiana Bertoni plant, a member of the Asteraceae (Compositae) family that is native to South America, where it has been used for its sweet properties for hundreds of years. With continued increasing rates of obesity, diabetes, and other related comorbidities, in conjunction with global public policies calling for reductions in sugar intake as a means to help curb these issues, low-and no-calorie sweeteners (LNCSs, also known as high-potency sweeteners) such as stevia are gaining interest among consumers and food manufacturers. This appeal is related to stevia being plant-based, zero calorie and with a sweet taste that is 50-350 times sweeter than sugar, making it an excellent choice for use in sugar-and calorie-reduced food and beverage products. Despite the fact that the safety of stevia has been affirmed by several food regulatory and safety authorities around the world, insufficient education about stevia's safety and benefits, including continuing concern with regard to the safety of LNCSs in general, deters health professionals and consumers from recommending or using stevia. Therefore, the aim of this review and the stevia symposium that preceded this review at the ASN's annual conference in 2017 was to examine, in a comprehensive manner, the state of the science for stevia, its safety and potential health benefits, and future research and application. Topics covered included metabolism, safety and acceptable intake, dietary exposure, impact on blood glucose and insulin concentrations, energy intake and weight management, blood pressure, dental caries, naturality and processing, taste and sensory properties, regulatory status, consumer insights, and market trends. Data for stevia are limited in the case of energy intake and weight management as well as for the gut microbiome; therefore, the broader literature on LNCSs was reviewed at the symposium and therefore is also included in this review.

The glucosyltransferase UGT76G1 from Stevia rebaudiana is a chameleon enzyme in the targeted biosynthesis of the next-generation premium stevia sweeteners, rebaudioside D (Reb D) and rebaudioside M (Reb M). These steviol glucosides carry five and six glucose units, respectively, and have low sweetness thresholds, high maximum sweet intensities and exhibit a greatly reduced lingering bitter taste compared to stevioside and rebaudioside A, the most abundant steviol glucosides in the leaves of Stevia rebaudiana. In the metabolic glycosylation grid leading to production of Reb D and Reb M, UGT76G1 was found to catalyze eight different reactions all involving 1,3-glucosylation of steviol C 13 -and C 19 -bound glucoses. Four of these reactions lead to Reb D and Reb M while the other four result in formation of side-products unwanted for production. In this work, side-product formation was reduced by targeted optimization of UGT76G1 towards 1,3 glucosylation of steviol glucosides that are already 1,2-diglucosylated. The optimization of UGT76G1 was based on homology modelling, which enabled identification of key target amino acids present in the substrate-binding pocket. These residues were then subjected to site-saturation mutagenesis and a mutant library containing a total of 1748 UGT76G1 variants was screened for increased accumulation of Reb D or M, as well as for decreased accumulation of side-products. This screen was performed in a Saccharomyces cerevisiae strain expressing all enzymes in the rebaudioside biosynthesis pathway except for UGT76G1. Conclusions: Screening of the mutant library identified mutations with positive impact on the accumulation of Reb D and Reb M. The effect of the introduced mutations on other reactions in the metabolic grid was characterized. This screen made it possible to identify variants, such as UGT76G1 Thr146Gly and UGT76G1 His155Leu , which diminished accumulation of unwanted side-products and gave increased specific accumulation of the desired Reb D or Reb M sweeteners. This improvement in a key enzyme of the Stevia sweetener biosynthesis pathway represents a significant step towards the commercial production of next-generation stevia sweeteners.

Introduction
The Fe iron significantly affects the quantity and quality of agricultural production. Factors affecting the absorption of this element increase its efficiency. Meanwhile, the pH of the nutrient solution plays an important role in iron absorption. Iron is one of the essential elements for plant growth and plays an essential role in chloroplasts. Due to iron deficiency, the activity of several enzymes such as catalase, cytochrome oxidase, and ferroxin is significantly reduced. The amount of iron in the soil is high, but plants only absorb two-capacity of soluble soil, which is negligible compared to the total iron. Soil environmental conditions affect the amount of iron by the plant, so it is difficult to control the uptake of iron by the plant. It has the highest ability to absorb iron and manganese at a pH less than six. For this reason, acetic acid was used to stabilize the pH of the solution. Acetic acid has all the properties of an acid. Acetic acid is a polar solvent and an organic compound. The use of inexpensive organic acid and citric acid in agriculture, despite its positive effects on calcareous soils and their reasonable price, is still not common in Iran. Acetic acid has a carboxylic group and therefore has all the properties of an acid.

Materials and Methods
The pH solution plays a role in absorbing iron elements. This experiment was carried out in the form of a split-split plot in a randomized complete block design on a strawberry plant of diamant cultivar in the research station of the University of Mohaghegh Ardabili during the years 2015-2017. Ardabil City in northwest Iran is located between Sablan and Baghrou mountain ranges. Due to its high altitude and mountainous nature, this city is colder than other cities in Iran and is considered one of the semi-arid cold regions. The average rainfall of this city is reported to be 400 mm. Factors included acetic acid (zero, one, two, and three percent), iron in two levels (sequestration 25 gr and nano one gr), and two levels of agricultural soap agents (zero and 7.5 %) as foliar spraying. Foliar application of pH nutrient solution from the three-leaf stage of the plant (mid-April) to the end of May, a total of five times 10 days apart in both years, was done. Two weeks after the last foliar application (June of the second year) plant growth indices included leaf fresh and dry weight, root fresh and dry weight. Root length, flower-to-fruit ratio, chlorophyll greenery a, b, and total, leaf iron content and fruit yield per plant were measured.

Results and Discussion
The results of the analysis of variance showed that the three-way effect of treatments in all studied traits except root length and yield at one percent probability level on leaf fresh weight, leaf dry weight, and chlorophyll a, b and total at five percent probability level on fresh weight and root dryness, flower to fruit ratio and leaf iron content were statistically significant. The three-way interaction of acetic acid, iron, and agricultural soap data showed that the highest leaf fresh weight, leaf dry weight, root fresh weight, root dry weight, ratio of flowers to fruits, chlorophyll b, total chlorophyll and the amount of leaf iron related to the treatment acetic acid two percent of sequestration iron in combination with agricultural soap (7.5 %) and in chlorophyll a, it was related to the treatment of acetic acid one percent of sequestrin iron in combination with agricultural soap. The lowest amount of fresh and dry weight of leaves, fresh root weight and dry root, flower-to-fruit ratio, and leaf iron content were related to the treatment of acetic acid zero with Nano iron with agricultural soap (control). The lowest Chlorophyll b and total chlorophyll were related to the treatment of acetic Acid three percent with Nano iron with agricultural soap (control). In addition, the interaction between acetic acid and iron at the level of five percent probability on root length and yield was significant.

Conclusion
It can be concluded that acetic acid two percent on the absorption of iron fertilizer, along with agricultural soap (7.5 %) application, has better results than other treatments and qualitative traits of strawberry fruit. In general, it can be concluded that the pH of the nutrient iron solution can improve plant growth and crop yield. The iron level of sequestration iron with two percent acetic acid with agricultural soap 7.5 % due to increasing the bicarbonate of the soil in the Ardabil region has caused the absorption of nutrient elements, especially iron by strawberries. The shelf life of the solution on the leaf surface had a more favorable effect on most growth indices in the strawberry Diamant cultivar in Ardabil soil conditions.

A moringa pod powder (MPP) fortified Burfi was developed with the aim to enhance its nutritional and functional profile. Response Surface Methodology (RSM) employing a central composite rotatable design was used to optimize the variable levels of MPP (0.50-1.00%) and sugar (5.00-7.00%), against sensory responses (colour & appearance, flavour, sweetness, body & texture, overall acceptability) and instrumental hardness. The optimized formulation, comprising 0.69% MPP and 6.02% sugar, achieved a desirability of 1.00. The predicted values for sensory scores and hardness closely matched the actual values obtained from seven independent trials, with all differences found non-significant (p<0.05). The optimized MPP Burfi had a moisture content of 19.80%, fat 17.46%, protein 15.40%, ash 3.16% and total carbohydrates 44.64%, reflecting a nutritionally balanced product. Additionally, it had acidity (0.30% LA), FFA (0.21% oleic acid), tyrosine value (12.20 µg/5 ml filtrate), water activity (0.84) and hardness (16.10 N), indicating good stable texture. Thus, the incorporation of MPP resulted in a valueadded Burfi with superior protein content, functional health properties of milk and moringa constituents with desirable sensory characteristics, effectively meeting the growing consumer demand for functional dairy sweets.

In this study, the Binary Puma Optimizer (BPO) is introduced as a novel binary metaheuristic. The BPO employs eight Transfer Functions (TFs), consisting of four S-shaped and four V-shaped mappings, to convert the continuous search space of the original Puma Optimizer into binary form. To evaluate its effectiveness, BPO is applied to two wellknown combinatorial optimization problems: the 0-1 Knapsack Problems (KPs) and the Uncapacitated Facility Location Problem (UFLP). The solver tailored for KPs is referred to as BPO1, while the solver for the UFLP is denoted as BPO2. In the UFLP experiments, only TFs are integrated into the solutions. Conversely, in the 0-1 KPs experiment, the additional mechanisms are (i) greedy-based population strategies; (ii) a crossover operator; (iii) a penalty algorithm; (iv) a repair algorithm; and (v) an improvement algorithm. Unlike KPs, the UFLP has no infeasible solutions, as facilities are assumed to be uncapacitated. Unlike KPs, the UFLP has no capacity constraints, as facilities are assumed to be uncapacitated. Thus, violations cannot occur, making improvement strategies unnecessary, and the BPO2 depends solely on TFs for binary adaptation. The proposed algorithms are compared with binary optimization algorithms from the literature. The experimental framework demonstrates the versatility and effectiveness of BPO1 and BPO2 in addressing different classes of binary optimization problems.

Neoerysiphe cumminsiana (U. Braun), the etiological agent of powdery mildew in diverse asteraceae plants, was recovered from symptomatic Stevia rebaudiana (Bertoni) plantlets grown under greenhouse conditions in Mérida, Venezuela. The etiology of the disease was confirmed by pathogenicity tests, and the identity of the fungus was performed using both morphologically and by sequencing and analysing the nuclear rRNA locus ITS4. Neoerysiphe cumminsiana (U. Braun) causante de mildiu polvoriento en distintas Asteráceas fue observado en plantas de Stevia rebaudiana (Bertoni) obtenidas por cultivo in vitro, que se encontraban creciendo en condiciones de invernadero en el estado MéridaVenezuela. El hongo en este hospedador crece ectofíticamente y aparece como un crecimiento blanco y polvoriento que lleva a la necrosis del tejido. Se realizaron pruebas de infección y patogenicidad que resultaron positivas y se confirmó su identidad por medio del análisis morfológico y de la comparación de se...

The leaves of Stevia rebaudiana, from Asteraceae family, are a source of steviol glycosides, which are utilized as a sweeter, more favorite addition. Among the most important anatomical results that were reached through the research and which distinguish the plant from an anatomical point of view are uniseriate-multicellular of non-glandular trichomes that are covered the epidermis of stem and leaves. Also, the vertical walls of the ordinary epidermis cells in the upper and lower epidermis that are heavily sinuous and the stomata scattered in both of it, the type of the stomata anomocytic. In addition, the mesophyll which differentiated into 2-3 layers of palisade cells under the upper epidermis and the spongy cells after it and the cross-section of midrib region of the leaf as crescent shape and the vascular bundle central, crescent shape. .

Goldenseal (Hydrastis canadensis L.) is a medicinal plant that is commonly used in traditional medicine and as a dietary supplement. This plant's rhizomes/roots have been used to treat a wide range of conditions. Berberine and hydrastine are the most active alkaloids in goldenseal. Goldenseal extract demonstrated several therapeutic benefits, including anti-diabetic, anti-inflammatory, antimicrobial, hypolipidemic, neuroprotective, cardioprotective, and digestive protection. Numerous studies have demonstrated that goldenseal extracts and components have health benefits. This study investigated the anti-diabetic and weight reduction effects of goldenseal root extract in Wistar albino rats. Diabetes was induced by administering streptozotocin intraperitoneally at a dose of 35 mg/kg body weight. The diabetic rats were given goldenseal root extract at various doses (150, 250, 350 and 450 ppm/kg body weight), or none, for 8 weeks. Diabetic groups were compared with a non-diabetic control group. After 8 weeks of application in diabetic rats, the goldenseal root extract was associated with a reduction in feed intake (11%; ***p<0.001) and body weight (15%; ***p<0.001), and significant decreases in random blood glucose (33.2%; **p<0.01), fasting blood glucose (45.6%; ***p<0.001), and glycosylated haemoglobin (HbA1c) (29.5%; ***p<0.001), while insulin (1.15 times; ***p<0.001) and liver glycogen (1.7 times; **p<0.01) levels significantly improved compared to untreated diabetic rats. The study found that goldenseal root extract has anti-diabetic and weight-management properties in Wistar albino rats. Therefore, it appears to be an effective alternative treatment for type 2 Diabetes and weight management.

Introduction: Stevia rebaudiana bertoni is an herbaceous plant widely distributed and used in Paraguay and Brazil, known for its sweetener character. Stevioside (E), a diterpenoid glycoside, is the main non-caloric sweetener extracted from Stevia leaves. Several papers have been published about stevia and its properties as antihypertensive, insulinotropic, antihyperglycemic and antitumor.  However, there are few documented findings on cardiovascular effects mediated by stevioside. In previous studies carried out in our laboratory, we demonstrated that oral administration of E improved the recovery of contractile activity in hearts subjected to I-R and decreased the infarct size. These effects were, at least in part, reverted by the administration of Wortmannin (W), an upstream inhibitor of Akt. Objective: The aim of the present study was to investigate the mitochondrial morphology and the rate of ATP synthesis after oral administration of E (168 mg/kg for 15 days) and its relation w...

Stevia rebaudiana (Bertoni) is a Paraguayan perennial herb of the family Asteraceae. The leaves contain a great amount of secondary metabolites with a wide range of important biological activities commonly known as steviol glycosides which differ in their molecular configuration, power of sweetness and their taste profile. Out of various steviol glycosides, the main compounds of interest are diterpenoid glycosides of ent-kaurene type extracted from the leaves of this plant as non-toxic, thermally stable, low-calorie natural sweeteners stevioside and rebaudioside A. These glycosides are a high-quality sugar substitute or dietary supplement with diverse applications in the medicinal world along with the food and beverage industry. This review article is aimed at the chemistry of stevioside and rebaudioside A, possible biosynthetic pathways, their metabolism and acceptable daily intake along with a broad spectrum of pharmacological and therapeutic applications.

Egyptian Jallab (EJ) is a conical candy (light to dark brown), manufactured from a part of sugar cane juice, that is used in the black honey industry. EJ is considered an unrefined sugar or a noncentrifugal form of sugar. The traditional use of Jallab is as candy, but it can also be used for making ice cream, cupcakes, biscuits, and toffee, as well as being used in other food applications. In this study, EJ was used as a sugar substitute in ice cream at 0, 25, 50, 75, and 100%. Total solids, titratable acidity, pH, protein, ash, fat, specific gravity, weight per gallon, viscosity, color attributes, total antioxidant activity, total phenolic content, and total flavonoid contents, as well as microbiological analyses, were tested. The total solids, protein, and ash in the Egyptian Jallab ice cream (EJIC) increased from 39.30, 4.85, and 0.87 to 41.19, 6.36, and 1.42, respectively. The gradual sugar substitution led to a significant increase in specific gravity and weight per gallon in pounds. The lightness (L*) of the ice cream decreased significantly due to the substitution of EJ for sugar. Moreover, there was a significant increase in a* (from 0.147 in control samples to 5.52 in treatment 4, which had 100% EJ). The changes in the b* values of Jallab ice cream samples were significantly increased due to the substitution of EJ for sugar. The control samples had a low value of antioxidant activity (21.53%) when compared with the treatment, which has EJ (88.82, 89.96, 91.98, and 92.14%) for EJIC1, EJIC2, EJIC3, and EJIC4, respectively. The total phenolic contents are 2.07, 3.03, 4.14, and 4.68 fold higher in the treatments with EJ substituted for sugar than in the control samples. Total flavonoid contents increased from 5.73 mg QE g -1 in control samples (TC) to 14.68, 21.54, 30.48, and 34.15 mg QE g -1 in EJIC1, EJIC2, EJIC3, and EJIC4 mg QE g -1 in ice cream samples, respectively.

This research was conducted to determine the effect of adding stevia leaf powder on the anti-diabetic properties of stevia-black tea drinks. A design is needed to find the best proportion of stevia black tea to get the best anti-diabetic properties. The research design used was a response surface method completely, with a central composite, 2 factors, and 1 replication. The factor was stevia and black tea, with varying weights according to the design proposed by the RSM program. The combination of stevia and black tea was put into a tea bag, weighing 2 g. Tea was brewed by steeping every 2 g of stevia black tea in 200 mL of hot water at 94 ± 5 °C for 4 ± 1 min. And then cool the infusion to room temperature for 15 min, and the tea was filled in a glass of dark brown color, to be analyzed one hour later. The parameter analyzed for anti-diabetic properties was the inhibition of the alpha-amylase enzyme. The data obtained from testing this parameter was then analyzed using RSM design Minitab 17. The results of the optimization show that stevia has a direct and positive effect on amylase inhibitory ability, while black tea has an inverse and negative effect.

Currently the consumer needs for anti-diabetic products are very large and require answer from healthy food provider answers. Products that are commonly known are tea and stevia. Tea and stevia have advantages as an antioxidant and anti-diabetic. The use of commercially available products is very depended on sensory characteristics which can be accepted by consumers. How much is consumed (food intake) depends on the sensory sensitivity of the consumer. Tea and stevia products are products that contain poly phenols which give a sense of briskness or bitterness. At the time of serving alone, stevia tea and steeping have a certain value, which will be different if brewed simultaneously. The study aims to determine the sensory acceptance threshold value, to steeping a mixture of black tea and dried stevia leaves. The study was conducted using the absolute threshold test, using a proportion of the concentration of stevia black tea between 20:1 to 4:1(w/w) and maximum weight of 2 grams. The mixture was brewed using 200 mL hot water with a temperature of 90 0 Celsius for 5 minutes. The steeped product was then tested by 80 untrained panelists to find out the absolute threshold for sweet and bitter taste. From the research it was known that the absolute threshold value for sweetness of stevia-black tea was 0.39% (w/v) and bitterness ofstevia -black tea was 0.42% (w/v).

Liver fibrosis is a natural process that initiates after liver injury to repair the damaged tissue. The liver has a significant capacity for self-repair of the damaged tissue. To a great extent, the miscellaneous interactions of immune cell subtypes manage these repair procedures like fibrosis and wound healing. Objectives: To assess the hepatoprotective and anti-inflammatory potential of Euphorbia pilulifera through modulating the NF-KB/Nrf2/AKT/TGF-β1 pathway. Methods: Euphorbia pilulifera methanolic extract was primarily assessed for its cytotoxic potential against HepG2 cells. Methanolic extract of E. pilulifera showed 90% hepatoprotective activity against CCl4- induced toxicity in HepG2 cells. The methanolic extract downregulated the NF-κB gene by 90%; the AKT, gene by 14%, and the TGF-β1 gene expression by 69% at the concentration of 50µg/ml at the mRNA level. On the other hand, methanolic extract of E. pilulifera increased the expression of the Nrf2 gene by 44% at 50µg/ml con...

Stevia Rebaudiana leaves have functional and sensory properties superior to those of many other high-potency sweeteners. Dental caries is an infectious microbiological disease that affects almost every human. There is a linear correlation of sugar consumption with dental caries in populations worldwide. Present invention was directed to design and develop Stevia based polyherbal formulation with enhanced performance in oral care as Stevia inhibits the growth of dental caries causing microorganisms. In the present study, twelve pure isolates grown on Snyder agar medium and five MTCC strains i.e. Streptococcus mutans, Staphylococcus aureus, Curvularia lunata, Escherichia coli and Bacillus subtilis were screened for antimicrobial activity through agar well diffusion method. Metabolites from plant leaves were extracted using six different solvents and investigated for phytochemical properties. Few cultures were found to be belonging to Staphylococcus species, Bacillus sp., Clostridium sp., Neisseria sp., and Escherichia. coli. On the basis of maximum zone of inhibition produced against cultures, diethyl ether extracts of plant were effective against bacteria and fungi, followed by petroleum ether and chloroform and finally minimum inhibitory concentration and minimum bactericidal concentration of extracts were performed. The phytochemical screening of different extracts of Stevia showed the presence of various secondary metabolites. Herbal formulation prepared in diethyl ether extract displayed significant bioactivity against all twelve isolated cultures and standard cultures. The study holds importance as one needs to be familiarized with alternatives to sugar and type of food products that are available with substitute sweetening agents, as we have the responsibility to reduce the incidence of dental caries by emphasizing the role of sugars in caries formation.

under drip irrigation system during the successive season, 2011. Experiment was carried out to evaluate the response of stevia herb yield and sweeteners content to cobalt supplement. Stevia seedlings were irrigated with cobalt once at concentrations: 0.0, 2.5, 5.0, 7.5, 10.0, 12.5, 15.0 and 17.5 ppm. The obtained results are summarized in the following: Cobalt has a significant promotive effect on all growth and yield parameters as well as nutritional and chemical content compared with control. Cobalt significantly increase stevia leaves content of sweetners such as glycosids, steviosides and rebaudiosides compared with untreated plants. Cobalt at 12.5 ppm recorded the greatest figures. Cobalt at 12.5 ppm increased glycoside to 34.11 and 37.0%, steviosides to 27.6 and 31.6%, rebaudiosides to 44.5 and 45.1% respectively in two cuts. Increasing cobalt in plant media more than 12.5 ppm the promotive effect decreased.

The aim of the present investigation was to study the effect of auxins for in-vitro callus induction and plant regeneration of an anti-diabetic medicinal plant, Stevia rebaudiana which has gained high demand in Bangladesh. Three different types of explants (nodal, inter-nodal and leaf segments) from in vitro raised shoots were cultured in Murashige and Skoog (MS) medium fortified with different auxins 2,4-dicholorophenoxy acetic acid (2,4-D), a-naphthaleneacetic acid (NAA) and indole-3-acetic acid (IAA) at various concentrations either singly or in combination. The presence of 2, 4-D [0.5 mg/l and 1.0 mg/l] in MS medium showed greater influence on nodal segments; excellent callus induction was observed when leaves were cultured in MS containing 2, 4-D [3.0 mg/l]. Moderate callogenic response was obtained with NAA or IAA in combination with 2, 4-D. On the other hand, NAA [0.5 mg/l] alone showed simultaneous shoot proliferation and root induction from nodes. The highest length of micro-shoot was 5.46 cm at 0.5 mg/l NAA and mean root numbers were 5.63/shoot. Maximum shoot multiplication observed at 1.5mg/l of NAA was 2.75. The plantlets showed 70% survival rate after acclimatization. This investigation could play a role towards cost effective production of callus and plant of Stevia rebaudiana.

The sweet flavor of Stevia rebaudiana (Bert.) leaf extract is well known and has raised the interest of huge food companies due to its natural bid. The extraction of their main glycosides stevioside and rebaudioside A is an important step on the preparation of final Stevia granules. The aim of the work reported here was to study and optimize the dynamic maceration of Stevia leaves using water and ethanol as green solvents. For instance, a fractional factorial design was chosen to evaluate the individual effects of the drug powder size, weight ratio of drug to solvent, temperature, agitation, and time on the yield of these glycosides. The glycosides were quantified by high pressure liquid chromatography. An exhaustive extraction by successive maceration steps showed that ethanol 70% was superior to water and ethanol 90% for stevioside and rebaudioside extraction. The liquid extract composition in dry basis and the yield of stevioside and rebaudioside A were significantly affected by the drug to solvent weight ratio, showing that larger volumes of solvent should be used. Furthermore, increasing solvent volume favors the extraction of the stevioside by a twofold factor as compared to rebaudioside A. Among the other factors, only drug powder size affected the yield of rebaudioside A significantly. The optimal solution for S. rebaudiana leaves dynamic extraction estimated by desirability functions methodology led to a condition which allows obtaining extraction yields of 2.31 and 1.24% for stevioside and rebaudioside A and their concentrations in dried extract corresponding to 8.38 and 4.51%, respectively. These high yields were obtained with drug to solvent ratio (1:10, w/w) much higher than previous works, thus resulting in a more sustainable and green process.

Stevia rebaudiana (Bertoni) Bertoni, Asteraceae, leaf extract has recently called the attention of food industry as a proposal for natural sweetener. The sweet flavor is attributed to the glycosides, in especial stevioside and rebaudioside A, which are the plant main chemical markers. The aim of the work reported here was to optimize the turbo-extraction of stevia leaves using water, ethanol 70% and 90% (w/w) as green solvents. A 2 5-2 factorial design was applied to study the linear effects of the drug size, solvent to drug ratio, temperature, time and also the turbolysis speed on the extraction of glycosides. The glycosides exhaustive extraction showed that ethanol 70% gave better results and was used for turbo-extraction. The stevioside and rebaudioside A contents were quantified by a validated method by high performance liquid chromatographic with photodiode array detector. The contents of stevioside and rebaudioside A in fluid extract increased with the drug size, but decreased at high shearing speeds and solvent to drug ratio, while their yields decreased at higher temperature and were not affected by turbo speed. An increase in solvent to drug ratio reduced significantly the glycosides percent in dried extract. Optimal solution for S. rebaudiana leaves turbo-extraction was determined by desirability functions. The optimal extraction condition corresponded to drug size of 780 m, solvent to drug ratio of 10, extraction time of 18 min; temperature of 23 • C and turbo speed of 20,000 rpm, resulting in yields of 4.98% and 2.70%, for stevioside and rebaudioside A, respectively. These yields are comparable to the ones recently published for dynamic maceration, but with the advantage of shorter extraction times. This work demonstrates that turbolysis is promising for S. rebaudiana glycosides extraction and stimulate new research on the purification of these extracts, which may become an interesting source of income for developing countries such as India and Brazil.

Stevia leaves have the potential to be developed as a natural sweetener, a companion to cane sugar, especially for those who need lowcalorie intake. Increasing the productivity of stevia plants is approached by increasing plant biomass accumulation through balanced nitrogen fertilization. This research aims to obtain optimal nitrogen fertilization in the cultivation of stevia plants in which the nitrogen derived from various sources. The experiment was arranged in a factorial Randomized Block Design (RBD) with two factors and three replications. The first factor was organic fertilizer consisting of 4 treatments (C1: Without Crotalaria juncea; C2: Crotalaria juncea 6 ton/ha; C3: Cow-dung manure 6 tons/ha; C4: Crotalaria juncea L. 3 ton/ha + cow-dung manure 3 tons/ha). The second factor was the dose of N fertilizer which consisted of 4 treatments (D0: 0 kg N ha-1 fertilizer; D1: 50 kg N ha-1 ; D2: 100 kg N ha-1 ; D3: 200 kg N ha-1). The study result showed that the optimum dose of N fertilization was at a level of N 50 kg ha-1. The leaf weight ratio was higher than the stem weight in all N fertilization treatments. Application of organic material derived from Crotalaria Juncea 6 tons' ha-1 gave the highest yield of stevia biomass, followed by the application of organic matter with a combination of crotalaria 3 tons' ha-1 + manure 3 tons' ha-1. The highest leaf ratio was achieved in the treatment of organic matter derived from Crotalaria juncea, namely 61% of the total weight of the biomass. The application of organic fertilizer derived from Crotalaria juncea would reduce the use of inorganic N fertilizers which leads to reduced stevia farming costs. Moreover, applying a combination of organic and inorganic fertilizers will provide benefits not only for plant growth and yields but for soil health as well.

Disinfection of Enterococcus faecalis bacteria in dental root canal treatment using NaOCL 0.5-2.5% is still a challenge. There is still very little data on the effectvity of Stevia rebaudiana Bertoni leaf extract irrigation solution against the growth of Enterococcus faecalis. This study evaluated the effectiveness of Stevia rebaudiana Bertoni leaf extract irrigation solution against Enterococcus faecalis ATCC 29212 through MIC (Minimal Inhibitory Concentration) and MBC (Minimal Bactericide Concentration) values. A number of 100 µl of E. faecalis suspension was inoculated into 96 microtitration wells, each containing 100 µl of 100%, 50%, 25%, 12.5%, 6.25%, 3.125%, 1.56% of Stevia leaf extract irrigation solutions, positive (NaOCL 2.5%) and negative (distilled water) control. Incubation was carried out for 24 hours at 37°C then optical density was measured with a microplate reader at 600 nm. Five µl of 10 -9 dilution of each group was dripped and spread onto nutrients agar plate. The number of Enterococcus faecalis colonies was counting manually. There was no significant difference in average colony count of Enterococcus faecalis between the 100% and 50% Stevia leaf extract irrigation solution groups and the positive control (p>0.05). However, the 25%, 12.5%, 6.25%, 3.125%, and 1.56% Stevia leaf extract irrigation solution groups showed significant differences (p<0.05). Stevia leaf extract irrigation solution suppressed growth at MIC 6.25% by turbidimetry and MIC 1.56% by total plate count method using nutrient agar and than killed Enterococcus faecalis at MBC 50%. The bactericidal ability of 100% and 50% Stevia leaf extract irrigation solution is equal to NaOCL 2.5%.

The aim of the present investigation was to improve steviol glycosides (SGs) of three stevia varieties; Sponti (SPNT), China1 (CHN) and EGY1 (EGY) by obtaining new genotypes with high ratio of rebaudioside A (rebA)/stevioside (stv) using five different doses of colchicine(0, 0.025, 0.05, 0.1 and 0.2%), during 2018 and 2019 seasons at ElSabahya Research Station, Alexandria Governorate. Plants with a significant large leaf area/plant and significantly high number of leaves/plant, as well as dry matter content from the three commercial varieties, were selected and evaluated for their steviol glycosides content by using high performance liquid chromatography (HPLC). Results of buds survival rate, showed that SPNT and EGY varieties performed better than CHN. SPNT-22, CHN-31, EGY-22, EGY-23and EGY-31 mutated clones showed a significant superiority for most desirable morphological characters (leaf area/plant, number of leaves/plant as well as dry matter content)compared to other tested ge...

La parasitosis gastrointestinal es una enfermedad que se da preferentemente en la niñez y en adultos inmunocomprometidos. Los tratamientos actuales son, en general, eficaces, pero la creciente resistencia a los antiparasitarios obliga a la búsqueda de nuevas drogas con urgencia. Este estudio tiene el propósito de evaluar preliminarmente la efectividad de Stevia rebaudiana Bertoni como antiparasitário. Se realizó un ensayo clínico aleatorizado a ciego simple, con 22 adultos y 134 niños (6-18 años), todos parasitados con helmintos y/o protozoarios. Los tratamientos asignados al azar consistieron en comprimidos de S. rebaudiana (SRB) o mebendazol-tinidazol (MBZ-TNZ), ambos en preparaciones farmacéuticas comerciales. En los adultos SRB y MBZ-TNZ redujeron en forma similar la infección por protozoarios y helmintos, y el número de eventos adversos informados para el tratamiento convencional fue mayor que para SRB (p<0,02). Atendiendo a esos resultados se amplió la evaluación a 134 niños portadores de parásitos intestinales, de los que 113 (99,3%) presentaron protozoarios y 28 (20,8%) helmintos, mayoritariamente en co infección. S. rebaudiana redujo significativamente (p<0,001) la frecuencia de global de infección por protozoarios (74,5%) en los niños frente a lo obtenido con MBZ-TNZ (40,7%), destacando la negativización de Giardia lamblia.que resultó el doble con SRB de lo observado con MBZ-TNZ (p<0,001). SRB redujo la frecuencia de helmintos en 70,6% de manera similar a MBZ-TNZ (p>0,05). No se registraron efectos adversos asociados a los tratamientos a los niños/as. Los resultados animan a proseguir la evaluación de SRB como potencial agente antiparasitario.

Preliminary clinical evaluation of the antiparasite effect of Stevia rebaudiana Bertoni (ka`a he ê) in adults and children Achucarro C, Ferro EA, Richer Y, Salazar ME, Ciciolli S3, Ortiz I4, Campos S, Sckell C, Samudio M, Alborno RM, Meza B, Varela I, Losanto J, Pedrozo JR 1 Dirección General de Investigación Científica y Tecnológica , Universidad Nacional de Asunción; 2 Cátedra de Bioquímica, Facultad de Ciencias Médicas, UNA; 3 Laboratorio Central del Hospital de Clínicas, Facultad de Ciencias Médicas, UNA; 4 Primera Cátedra de Clínica Médica, Facultad de Ciencias Médicas, UNA; 5 Instituto de Investigación en Ciencias de la Salud, UNA; 6 Primera Cátedra de Patología Médica, Facultad de Ciencias Médicas, UNA ; 7 Facultad de Ciencias Médicas, UNA.

Candida albicans merupakan jamur oportunistic yang sering dijumpai pada infeksi saluran akar gigi. Kemampuannya membentuk biofilm pada dentin dinding saluran akar gigi, menyebabkan peningkatan virulensi dan resistensinya terhadap NaOCL sebagai gold standard larutan irigasi. Ekstrak daun Stevia rebaudiana Bertoni diketahui bersifat antijamur terhadap Candida albicans, namun data penelitian aktivitas terhadap pembentukan biofilm Candida albicans sangat terbatas. Penelitian ini bertujuan menganalisis aktivitas hambatan biofilm larutan irigasi ekstrak daun Stevia rebaudiana Bertoni terhadap jamur Candida albicans. Penelitian eksperimental laboratorium in vitro dengan post-test only with control group design. Serial dilusi dilakukan untuk menyiapkan ekstrak daun Stevia 100%,50%,25%,12,5%, 6,25%, 3,125% dan 1,56%, dengan larutan NaOCL 2,5% sebagai kontrol positif dan media Sabouraud Dextrose Broth (SDB) sebagai kontrol negatif. Metode crystal violet assay untuk menilai aktivitas hambatan biofilm melalui pengukuran nilai densitas optik (OD) pada panjang gelombang 490nm. Uji Kruskal-Wallis menunjukkan perbedaan signifikan (p=0.000) antara rata-rata nilai OD eksperimen. Uji Mann-Whitney U terdapat rata-rata nilai OD eksperimen seluruh larutan irigasi ekstrak daun Stevia lebih kecil signifikan dibandingkan kontrol negatif dan lebih besar signifikan dibandingkan kontrol positif (p<0,05). Larutan irigasi ekstrak daun Stevia menunjukkan rata-rata nilai persentase hambatan biofilm C.albicans berkisar 34,24%-69,70%, sedangkan kontrol positif sebesar 96,87%. Aktivitas hambatan biofilm larutan irigasi ekstrak daun Stevia konsentrasi 100%, 50%, 25%, 12,5%, 6,25%, 3,125% terhadap C.albicans lebih kecil signifikan dibandingkan NaOCL 2,5% namun lebih besar signifikan dibandingkan kontrol negatif dengan rata-rata persentase aktivitas hambatan biofilm sebesar 34,24%-69,70%.

Introduction: Increased blood glucose levels are caused by a lifestyle that involves excessive consumption of foods and drinks containing sweeteners. Stevia Rebaudiana Bertoni leaves are a natural ingredient that can be used as a natural sweetener because they contain stevioside and rebaudioside A, with a sweet taste effect that is 200 to 300 times higher than the sucrose found in granulated sugar. However, few studies still report comparative data on fasting blood glucose levels after consuming natural Stevia and granulated sugar as a sweetener in healthy individuals. Purpose: To compare the increase in fasting blood glucose levels after consuming Stevia and granulated sugar as sweeteners in healthy individuals. Methods: Experimental research with a pretest-posttest with control group design was carried out using purposive sampling. The research subjects were 40 people aged 17-23 years who were asked to fast for 8-10 hours. Fasting blood glucose levels were measured before consuming powdered and liquid Stevia, granulated sugar, and mineral water as a control, and further measurements were carried out 45 minutes after treatment. Results: The highest increase in fasting blood glucose levels occurred in subjects who consumed granulated sugar solution, namely 15.8 mg/dL, followed by subjects who consumed liquid Stevia solution at 6.6 mg/dL, and the lowest in subjects who consumed powdered Stevia solution. Powdered Stevia increased blood glucose by 5.6 mg/dL. Conclusion: There is a significant difference in blood glucose levels before and after the treatment of consuming powdered Stevia, liquid Stevia, and granulated sugar sweetener. The increase in fasting blood glucose levels after consuming powdered or liquid Stevia sweetener was significantly lower than that of the granulated sugar sweetener.

Stevia rebaudiana (Bertoni) leaves consist of dietetically important diterpene steviol glycosides (SGs): stevioside (ST) and rebaudioside-A (Reb-A). ST and Reb-A are key sweetening compounds exhibiting a sweetening potential of 100 to 300 times more intense than that of table sucrose. Ultrasound-assisted extraction (UAE) of SGs was optimized by effective process optimization techniques, such as response surface methodology (RSM) and artificial neural network (ANN) modeling coupled with genetic algorithm (GA) as a function of ethanol concentration (X1: 0–100%), sonication time (X2: 10–54 min), and leaf–solvent ratio (X3: 0.148–0.313 g·mL−1). The maximum target responses were obtained at optimum UAE conditions of 75% (X1), 43 min (X2), and 0.28 g·mL−1 (X3). ANN-GA as a potential alternative indicated superiority to RSM. UAE as a green technology proved superior to conventional maceration extraction (CME) with reduced resource consumption. Moreover, UAE resulted in a higher total extra...

Today, plant-based therapies have been attracted attention to overcome diabetes complications. This study was an attempt to evaluate whether antidiabetic and nephroprotective effects of Stevia Rebaudiana Bertoni (SRB) can be exerted via upregulation of GLUT-4, SNAP23, and Stx4 in skeletal muscles or modulation of AQP2 mRNA expression and antioxidant signaling pathway activity (Nrf2/Keap1) in kidneys. To achieve this aim, diabetes was induced via STZ-nicotinamide (STZ-NA). Diabetes increased the level of Blood Urea Nitrogen (BUN), serum creatinine, Fasting Blood Sugar (FBS), and Keap1 mRNA expression, which was coincide with reduction in mRNA levels of Nrf2, GLUT4, SNAP23, and Stx4. SRB and metformin compensate mentioned variables. However, SRB extract was more effective than metformin to increase the levels of GLUT4 and Nrf2 mRNA. It seems that SRB might attenuate the diabetic complications via manipulating the glucose uptake components in peripheral tissues and might exert the nephroprotective effects by modulation of AQP2, and Nrf2/Keap1 mRNA expression. Synthetic antidiabetic drugs have been only partially successful in controlling the diabetic complications. Moreover, use of these drugs is associated with a number of adverse effects. Over the past few years, a renewed attention has been paid to the prevention and treatment of diabetes using medicinal plants and functional foods. SRB that have been known as natural sweetener for centuries, is a such natural agent that has high source of various phytochemicals with antidiabetic, renal protective, antitumor, and antioxidant properties. In the current study, possible molecular mechanisms of insulin-mimetic and nephroprotective effects of SRB extract was evaluated in diabetic rats. Due to powerful antihyperglycemic and nephroprotective effects of SRB extract that were showed in this study and previous studies, hence the fact that SRB is to be highlighted for future research as a new therapeutic agent for diabetes.

Egyptian Journal of Agronomy Short Note 12 A N EFFICIENT and simple regeneration protocol using leaf explants was described on MS medium in the presence of various plant growth regulators. After four weeks of culture, the highest induction of adventitious shoot buds from leaf culture (90%) was achieved on MS medium with 1.0mg/L BA. The largest number of shoot buds (3.02) per leaf explant with no callus formation was achieved on MS medium with 2.0mg/L BA. The best combination for the induction of numerous shoot buds was found to be TDZ + BA + IBA (0.5, 1.0, and 0.5mg/L, respectively), which presented 10.4 shoots per explant with 3.15cm in length. For in vitro root formation stage, Healthy shoots were collected and cultivated on a half-strength MS with various concentrations of NAA and IAA alone or in combination with 2,4-D or IBA. Furthermore, the highest roots (8.17) per shoot with root formation (100 %) was observed using a half-strength MS medium in the presence of 2.0mg/L IAA. Therefore, healthy in vitro rooted plantlets were acclimatized successfully after four weeks in the greenhouse.

The crude methanolic extract and various fractions of leaves of Stevia rebaudianawere investigated for antibacterial, phytotoxic and haemagglutinationactivities. The biological investigation revealed that crude extract showed good activity against E. coli (69%), moderate activity against B. subtilus (51.8%) and S. aureus (50%). No considerable phytotoxic activity was shown by S. rebaudiana against Lemnaminor. S. rebaudiana showed nohaemagglutination activity against all blood groups indicating that the herb lacks phytolectins. The test samples showed significant antifungal activity, negligible phytotoxic activity and no haemagglutination activity.

Introduction: Stevia rebaudiana Bertoni, is a plant with hypoglycemic and anti-hyperlipidemic properties. Therefore, it has become a complement in the diabetes treatment and other metabolic disorders, such as hypertension, dyslipidemia and cardiovascular ills. However, It is unknown the effect on pancreas of people that consume frequently Stevia without history of metabolic disorders. Objective: To investigate the effect of chronic administration of Stevia on pancreas in Wistar rats with normocaloric feeding. Materials and methods: Wistar rats of one-month of age were administered with an intraperitoneal dose of Stevia extract (1 g/kg/day) for 60 days. Just prior to finish, the rats received an oral glucose load (1.75 g-glucose/kg). Under anesthesia, the whole blood (200 L) was drawn via cardiac puncture at 0, 30, 60 and 90 min. Then, glucose and insulin were measured. In serum (basal), triglycerides and HDL-Chol were measured. Subsequently, rats were perfused and a pancreatectomy was realized. The oxidative stress and histological changes were evaluated in pancreas. Results: After 2-months of Stevia administration, low levels of glucose, triglycerides and HDL-Chol were observed. Glucose tolerance was impaired, and rats showed hyperinsulinemia and insulin resistance. In regard to pancreatic redox state, GSH and redox index showed significant increases; however, SOD activity was depleted. Lipoperoxidation was reduced and nitrite level did not show changes. Surprisingly, pancreatic insulin production showed an important increase, and also hyperplasic changes were observed in Langerhans islets. Conclusions: Despite the antioxidant features, hypoglycemic and hypolipidemic effects, the chronic administration of Stevia generated toxicity, producing hyperinsulinemic states in rats, and promoting an insulin resistance. At tissue level, Langerhans islets hyperplasic were observed with over-production of insulin, which could bring a ␤-cell exhaustion. Financial support: Authors thank VIEP projects.

The increasing demand for natural and low-calorie sweeteners has intensified interest in Stevia rebaudiana-derived glycosides, particularly rebaudioside A and steviolbioside. However, conventional plant extraction is inefficient and unsustainable at industrial scales. This study explores a synthetic biology approach utilizing genetically engineered Saccharomyces cerevisiae for the heterologous production of steviol glycosides. We developed a yeast strain with optimized metabolic flux and co-factor regeneration for enhanced glycoside biosynthesis, using gene clusters from Stevia rebaudiana. Results show significantly higher titers of targeted glycosides under controlled fermentation conditions. This biotechnological platform demonstrates potential for scalable, cost-effective, and environmentally friendly production of Stevia-based sweeteners.

The UDP-glycosyltransferase 76G1 (UGT76G1) is responsible for the conversion of stevioside to rebaudioside A. Four single guide RNAs (sgRNAs) were designed from the UGT76G1 proximal promoter region of stevia by using the online-based tool, benchling. The dCas9 fused with VP64 as a transcriptional activation domain (TAD) was produced and purified for the formation of ribonucleoproteins (RNPs) by mixing with the in vitro transcribed sgRNAs. Protoplast yield was the highest from leaf mesophyll of in vitro grown stevia plantlets (3.16 × 106/g of FW) using ES5 (1.25% cellulase R-10 and 0.75% macerozyme R-10). The RNPs were delivered into the isolated protoplasts through the Polyethylene glycol (PEG)-mediated transfection method. The highest endogenous activation of the UGT76G1 gene was detected at 27.51-fold after 24 h of transfection with RNP30 consisting of CRISPR/dCas9-TAD with sgRNA30 and a similar activation level was obtained using RNP18, RNP33, and RNP34, produced using sgRNA18, s...

A plant tissue culture protocol from stevia was optimized for the production of planting materials and the natural sweetener, rebaudioside A. The highest survivability (88.90% ± 5.55) of explants was achieved at 15 and 30 days after culture initiation (DACI) on Murashige and Skoog (MS) media by sterilization with 30% Clorox (5 min) and 10% Clorox (10 min), respectively. Supplementation of MS with 0.50 mg/L 2,4-Dichlorophenoxyacetic acid (2,4-D) and 0.10 mg/L zeatin produced 50% callus at 15 DACI while 1.50 mg/L 2,4-D and 0.10 mg/L zeatin at 30 DACI increased callus production to 76.67%. The highest shoot proliferation per callus was achieved with 10.00 mg/L 6-benzyl amino purine (BAP) in MS at 15 DACI (5.80) and 30 DACI (12.33). The longest shoots of 4.31 cm and 6.04 cm at 15 and 30 DACI, respectively, were produced using BAP (10.00 mg/L) and 1.00 mg/L naphthalene acetic acid (NAA). MS media (0.50 strength) induced 2.86 and 6.20 roots per shoot and produced 3.25 cm and 7.82 cm long ...

Special diets are used to mitigate many human diseases. When these diets require changes in carbohydrate content, then sweetness becomes an important characteristic. The range of low-calorie sweeteners available to the food industry is expanding. It is essential to have an exact knowledge of the relative sweetness of various sweeteners in relation to different sucrose concentrations. The objective of this study was to determine the variation on the relative sweetness of aspartame (APM), stevia [Stevia rebaudiana (Bert.) Bertoni] leaf extract (SrB) and the mixture cyclamate/saccharin -two parts of cyclamate and one part of saccharin -(C/S) with the increase in their concentrations, and in neutral and acid pH in equisweet concentration to 10% sucrose, using magnitude estimation. Sweetness equivalence of SrB in relation to sucrose concentrations of 20% or higher and of APM and C/S to sucrose concentrations of 40% or higher could not be determined, because a bitter taste predominated. The potency of all sweeteners decreased as the level of sweetner increased. In equi-sweet concentration of sucrose at 10%, with pH 7.0 and pH 3.0, the potency was practically the same for all sweeteners evaluated.

Stevia (Stevia rebaudiana Bertoni) belongs to the family Asteracae and the intensive sweet taste of its leaves provided the name „new age sugar“. Stevia is cultivated for the sweet taste leaves, which are, after drying and grinding, used as a substitute for artificial sweeteners. The Stevia products are widespread as food additives and a natural calorie free sweetener. An intensive sweetness originates from glycosides, i.e. termostable chemical compounds well tolerated the low pH values. The subject of the whole interdisciplinary study was to determine the effect of fertilizer doses and the soil type (pH) on the fresh leaves yield. Furthermore, content of glycosides (stevioside and steviol) and mineral composition of Stevia leaves were determined. Plants were grown in a 5 L pots on the two types of acid soil (pH 4.0 and 5.0), which were different according to their supply with major nutrients. Five doses (from 0 up to 1.6 g per plant) of Multi-Comp Base mineral fertilizer 14:13:20+2...

Pressurised fluid extraction using water or methanol was employed for the extraction of stevioside from Stevia rebaudiana Bertoni. The extraction method was optimised in terms of temperature and duration of the static or the dynamic step. Extracts were analysed by liquid chromatography followed by UV and mass-spectrometric (MS) detections. Thermal degradation of stevioside was the same in both solvents within the range 70-160 °C. Methanol showed better extraction ability for isolation of stevioside from Stevia rebaudiana leaves than water within the range 110-160 °C. However, water represents the green alternative to methanol. The limit of detection of stevioside in the extract analysed was 30 ng for UV detection and 2 ng for MS detection.