Influence of Environmental Parameters, Pinching, and Ethephon Application on Growth and Branching of Potted Stevia

Stevia (Stevia rebaudiana) is an herb grown commercially for the extraction of intensely sweet-tasting, non-caloric, steviol glycosides produced primarily in the leaves and used as a sugar substitute. While most stevia production occurs as an industrial field crop, more recently, consumer demand for stevia for home gardens and patio containers has increased. Research on how environmental inputs impact growth, branching, and flowering of stevia under greenhouse conditions for potted plant production is currently lacking. A series of experiments was conducted to quantify how methods to promote branching, fertilizer concentration, photoperiod and temperature impact branch production, growth and development, and flowering of stevia. Both manual decapitation and ethephon application increased lateral branch production, though hard pinching (cutting plants back to leave four nodes) yielded a more desirable plant architecture. Neither temperature nor fertilizer concentration impacted the number of branches produced by plants given a hard pinch. Shoot dry biomass was similar at fertilizer concentrations (applied at each watering) of 50, 100, and 200 mg⋅L−1 N, but decreased at 300 or 400 mg⋅L−1 N. Stevia responded to photoperiod as a facultative short-day plant, with earliest flowering occurring, both in days to flower and the number of nodes produced before flowering, at photoperiods <13 hours. The number of nodes produced on the longest branch increased as temperature increased from 17 to 26 °C. Plant height and longest branch length were shorter at 17 °C than at higher temperatures. The results of these studies indicate that for potted plant production, stevia should be grown under a photoperiod of 14 hours or longer with moderate nutrient levels, a minimum temperature of 20 °C, and plants should receive one or more manual pinches to promote branching.

Macroelements and Microelements in the Soil and Their Relationship with the Content of Steviol Glucosides in Stevia rebaudiana Bert from Five Regions of Colombia

This study was conducted to determine the effect of edaphic environmental conditions in the concentration of principal steviol glycosides and Stevia rebaudiana Bert yield, utilizing leaves from five Colombian regions. The structure of the experiment was a randomized complete block design with two treatments in a 5 × 2 factorial arrangement (5 locations × 2 radiation levels). In each experimental unit (UE), five healthy plants of similar physiological growth age were selected for the extraction of total glycosides (GT), stevioside (Stv), rebaudioside A (Rb-A),and leaf yield. Results were analyzed with the SAS statistical package (version 9.1). Concentrations of total glycosides and rebaudioside A showed a positive effect with the increase of nitrogen (N), phosphorus (P), magnesium (Mg), and copper (Cu). Therefore, they are important in the available phase of the soil to obtain an increase in these glycosides. Meanwhile, boron (B) presented a negative correlation under these conditions. For the production of stevioside, N, Mg, manganese (Mn) had a positive correlation, and calcium (Ca) and sodium (Na) had a negative correlation. Similarly, for leaf yields by locality, it was found that N, Ca, Mg, and B have a positive correlation with leaf production, while Mg, Mn, and iron (Fe) negatively correlate with biomass gain.The cultivation of stevia can be established in different soil conditions, precipitation and solar radiation in Colombia. Therefore, it is necessary to advance fertilization plans with these nutrients, considering the response of these metabolites to their application.

Stevia rebaudiana Bertoni: The Interaction of Night Interruption on Gas Exchange, Flowering Delay, and Steviol Glycosides Synthesis

The Stevia market is estimated to be USD 1.14 billion in 2028 due to its acceptance in the food and beverage industry. Stevia rebaudiana and its two more relevant edulcorants: stevioside (St) and rebaudioside A (Reb-A) can reach 450-fold sweeter than sucrose. The species is considered a long night plant, promoting flowering and shortening vegetative growth. Thus, to increase the leaf area and St and Reb-A increase, we broke the long night with a short light pulse, here called night interruption (NI). In this study, three NI times and two S. rebaudiana genotypes were tested to promote larger vegetative growth, flowering delay, and higher synthesis of steviol glycosides (SvGly). The main goal of this study was to demonstrate that NI increased net photosynthesis (9% to 20%), the internode length (59%), the leaf area (25%), while delays in 4 to 10 days of the flowering phase, impacting in 17% to 25% more St and Reb-A, respectively. Here we describe an inexpensive flowering delay, elongation of vegetative growth, allowing extended harvesting, which could yield four to five annual harvesting of leaves, increasing the production in 21% to 24% more St and Reb-A yield (kg ha−1).

Catalytic flexibility of rice glycosyltransferase OsUGT91C1 for the production of palatable steviol glycosides

Steviol glycosides are the intensely sweet components of extracts from Stevia rebaudiana. These molecules comprise an invariant steviol aglycone decorated with variable glycans and could widely serve as a low-calorie sweetener. However, the most desirable steviol glycosides Reb D and Reb M, devoid of unpleasant aftertaste, are naturally produced only in trace amounts due to low levels of specific β (1–2) glucosylation in Stevia. Here, we report the biochemical and structural characterization of OsUGT91C1, a glycosyltransferase from Oryza sativa, which is efficient at catalyzing β (1–2) glucosylation. The enzyme’s ability to bind steviol glycoside substrate in three modes underlies its flexibility to catalyze β (1–2) glucosylation in two distinct orientations as well as β (1–6) glucosylation. Guided by the structural insights, we engineer this enzyme to enhance the desirable β (1–2) glucosylation, eliminate β (1–6) glucosylation, and obtain a promising catalyst for the industrial production of naturally rare but palatable steviol glycosides.

The Sharp Rise in the Use of Low- and No-Calorie Sweeteners in Non-Alcoholic Beverages in Slovenia: An Update Based on 2020 Data

Reducing added sugars in non-alcoholic beverages is an important public health goal, which can result in increased use of low- and no-calorie sweeteners (LNCS). The aim of this study was to investigate recent changes in the use of LNCS in non-alcoholic beverages in the Slovenian food supply. The national branded foods dataset was updated with beverages available in 2020, and compared with previous datasets. The data were extracted from food labels. In 2020, N = 1,650 unique beverages were found in shops from five different retailers, covering the majority of the national market. The use of LNCS increased from 13.2% in 2017 and 15.5% in 2019 to 20.2% in 2020, with a major growth in soft drinks (16.8, 19.6, and 26.7%, respectively). We observed a significant growth of beverages containing both LNCS and added sugar. Results were also consistent with sales data, which showed that increased offer of beverages with LNCS also resulted in similarly increased sales of such beverages. The average energy and total sugar content in non-alcoholic beverages decreased, which reflects both the higher percentage of beverages with LNCS, and also the reduction of the sugar content in beverages with only added sugar. Analyses of product-specific reformulation practices highlighted reduced sugar content in 16.8% of products, and in 3.6% with the use of LNCS. The most commonly used LNCS are acesulfame K, sucralose, and aspartame. Typically, combinations are used, however steviol glycosides, sucralose and saccharin are also used alone, in most cases combined with added sugar. The results indicated rapid changes in the use of LNCS in non-alcoholic beverages in the Slovenian food supply, making further monitoring of this area highly relevant.

GlucoMedix ®, an Extract of Stevia Rebaudiana and Uncaria Tomentosa, Reduces Hyperglycemia, Hyperlipidemia, and Hypertension in Rat Models Without Toxicity: A Treatment For Metabolic Syndrome

Background: The objective of this in vivo study is to evaluate in five rat models the pharmacologic effects and toxicity of a commercial hydro-alcoholic extract, GlucoMedix®, derived from Stevia rebaudiana and the pentacyclic chemotype of Uncaria Tomentosa (Willd.) DC, for use as a treatment for Metabolic Syndrome. The extract contains phytochemicals of Stevia (e.g., steviol glycosides) and Uncaria (e.g., pentacyclic oxindole alkaloids, but lacks tetracyclic oxindole alkaloids).Methods: The pharmacologic assessments in three rat models include reductions in chemically induced hyperglycemia, hyperlipidemia (cholesterol and triglycerides), and hypertension, all of which are comorbidities of Metabolic Syndrome. Acute toxicity and 28-day subacute toxicity were assessed in rat models at doses higher than those used in the efficacy models.Results: The acute oral toxicity was evaluated in Holtzman rats and the extract did not produce acute toxic effects or lethality, with the LD50 > 5000 mg/kg (extract wet weight). Furthermore, subacute oral toxicity was evaluated in rats for 28 days at daily doses as high as 2000 mg/kg without toxicity or abnormal clinical chemistry or hematological effects. Daily oral doses of 250 - 1000 mg/kg were used to evaluate the treatment effects in hyperglycemic (alloxan-induced and glibenclamide-controlled), hyperlipidemic (cholesterol-induced and atorvastatin-controlled), and hypertensive (L-NAME-induced and enalapril-controlled) rat models. Alloxan-induced hyperglycemia was reduced in a dose-dependent manner within 28 days or less. Cholesterol-induced hyperlipidemic rats exhibited dose-dependent reductions in cholesterol and triglycerides at 21 days. Furthermore, GlucoMedix® produced a dose-dependent decrease in systolic and diastolic arterial blood pressure in L-NAME-induced hypertensive rats at 28 days.Conclusions: The five in vivo rat models revealed that the all-natural phytotherapy GlucoMedix® is a safe and effective treatment for hyperglycemia, hyperlipidemia, and hypertension. This extract is expected to affect multiple comorbidities of Metabolic Syndrome, without any acute or subacute oral toxicity in humans. Although multiple prescription drugs are well known for the treatment of individual comorbidities of Metabolic Syndrome, no drug monotherapy concurrently treats all three comorbidities.

Characterization of Microbial Degradation Products of Steviol Glycosides

Steviol glycosides were subjected to bacteria present in a soil sample collected from a Stevia plantation in Paraguay. During the incubation experiments, next to the aglycon steviol, steviol degradation products were also formed. X-ray analysis and NMR methods in combination with chemical synthesis and GIAO NMR calculations were used to fully characterize the structure of these compounds as a tricyclic ketone and the corresponding reduced form. They were nicknamed monicanone and monicanol. The latter has the (S)-configuration at the alcohol site.