Stevia is a plant containing many active compounds, but usually propagated by stem cuttings because of low seed-yield-germination ability. The aim of this study was to investigate the impact of plant-growth regulators on stevia callus induction and growth from somatic tissue, as well as to determine the effect α-naphthalene acetic acid (NAA) and proline (PRO) on the amount of stevioside, rebaudioside A, phenols, flavonoids, and antioxidant activity. Stem and leaf segments were inoculated on a Murashige and Skoog (MS) medium supplemented with different concentrations of NAA and 6-benzylaminopurine (BAP) for callus genesis. The amount of steviol glycosides (SGs) was evaluated using high-performance liquid chromatography (HPLC), and the amounts of total phenols, flavonoids, and antioxidant activity by spectrophotometric methods. The highest callus-induction frequency and callus-mass increase were obtained from the leaf explants in MS medium supplemented with 2.0 μM NAA. The highest amount of SGs, phenols, and flavonoids, and stronger antioxidant activity were determined in the cellular compounds of callus from leaf explant. PRO reduced the amount of SGs and flavonoids. The significantly highest amount of total phenolic compounds was obtained in the callus from leaf explants in the medium supplemented with 2.0 µM NAA and 2.0 µM PRO.
Allergic rhinitis (AR) is a common and complex disease. It is associated with environmental as well as genetic factors. Three recent genome-wide association studies (GWAS) reported altogether 47 single nucleotide polymorphisms (SNPs) associated with AR or allergic sensitization (AS) in Europeans and North Americans. Two follow up studies in Swedish and Chinese replicated 15 associations. In these studies individuals were selected based on the self-reported AR, or AR/AS diagnosed using blood IgE test or skin prick test (SPT), which were performed often without restriction to specific allergens. Here we performed third replication study in Lithuanians. We used SPT and carefully selected set of allergens prevalent in Lithuania, as well as Illumina Core Exome chip for SNP detection. We genotyped 270 SPT-positive individuals (137 Betulaceae -, 174 Poaceae-, 199 Artemisia-, 70 Helianthus-, 22 Alternaria-, 22 Cladosporium-, 140 mites-, 95 cat- and 97 dog dander-sensitive cases) and 162 SPT-negative controls. We found altogether 13 known SNPs associated with AS (p ≤0.05). Three SNPs were found in Lithuanians sensitive to several allergens, and 10 SNPs were found in Lithuanians sensitive to a certain allergen. For the first time, SNP rs7775228:C was associated with patient sensitivity to dog allergens (F_A=0,269, F_U=0.180, P=0.008). Thus, careful assessment of AS allowed us to detect known genetic variants associated with AS/AR in relatively small cohort of Lithuanians.
The aim of this research was to investigate the effect of explant type and cytokinin type and concentration on stevia direct organogenesis in vitro, and to determine the effect of cytokinin type on stevioside and rebaudioside A content in regenerated shoots. Stem and leaf segments of stevia (Stevia rebaudiana Bertoni), taken from 6-week-old in vitro grown plants, were inoculated on Murashige and Skoog (MS) medium supplemented with different (1.0-6.0 µM) concentrations of 6-benzylaminopurine (BAP), thidiazuron (TDZ) and kinetin (KIN) for shoot regeneration. The content of steviol glycosides (stevioside and rebaudioside A) in S. rebaudiana donor plants and regenerated shoots was evaluated using high-performance liquid chromatography (HPLC). Results revealed a significant effect (p < 0.01) of explant type, cytokinin type and cytokinin concentration on shoot regeneration frequency. The mean value of regeneration frequency from stem segments was significantly higher in comparison with leaf segments. Thidiazuron significantly promoted the mean regeneration frequency in comparison to 6-benzylaminopurine and kinetin. The highest mean shoot formation frequency was obtained on the medium supplemented with 3.0 µM of TZD. Incorporation of 4.0 µM BAP and 4.0 µM TZD into culture media resulted in significant increase in rebaudioside A and stevioside ratio, while a decrease in rebaudioside A and stevioside ratio was recorded when the medium was supplemented with 4.0 µM KIN. Rebaudioside A and stevioside ratio in the medium supplemented with 4.0 µM TZD was significantly higher in comparison with 4.0 µM BAP and 4.0 µM KIN.
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