Exposure of stevia (Stevia rebaudiana B.) to silver nanoparticles in vitro: transport and accumulation.

The impact of nanotechnology in the field of agricultural sciences creates the need to study in greater detail the effect of products offering nanoparticles for application in plant species of agricultural interest. The objective of this study was to determine the response of stevia (Stevia rebaudiana B.) in vitro to different concentrations of AgNPs (silver nanoparticles), as well as to characterize and identify their absorption, translocation and accumulation mechanisms. Nodal segments of stevia grown in MS medium supplemented with AgNPs (0,12.5, 25, 50,100 and 200 mg L-1) were used. After 30 days of in vitro shoot proliferation, the number of shoots per explant, shoot length, chlorophyll content, dry matter content and the metallic silver (Ag) content of the plants were quantified. In addition, characterization, transport and accumulation of silver nanoparticles were performed by microscopic analysis. AgNPs were shown to be present in epidermal stem cells, within vascular bundles and in intermembrane spaces. In leaves, they were observed in ribs and stomata. The current and future use of AgNPs in agricultural sciences opens up the possibility of studying their effects on different plant species.

Post-seminal development and morphoanatomy of vegetative and reproductive organs in Stevia rebaudiana (Bert.) Bertoni (Asteraceae).

Stevia rebaudiana belongs to the Asteraceae family with high economic and medicinal potential. This article describes and illustrates morphological and histological aspects of leaves and reproductive organs, and the germination process, to provide detailed information on this species and to contribute to taxonomic, phylogenetic and pharmacobotanical projects. The fruit is a cypsela, small, simple, dry, indehiscent, monospermic, light or dark colored, with aristate pappus, and the seed presents a spatulate axile embryo. Germination is phaneroepigeal with a pivotal root system and many absorbing root hairs. The leaves are simple, elliptical to obovate, with two types of trichomes (glandular and tector), with a short petiole, exhibiting an opposite decussate phyllotaxy. Our results showed 37.5% germination after 12 days, only in the dark cypsela, the light colored being considered unviable. The inflorescence is paniculate and the florets are grouped in capitula with isomorphic ends, monoclinous (bisexual), dichlamydeous, heterochlamydeous, pentamerous calyx and corolla, gamossepalous and gamopetalous. The androecium is gamostemone comprised of five stamens with free filaments, isodynamous and epipetalous stamens, synandrous and rimose anthers. The flower presents an inferior ovary, bicarpelar, unilocular and ovules with a basal placentation. The pollen grains are small, isopolar, radial symmetry, tricolporate, with echinate ornamentation.

Comparative Transcriptomics Unravel Biochemical Specialization of Leaf Tissues of Stevia for Diterpenoid Production.

Stevia (Stevia rebaudiana) produces not only a group of diterpenoid glycosides known as steviol glycosides (SGs), but also other labdane-type diterpenoids that may be spatially separated from SGs. However, their biosynthetic routes and spatial distribution in leaf tissues have not yet been elucidated. Here, we integrate metabolome and transcriptome analyses of Stevia to explore the biosynthetic capacity of leaf tissues for diterpenoid metabolism. Tissue-specific chemical analyses confirmed that SGs were accumulated in leaf cells but not in trichomes. On the other hand, Stevia leaf trichomes stored other labdane-type diterpenoids such as oxomanoyl oxide and agatholic acid. RNA sequencing analyses from two different tissues of Stevia provided a comprehensive overview of dynamic metabolic activities in trichomes and leaf without trichomes. These metabolite-guided transcriptomics and phylogenetic and gene expression analyses clearly identified specific gene members encoding enzymes involved in the 2-C-methyl-d-erythritol 4-phosphate pathway and the biosynthesis of steviol or other labdane-type diterpenoids. Additionally, our RNA sequencing analysis uncovered copalyl diphosphate synthase (SrCPS) and kaurene synthase1 (SrKS1) homologs, SrCPS2 and KS-like (SrKSL), which were specifically expressed in trichomes. In vitro and in planta assays showed that unlike SrCPS and SrKS1, SrCPS2 synthesized labda-13-en-8-ol diphosphate and successively catalyzed the formation of manoyl oxide and epi-manoyl oxide in combination with SrKSL. Our findings suggest that Stevia may have evolved to use distinct metabolic pathways to avoid metabolic interferences in leaf tissues for efficient production of diverse secondary metabolites.