Fig. 1
Middle vein (MV) in cross-sections of leaves fully expanded of “arnicas”. A–J. Light microscopy. A. Calea uniflora; B. Chaptalia nutans; C. Lychnophora diamantinana; D. Lychnophora ericoides; E. Lychnophora pinaster; F. Lychnophora salicifolia; G. Porophyllum ruderale; H. Pseudobrickellia brasiliensis; I. Sphagneticola trilobata; J. Solidago chilensis. A. Staining with ruthenium red; B–D, F–J. Staining with toluidine blue; E. Staining with Sudan. Abbreviations: COL, collenchyma; XI, xylem; FLO, phloem; PAR, regular parenchyma; ESC, sclerehydes; CV, internal secretory cavity. A, J, I. presence of CV; E. presence of ESC. C–F. possible sclerenchyma fibers below the phloem. Findings: A, B, J. MV with biconvex form; C, F. adaxial face with central depression and abaxial face with rounded projection to the MV form; H. undefined MV; G. convex MV form; I. concave-convex MV form; A–J. uniseriate epidermal cells, covered by cuticle; A. open arch vascular bundles form; B. cord format vascular bundles form; C–F. three free bundles with cilindric form; G. small and straight vascular pattern with extension of the bundle sheath; H. undefined vascular bundles form; I. cord format vascular bundles form; J. conical and small dimensions vascular bundles form.
Fig. 2
View of stomata and epidermal cells in abaxial surface of leaves fully expanded of “arnicas”. A–F. Light microscopy; A. Calea uniflora; B. Chaptalia nutans; C. Porophyllum ruderale; D. Pseudobrickellia brasiliensis; E. Sphagneticola trilobata; F. Solidago chilensis; A–B. Staining with toluidine blue. C–F. Staining with safranine. Findings: A–F. anomocytic characteristics by stomata.
Fig. 3
View of stomata in abaxial surface of leaves fully expanded of “arnicas” by SEM. A. Calea uniflora; B. Chaptalia nutans; C. Lychnophora ericoides; D. Lychnophora pinaster; E. Lychnophora salicifolia; F. Porophyllum ruderale; G. Pseudobrickellia brasiliensis; H. Sphagneticola trilobata; I. Solidago chilensis. Findings: A, B, H. regular level of stomata in relation to the epidermis. C, F–G. below of the epidermis level. D, E, I. above the epidermis level.
Fig. 4
Epidermis cells (adaxial vs. abaxial surface) of leaves fully expanded of “arnicas” by SEM. Adaxial face: A. Calea uniflora; B. Chaptalia nutans; C. Lychnophora diamantinana; D. Lychnophora ericoides; E. Lychnophora pinaster; F. Lychnophora salicifolia; G. Porophyllum ruderale; H. Pseudobrickellia brasiliensis; I. Sphagneticola trilobata; J. Solidago chilensis. Abaxial face: A′. Calea uniflora; B′. Chaptalia nutans; C′. Lychnophora diamantinana; D′. Lychnophora ericoides; E′. Lychnophora pinaster; F′. Lychnophora salicifolia; G′. Porophyllum ruderale; H′. Pseudobrickellia brasiliensis; I′. Sphagneticola trilobata; J′. Solidago chilensis. Findings: A–J and A′–J′. uniform epidermis of the leaf surface (both faces); A, B, I and A′, B′, I′. sinuous walls of the epidermal cells; G, H, J and G′, H′, J′. straight wall of the epidermal cells; C–F and C′–F′. straight wall of the epidermal cells in the adaxial face (C–F) being more rounded and the abaxial cells (C′–F′) more elongated.
Fig. 5
Glandular trichome (GT) in cross-sections of leaves fully expanded of “arnicas”. Light microscopy: A. Calea uniflora; B. Lychnophora diamantinana; C. Lychnophora ericoides; D. Lychnophora pinaster; E. Lychnophora salicifolia; F, F′, F″. Solidago chilensis; G, G′. Sphagneticola trilobata; A, F′, F″, G and G′. staining with safranine; F. staining with toluidine blue; B, C, D, E. uncolored. Findings: A. GT inserted in small depression and multicellular and bicellular or capited with unicellular pedicel and globoid head. B–D. GT inclosed in crypts; F. GT multicellular plurisseriate and/or capitate with unicellular pedicel with elongated terminal cell; G. GT bisseried in depression and/or multicellular unisseriate with elongated terminal cell.
Fig. 6
Glandular trichome (GT) of leaves fully expanded of “arnicas” by SEM. A. Calea uniflora; B. Lychnophora diamantinana; C. Lychnophora ericoides; D. Lychnophora pinaster; E. Lychnophora salicifolia; F. Sphagneticola trilobata; G. Solidago chilensis. Abbreviations: EP, epidermis; ST, stoma; GT, glandular trichome; NGT, non glandular trichome. Findings: A. GT inserted in small depression and multicellular and bicellular or capited with unicellular pedicel and globoid head; B–D. GT inclosed in crypts; F. GT multicellular plurisseriate and/or capitate with unicellular pedicel with elongated terminal cell; G. GT bisseried in depression and/or multicellular unisseriate with elongated terminal cell.
Fig. 7
Non glandular trichome (NGT) in cross section of leaves fully expanded of “arnicas”. A. Calea uniflora; B, B′. Chaptalia nutans; C. Lychnophora diamantinana; D. Lychnophora ericoides; E. Lychnophora pinaster; F. Lychnophora salicifolia; G, G′. Solidago chilensis; H. Sphagneticola trilobata; Findings: A, G, G′, H. staining with safranine; B, C, D, E, F. uncolored; A, G, G′, H. multicellular (5–8 cells) and unisseriate NGT; B. long and numerous NGT; C, F. starred trichomes (3–5 arms); G, G′. elongated TNG, with slightly enlarged base and slim terminal cell; H. NGT with the basal cell of verrucous cuticle.
Fig. 8
Non-glandular trichome (NGT) of leaves fully expanded of “arnicas” by SEM. A, A′. Calea uniflora; B, B′. Chaptalia nutans; C, C′. Lychnophora diamantinana; D. Lychnophora ericoides; E. Lychnophora pinaster; F. Lychnophora salicifolia; G. Solidago chilensis; H, H′, H″. Sphagneticola trilobata; Abbreviations: EP, epidermis; ST, stoma; GT, glandular trichome; NGT, non glandular trichome. Findings: A, G–H. multicellular (5–8 cells) and unisseriate NGT; B. long and numerous NGT; C–F. starred trichomes (3–5 arms); G. elongated TNG, with slightly enlarged base and slim terminal cell; H. NGT with the basal cell of verrucous cuticle.
Fig. 9
Intermediate region of mesophyll (IRM) in cross section of leaves fully expanded of “arnicas”. A. Calea uniflora; B. Chaptalia nutans; C. Lychnophora diamantinana. D. Lychnophora ericoides; E. Lychnophora pinaster; F. Lychnophora salicifolia; G. Porophyllum ruderale; H. Pseudobrickellia brasiliensis; I. Solidago chilensis; J. Sphagneticola trilobata. Abbreviations: CUT, cuticule; PP, palisade parenchyma; LP, lacunar parenchyma; AP, aquiferous parenchyma; PR, regular parenchyma; VB, vascular bundle blond; ESC, sclerehyde; EP, epidermis; GT, glandular trichome; NGT, non-glandular trichome; IC, internal cavity. Findings: A–F. staining with ruthenium red; G, J. staining with toluidine blue; A, B, G, J. dorsiventral parenchyma; C–F. palisade (adaxial), spongy (abaxial) and aquiferous parenchyma strata close to the vascular bundles with extension of the bundle sheath; H. homogenous mesophyll; I. isobilateral mesophyll and aquiferous parenchyma (interconnecting the vascular bundles); A–J. collateral type of the bundles.
Fig. 10
Leaf margin in cross section of leaves fully expanded of “arnicas”. A. Calea uniflora; B. Chaptalia nutans; C, C′. Lychnophora diamantinana; D. Lychnophora ericoides; E. Lychnophora pinaster; F. Lychnophora salicifolia; G, G′. Porophyllum ruderale; H. Pseudobrickellia brasiliensis; I. Solidago chilensis; J. Sphagneticola trilobata; A, B, F. staining with ferric chloride; C–E and G–I. staining with ruthenium red; J. toluidine blue. Findings: A–F, J. rounded and flexed sheet edge for the abaxial face; C–F. flexion with a final margin narrowing. I. discrete flexion of the margin with epidermal projections to the outside of the leaf; G–H. straight margin, without flexion or little flexure. G, G′. large secretory cavity on the margin of leaf.
Fig. 11
Analyses of leaves fully expanded of Arnica montana by SEM. A. adaxial face; B. abaxial surface; C. stomata; D. base of the NGT; E. peduncle of NGT; F. apex of NGT. G. GT.
Fig. 12
Flowchart to quality control of the “arnicas” evaluated using the leaves fully expanded, through the middle vein. Orange, anatomical parameters; Blue, anatomical definition and target; Green, specie confirmation.
Fig. 13
Flowchart to quality control of the “arnicas” evaluated using the leaves fully expanded, through the stomata. Orange, anatomical parameters; Blue, anatomical definition and target; Green, specie confirmation.
Fig. 14
Flowchart to quality control of the “arnicas” evaluated using the leaves fully expanded, through the trichomes. Orange, anatomical parameters; Blue, anatomical definition and target; Green, specie confirmation.
Fig. 15
Histochemical reaction for lipophilic substances in cross-sections of leaves of “arnicas”, highlighted by Sudan. Light microscopy. The figure shows positive reactions (red) for lipophilic substances with accumulation of metabolites in glandular trichomes. A, A′. Calea uniflora; B, B′. Lychnophora diamantinana; C. Lychnophora ericoides; D. Lychnophora pinaster; E. Lychnophora salicifolia; F. Solidago chilensis; G. Sphagneticola trilobata.
Fig. 16
Histochemical reaction for lipophilic substances in cross-sections of leaves of “arnicas”, highlighted by Sudan. Light microscopy. The figure shows positive reactions for lipophilic substances with accumulation and secretion of metabolites in the epidermal cells and other secretory structures of leaves. A, A′. Calea uniflora; B. Lychnophora diamantinana; C. Lychnophora ericoides; D. Lychnophora pinaster; E. Lychnophora salicifolia; F. Solidago chilensis; G. Sphagneticola trilobata. Findings: A, B″, C′, D′, I′, F′. midvein with accumulation of metabolites in parenchyma cells. A′, B, B′, C, D, D′, E, F′, G, H, I, J. accumulation of metabolites in the epidermis and/or chlorophyll parenchyma of the mesophyll; A, A′, G, I, I′. accumulation of metabolites in the internal secretory structure. Reaction pattern at the tip of the arrow (red).
Fig. 17
Histochemical reaction for screening of sesquiterpene lactones in cross-sections of leaves of “Arnicas”, evidenced by sulphuric acid. Light microscopy. The figure shows positive reactions (brown/orange) for screening of sesquiterpene lactones with accumulation of metabolites in glandular trichomes. A. Calea uniflora; B. Lychnophora diamantinana; C. Lychnophora ericoides; D. Lychnophora pinaster; E. Lychnophora salicifolia; F. Solidago chilensis; G, G′. Sphagneticola trilobata.
Fig. 18
Histochemical reaction for screening of sesquiterpene lactones in cross-sections of leaves of “arnicas”, evidenced by sulphuric acid. Light microscopy. The figure shows positive reactions for screening of sesquiterpene lactones with accumulation and secretion of metabolites in the epidermal cells and other secretory structures of leaves. A. Calea uniflora; B. Lychnophora diamantinana; C. Lychnophora ericoides; D. Lychnophora pinaster; E. Lychnophora salicifolia; F. Solidago chilensis; G. Sphagneticola trilobata. Findings: B, D, E, F′, H. midvein with accumulation of metabolites in parenchyma cells; A, C, F, G, H, I. accumulation of metabolites in the epidermis and/or chlorophyll parenchyma of the mesophyll; F, G, H. accumulation of metabolites in the internal secretory structure. Reaction pattern at the tip of the arrow (brown/orange).
Fig. 19
Histochemical reaction for phenolic substances in cross-sections of leaves of “arnicas”, highlighted by ferric chloride. Light microscopy. The figure shows positive reactions (brown) for phenolic substances with accumulation of metabolites in glandular trichomes. A. Calea uniflora; B. Lychnophora diamantinana; C. Lychnophora ericoides; D. Lychnophora pinaster; E. Lychnophora salicifolia; F. Solidago chilensis; G. Sphagneticola trilobata.
Fig. 20
Histochemical reaction for phenolic substances in cross-sections of leaves of “arnicas”, highlighted by ferric chloride. Light microscopy. The figure shows positive reactions for phenolic substances with accumulation and secretion of metabolites in the epidermal cells and other secretory structures of leaves. A. Calea uniflora; B. Lychnophora diamantinana; C. Lychnophora ericoides; D. Lychnophora pinaster; E. Lychnophora salicifolia; F. Solidago chilensis; G. Sphagneticola trilobata. Findings: A′, B′, C, D, E, F, F′, I, J′. midvein with accumulation of metabolites in parenchyma cells; A, B, C, D′, G, G′, H, I, J. accumulation of metabolites in the epidermis and/or chlorophyll parenchyma of the mesophyll; A′, G, I′, J. accumulation of metabolites in the internal secretory structure. Reaction pattern at the tip of the arrow (brown).
Fig. 21
Main classes of metabolites reported in each of the “arnicas” used in Brazil.