Leaf Venation Pattern to Recognize Austral South American Medicinal

G Model BJP 330 1–4 ARTICLE IN PRESS Revista Brasileira de Farmacognosia xxx (2016) xxx–xxx ww w.elsevier.com/locate/bjp Original Article 1 Leaf venation pattern to recognize austral South American medicinal 2 species of “cow’s hoof” (Bauhinia L., Fabaceae) a b c d,∗ 3 Q1 Renée H. Fortunato , Beatriz G. Varela , María A. Castro , María J. Nores a 4 Instituto de Recursos Biológicos, Instituto Nacional de Tecnología Agropecuaria – Consejo Nacional de Investigaciones Científicas y Técnicas, Facultad de Agronomía y Ciencias 5 Q2 Agroalimentarias, Universidad de Morón, Buenos Aires, Argentina b 6 Facultad de Farmacia y Bioquímica, Universidad Nacional de Buenos Aires, Buenos Aires, Argentina c 7 Facultad de Ciencias Exactas y Naturales, Universidad Nacional de Buenos Aires, Buenos Aires, Argentina d 8 Instituto Multidisciplinario de Biología Vegetal, Universidad Nacional de Córdoba – Consejo Nacional de Investigaciones Científicas y Técnicas, Facultad de Ciencias Exactas, Físicas 9 y Naturales, Universidad Nacional de Córdoba, Córdoba, Argentina 10 a b s t r a c t 11 a r t i c l e i n f o 12 13 Article history: The leaves extracts of some species of Bauhinia L. s.l. are consumed to treat diabetes, inflammation, 14 Received 6 September 2016 pains and several disorders in traditional medicine in austral South America. Despite its wide use and 15 Accepted 28 October 2016 commercialization, sale is not controlled, and botanical quality of samples is not always adequate because 16 Available online xxx of plant misidentification and adulteration. Here, we characterized leaf vein pattern in nineteen taxa to 17 contribute to the recognition and commercial quality control of plant material commercially available. 18 Keywords: The vein characters intercostal tertiary and quinternary vein fabric, areole development and shape, free 19 Areolation ending veinlet branching and marginal ultimate venation allowed to distinguish the main medicinal 20 Botanical identification species in the region. 21 Freely ending veinlets 22 Leaf venation pattern © 2016 Sociedade Brasileira de Farmacognosia. Published by Elsevier Editora Ltda. This is an open 23 Quality control access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). 24 Schnella 25 Introduction Gadotti et al., 2005; Luna et al., 2005; Nakahata et al., 2006; Cechinel 44 Filho, 2009; Paula et al., 2014; Rozza et al., 2015; Martins-Olivera 45 26 The leaves extracts of some species of Bauhinia L., Fabaceae, so et al., 2016). 46 27 called cow’s hoof, “pata-de–vaca”, cow’s paw or orchid trees, are These taxa are extensively employed and sold in native Amer- 47 28 consumed to treat diabetes, inflammation, pains and several dis- ican, rural and urban communities in the region and surrounding 48 29 orders in traditional medicine in austral South America (Oliveira areas (Bortoluzzi et al., 2006; Barboza et al., 2009; Hurrell et al., 49 30 and Saito, 1989; Bortoluzzi et al., 2006; Barboza et al., 2009; 2011; Ibarrola and Degen de Arrúa, 2011). Despite its wide use and 50 31 Pizziolo et al., 2011; Fortunato, 2012). With 300 species worldwide, commercialization, sale is not controlled, and botanical quality of 51 32 Bauhinia s.l. is represented by fifteen species in Argentina, Paraguay, samples is not always adequate because of plant misidentification 52 33 Uruguay and Santa Catarina, Paraná and Rio Grande do Sul states and adulteration. Commercial samples – crude herbs or industrial- 53 34 of Brazil (Fortunato, 1986, 1996; Fortunato et al., 2008; Vaz et al., ized herbal medicines – include entire or broken dried leaves, stems 54 35 2010). and often pods. In general, plants are harvested from their natural 55 36 Bauhinia bauhinioides (Mart.) J.F.Macbr., B. microstachya (Raddi) habitats, being popularly recognized by their bilobed or bifoliolate 56 37 J.F.Macbr., B. ungulata L., B. holophylla (Bong.) Steud., B. forficata leaves. Species are difficult to identify, especially from vegetative 57 38 Link, B. cheilantha (Bong.) Steud. and B. rufa (Bong.) Steud. are stages or in fragmented material, and sample labeling and trading 58 39 the main species for which pharmacological activities have been are mostly accomplished by using common names, generic name 59 40 reported. Hypoglycemic, antioxidant, analgesic, antiinflammatory or incorrect names (Melo et al., 2004). These often result in adul- 60 41 and larvicidal activities along with antitumor potentiality let them terated, falsified or substitute samples, which would interfere with 61 42 auspicious targets for new bioactive substances researching and the effectiveness and even the safety of the product (Ferreres et al., 62 43 phytotherapy (Silva and Cechinel Filho, 2002; Sousa et al., 2004; 2012). Leaf anatomical features are useful for identification and 63 adequate sample botanical quality, in Bauhinia s.l., venation pattern 64 has been utilized in taxonomy, phylogenetic analyses and even in 65 ∗ palaeobotany (Vaz, 1979; Fortunato, 1986; Zhang, 1994; Lin et al., 66 Corresponding author. 2015). 67 E-mail: [email protected] (M.J. Nores). http://dx.doi.org/10.1016/j.bjp.2016.10.007 0102-695X/© 2016 Sociedade Brasileira de Farmacognosia. Published by Elsevier Editora Ltda. This is an open access article under the CC BY-NC-ND license (http:// creativecommons.org/licenses/by-nc-nd/4.0/). Please cite this article in press as: Fortunato, R.H., et al. Leaf venation pattern to recognize austral South American medicinal species of “cow’s hoof” (Bauhinia L., Fabaceae). Revista Brasileira de Farmacognosia (2016), http://dx.doi.org/10.1016/j.bjp.2016.10.007 G Model BJP 330 1–4 ARTICLE IN PRESS 2 R.H. Fortunato et al. / Revista Brasileira de Farmacognosia xxx (2016) xxx–xxx Q5 Fig. 1. Leaf venation in southern South American Bauhinia. (a and b) General vein pattern. (c–g, i–p, r and s) Areolation. (h and q) Looped marginal ultimate venation. (a) B. bauhinioides. (b) B. hagenbeckii. (c) Bauhinia affinis. (d) B. angulosa. (e) B. argentinensis. (f) B. bauhinioides. (g and h) B. campestris. (i) B. cheilantha. (j) B. forficata ssp. pruinosa. (k) B. hagenbeckii. (l) B. holophylla. (m) B. microstachya. (n) B. mollis var. mollis. (o) B. mollis var. notophila. (p and q) B. rufa. (r) B. ungulata var. ungulata. (s) B. uruguayensis. Arrowhead: basal veins. Arrow: agroaphic veins. Scale bar: a and b: 3 mm; c–g, i–p, r and s: 300 ␮m; h, q: 1 mm. 68 In this work, we contribute to the recognition and commer- differentiating three main groups, some of them, but not all, taxo- 91 69 cial quality control of austral South American medicinal Bauhinia nomically related. 92 70 through a study of leaf vein pattern. The first group is characterized by a good areole development 93 that is found in some medicinal members of sect. Pauletia ser. 94 Cansenia, in the liana Bauhinia angulosa (= S. angulosa) and in the 95 71 Materials and methods spiny tree B. uruguayensis (Fig. 1d, l, p–s). The medicinal B. holo- 96 phylla and B. rufa both display quadrangular and pentagonal, rarely 97 72 We analyzed 19 austral South American taxa (Appendix 1). The triangular areoles, with absent and unbranched freely ending vein- 98 73 study of leaf venation pattern was carried out following Ellis et al. lets (FEV), the first one can be further recognized since FEV in 99 74 (2009). For this, fresh or herbarium leaves were cleared accord- general cross the areola and marginal ultimate venation and quin- 100 75 ing to Dizeo de Strittmatter (1973) and stained in safranin/80% ternary vein fabric venation are absent (Fig. 1l). B. rufa, instead, 101 76 ethanol. Samples were mounted in DPX (Aldrich Chemical Com- presents looped marginal ultimate venation and quinternary vein 102 77 pany, Gillingham, UK) and examined using Zeiss Stereo Microscope fabric present (Fig. 1p and q). The medicinal B. ungulata has absent, 103 78 a Zeiss Axiolab light microscope (Carl Zeiss, Germany) with a trinoc- unbranched and one branched FEV and areola shape more vari- 104 79 ular phototube and a color digital imaging camera. Some venation able (Fig. 1r). Some similar features have been described in others 105 80 data were extracted from previous work (Fortunato, 1986, 1996). taxa from ser. Cansenia, and Zhang (1994) named it as the Canse- 106 nia venation type. B. angulosa is easily distinguished because of the 107 108 81 Results and discussion very small areoles lacking FEV and marginal ultimate vein absent (Fig. 1d). B. uruguayensis is the only taxon that presents paxillate 109 110 82 All taxa analyzed present basal actinodromous primary vein areole development without FEV, or just very few FEV (Fig. 1s), 111 83 framework, with no naked basal veins, 3–5 (7) basal veins per lobe these features are similar to the Elayuna type described by Zhang 112 84 or leaflet and agrophic veins simple (Fig. 1a and b). Previously, some (1994). 113 85 of these taxa have been described as acrodromous (Fortunato, 1986, By the other side, the second group, composed by the prickly 114 86 1996) or campylodromous (Vaz, 1979), and recently both American trees and shrubs from sect. Pauletia ser. Aculeatae and the suffru- 115 87 Bauhinia s. s. and Schnella are considered actinodromous or acro- tices and shrubs from ser. Pentandrae, possesses moderate areole 116 88 dromous by Lin et al. (2015). Main vein characters (Fig. 1, Table 1) development along with marginal ultimate venation absent and 117 89 provide a criterion to identification of medicinal species and detec- quinternary vein fabric present (Fig. 1b–e, j–k, m–o). Compara- 118 90 tion of falsifications or adulteration. The areole development allows tively, areoles are larger and more variable in size than in previous Please cite this article in press as: Fortunato, R.H., et al.

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