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Revista Brasileira de Farmacognosia

Print version ISSN 0102-695XOn-line version ISSN 1981-528X

Rev. bras. farmacogn. vol.27 no.2 Curitiba Mar./Apr. 2017

http://dx.doi.org/10.1016/j.bjp.2016.10.007 

Original articles

Leaf venation pattern to recognize austral South American medicinal species of "cow's hoof" (Bauhinia L., Fabaceae)

Renée H. Fortunatoa 

Beatriz G. Varelab 

María A. Castroc 

María J. Noresd  * 

aInstituto de Recursos Biológicos (Instituto Nacional de Tecnología Agropecuaria – Consejo Nacional de Investigaciones Científicas y Técnicas) and Facultad de Agronomía y Ciencias Agroalimentarias, Universidad de Morón, Buenos Aires, Argentina

bFacultad de Farmacia y Bioquímica, Universidad Nacional de Buenos Aires, Buenos Aires, Argentina

cFacultad de Ciencias Exactas y Naturales, Universidad Nacional de Buenos Aires, Buenos Aires, Argentina

dInstituto Multidisciplinario de Biología Vegetal (Universidad Nacional de Córdoba – Consejo Nacional de Investigaciones Científicas y Técnicas) and Facultad de Ciencias Exactas, Físicas y Naturales, Universidad Nacional de Córdoba, Córdoba, Argentina

ABSTRACT

The leaves extracts of some species of Bauhinia L. s.l. are consumed to treat diabetes, inflammation, pains and several disorders in traditional medicine in austral South America. Despite its wide use and commercialization, sale is not controlled, and botanical quality of samples is not always adequate because of plant misidentification and adulteration. Here, we characterized leaf vein pattern in nineteen taxa to contribute to the recognition and commercial quality control of plant material commercially available. The vein characters intercostal tertiary and quinternary vein fabric, areole development and shape, free ending veinlet branching and marginal ultimate venation allowed to distinguish the main medicinal species in the region.

Keywords: Areolation; Botanical identification; Freely ending veinlets; Leaf venation pattern; Quality control; Schnella

Introduction

The leaves extracts of some species of Bauhinia L., Fabaceae, so called cow's hoof, "pata-de–vaca", cow's paw or orchid trees, are consumed to treat diabetes, inflammation, pains and several disorders in traditional medicine in austral South America (Oliveira and Saito, 1989; Bortoluzzi et al., 2006; Barboza et al., 2009; Pizziolo et al., 2011; Fortunato, 2012). With 300 species worldwide, Bauhinia s.l. is represented by fifteen species in Argentina, Paraguay, Uruguay and Santa Catarina, Paraná and Rio Grande do Sul states of Brazil (Fortunato, 1986, 1996; Fortunato et al., 2008; Vaz et al., 2010).

Bauhinia bauhinioides (Mart.) J.F.Macbr., B. microstachya (Raddi) J.F.Macbr., B. ungulata L., B. holophylla (Bong.) Steud., B. forficata Link, B. cheilantha (Bong.) Steud. and B. rufa (Bong.) Steud. are the main species for which pharmacological activities have been reported. Hypoglycemic, antioxidant, analgesic, antiinflammatory and larvicidal activities along with antitumor potentiality let them auspicious targets for new bioactive substances researching and phytotherapy (Silva and Cechinel Filho, 2002; Sousa et al., 2004; Gadotti et al., 2005; Luna et al., 2005; Nakahata et al., 2006; Cechinel Filho, 2009; Paula et al., 2014; Rozza et al., 2015; Martins-Olivera et al., 2016).

These taxa are extensively employed and sold in native American, rural and urban communities in the region and surrounding areas (Bortoluzzi et al., 2006; Barboza et al., 2009; Hurrell et al., 2011; Ibarrola and Degen de Arrúa, 2011). Despite its wide use and commercialization, sale is not controlled, and botanical quality of samples is not always adequate because of plant misidentification and adulteration. Commercial samples – crude herbs or industrialized herbal medicines – include entire or broken dried leaves, stems and often pods. In general, plants are harvested from their natural habitats, being popularly recognized by their bilobed or bifoliolate leaves. Species are difficult to identify, especially from vegetative stages or in fragmented material, and sample labeling and trading are mostly accomplished by using common names, generic name or incorrect names (Melo et al., 2004). These often result in adulterated, falsified or substitute samples, which would interfere with the effectiveness and even the safety of the product (Ferreres et al., 2012). Leaf anatomical features are useful for identification and adequate sample botanical quality, in Bauhinia s.l., venation pattern has been utilized in taxonomy, phylogenetic analyses and even in palaeobotany (Vaz, 1979; Fortunato, 1986; Zhang, 1994; Lin et al., 2015).

In this work, we contribute to the recognition and commercial quality control of austral South American medicinal Bauhinia through a study of leaf vein pattern.

Materials and methods

We analyzed 19 austral South American taxa (Appendix 1). The study of leaf venation pattern was carried out following Ellis et al. (2009). For this, fresh or herbarium leaves were cleared according to Dizeo de Strittmatter (1973) and stained in safranin/80% ethanol. Samples were mounted in DPX (Aldrich Chemical Company, Gillingham, UK) and examined using Zeiss Stereo Microscope a Zeiss Axiolab light microscope (Carl Zeiss, Germany) with a trinocular phototube and a color digital imaging camera. Some venation data were extracted from previous work (Fortunato, 1986, 1996).

Results and discussion

All taxa analyzed present basal actinodromous primary vein framework, with no naked basal veins, 3–5 (7) basal veins per lobe or leaflet and agrophic veins simple (Fig. 1A and B). Previously, some of these taxa have been described as acrodromous (Fortunato, 1986, 1996) or campylodromous (Vaz, 1979), and recently both American Bauhinia s. s. and Schnella are considered actinodromous or acrodromous by Lin et al. (2015). Main vein characters (Fig. 1, Table 1) provide a criterion to identification of medicinal species and detection of falsifications or adulteration. The areole development allows differentiating three main groups, some of them, but not all, taxonomically related.

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 µm; H, Q: 1 mm. 

Table 1 Leaf venation pattern in austral South American Bauhinia. FEV, freely ending veinlets; P, pentagonal; PE, percurrent; Q, quadrangular; R, reticulate; T, triangular. 

Species Intercostal tertiary vein fabric Quinternary vein fabric Areole development Free ending veinlet (FEV) branching Areole shape Marginal ultimate venation Fig. 1
B. affinis PE Present Moderate Unbranched and one or two branched; rarely absent variable Absent C
B. amambayensis R at apex, PE at base Present Moderate Unbranched and with one or more branches; sometimes absent variable Absent
B. angulosa (= S. angulosa) R at apex, PE at base Absent Good Absent; rarely unbranched Q and P, rarely T Absent D
B. argentinensis R at apex, PE at base Present Moderate Mostly with one or more branches; sometimes unbranched Variable Absent E
B. bauhinioides R Absent, to poor distinct Moderate to good Mostly unbranched and one branched; rarely two branched or absent Variable Absent A, F
B. campestris PE to R Absent Moderate, rarely good Mostly with one or more branches; sometimes unbranched; rarely absent Variable Looped G and H
B. cheilantha PE Absent Good to moderate Absent, unbranched and one branched; rarely two branched. Variable Absent I
B. forficata PE, notably at base Present Moderate Mostly with one or more branches; sometimes unbranched Variable Absent J
B. hagenbeckii R at apex, PE at base Present Moderate Mostly with one and more branches; rarely unbranched Variable Absent B, K
B. holophylla PE Absent Good Absent and unbranched (FEVs mostly crossing areoles); rarely one branched Q and P, rarely T Absent L
B. microstachya (= S. microstachya) R at apex, PE at base Absent, to poor distinct Moderate Mostly unbranched and one branched; sometimes with more branches Variable Looped and absent M
B. mollis var. mollis PE to R Present Moderate Mostly unbranched and one branched; sometimes with more branches or absent Variable Absent N
B. mollis var. notophila PE Poor distinct Moderate Mostly unbranched and with one or more branches; rarely absent Variable Absent O
B. rufa PE Present Good Absent and unbranched Q and P, rarely T Looped P and Q
B. ungulata var. cuiabensis PE to R at apex Absent Good Absent, unbranched and one branched Variable to Q, P, T Looped and absent
B. ungulata var. ungulata PE to R at apex Absent, to poor distinct Good Absent, unbranched and one branched Variable to Q, P, T Looped R
B. uruguayensis R at apex, PE at base Present Good/paxillate Absent; rarely unbranched and one branched Q and P, rarely T Looped and in part absent S

The first group is characterized by a good areole development that is found in some medicinal members of sect. Pauletia ser. Cansenia, in the liana Bauhinia angulosa (= S. angulosa) and in the spiny tree B. uruguayensis (Fig. 1D, L, PS). The medicinal B. holophylla and B. rufa both display quadrangular and pentagonal, rarely triangular areoles, with absent and unbranched freely ending veinlets (FEV), the first one can be further recognized since FEV in general cross the areola and marginal ultimate venation and quinternary vein fabric venation are absent (Fig. 1L). B. rufa, instead, presents looped marginal ultimate venation and quinternary vein fabric present (Fig. 1P and Q). The medicinal B. ungulata has absent, unbranched and one branched FEV and areola shape more variable (Fig. 1R). Some similar features have been described in others taxa from ser. Cansenia, and Zhang (1994) named it as the Cansenia venation type. B. angulosa is easily distinguished because of the very small areoles lacking FEV and marginal ultimate vein absent (Fig. 1D). B. uruguayensis is the only taxon that presents paxillate areole development without FEV, or just very few FEV (Fig. 1S), these features are similar to the Elayuna type described by Zhang (1994).

By the other side, the second group, composed by the prickly trees and shrubs from sect. Pauletia ser. Aculeatae and the suffrutices and shrubs from ser. Pentandrae, possesses moderate areole development along with marginal ultimate venation absent and quinternary vein fabric present (Fig. 1BE, JK, MO). Comparatively, areoles are larger and more variable in size than in previous group. The medicinal B. forficata (both subspecies), B. argentinensis and B. hagenbeckii show almost the same pattern, with mostly branching FEV (one or more branches) and less frequently, unbranched FEV (Fig. 1J, E, K). B. forficata can be distinguished by the percurrent tertiary vein fabric, while the others are percurrent at the base and reticulate at apex. B. affinis (Fig. 1C), B. amambayensis (not shown) and B. mollis (Fig. 1N, O) have in general both unbranched and branched FEV, in different frequency. The medicinal liana B. microstachya (= S. microstachya) possesses also moderate areole development, mostly unbranched and one branched FEV, but is recognized because of the presence of both looped and absent marginal ultimate venation (Fig. 1M), this report is similar to the one by Fortunato (1986) but differs from previous (Zhang, 1994).

Finally, the medicinal B. cheilantha and B. campestris (sect. Pauletia ser. Cansenia) present areole development intermediate between good and moderate. B. cheilantha alternates between absent, unbranched and one branched FEV and ultimate venation absent while B. campestris can be distinguished because FEV are in general branched and ultimate venation is looped (Fig. 1I, G and H). The medicinal B. bauhinioides (sect. Pauletia ser. Perlebia) also possess areole development moderate to good but areoles are mostly unbranched and one branched (Fig. 1F).

Interestingly, all species present prismatic crystals of calcium oxalate associated to vascular bundles and often druses in mesophyll cells, more notable in B. affinis, B. argentinensis, B. mollis and B. ungulata var. cuiabensis.

Here we provide a helpful tool for the recognition of the main regional medicinal plants through the analysis of the leaf vein pattern based on the intercostal tertiary and quinternary vein fabric, areole development and shape, free ending veinlet branching and marginal ultimate venation characters.

Acknowledgements

We thank Cristo S. for technical assistance. CONICET (PIP 112-201101-00250), INTA (PNHFA-1106094) and SECyT-UNC (32720160200161CB) are acknowledged for financial support.

Appendix A

Supplementary data

Supplementary data associated with this article can be found, in the online version, at doi:10.1016/j.bjp.2016.10.007.

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Received: September 06, 2016; Accepted: October 28, 2016

* Corresponding author. E-mail:jnores@imbiv.unc.edu.ar (M.J. Nores).

Authors' contributions

RHF collected and identified the plants, performed the laboratory work and data analysis and participated in the results discussion and the manuscript final writing. MAC participated in the laboratory work and data analysis. BGV helped perform the analysis with constructive discussions. MJN drafted the manuscript.

Conflicts of interest

The authors declare no conflicts of interest.

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