A safer and more sustainable methodology for herbarium specimen rehydration in morphoanatomical studies

Autores

  • Ítalo Antônio Cotta Coutinho Universidade Federal do Ceará
  • Marcus Levy Sanders Fernandes Universidade Federal do Ceará
  • Lara Luiza Sales Alves Universidade Federal do Ceará
  • Antonia Mirelle Lopes Marques Universidade Federal do Ceará
  • Valdnéa Casagrande Dalvi Instituto Federal de Educação, Ciência e Tecnologia Goiano

DOI:

https://doi.org/10.36560/19520262249

Palavras-chave:

anatomical techniques, leaf, light microscopy, plant anatomy, sustainability

Resumo

Morphoanatomical studies based on herbarium material depend on rehydration procedures, in which the use of 2% potassium hydroxide (KOH) after boiling is common practice. Although this reagent facilitates tissue rehydration, it is a strong and corrosive base, capable of damaging sensitive tissues besides being associated with environmental concerns. Therefore, the adoption of methodologies that reduce or even eliminate its use is desirable. In this study, we evaluated the suppression of KOH during the rehydration of herbarium specimens. Leaf samples of different types (i.e. papery, coriaceous and succulent leaves, either glabrous or pubescent) were dried and subsequently rehydrated without alkaline treatment. Freehand sections were clarified with 50% sodium hypochlorite, stained with Astra blue and safranin, mounted on temporary slides, and examined under light microscopy. The material exhibited structural integrity, allowing detailed visualization of leaf anatomical features. The elimination of KOH proved to be effective, resulting in a simpler, faster, and more environmentally friendly procedure

Referências

AGOURAKIS, D. C.; CAMARGO, I. M. C. de; COTRIM, M. B. & FLUES, M. Comportamento de zinco e manganês de pilhas alcalinas em uma coluna de solo. Química Nova, v.29, p. 960–964, 2006. DOI: https://doi.org/10.1590/S0100-40422006000500014

AMORIM, E. P. et al. Ash content and chemical elements in 10 clones of Hevea by scanning electron microscopy and energy dispersive spectroscopy SEM/EDS. Scientific Electronic Archives, v. 13, n. 8, p. 66-70, 2020. DOI: https://doi.org/10.36560/13820201175

ATKINS, P. & JONES, L. Chemical Principles: The Quest for Insight. 4th. ed. New York, USA: W. H. Freeman and Company, 2008.

BOULAMANTI, A. & MOYA, J. A. Energy efficiency and GHG emissions: prospective scenarios for the chemical and petrochemical industry: Joint Research Centre (JRC), European Commission, 2017.

CARVALHO, R. R. Caracterização anatômica e histoquímica dos órgãos vegetativos de Adenium obesum (Forssk.) Roem. & Schult, Apocynaceae. 2022. Master’s Dissertation - Universidade Federal de Pernambuco, 2022.

COUTINHO, Í. A. C.; RANDO, J. G.; CONCEIÇÃO, A. de S. & MEIRA, R. M. S. A. A study of the morphoanatomical characters of the leaves of Chamaecrista (L.) Moench sect. Apoucouita (Leguminosae-Caesalpinioideae). Acta Botanica Brasilica, v.30, p.205–221, 2016. DOI: https://doi.org/10.1590/0102-33062016abb0035

DALVI, V. C.; MEIRA, R. M. S. A. & AZEVEDO, A. A. Extrafloral nectaries in neotropical Gentianaceae: occurrence, distribution patterns, and anatomical characterization. American Journal of Botany, v.100, p.1779–1789, 2013. DOI: https://doi.org/10.3732/ajb.1300130

DALVI, V. C.; MEIRA, R. M. S. A.; FRANCINO, D. M. T.; SILVA, L. C. & AZEVEDO, A. A. Anatomical characteristics as taxonomic tools for the species of Curtia and Hockinia (Saccifolieae – Gentianaceae Juss.). Plant Systematics and Evolution, v.300, p.99–112, 2014. DOI: https://doi.org/10.1007/s00606-013-0863-1

DEHAUT, A. et al. Microplastics in seafood: benchmark protocol for their extraction and characterization. Environmental Pollution, v.215, p.223–233, 2016. DOI: https://doi.org/10.1016/j.envpol.2016.05.018

DIAS, K. N. L.; SILVA, A. N. F.; GUTERRES, A. V. F.; LACERDA, D. M. A. & ALMEIDA JR., E. B. A importância dos Herbários na construção de conhecimentos sobre a diversidade vegetal. Revista Trópica - Ciências agrárias e biológicas, v.11, 25–35, 2019.

DUARTE, M. R. & LOPES., J. F. Stem and leaf anatomy of Plectranthus neochilus Schltr., Lamiaceae. Revista Brasileira de Farmacognosia, v.17, p.549–556, 2007. DOI: https://doi.org/10.1590/S0102-695X2007000400013

EL-TAHER, A. M.; GENDY, A. E.-N. G. El; ALKAHTANI, J.; ELSHAMY, A. I. & ABD-ELGAWAD, A. M. Taxonomic implication of integrated chemical, morphological, and anatomical attributes of leaves of eight Apocynaceae taxa. Diversity, v.12, p.334, 2020. DOI: https://doi.org/10.3390/d12090334

FIDALGO, O. & BONONI, V. Técnicas de coleta, preservação e herborização de material botânico. Vol. 4. São Paulo: Instituto de Botânica, 1984.

GADELHA NETO, P. da C.; LIMA, J. R. de; BARBOSA, M. R. de V; BARBOSA, M. de A.; MENEZES, M.; PÔRTO, K. C.; WARTCHOW, F. & GIBERTONI, T. B. Manual de Procedimentos para Herbário: Editora Universitária UFPE, 2013.

GALÃO, Ana Tereza Durão et al. Wood hydraulic characteristics of pioneers/early secondary and non-pioneer species. Scientific Electronic Archives, v. 14, n. 7, p. 45-52, 2021. DOI: https://doi.org/10.36560/14720211370

GARCIA-HERRERO, I.; MARGALLO, M.; ONANDÍA, R.; ALDACO, R. & IRABIEN, A. Environmental challenges of the chlor-alkali production: Seeking answers from a life cycle approach. Science of The Total Environment, v.580, p.147–157, 2017. DOI: https://doi.org/10.1016/j.scitotenv.2016.10.202

GONÇALVES, J. R.; DE PAULA-SOUZA, J.; KUSTER, V. C. & DALVI, V. C. Leaf anatomy of species from the Pombalia lanata complex (Violaceae) with implications for taxonomy and ecology. Flora, v.312, p.152471, 2024. DOI: https://doi.org/10.1016/j.flora.2024.152471

HODGE, E. The use of alcohol in plant collecting. Rhodora, v.49, p.207–210, 1947.

IBGE. Manual técnico da vegetação brasileira: sistema fitogeográfico: inventário das formações florestais e campestres: técnicas e manejo de coleções botânicas: procedimentos para mapeamentos. 2a edição ed. Rio de Janeiro, Brasil. 2012.

INCEER, H. & OZCAN, M. Taxonomic evaluations on the anatomical characters of leaf and achene in Turkish Tripleurospermum with its relative Matricaria (Asteraceae). Flora, v.275, p.151759, 2021. DOI: https://doi.org/10.1016/j.flora.2020.151759

KOLB, K. L.; GOMES, S. M. A. & LOMBARDI, J. A. Leaf anatomy as a taxonomy tool for the identification of Brazilian native species of Chionanthus (Oleaceae). Flora, v.266, p.151590, 2020. DOI: https://doi.org/10.1016/j.flora.2020.151590

KORSAGA-SOMÉ, N.; TAPSOBA, P.; OUÉDRAOGO, M. S.; BAOWENDSOM ILBOUDO, L.; BARRO-TRAORÉ, F.; NIAMBA, P. & TRAORÉ, A. Nécrolyse épidermique liée à l’application cutanée d’une solution d’hydroxyde de potassium. Pan African Medical Journal, v.21, p.239–245, 2015. DOI: https://doi.org/10.11604/pamj.2015.21.299.7673

KRAUS, J. E. & ARDIUN, M. Manual básico de métodos em morfologia vegetal. Seropédica, RJ - Brazil: EDUR, 1997.

LIANG, X. M.; XING, Y. Z.; LI, L. T.; YUAN, W. K. & WANG, G. F. An experimental study on the relation between friction force and real contact area. Scientific Reports, v.11, p.20366, 2021. DOI: https://doi.org/10.1038/s41598-021-99909-2

LOPES, Olívia Pereira et al. Hydraulic structure and wood density from root to the crown in in three native Brazilian tree species. Scientific Electronic Archives, v. 15, n. 4, 2022. DOI: https://doi.org/10.36560/15420221519

MAURO, C.; SILVA, C. de P.; MISSIMA, J.; OHNUKI, T.; RINALDI, R. B. & FROTA, M. Estudo anatômico comparado de órgãos vegetativos de Plectanthrus ornatus Codd. e malvariço, Plectranthus amboinicus (Lour.) Spreng. - Lamiaceae. Revista Brasileira de Farmacognosia, v.18, p.608–613, 2008. DOI: https://doi.org/10.1590/S0102-695X2008000400019

MEIRA, R. M. S. A. & MARTINS, F. M. Inclusão de material herborizado em metacrilato para estudos de anatomia vegetal. Revista Árvore, v.27, p.109–112, 2003. DOI: https://doi.org/10.1590/S0100-67622003000100015

MONTEIRO, S. S. & SIANI, A. C. A Conservação de exsicatas em herbários: contribuição ao manejo e preservação. Revista Fitos, v.4, p.24–37, 2009. DOI: https://doi.org/10.32712/2446-4775.2009.95

MOREIRA, N. S.; NASCIMENTO, L. B. S.; LEAL-COSTA, M. V. & TAVARES, E. S. Comparative anatomy of leaves of Kalanchoe pinnata and K. crenata in sun and shade conditions, vas a support for their identification. Revista Brasileira de Farmacognosia, v.22, p.929–936, 2012. DOI: https://doi.org/10.1590/S0102-695X2012005000056

MOUZELLA, C. P.; SOUSA, A. A.; CARUZO, M. B. R.; MONTERO-MUÑOZ, I.; MEIRA, R. M. S. A. & RIINA, R. Laticifers are present in Acalyphoideae after all: new insights from leaf anatomy with implications for the systematics and evolution of Euphorbiaceae. AoB PLANTS, v.17, 2025. DOI: https://doi.org/10.1093/aobpla/plaf006

OMINO, E. A contribution to the leaf anatomy and taxonomy of Apocynaceae in Africa : the leaf anatomy of Apocynaceae in East Africa; a monograph of Pleiocarpinae (series of revisions of Apocynaceae 41). 1996. - Wageningen University, Wageningen, The Netherlands, 1996.

PINTO-SILVA, N.P.; DE SOUZA, K.F.; MARQUES SILVA, O.L.; DA PAIXÃO NORONHA PEREIRA, A.; SOARES, D.A.; RIINA, R.; CARUZO, M.B.R. & MEIRA, R.M.S.A. Trichomes in the megadiverse genus Croton (Euphorbiaceae): a revised classification, identification parameters and standardized terminology. Botanical Journal of the Linnean Society, v.203, p.37–49, 2023. DOI: https://doi.org/10.1093/botlinnean/boad008

ROSTRUP‐NIELSEN, J. R.; SEHESTED, J. & NOERSKOV, J. K. Hydrogen and synthesis gas by steam‐ and CO2 reforming. ChemInform, v.34, 2003. DOI: https://doi.org/10.1002/chin.200317288

ROTTA, E.; CARVALHO, L. C. & ZONTA, B. M. Manual de prática de coleta e herborização de material botânico. Documentos 173. EMBRAPA Florestas, Colombo, PR, Brazil, 2008.

SCHULTES, R. E. The use of formaldehyde in plant collecting. Rhodora, v.49, p.54–60, 1947.

SIBBR. Sistema de Informação Brasileiro sobre a Biodiversidade. 2026. Available at: https://sibbr.gov.br/. Accessed on 02 June. 2026.

SILVA, H. C. C. Caracterização anatômica e histoquímica comparativa de espécimes de Cucurbita pepo L. Master's dissertation, Universidade Federal de Pernambuco, 2023.

SILVA, M. M. B.; SANTANA, A. S. C. O.; PIMENTEL, R. M. M.; SILVA, F. C. L.; RANDAU, K. P. & SOARES, L. A. L. Anatomy of leaf and stem of Erythrina velutina. Brazilian Journal of Pharmacognosy, v.23, p.200–206, 2013. DOI: https://doi.org/10.1590/S0102-695X2013005000013

SMITH, F. H. & SMITH, E. C. Anatomy of the inferior ovary of Darbya. American Journal of Botany, v.29, p.464–471, 1942. DOI: https://doi.org/10.1002/j.1537-2197.1942.tb10236.x

SOUSA, R.; SOUSA, V.; FRANÇA DE SÁ, S. & SEQUEIRA, S. O. Understanding pith paper: anatomical characteristics and ageing of a challenging cultural heritage support. Heritage, v.8:542, 2025. DOI: https://doi.org/10.3390/heritage8120542

SOUZA, V. C.; FLORES, T. B. & LORENZI, H. Introdução à botânica: morfologia. Nova Odessa, Brazil: Instituto Plantarum de Estudos da Flora, 2019.

STOREY, R. & LEIGH, R. A. Processes modulating calcium distribution in citrus leaves. An investigation using x-ray microanalysis with strontium as a tracer. Plant Physiology, v.136, p.3838–3848, 2004. DOI: https://doi.org/10.1104/pp.104.045674

VICENTIN, Patricia Gurgel et al. Longitudinal variation of wood basic density and anatomy of Curatella americana L. Scientific Electronic Archives, v. 14, n. 10, 2021. DOI: https://doi.org/10.36560/141020211396

Vital, F. A. Z., Nakamura, A. T., & Terra, V. (2018). Anatomy of Extrafloral Nectary on the Petiole of Senegalia angico (Mart. in Colla) Seigler & Ebinger (Leguminosae - Mimosoideae). Scientific Electronic Archives, 11(3), 64–69. https://doi.org/10.36560/1132018507

VITARELLI, N. C.; RIINA, R.; CARUZO, M. B. R.; CORDEIRO, I.; FUERTES-AGUILAR, J. & MEIRA, R. M. S. A. Foliar secretory structures in Crotoneae (Euphorbiaceae): Diversity, anatomy, and evolutionary significance. American Journal of Botany, v.102, p.833–847, 2015. DOI: https://doi.org/10.3732/ajb.1500017

ZAGOTTA-VITAL, Flávio Antônio. Comparative occurrence of Prestonia R. Br.(Apocynaceae: Apocynoideae). Scientific Electronic Archives, 2020 DOI: https://doi.org/10.36560/1312020757

Publicado

2026-08-17

Como Citar

Cotta Coutinho, Ítalo A., Fernandes, M. L. S., Alves, L. L. S., Marques, A. M. L., & Dalvi, V. C. (2026). A safer and more sustainable methodology for herbarium specimen rehydration in morphoanatomical studies. Scientific Electronic Archives, 19(5), 1–9. https://doi.org/10.36560/19520262249

Edição

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Ciências Biológicas

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