Biologia 68/3: 433—438, 2013 Section Cellular and Molecular Biology DOI: 10.2478/s11756-013-0185-3 Cytogenotoxic effects of ethanolic extracts of Annona crassiflora (Annonaceae) Lília R. Ribeiro 1,2 , Maria F. Santos 1 , Qu˛ enia M. Silva 1 , Marcel J. Palmieri 2 , Larissa F. Andrade-Vieira 3 & Lisete C. Davide 2 1 Departamento de Ci˛ encias Biológicas, Centro Universitário de Formiga, Água Vermelha, 35570-000, Formiga, MG, Brazil; e-mail: lilia@uniformg.edu.br 2 Departamento de Biologia, Universidade Federal de Lavras, 37200-000, Lavras, MG, Brazil 3 Departamento de Produ¸ c˜ ao Vegetal, Centro de Ci¸ encias Agrárias, Universidade Federal do Espírito Santo, Alegre, 29500- 000, Espírito Santo, Brazil Abstract: Infusions of the leaves and seeds of Annona crassiflora Mart. are commonly employed in the treatment of diarrhoea, snakebites, tumours and disorders of the hair and scalp. The aim of the present study was to investigate the cytotoxic and genotoxic properties of ethanolic extracts of A. crassiflora by evaluating their effects on germination, root elongation, chromosome structure and the cell division of Lactuca sativa (lettuce). The experiment followed a randomized design involving the treatment of L. sativa seeds with ethanolic extracts from leaves and seeds of A. crassiflora applied at ten concentrations (0.0125, 0.025, 0.05, 0.1, 0.2, 0.4, 0.6, 0.8, 1.0, and 1.2 mg/L) and with five repetitions per treatment. Seeds of L. sativa exposed for 48 h to A. crassiflora leaf extract at concentrations ≥ 0.1 mg/L, or to seed extracts at concentrations ≥ 0.2 mg/L, showed germination percentages that were significantly (p < 0.05) lower than those of seeds exposed to aqueous ethanol control. Exposure of L. sativa seedlings to leaf (but not seed) extracts of A. crassiflora produced significant (p < 0.05) reductions in the mitotic indices of root meristem cells of lettuce and induced chromosome and nuclear abnormalities in the root cells. The presence of chromosome stickiness, bridges, fragments, laggard chromosomes and nuclear condensation were also observed. The cytogenetic effects observed suggest that folkloric medicines prepared with extracts of the leaves or seeds of A. crassiflora should be employed with caution. Key words: Annona crassiflora; araticum; Lactuca sativa; germination; root-tip test; chromosomal abnormalities. Abbreviations: BOD, biochemical oxygen demand; MG, state of Minas Gerais; SD, standard deviation; IC50, 50% inhi- bition concentration. Introduction The World Health Organization (WHO) asserts that approximately 80% of the population in developing countries rely on traditional practices for basic health care, and that 85% of folklore medicines employed in the treatment of domestic and community ailments are prepared using plant materials (Ministério da Saúde do Brasil 2006). However, medicinal plants may synthesize toxic compounds as part of their natural defence mech- anisms against phytopathogens, insects and herbivores (Negrelle & Gomes 2007). On this basis, studies relating to toxicity and mutagenesis are indispensable in order to ensure that plant resources are employed in a safe manner (Lubini et al. 2008). Although numerous plants from the native Brazil- ian flora are ingested as food or for medicinal pur- poses, scientific evidence relating to their pharmacologi- cal or toxicological properties is generally scarce or non- existent, a situation that constitutes a serious public health problem (Veiga Júnior et al. 2005). In this con- text, many species of the family Annonaceae possess im- portant biological properties, including cytotoxic, anti- tumour, abortive, immunosuppressant, antiemetic, ver- micidal, antimicrobial, antimalarial and appetite sup- pressant activities, and some are employed as natural pesticides (Nascimento et al. 2003). In the case of An- nona crassiflora Mart., a plant that is endemic to the Brazilian Cerrado, where it is known as “araticum” or “marolo”, the fruit is edible and much appreciated by the local population. Moreover, the pulp of the ripe fruit is employed as an astringent and a tonic, while in- fusions of the leaves and triturated seeds are used in the treatment of diarrhoea, snakebites, tumours and disor- ders of the hair and scalp, and as emmenagoguic agents (Pereira et al. 2008; Soares et al. 2009). Despite the numerous potential therapeutic appli- cations of the species and its common use in traditional medicine, few studies have been performed regarding its cytotoxicity. However, Vilar et al. (2008) demonstrated that ethanolic extracts of leaves from A. crassiflora pre- sented cytotoxic and antimutagenic activities accord- ing to the micronucleus test in mice, while Santos et al. (1996) showed that araticulin, a compound isolated c 2013 Institute of Molecular Biology, Slovak Academy of Sciences Unauthenticated Download Date | 9/22/17 6:55 AM