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Redalyc.Phytotoxicity and Cytotoxicity of Lepidaploa Rufogrisea

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Redalyc.Phytotoxicity and Cytotoxicity of Lepidaploa Rufogrisea Revista de Biología Tropical ISSN: 0034-7744 [email protected] Universidad de Costa Rica Costa Rica Barroso Aragão, Francielen; Tebaldi Queiroz, Vagner; Ferreira, Adésio; Vidal Costa, Adilson; Fontes Pinheiro, Patricia; Tavares Carrijo, Tatiana; Fonseca Andrade-Vieira, Larissa Phytotoxicity and cytotoxicity of Lepidaploa rufogrisea (Asteraceae) extracts in the plant model Lactuca sativa (Asteraceae) Revista de Biología Tropical, vol. 65, núm. 2, junio, 2017, pp. 435-443 Universidad de Costa Rica San Pedro de Montes de Oca, Costa Rica Available in: http://www.redalyc.org/articulo.oa?id=44950834001 How to cite Complete issue Scientific Information System More information about this article Network of Scientific Journals from Latin America, the Caribbean, Spain and Portugal Journal's homepage in redalyc.org Non-profit academic project, developed under the open access initiative Phytotoxicity and cytotoxicity of Lepidaploa rufogrisea (Asteraceae) extracts in the plant model Lactuca sativa (Asteraceae) Francielen Barroso Aragão 1, Vagner Tebaldi Queiroz 2, Adésio Ferreira 3, Adilson Vidal Costa 2, Patricia Fontes Pinheiro 2, Tatiana Tavares Carrijo 4 & Larissa Fonseca Andrade-Vieira 5* 1. Department of Biological Sciences, Center of Human and Natural Sciences, Universidade Federal do Espírito Santo, 29075-910 Vitória – ES, BR; [email protected] 2. Department of Chemistry and Physics, Center of Agronomical Sciences, Universidade Federal do Espírito Santo, 29500-000 Alegre – ES, BR; [email protected], [email protected], [email protected] 3. Department of Plant Production, Center of Agricultural Sciences, Universidade Federal do Espírito Santo, 29500-000 Alegre – ES, BR; [email protected] 4. Department of Biology, Center of Agronomical Sciences, Universidade Federal do Espírito Santo, 29500-000 Alegre – ES, BR; [email protected] 5. Department of Biology, Universidade Federal de Lavras, 37200-000 Lavras – MG, BR; [email protected] * Correspondence Received 22- VII -2016. Corrected 06- II -2017. Accepted 02- III -2017. Abstract: Lepidaploa rufogrisea is a promising candidate in the search of natural herbicides, with compounds (sesquiterpene lactones-glaucolide B) which have been reported to present phytotoxic potential against weeds. Thus, this work aimed to accomplish the phytochemical investigation of L. rufogrisea as well as the evaluation of its biological effects on the plant model Lactuca sativa . To this end, different extracts (aqueous, hydroalco - holic and hexane) of L. rufogrisea at different concentrations [0.25, 0.5, 1.0, 2.0 and 4.0 % (m/v)] were evaluated from macroscopic (percentage of germinated seeds, germination speed index and root length) and microscopic analysis (mitotic index and cell cycle alterations) in plantlets (150 per treatment) and meristematic cells (10 000 per treatment) of L. sativa var. Cristina. The extracts decreased the germination speed index and also the root growth of L. sativa plantlets. Cell proliferation was not significantly affected by the extracts. However, some chromosome and nuclear alterations in the mitotic spindle were observed, due to the interference of allelochemi - cals present in L. rufogrisea . Moreover, the plant model L. sativa demonstrated to be efficient for this kind of studies. Rev. Biol. Trop. 65 (2): 435-443. Epub 2017 June 01. Key words: cytotoxicity, phytotoxicity, root growth, allelopathic potential, cell cycle alterations, plant bioassays. The continuous use of pesticides may to the control of pests, diseases and invading bring negative effects to human health and plants in crops (Isman, 2006; Kato-Noguchi, the environment (Souza Filho, Lobo, & Arru - Takeshita, Kimura, Ohno, & Suenaga, 2013). da, 2005). Besides environmental pollution, Lepidaploa rufogrisea (A. St.-Hil.) H. Rob. synthetic herbicides, could increase the inci - (family Asteraceae) has stood out in previous dence of weeds (Narwal, 1999). Thus, there is studies for its allelopathic potential, due to the an incentive to seek alternative strategies for sesquiterpene lactones (glaucolide B) (Padolina weeds control and management. To achieve et al., 1974), which showed phytotoxic activity this goal, systems of sustainable agriculture against various weeds (Macias, Varela, Torres, have been developed. It is based on plants or Oliva, & Molinillo, 1998; Wedge, Galindo, & plant-derived products that are used as bio - Macias, 2000). The allelopathic activity of a herbicides, less hazardous to the environment, plant-derived product is of research interest Rev. Biol. Trop. (Int. J. Trop. Biol. ISSN-0034-7744) Vol. 65 (2): 435-443, June 2017 435 as they are sources of bioactive chemicals for Fina Ltda. (Duque de Caxias, Rio de Janeiro, bioherbicides formulation. Brazil). Tween ® 80 and acetone were purcha - In this context, seed germination and plant - sed from Isofar Indústria e Comércio de Produ - let growth, parameters accessed by macrosco - tos Químicos Ltda. (Duque de Caxias, Rio de pic analysis using a model plant species, are Janeiro, Brazil). The other chemicals used in the bioassays most commonly used for study this study were purchased from Sigma-Aldrich and identification of the allelopathic poten - (Minas Gerais, Brazil). tial of allelochemicals extracted or present in plants (Rice, 1984). Although these assays Materials: The collection of L. rufogrisea provide general information about the biolo - samples was carried out in October 2011 at the gical effects of such allelochemicals, they do Natural Reserve of Vale do Rio Doce, Linhares not inform about their mechanisms of action (ES, Brazil), in Mussununga-type vegetation. in the target plant (Campos, Davide, Soares, The plant material in the field was flowering. & Viccini, 2008). Therefore, the evaluation of A young branch, with a flower, was collected, microscopic parameters via cytogenetic assay dried, and the exsiccate (T.T. Carrijo n. 1406) is a strategy that has been developed in addition was mounted and deposited in the herbarium of to macroscopic analyses (Andrade, Davide, & Federal University of Espírito Santo VIES, sec - Gedraite, 2010). Cytogenetic assays are based tion of Jerônimo Monteiro (voucher no. 537), on the evaluation of the cell cycle, allowing with duplicates in the herbarium of the Natural the observation of alterations throughout the Reserve of Vale (CVRD). mitosis. It also enables to identify whether the Seeds of L. sativa L. (2n = 2x = 18) var. extract in question presents cytotoxic, geno - “Cristina” (Feltrin Seeds), purchased at agri - toxic, mutagenic or even carcinogenic action business shops, were used as test system to (Leme & Marin-Morales, 2009). evaluate the phytotoxicity and cytogenotoxic Among the plant models used for alle - potential of the different extracts obtained lopathic studies, the species Lactuca sativa from L. rufogrisea . L. can be highlighted (Campos et al., 2008). Besides being highly sensitive and enabling the Phytochemical investigation: The aerial simultaneous analysis of various compounds parts (young stem and leaves) of L. rufogrisea (Grant, 1994), it also exhibits high proliferation were dried in forced air circulation dehydrator rate, rapid germination (up to 24 h), and has (S480AD, Biopar, Porto Alegre- RS, Brazil) for been useful in the analysis of cytogenetic para - a period of seven days at 40 ºC, then broken meters (Andrade et al., 2010; Andrade-Vieira, into smaller pieces and pulverized in cutting Botelho, Palmieri, Laviola, & Praça-Fontes, mill (MA-340, Marconi, Piracicaba - SP, Bra - 2014). Moreover, the laboratory procedures for zil). The obtained powder was stored in tightly these tests generally have low operational costs closed vessel protected from light incidence. and generate results within a short time. Con - The powder was used to prepare the aqueous, sidering all the above, the present work aimed hydroalcoholic and hexane extracts at 10 % to investigate the mechanisms of action of (m/v), and phytochemical screening was per - different extracts (aqueous, hydroalcoholic and formed. The phytochemical analyses to deter - hexane) of L. rufogrisea on the germination, mine the main chemical classes of secondary plantlet development and meristematic root tip metabolites (alkaloids, phenolic compounds, cells of L. sativa . terpenoids and steroids) were performed accor - ding to protocol described by Matos (2009). MATERIALS AND METHODS Preparation of extracts: The aqueous, Chemical products: Hexane, ethanol and hydroalcoholic and hexane extracts were obtai - acetic acid were purchased from Vetec Química ned by cold maceration of the L. rufogrisea 436 Rev. Biol. Trop. (Int. J. Trop. Biol. ISSN-0034-7744) Vol. 65 (2): 435-443, June 2017 powder (200 g) for 120 h at room temperature, in test solutions for 48 h. Roots were collected, using distilled water, 70 % ethanol (v/v) and fixed and the slides were prepared (two slides hexane, respectively. At the end of the extrac - per Petri dish) and evaluated as described Ara - tion, each mixture was centrifuged (6 000 gão et al. (2015). Ten slides were accessed for RPM, 20 min at 25 ºC) and the supernatants each treatment and about 10 000 meristematic were vacuum filtered. cells were counted per treatment. The mitotic The aqueous extract was transferred to index (MI), frequency of chromosomal (CA) volumetric flask (2 L) with the volume being and nuclear (NA) alterations were recorded as adjusted with distilled water. The hydroalco - described Aragão et al. (2015). holic and hexane extracts
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