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Annonaceae) in the CONTROL of BROWN STINK BUG

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Annonaceae) in the CONTROL of BROWN STINK BUG 581 Original Article POTENTIAL PHYTOINSECTICIDE OF Annona mucosa (JACQ) (Annonaceae) IN THE CONTROL OF BROWN STINK BUG POTENCIAL FITOINSETICIDA DE Annona mucosa (JACQ) (Annonaceae) NO CONTROLE DO PERCEVEJO-MARROM Leonardo Morais TURCHEN 1; Leilane Marisa HUNHOFF 2; Marcela Vieira PAULO 2; Camila Patrícia Ribeiro SOUZA 3; Mônica Josene Barbosa PEREIRA 4 1. Engenheiro Agrônomo, Universidade do Estado de Mato Grosso, Departamento de Agronomia, Tangará da Serra, MT, Brasil. [email protected]; 2. Graduando em Agronomia, Universidade do Estado de Mato Grosso, Departamento de Agronomia, Tangará da Serra, MT, Brasil; 3. Mestre em Ambiente e Sistema de Produção Agrícola, Universidade do Estado de Mato Grosso, Departamento de Agronomia, Tangará da Serra, MT, Brasil; 4. Doutor em Entomologia, Universidade do Estado de Mato Grosso, Departamento de Agronomia, Tangará da Serra, MT, Brasil. ABSTRACT : Plant extract have been recognized as important natural source of insecticide. Herein, we assessed the toxicity of leaves-extract and seeds-extract of Annona mucosa (Annonaceae) against brown stink bug, Euschistus heros (F.) (Hemiptera: Pentatomidae), a soybean pest in Neotropical America. For this, bioassays with nymphs and adults were conducted using the concentrations of 5, 10, 20, 40 and 80 mg.mL -1 of leaves and seeds extract of A. mucosa and DMSO (10%), as control. Ten replicates were conducted by treatment, with five nymphs (3 rd instar) by replicate (bioassay with nymphs) and one couple of adults (< 24h of old) by replicate (bioassay with adults). The number of nymphs death (mortality) and survival of adults were assessed. For adults the fecundity (number of eggs) and fertility (number of nymphs hatched) of females also were assessed. The nymphs bioassay has promising results in seeds extract, when compared to leaves extract. The mortality of nymphs treated with seeds extract was dependent-concentration, with increased in mortality in concentrations > 5.0mg.mL -1, while nymphs treated with leaves extract do not exhibit difference between treatments. In adults the survival was reduced, when were treated with seed extract of A. mucosa at concentrations higher than 20mg.mL -1, with reduction up to 92 days in contrast to control. A drastic reduction in the fecundity and fertility was observed, which highlight deleterious action of seed extract against reproductive potential of E. heros . This is the first report about toxicity A. mucosa (leaves and seed) on E. heros , and our results suggest that seeds extract of A. mucosa is promising for control of nymphs and adults of brown stink bug. KEYWORDS : Botanical insecticides. Euschistus heros . Toxicity. Nymphs. Adults. INTRODUCTION lethal to herbivorous insects (MITHOFER and BOLAND, 2012). Furthermore, they have different The Neotropical brown stink bug, modes of action (EL-WAKEIL, 2013), which makes Euschistus heros (F.) (Hemiptera: Pentatomidae) is them interesting for researchers aiming to develop pest in soybean crop (PANIZZI et al., 2012). The new insecticidal molecules. damage caused by this insect are directly in the In this context, Annonaceae family has been soybean pods, which reduces the yield and grain investigated to insect control (BERMEJO et al., quality (commercial product) (PANIZZI et al., 2005; CASTILLO-SÁNCHEZ et al., 2010; 2012; SILVA et al., 2012). In Mato Grosso, the state AMINIMOGHADAMFAROUJ et al., 2011; with greater soybean production is reported that E. COSTA et al., 2014; KRINSKI et al., 2014) with heros can cause damage of up to 30% in production promising results in the control of Spodoptera (VIVAN; DEGRANDE, 2011). frugiperda Smith (Lepidoptera: Noctuidae) pest in Insecticides are generally used for control of maize crop (BLESSING et al., 2010), Sitophilus arthropod pests, mainly due the efficiency of these zeamais Mots (Coleoptera: Curculionidae) pest of products. However, these are often used incorrectly, stored grain (RIBEIRO et al., 2013), Oncopeltus which can contribute to selection of resistant fasciatus Dallas (Hemiptera: Lygaeidae) cotton pest populations of insects (SOSA-GOMES and SILVA (COLOM et al., 2008) and also against E. heros , 2010) and suppression of natural enemies, such as soybean pest (SILVA et al., 2013; TURCHEN et egg parasitoids (TURCHEN et al., 2016). In al., 2014). contrast, plants with insecticidal properties have In Annonaceae family the specie Annona been investigated as a strategy for pest control, with mucosa (Jacq) have biologically active compounds, particular emphasis on substances of secondary such as acetogenins, alkaloids, amides and lignans metabolism, which are known to act as repellents, (CHEN et al., 1996; CHAVEZ et al., 1999; deterrents (oviposition or feeding) or may even be ESTRADA-REYES et al., 2002; LIAW et al., Received: 07/01/16 Biosci. J., Uberlândia, v. 32, n. 3, p. 581-587, May/June. 2016 Accepted: 20/03/16 582 Potential phytoinsecticide… TURCHEN, L. M. et al. 2003), which makes it interesting to search for organza. The pots were lined with filter paper and substances with insecticidal properties for the green-bean pods ( Phaseolus vulgaris ) were used for development of new insecticidal molecules. feeding the insects (COSTA et al., 1998) and However, few studies have been conducted with this changed every two days. The number of dead species to evaluate its potential in control of insect nymphs (15 days) and survival, fecundity (number pests. Therefore, we used as a model nymphs and of eggs/female) and fertility (number of eggs adults of E. heros to assess the insecticide potential viable/female) of adults until death were assessed. of A. mucosa . Statistical analysis MATERIAL AND METHODS Normality and homoscedasticity of the data obtained were checked using Shapiro-Wilk and Insect populations and extract vegetal: Bartlett tests, respectively. The number of nymph The populations of E. heros were collected dead (mortality), number of eggs (fecundity) and in soybean fields and reared in controlled number of nymphs hatched (fertility) were subjected environmental conditions at 26ºC ± 2ºC to analyses of deviance and general linear model temperature, 70% ± 10% relative humidity, and (GLM) with Poisson distribution and log as function L12:D12 photoperiod, the same conditions in which link. Significant treatments were compared by the bioassays were carried out. The brown stink contrasts (P < 0.05). The survival of adults over bugs were reared in natural diet, as described time were subjected to survival analyses using elsewhere (SILVA et al., 2008). The leaves and seed Kaplan-Meier (Gehan-Breslow) estimators and the of A. mucosa were collected in Tangará da Serra, survival curves compared using Holm-Sidak’s Mato Grosso (14° 29’S and 57°54’W), in December method (P < 0.05), always using the R software (2013) and January (2014). These plants were version. 3.1.1 (R-CORE TEAM, 2014). identified and deposited in the Tangará herbarium (TANG) (voucher number: 964). RESULTS For the preparation of the extract, the leaves and seed were dried in a drying oven with forced air Nymphs bioassay: circulation for 96 hours at 37°C, following the The mortality of the nymphs of E. heros methodology of Prista et al. (1981). After drying, was different between extracts (leaves or seeds) ( χ2 the leaves and seed were ground to a vegetable = 219.39; df. = 1,118; P < 0.001), with greater powder using a knife mill with a sieve with an mortality in the seeds extract (4.1 ± 1.75a), when aperture of 1 and 2mm of diameter, respectively. compared to the leaves extract (0.36 ± 0.63b). In This powder was added to the solvent (methyl seeds extract of A. mucosa the nymphs mortality alcohol) at a ratio of 500g to 1500mL and allowed was dependent of concentrations ( χ2 = 66.98; df. = to percolate for seven days. Thereafter, the mixture 5,54; P < 0.001), with mortality higher 90% of (solvent + powder) was filtered and the solvent was nymphs at concentrations of 5 mg.mL -1 and 80 evaporated by rotative evaporator to obtain the mg.mL -1, which differ of control (< 10% nymphs crude extract of leaves and seeds of A. mucosa . death) (see gray bars - Figure 1). Contrasting results were observed for nymph treated with leaf extract, Stink bug bioassay since that do not have difference between treatments Nymphs and adult bioassays were (χ2 = 2.06; df. = 5,54; P = 0.84), which indicated conducted in a completely randomized design at inactivity of leaves extract against E. heros nymphs -1 concentrations 0, 5, 10, 20, 40 and 80 mg.mL of (see white bars - Figure 1). the crude extract of leaves and seed of A. mucosa , Adults bioassay : with 10 replications/treatment. For the bioassay with The longevity of E. heros adults was rd nymphs, five 3 instar individuals were used per affected, when applied the seed extract of A. mucosa repetition (n=50 nymphs/treatment), and for the (Kaplan-Meier = 59.747; df. =5; P > 0.001). The bioassay with adults, one pair of E. heros up to 24 concentrations 5 and 10mg.mL -1 not exhibit hours old was used per repetition (n=20 reduction in longevity of adults. However, the adults/treatment). All bioassays were conducted concentrations of 20, 40 and 80mg.mL -1 following the methodology described by Piton et al. significantly differ, with reduction up to 92 days in (2014). The nymphs and adults were treated by survival of adults, when compared to control. In topic application of l µL and 5 µL treatments, addition, seeds extract also can affected fecundity respectively. After treatment the insects were kept in (χ2 = 4386.4; df. = 5,54; P < 0.001) and fertility ( χ2 plastic pots (145mL) with the opening sealed with = 5749.1; df. = 5,54; P < 0.001) of female. The Biosci. J., Uberlândia, v. 32, n. 3, p. 581-587, May/June. 2016 583 Potential phytoinsecticide… TURCHEN, L. M. et al. fecundity varied from 72.70 to 7.10 eggs/female at unviable eggs at concentrations 40 and 80 mg.mL -1.
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