Journal of Global Biosciences ISSN 2320-1355 Volume 5, Number 6, 2016, pp. 4166-4171 Website: www.mutagens.co.in E-mail: [email protected] [email protected] Research Paper ALLELOPATHIC ACTIVITY OF edulis Rich. LEAF EXTRACT Vanessa Damasceno Gonçalves 1, Maria de Fatima Barbosa Coelho 1, Elisangela Clarete Camili 1, Carla Maria Abido Valentini 2, Rodrigo Aleixo Brito deAzevedo 3 and Ludmila Porto Piton 1

1Universidade Federal de Mato Grosso (UFMT), Faculdade de Agronomia, Medicina Veterinária e Zootecnia (FAMEVZ), Programa de Pós-Graduação em Agricultura Tropical (PPGAT), Av. Fernando Correa da Costa, nº 2367, Cidade Universitária, 78060-900, Cuiabá, MT. 2Instituto Federal de Mato Grosso, Campus Cuiabá - Bela Vista, Avenida Juliano Costa Marques, s/n, Bela Vista, 78050-560 - Cuiabá, MT. 3Universidade da Integração Internacional da Lusofonia Afro-brasileira (UNILAB), Instituto de Desenvolvimento Rural Avenida da Abolição, 7. 62790-000, Redenção, CE.

Abstract The aim of this study was to evaluate the allelopathic potential Alibertia edulis leaves in Lactuca sativa L. germination. Were used concentrations of the leaf extract of 0 (control), 25, 50, 75 and 100%. The bioassay was conducted in a completely randomized design with five levels and four replicates of 50 seeds of L. sativa . There was no difference between the extracts for germination ranged from 99.5 to 100%. The extract concentrations affected the germination rate, root length, shoot lenght and fresh weight of seedling. The pH and osmotic potential remained within acceptable standards for the germination and initial growth of lettuce, suggesting that the presence of compounds in the extracts have caused the observed effects. In the presence of the extract all the seedlings were abnormal indicating potential herbicide. Key words: Marmelada bola, seedling vigor, germination, morphological changes, Cerrado.

INTRODUCTION Alibertia edulis Rich. () is known as purui, marmelada, goiaba preta, apuruí, marmelada nativa, ajuruí, and marmelada bola in Mato Grosso. In Brazil it occurs in the states of Amazonas, Amapá, Bahia, Distrito Federal, Goiás, Maranhão, Mato Grosso, Mato Grosso do Sul, Minas Gerais, Pará, São Paulo, Tocantins [1]. The is arboreal in the Brazilian cerrado and can reach up to eight meters high, with beautiful foliage and white flowers [2].In addition to the ornamental potential, the fruits of A. edulis are an important food source for the local population and are part of the native fauna of the cerrado fauna diet [3]. The leaves, the fruit and the root of A. edulis are used in folk medicine. The tea is prepared with the leaves and used as a tranquilizer [4]. Journal of Global Biosciences Vol. 5(6), 2016 pp. 4166-4171 ISSN 2320-1355

The fruit and root are indicated for use in cases of pneumonia and syrup fruit is commonly used in the Amazon [5].The macerated fruits are estomáquicos and are also used against chickenpox [6].In pharmacological studies with Alibertia edulis compounds were isolated in 10 triperpenicos leaf extract [7].and four iridoid by chromatographic fractionation of the butanolic extract of the stems [2]. The presence of several secondary compounds in Alibertia edulis can have allelopathic activity on other species. The positive or negative interference of secondary metabolic compounds produced by a and released in the medium is known as "allelopathy" and the effect on the development of another plant can be indirect, by processing these substances in soil and microorganisms activity [8]. The presence of secondary compounds or allelochemicals has been found in all plant organs, with a tendency to accumulate in the leaves, and the release of these compounds can occur by root exudation, volatilization or leaching [9].The leaf is the most active plant body metabolically, reasonable being to present more diverse allelochemicals [10]. The possibility of controlling weed species at allelochemicals was demonstrated by [11, 12,13]. The natural compounds have several advantages over synthetic compounds, such as increased water solubility , absence of halogenated molecules and smaller half- life [14]. Therefore, the allelochemicals reveal themselves as natural herbicides, which would be free from the harmful effects caused by synthetic herbicides. The aim of this study was to evaluate the allelopathic potential of leaf extracts of Alibertia edulis under the lettuce germination.

MATERIAL AND METHODS The experiment was conducted at the Faculty of Agronomy Seed Laboratory, Veterinary Medicine and Animal Science (FAMEVZ) of the Federal University of Mato Grosso (UFMT) Campus Cuiabá. The leaves of Alibertia edulis were collected from adults in Chapada dos Guimarães (MT) between the geographic coordinates 15º10'- 15º30' south latitude and 55º40'-56º00 west longitude. The climate is Aw (Cerrado Climate), according to Köppen. The leaves were placed for 5 min in containers containing 10 ml of sodium hypochlorite diluted in 500 ml of distilled water to be cleaned and then washed in water and dried with a paper towel. The leaves were ground in a blender at a ratio of 50g leaves/500ml of distilled water. The extract was filtered through filter paper and stored in glass bottle. For bioassay of germination was used completely randomized design with five treatments consisting of the concentration of leaf extract obtained by dilution in distilled water (0 - control, 25, 50, 75 and 100%), with four replications of 50 lettuce seeds ( Lactuca sativa ). The pH determination was made pH meter and the electrical conductivity (EC) was measured by conductivity. From the EC values determined the osmotic potential (OP) according to the formula proposed by [15]: osmotic potential in the atmosphere (ATM) = - 0.36 * CE. The data were transformed to ATM (Mpa). The seeds were placed in transparent plastic boxes gerbox (11x11x3cm) on two sheets of blotting paper moistened with their treatment, in the amount of 2.5 times the mass of the substrate. The boxes were covered and sealed with plastic wrap and kept in BOD incubator at 30 °C during the day and 20 °C overnight in a photoperiod of 12 hours for seven days. Every 24 hours was noted the number of germinated seeds. The criterion to assess the germinability was based on physiological germination concept cited by [16],which indicates the beginning of germination to seed soaking and its end with radicle protrusion. Seven days after treatment application, lettuce seeds were evaluated as to

http://mutagens.co.in 4167 Journal of Global Biosciences Vol. 5(6), 2016 pp. 4166-4171 ISSN 2320-1355 the length of the shoot: root transition region to the insertion of the cotyledons and root length: transition region of the aerial part to the apex of the root. The dry weight of the seedling was obtained after drying the seedlings in a greenhouse at 80 °C for 24h to give the amount in grams. Seedlings were classified as normal or abnormal according to the specifications of [17].Were considered abnormal those that showed no potential to continue their development and normal seedlings with little defects such as small or limited damage, growth retardation in the root system. Thus, it was considered abnormal, seedlings with root systems or rotted air, absent totally atrophied. The germination rate was determined according to [18]calculated by the expression: GRI = (G 1/N 1) + (G 2/N 2) + ... + (G n/N n), where: G 1 = number of germinated seeds in the first count N 1 = number of days until the first count, G 2 = number of germinated seeds in the second count,N 2 = number of days until the second count, n = last count. The variables were subjected to analysis of variance and regression when significant models were selected with higher R 2.

RESULTS AND DISCUSSION There was pH variation in the control, with acidity in the extracts, but remained within acceptable standards for germination and seedling growth in allelopathic tests (Table 1). [19]state that pH effects on allelopathic tests are caused by pH below 3 and above 11. As the osmotic potential, the values were lower than -0.2 MPa and is therefore not responsible for possible changes in germination behavior of lettuce according [20].

Table 1 -Physico-chemical characteristics of aqueous extracts of Alibertia edulis leaves Cuiabá, MT. 2015. pH Concentration of leaf extract(%) Osmotic Potential(Mpa) 0 7.60 -0.000291793 25 4.53 -0.010285714 50 4.56 -0.010759878 75 4.49 -0.026042553 100 4.51 -0.034066869

There was no difference in the percentage of germination of lettuce, which ranged from 99.5 to 100%. In general, the studies of the action of allelopathy allelochemicals is not evident in the total percentage of germination, but the characteristics related to the seedling development [21]. Germination is less sensitive to secondary metabolites than seedling growth, as this parameter is influenced by these substances can lead to the appearance of abnormal seedlings, and necrosis as a common symptom [22]. The extract concentrations affected the GRI, length of root and shoot fresh weight and seedling (Figure 1). The most striking effect was compared with the percentage of abnormal seedlings in all extract concentrations were 100% abnormal seedlings. These seedlings showed absence of the absorbent at the root, roots necrotic, darkening and hardening of the root apex, negative geotropism, atrophied and defective primary roots, short roots and disproportionate to shoot.

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60 3 50 2.5 y = 2.3 - 6x + 4.3x 2 40 2 R² = 0.90 30 y = + 47.5+ 18.6x-30.08x 2 1.5 R² = 0.97

GRI 20 1

10 lenght Root 0.5 B 0 0 0.00 0.25 0.50 0.75 1.00 0.00 0.25 0.50 0.75 1.00 Concentration of the extract Concentration of the extract

3 0.06 2.5 0.05 2 0.04 1.5 0.03 1 0.02 2

Shoot lenght Shoot y = 0.007x - 0.03x + 0.05 2 0.5 CC y = 1.5 - 0.2x - 0.3x 0.01 D R² = 0.92 R² = 0.85 0 0

0.00 0.25 0.50 0.75 1.00 seedlings of Freshweight 0.00 0.25 0.50 0.75 1.00 Concentration of the extract Concentration of the extract

Figure 1 -Germination rate index (A), root length (B), shoot length (C), fresh weight of seedlings (D) of lettuce in different concentrations Alibertia edulis extract. UFMT, Cuiabá, MT. 2015.

The hardening and darkening of the root tips are evidence of morphological and anatomical changes caused by phytotoxins [23].The disproportion between the roots and shoots of seedlings, according to [24] is due to the fact that there is more contact between the roots and the extract (allelochemicals) than other structures of seedlings. Several authors also noted these effects in lettuce seeds, such as, with Erithryna mulungu extract [25, 26], with extracts of bark of Mimosa tenuiflora [27]with aqueous extracts of Casearia sylvestris [28].

CONCLUSIONS The leaf extract Alibertia edulis in different concentrations did not affect the porcentage of lettuce seed germination. The allelopathic potential was seen in developmental characteristics of lettuce seedlings, with 100% of abnormal seedlings.The pH and osmotic potential remained within acceptable standards for the germination and initial growth of lettuce, suggesting that the presence of compounds in the extracts have caused the observed effects.

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