Allelopathic potential of Inga laurina leaf extract on

Jaboticabal
ISSN: 1984-5529
v.44, n.3, p.333–337, 2016
http://dx.doi.org/10.15361/1984-5529.2016v44n3p333-337
Allelopathic potential of Inga laurina leaf extract on lettuce seed
germination
Potencial alelopático de extratos de folhas de Inga laurina na germinação de
sementes de alface
Vanessa Damasceno GONÇALVES1; Maria de Fátima Barbosa COELHO 2*; Elisangela Clarete
CAMILI3; Carla Maria Abido VALENTINI4;
1
Engenheira Agrônoma, Programa de Pós-Graduação em Agricultura Tropical (PPGAT), Av. Fernando Correa da Costa,
2367, Cidade Universitária, 78060-900, Cuiabá, MT, Brazil. E-mail:[email protected]
2 Profa. Titular Doutora, Programa de Pós-Graduação em Agricultura Tropical (PPGAT), Av. Fernando Correa da Costa,
2367, Cidade Universitária, 78060-900, Cuiabá, MT, Brazil. *Corresponding author: [email protected]
3 Profa. Doutora, Universidade Federal de Mato Grosso (UFMT), Faculdade de Agronomia, Medicina Veterinária e
Zootecnia (FAMEVZ), Av. Fernando Correa da Costa 2367, Cidade Universitária, 78060-900, Cuiabá, MT, Brazil. E-mail:
[email protected]
4 Profa. Doutora, Instituto Federal de Mato Grosso, Campus Cuiabá - Bela Vista. Avenida Juliano Costa Marques, s/n Bela
Vista, 78050-560 - Cuiaba, MT, Brazil E-mail: [email protected]
Recebido em: 02-03-2016; Aceito em: 18-05-2016
Abstract
The aim of this study was to evaluate the allelopathic potential of Inga laurina leaves on the germination and
growth of Lactuca sativa L. seedlings. The extract was obtained by blending 50 g of leaves in 500 ml of distilled
water. Extracts were obtained by diluting the leaf homogenate in water to concentrations of 0 (control), 25, 50,
75 and 100%. The bioassay was conducted in a completely randomized design with five treatments and four
repetitions of 50 seeds of L. sativa. The characteristics observed were germination and abnormal seedlings,
germination rate, shoot length and root length, dry weight of seedlings and allelopathy index. There was no
difference between extracts for germination ranging from 98%to 100%. The speed of germination rate and
seedling root length decreased with increasing concentration and could be explained by second degree
polynomial regression models with R2 above 98%. The fresh mass increased with higher concentrations of the
extract. The allelopathic index was negative, indicating inhibition, and increased with increasing extract
concentration. The leaf extract of Inga laurina at different concentrations did not affect lettuce seed germination.
Allelopathic potential was seen in the developmental characteristics of lettuce seedlings.
Additional keywords: allelopathic index; Fabaceae; germination rate; Ingá.
Resumo
O objetivo deste estudo foi avaliar o potencial alelopático de Inga laurina na germinação e no crescimento de
plântulas de alface. O extrato foi obtido por trituração de 50 g de folhas em 500 ml de água destilada. A partir
deste extrato obteve-se por diluição em água as concentrações de 0,(controle);25; 50; 75 e 100%. O ensaio foi
realizado em delineamento inteiramente casualizado, com cinco níveis e quatro repetições de 50 sementes de L.
sativa. As características observadas foram germinação e plântulas anormais, índice de velocidade de
germinação, comprimento da parte aérea e comprimento de raízes, massa seca de plântulas e índice de
alelopatia. Não houve diferença para a germinação que variou de 98 a 100% nas concentrações dos extratos. A
velocidade de germinação e comprimento de raiz diminuiu com o aumento da concentração e foi explicada por
regressão de modelos polinomiais de segundo grau com R2 acima de 98%. A massa fresca aumentou com as
maiores concentrações do extrato. O índice alelopático foi negativo indicando inibição e aumentou com a
concentração. O extrato de folhas de Inga laurina em diferentes concentrações não afetou a germinação de
sementes de alface. O potencial alelopático ocorreu em características de desenvolvimento das plântulas de
alface.
Palavras-chave adicionais: Fabaceae; índice alelopático; Ingá; velocidade de germinação.
Introduction
Allelopathy can be defined as positive or negative interference of secondary metabolic compounds
produced by a plant (allelochemicals) and released
into the environment. Interference in the growth of
another plant can be indirect, by transforming these
substances into the soil by microorganism activity
(Inderjit et al., 2011).
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Científica, Jaboticabal, v.44, n.3, p.333-337, 2016
This phenomenon influences the formation of
plant communities in natural environments and also
serves as an important tool in agronomy because it
can be used to control undesirable plants, maintaining
healthy cultures (Silva, 2012). Furthermore,
allelopathic bioassays contribute to the identification
of possible sources of biocide compounds that can be
used to combat pests and diseases, and the
resistance of weeds to synthetic herbicides has led to
the search for new alternatives, which are less
harmful to the environment, as is the case of natural
allelochemicals (Matsumoto et al., 2010).
Studies with native Cerrado species showed
the present allelopathic potential to be as yet unexplored (Oliveira et al., 2004; Gatti, 2007). Several
authors (Ribeiro et al, 2009; Centenaro et al, 2009;
Matsumoto et al, 2010; Oliveira et al., 2014) have
demonstrated the ability to control invasive species
through allelochemicals. A first study, conducted with
the genera Inga by Costa et al. (2015), found that
aqueous extracts of green leaves and dried roots of
Inga vera Willd. subsp. affinis (DC.) T. D. showed no
allelopathic effect on the germination and early
growth of Schizolobium parahyba (Vell.) SF Blake,
Piptadenia gonoacantha Mart. Macbr., Chorisia
speciosa A. St.-Hil. and Bixaorellana L., but inhibited
the hypocotyl growth of Bixaorellana L. seedlings.
Among species of the Brazilian Cerrado
occurs Inga laurina (Sw.) Willd., popularly known as
ingá-branco, ingá-de-macaco, ingá-de-praia, ingá-mirim or ingaí is distributed in riparian forests. The
species Inga laurinais important to the flora forits
ornamental properties and forest value because its
fruits are a food source for wild animals (Souza &
Lorenzi, 2008). It is an ideal species for urban forestry
because it adapts excellently to urban environments
and keeps its leaves during the entire dry period, thus
its crown provides considerable shade. In addition,
ingá-mirim grows white-pulp fruits that are consumed
by the fauna and even by humans (Leão et al., 2012).
Furtado (2014) considers it important to develop
studies with more species because of the action of
essential oils and leaf extracts against bacteria and
as an antioxidant.
Since allelopathic action occurs in Inga vera,
this action is also expected to be present in Inga
laurina on germination and initial growth. The
objective of this study was to evaluate the allelopathic
potential of Inga laurina leaves on the germination
and seedling development of Lactuca sativa L.
Material and methods
The experiment was conducted in the Faculty
of Agronomy Seed Laboratory Animal Science and
Veterinary Medicine (FAMEVZ) of the Federal
University of Mato Grosso (UFMT) Campus Cuiabá.
The leaves of Inga laurina were collected from adult
plants in Chapada dos Guimarães (MT) between the
geographic coordinates 15º10'15º30' south latitude
ISSN: 1984-5529
and 55º40'56º00 west longitude. The climate is Aw
(savanna climate), according to Köppen. The trees
were in the phenological stage of fruiting, the
collected leaves were fully developed and the same
size, healthy and taken from different branches of five
different individuals.
The leaves were placed for 5 min in
containers containing 10 mL of sodium hypochlorite
diluted in 500 mL of distilled water so that they were
cleaned, then rinsed in tap water and dried with a
paper towel. The leaves were ground in a blender at a
ratio of 50 g leaves to 500 mL of distilled water and
the extract was filtered on filter paper and stored in
glass according Silveira et al. (2012). Souza Filho et
al. (2010) presented a critical review that includes this
methodology.
For bioassay of germination, a completely
randomized design was used with five treatments
consisting of the concentrations 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 and electrical conductivity
(EC) determinations were made using a pH meter and
an EC meter, respectively. From the EC values (µS
cm-1 transformed to dS m-1), the osmotic potential (PO)
was determined according to the formula proposed by
Ayers & Westcot (1999): osmotic potential in
atmosphere (atm) = -0.36 × EC. The data in atm were
transformed to MPa.
The seeds were placed in transparent gerbox
plastic boxes (11x11x3 cm) on two sheets of blotting
paper moistened with treatment solution in the amount
of 2.5 times the mass of the substrate. The boxes were
covered, sealed with plastic wrap and kept in a BOD
incubator at 30 °C during the day and 20 °C overnight
with a 12-h photoperiod for seven days.
The number of germinated seeds was noted
every 24 h. The criterion for the assessment of seed
germination was based on the concept of
physiological germination cited by Marcos Filho
(2015), which describes the beginning of germination
to seed soaking and its end with the protrusion of the
radicle. Seven days after treatment application,
lettuce seeds were evaluated as to the length of the
shootroot transition region to the insertion of the
cotyledons and root length: transition region of the
shoot to the apex of the root. The seedlings were
placed in brown paper bags and exposed to drying in
a forced ventilation oven at 70 °C ± 2 °C for 72 h, to
obtain the dry weight.
The seedlings were classified as normal or
abnormal according to the specifications of BRASIL
(2009). Those that showed potential to continue their
development, as well as normal seedlings with minor
defects such as limited or minor damage, or growth
retardation in the root system, were considered
abnormal.
The germination rate was determined
according to Maguire (1962) and calculated by the
expression: IVG = (G1/N1) + (G2/N2) + ... + (Gn / Nn),
where: G1 = number of germinated seeds in the first
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Científica, Jaboticabal, v.44, n.3, p.333-337, 2016
count, N1 = number of days until the first count,
G2 = number of seeds germinated in the second count,
N2 = number of days elapsed until the second count
and n = last count.
The allelopathic effect index (RI) according to
the following formula: RI = 1 - C/T (T ≥ C) or RI = T/C - 1
(T < C) Where: C = speed of germination control and
T = speed of germination treatment (Gao et al., 2009).
The variables were subjected to analysis of
variance and regression when significant models were
selected with higher R2.
ISSN: 1984-5529
There was no difference in the percentage of
lettuce germination (98%100%) in the dry mass of
seedlings and percentage of abnormal seedlings. The
extract concentrations affected the IVG, root length,
shoot length and seedling fresh weight (Figure 1).
According to Ferreira & Borghetti (2005), the
allelopathic effect often occurs not by reducing final
germination, but by influencing the germination speed
and other parameters related to development.
Table 1- Physical and chemical characteristics of
aqueous extracts of leaves of Inga laurina, Cuiabá,
MT. 2015.
Results and discussions
The physicochemical characteristics presented
by extracts from Inga laurina leaves at different
concentrations are shown in Table 1. A narrow pH
range was observed in the extracts compared with the
control. According Gatti et al. (2004), in allelopathic
testing, the pH should be between 4 and 7, and the
osmotic potential should be less than -0.2 MPa.
Therefore, these features of the extracts are not
responsible for possible changes in the germination
behaviour of lettuce in this study.
Root lenght (cm)
IVG
1.00
2.00
y = 1.49 + 0.81x - 0.86x2
C
R² = 0.80
1.00
Allelophaty Index
-0.0002
-0.0100
-0.0196
-0.0279
-0.0353
y = 3.48 -3.99x + 1.31x2
R² = 0.99
3.00
2.00
1.00
Fresh mass seedling (mg)
Aerieal part lenght (cm)
6.08
5.88
5.77
5.91
5.90
Osmotic
potential (MPa)
B
3.00
0.25
0.50
0.75
Extract concentration
0%
25%
50%
75%
100%
0.00
0.00
4.00
0.00
0.00
pH
4.00
60.00
50.00
40.00
30.00
20.00
y = 49.87 - 23.48x
10.00
A R² = 0.98
0.00
0.00
0.25
0.50
0.75
Extract concentration
1.00
Extract
concentration
0
0.25
0.50
0.75
Extract concentration
1.00
0.05
0.04
0.03
0.02
0.01
y = 0,015 + 0.0191x + 0.0104x2
R² = 0.98
D
0.00
0.00
Extract concentration
0.25 0.5 0.75
0.25
0.50
0.75
1.00
Extract concentration
1
0.0%
-5.0%
-10.0%
-15.0%
-20.0%
E
-25.0%
Figure 1 - Germination speed index (A), root length (B), shoot length (C), fresh mass of seedling (C) and
allelopathic index (E) of lettuce in different concentrations of Inga laurina extract. Cuiabá, MT. 2015.
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Científica, Jaboticabal, v.44, n.3, p.333-337, 2016
The germination rate index (Figure 1A)
decreases linearly with increasing extract
concentration. Candido et al. (2010) evaluated the
allelopathic potential of Senna occidentalis L. and
observed a significant reduction in germination
speed index (GSI) of lettuce and tomato seeds
when they increased the concentration of the
extract. Tur et al. (2010) found a significant
reduction in IVG with increasing concentration of
the extract of fresh and dried leaves of Duranta
repens L. on Lactuca sativa L.
Borella et al. (2011a) tested the
allelopathic effect of nightshade (Solanum
americanum Mill.) on radish seed germination
(Raphanus sativus L.) and found that all of the
aqueous extracts of nightshade reduced the
average number of seeds germinated per day.
Reducing the speed of germination index results
in a reduction of seed vigour due to the toxic
effect of the extract.
The root length (Figure 1 B) also
decreases with increasing concentration of the
extract according to a second degree polynomial
model. These results are similar to those of Rickli
et al. (2011) who found that the extract of fresh
leaves of Azadirachta indica inhibited the growth
of the lettuce roots at all concentrations.
The length of the shoot follows a seconddegree polynomial model (Figure 1C). However,
the effect on the root was much more evident, and
the disproportion between the roots and shoots of
seedlings, according to Chung et al. (2001) is due
to the fact that there is more contact between the
roots and the extract (allelochemicals) than the
other structures of the seedlings.
The greatest extract concentrations
provide higher values of seedling fresh weight
(Figure 1D). Similar results were found by Carmo
et al. (2007) with Ocotea extracts (Vell.) Rohwer in
sorghum germination (Sorghum bicolor), where
the extracts of leaves and stem bark caused an
increase in the fresh weight of the root system of
sorghum, compared with seedlings in control
treatment under the action of root extracts.
From the data presented (Figure 1E), it
can be observed that all tested concentrations of
Inga laurina inhibited the germination of lettuce
seeds. The allelopathic effect index (RI) indicates
stimulus when it presents positive values in the
control and negative values indicate inhibition.
Borella et al. (2011b) found that soft Schinus
extracts caused inhibitory effects on the
germination of radish in proportion to the extract
concentration.
The allelopathic effect index was used by
Khong et al. (2002) and Abdelgaleil & Hashinaga
(2007) to demonstrate the allelopathic action of
the extracts in bioassays. The RI values varied
with the impact of the effects, either positive or
negative. Aqueous extracts of Hemisepta lyrata
ISSN: 1984-5529
Bunge caused inhibitory effects on the
germination of radish seeds, regardless of the
concentration used (Gao et al., 2009). Zhang et al.
(2010) also reported negative effects caused by
eucalyptol extracts on the radish.
Conclusions
The leaf extract of Inga laurina at different
concentrations did not affect the germination
percentage of lettuce seed. The lettuce germination
speed, root length and allelopathic index were
reduced with the highest concentrations of
extract.The leaf extract of Inga laurina must be
characterized and studied in weed species.
Acknowledgments
The authors wish to acknowledge the
Conselho Nacional de Desenvolvimento Científico e
Tecnológico (CNPq) for Productivity Grant.
References
Abdelgaleil SAM, Hashinaga F (2007) Allelopathic
potential of two sesquiterpene lactones from Magnolia
grandiflora L. Biochemical Systematics and Ecology
35:737-742.
Ayers RS, Westcot DW (1999) Water quality for
agriculture. Roma: FAO. 97p. (FAO Irrigation and
Drainage Paper, 29).
Borella J, Martinazzo EG, Aumonde TZ (2011b)
Atividade alelopática de extratos de folhas de
Schinus molle L. sobre a germinação e o crescimento
inicial do rabanete. Revista Brasileira de Biociências
9(3):398-404.
Borella J, Wandscheer ACD, Pastorini LH (2011a.)
Potencial alelopático de extratos aquosos de frutos
de Solanum americanum Mill.sobre as sementes de
rabanete. Revista Brasileira de Ciências Agrárias
6(2):309-313.
BRASIL (2009) Ministério da Agricultura e Reforma
Agrária. Regras para análise de sementes. Brasília:
SNAD/CLAV, 395p.
Candido ACS, Schmidt V, Laura VA, Faccenda O,
Hess SC, Simionatto E, Peres MTLP (2010) Potencial
alelopático da parte aérea de Senna occidentalis (L.)
Link (Fabaceae, Caesalpinioideae): bioensaios em
laboratório. Acta Botânica Brasílica 24(1):235-242.
Carmo FMS, Borges EEL, Takaki M (2007) Alelopatia
de extratos aquosos de canela-sassafrás (Ocotea
odorífera (Vell.) Rohwer). Acta Botânica Brasílica
21(3):697-705.
336
Científica, Jaboticabal, v.44, n.3, p.333-337, 2016
Centenaro C, Corrêa LGP, Karas MJ, Virtuoso S,
Dias JFG, Miguel OG, Miguel MD (2009)
Contribuição ao estudo alelopático de Erythrina
velutina Willd., Fabaceae. Revista Brasileira de
Farmacognosia 19(1B):304-308.
Chung IM, Ahn JK, Yoon SJ (2001) Assessment of
allelopathic potential of barnyard grass (Echinochloa
crus-gall) on rice (Oriza sativa L.) cultivars. Crop
Protection 20(10):921-928.
Costa SML, Ferreira MC, Pasin LAAP (2015)
Avaliação do potencial alelopático de ingá sobre o
desenvolvimento inicial de espécies arbóreas. Acta
Iguazu 4(1):1-13.
Ferreira AG, Borghetti F (2005) Germinação: do
básico ao aplicado. Porto Alegre: Artmed, 323p.
Furtado FB (2014) Estudo químico, analise do óleo
essencial e atividades biológicas de Inga laurina (S
W.) Willd.UFU (Dissertação de Mestrado em Química).
Gao X, Li M, Gao Z, Li C, Sun Z (2009) Allelopathic
effects of Hemistepta lyrata on the germination and
growth of wheat, sorghum, cucumber, rape, and
radish seeds. Weed Biology and Management
9(3):243–249.
Gatti AB, Perez SCJGA, Ferreira AG (2007)
Avaliação da atividade alelopática de extratos
aquosos de folhas de espécies de Cerrado.Revista
Brasileira de Biociências 5(2):174-176.
Gatti AB, Perez SCJG, Lima MIS (2004) Efeito
alelopático de Aristolochiae speranzae O. Kuntze na
germinação e no crescimento de Lactuca sativa L. e
Raphanus sativus L. Acta Botânica Brasílica
18(3):459-470.
ISSN: 1984-5529
Marcos Filho J (2015) Fisiologia de sementes de
plantas cultivadas. Piracicaba/FEALQ, 795p.
Matsumoto RS, Ribeiro JPN, Takao LK, Lima MIS
(2010) Potencial alelopático do estrato foliar de
Annona glaba L. (Annonaceae). Acta Botânica
Brasílica 24(3):631-635.
Oliveira AKM, Pereira KCL, Muller JAI, Matias R
(2014) Análise fitoquímica e potencial alelopático das
cascas de Pouteria ramiflora na germinação de
alface. Horticultura Brasileira 32(1):41-47.
Oliveira SCC, Ferreira AG, Borghetti F (2004) Efeito
alelopático de folhas de Solanum lycocarpum A. St.Hil. (Solanaceae) na germinação e crescimento de
Sesamum indicum L. (Pedaliaceae) sob diferentes
temperaturas. Acta Botânica Brasílica 18(3):401-406.
Ribeiro JPN, Matsumoto RS, Takao LK, Voltarelli VM,
Lima MIS (2009) Efeitos alelopáticos de extratos
aquosos de Crinum americanum L. Revista Brasileira
de Botânica 32(1):183-188.
Rickli HC, Fortes AMT, Silva PSS, Pilatti DM, Hutt DR
(2011) Allelopathic effect of aqueous extract of
Azadirachta indica A. Juss. on lettuce, soybeans,
maize, beans and Bidens pilosa. Semina: Ciências
Agrárias 32(2):473-484.
Silva PSS (2012) Atuação dos aleloquímicos no
organismo vegetal e formas de utilização da
alelopatia na agronomia. Biotemas 25(3):65-74.
Silveira PFS, Coelho MFB, Maia SSS (2012)
Potencial alelopático do extrato de folhas de Mimosa
tenuiflora (Willd.) Poir. na germinação de Lactuca
sativa L. Bioscience Journal 28(3):472-477.
Inderjit K, Wardle DA, Karban R, Callaway RM (2011)
The ecosystem and evolutionary contexts of
allelopathy. Trends in Ecology & Evolution 26(12):655662..
Souza Filho APS, Guilhon GMSP, Santos LS
(2010) Metodologias empregadas em estudos de
avaliação da atividade alelopática em condições de
laboratório: revisão crítica. Planta daninha 28(3):689697.
Khong C, Hu F, Xu X (2002) Allelopathic potenatial
and chemical constituents of volatiles from Ageratum
conyzoides under stress. Journal of Chemical
Ecology 28(6):1173-1182.
Souza VC, Lorenzi H. (2008) Botânica Sistemática:
Guia ilustrado para identificação das famílias de
angiospermas da flora brasileira, baseado em APGII.
3. Ed. Nova Odessa: Instituto Plantarum. 768p.
Leão JRA, Lima JPC, Pinto SN, Paiva AV. (2012)
Seed germination and initial growth of seedlings of
ingá-mirim - Inga laurina (S W.) Willd – used in urban
florestry of Rio Branco city, Acre State – Brazil.
Revista da Sociedade Brasileira de Arborização
Urbana 7(3):10-17.
Tur CM, Borella J, Pastorini LH (2010). Alelopatia de
extratos aquosos de Duranta repens sobre a
germinação e crescimento inicial de Lactuca sativa e
Lycopersicum
esculentum.
Revista
Biotemas
2(23):13-22.
Maguire JA (1962) Speed of germination: aid in
selection and evaluation for seedling emergence and
vigor. Crop Science 2:176-177.
Zhang C, Fu S (2010) Allelopathic effects of leaf litter
and live roots exudates of Eucalyptus species on
crops. Allelopathy Journal 26(1):91-100.
337