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Repellent Effect of Basil (Ocimum spp) on Pea Aphid (Acyrthosiphon pisum Harris) and Potential Use in Crops †


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Synthetic insecticides used for aphid control continue to be a threat to humans and the environment. Therefore, in order to reduce these problems, it is important to use less harmful, environmentally friendly agricultural practices. It is with this objective in mind that the choice behaviour of the pea aphid-Acyrthosiphon pisum Harris (Hemiptera: Aphididae-towards basil odors (Ocimum basilicum L. and Ocimum gratissimum L. (Lamiaceae)) and the broad bean-Vicia faba (Fa-baceae)-was studied using a Y-tube olfactometer. Pea aphid negatively responded to basil plants and spent less time there. The repellent activity and the possibility to use basil as an aphid-repellent plant are discussed in relation to kinds of crops and local conditions.
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Proceedings 2021, 68, x.
Repellent Effect of Basil (Ocimum spp) on Pea Aphid
(Acyrthosiphon pisum Harris) and Potential Use in Crops
Boni Barthélémy Yarou 1,2,3*, Thomas Bawin 2,††, Françoise Assogba-Komlan 3, Armel G.C. Mensah 3 and
Frédéric Francis 2
1 World Vegetable Center, West and Central Africa - Coastal & Humid Regions, 08 BP 0932 Tri Postal, Coto-
nou, Benin; / (B.B.Y)
2 Functional and Evolutionary Entomology, TERRA research center, Gembloux Agro-bio Tech, University of
Liège (ULiège), Passage des Déportés, 2 BE-5030 Gembloux, Belgium; (B.B.Y), (T.B), (F.F)
3 Institut National des Recherches Agricoles du Benin, Programme Cultures Maraîchères, 01 BP 884 Recette
Principale, Cotonou, Godomey, Route IITA, Benin ; B.B.Y : (B.B.Y) fassogbakom- (F.A-K), ( A.G.C.M)
* Correspondence: (B.B.Y)
†† Current address: Department of Arctic and Marine Biology, UiT The Arctic University of Norway, Framstre-
det 39, 9019 Tromsø, Norway; (T.B)
Presented at the 1st International Electronic Conference on Entomology (IECE 2021), 1–15 July 2021;
Available online:
Abstract: Synthetic insecticides used for aphid control continue to be a threat to humans and the
environment. Therefore, in order to reduce these problems, it is important to use less harmful, en-
vironmentally friendly agricultural practices. It is with this objective in mind that the choice behav-
iour of the pea aphid Acyrthosiphon pisum Harris (Hemiptera: Aphididae towards basil odors
(Ocimum basilicum L. and Ocimum gratissimum L. (Lamiaceae)) and the broad bean Vicia faba (Fa-
baceae) – was studied using a Y-tube olfactometer. Pea aphid negatively responded to basil plants
and spent less time there. The repellent activity and the possibility to use basil as an aphid-repellent
plant are discussed in relation to kinds of crops and local conditions.
Keywords: Aphid; basil; volatiles; alternative control; behaviour; pesticide plant
1. Introduction
The pea aphid, Acyrthosiphon pisum H. (1776) (Hemiptera: Aphididae), is one of the
most important crop pests in the world [1]. Due to its feeding behaviour (biting and suck-
ing), the pea aphid cause not only direct (leaf curling, discoloration and plant defor-
mation) but also indirect (transmission of phytovirus and growth of sooty molds) damage
on plant organs [2,3]. As a result, they can cause enormous yield loss on economically
important crops such as peas and beans. It was reported that the damage value reaches
up to hundreds of millions of dollars every year [2]. Synthetic chemical insecticides are
mainly used in agriculture to protect crops from pest attacks, including pea aphids [4,5].
This conventional approach is relatively effective and its ability to maintain pest popula-
tions under economically acceptable levels has been widely demonstrated [6,7]. However,
the intensive use of these synthetic pesticides was reported to induce resistance in several
populations of aphids [8-10]. In addition, synthetic pesticides produce negative effects on
non-target organisms and are very harmful to the environment and human health [10,11].
Environmental-friendly agricultural practices have become attractive in order to
solve these problems. An interesting approach in this respect is the use of pesticide plants
[12,13]. The biocidal activities of various plant families used as extracts, essential oils or
repellent odor sources have been demonstrated on various pest species including aphids
Citation: Yarou B.B., Bawin T., As-
sogba-Komlan F., Mensah A. G.C.
and Francis F. Repellent effect of
basil (
Ocimum spp) on pea aphid
(Acyrthosiphon pisum Harris) and po-
tential use in crops. Proceedings 2021,
68, x.
Published: 30 June 2021
Publisher’s Note: MDPI stays neu-
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Copyright: © 2021 by the authors.
Submitted for possible open access
publication under the terms and con-
ditions of the Creative Commons At-
tribution (CC BY) license (http://crea-
Proceedings 2021, 68, x FOR PEER REVIEW 2 of 8
[14-16]. For example, a lectin isolated from Phycella australis Ravenna (Amaryllidaceae) or
Allium porrum L. (Alliaceae) showed insecticidal activity on A. pisum [17,16]. Deterrent
activity of Laurelia sempervirens Ruiz & Pav. (Monimiaceae) and Drimys winteri Forst &
Forst (Winteraceae) essential oils was reported on the same aphid species [17]. In an agro-
ecological context, the spatial integration of some pesticide plants into main crops is in-
tended to push pests away (i.e., reduce their abundance) or attract natural enemies [18,19].
The repellent activity of Mentha piperita L., Hemizygia petiolata Ashby, Satureja hortensis L.
has been observed on both A. pisum and other aphid species including A. fabae, Sitobion
avenae F and B. brassicae – [20,21].
The genus Ocimum (Lamiaceae) is among pesticide plants with a great interest for
research both to their medicinal proprieties and its biocidal activity on various pests spe-
cies [22-24]. Some studies indicated the quality of Ocimum species as repellent or compan-
ion plants to decrease pest abundance on crops. For example [25], reported that intercrop-
ping Ocimum basilicum and Gossypium barbadense L. (Malvaceae) reduced pest abundance.
The repellent effect of O. americanum L. essential oil was also showed on Agrotis ipsilon
Hufnagel (Lepidoptera: Noctuidae) [26]. In greenhouse [27], demonstrated that the pres-
ence of O. basilicum significantly reduces Trialeurodes vaporariorum W. population on to-
mato plants. Similarly [28], demonstrated that Ocimum gratissimum and O. basilicum re-
duced Tuta absoluta oviposition (Lepidoptera: Gelechiidae) on tomato plants in laboratory
A repellent effect was also reported on cabbage pests including Phyllotreta sinuata
Steph. (Coleoptera: Chrysomelidae), Hellula undalis F. (Lepidoptera: Crambidea), Spodop-
tera litura F. (Lepidoptera: Noctuidae) and Spodoptera littoralis F. (Lepidoptera: Noctuidae)
when the crop was intercropped with Ocimum species [29]. On orchard ecosystem, it was
reported that planting Ocimum species between trees reduce pest level and also attract
natural enemies from the Coccinellidae, Syrphidae, Chrysopidae and Phytoseiidae fami-
lies [30,31]. Intercropping cabbage and O. basilicum [32], noted that there were less aphids
(Brevicoryne brassicae L) on cabbages plants compared to the pure cabbage plots. Repellent
activity of O. basilicum and O. gratissimum was also revealed on Aphis craccivora K., A. fabae
S. and Myzus persicae S. [24]. To our knowledge, very few studies have been carried out
on the biocidal activity of the genus Ocimum on A. pisum. Thus, this work aims to evaluate
the repellent activity of O. basilicum L. and O. gratissimum L. on the pea aphid under la-
boratory conditions using an Y-tube olfactometer.
2. Materials and Methods
2.1. Aphid Rearing
Acyrthosiphon pisum individuals were obtained from stock colonies maintained at the
Functional and Evolutionary Entomology Laboratory, Gembloux Agro-bio Tech, Univer-
sity of Liège (Belgium). Aphids were reared on Vicia faba L. plants under laboratory con-
ditions (25 ± 5 °C; 50-70% relative humidity; 16:8-h light: dark photoperiod) in net cages
(45×45×45 cm, BugDorm, MegaView Science, Taichung, Taiwan). Synchronized winged
aphids (one week old) were used for the experiments under the same laboratory condi-
2.2. Plant Materials
Basil – O. gratissimum and O. basilicum – seeds were provided by the Vegetable Crops
Program of the National Institute of Agricultural Research of Benin (INRAB), West Africa
[28]. Plants were individually grown under a greenhouse (25 ± 5°C, 50-70% relative hu-
midity, 16:8-h light: dark photoperiod) in plastic pots (8×8×9 cm) filled with potting soil
(VP113BIO, Peltracom, Belgium) [28]. The plants were watered every two days and used
in experiments once they reached four and five weeks after seeding for respectively O.
basilicum and O. gratissimum.
Proceedings 2021, 68, x FOR PEER REVIEW 3 of 8
2.3. Behavioral assays (olfactometer tests)
A Y-tube glass olfactometer – 15 cm long stem; 20 cm long arms; 1.5 cm internal di-
ameter (ID) was used to investigate A. pisum behavior under volatiles emitted by the
two basil species. To observe the trajectory of aphids in the olfactometer, black lines were
drawn at two centimeters from the bottom of the stem and from the bottom of the two
arms [33]. Each arm was randomly connected with Teflon® pipes to a sealed glass jar (4L,
20 cm ID) containing either a whole plant of V. faba, O. gratissimum or O. basilicum, or a pot
with soil only. In every case, pots were previously wrapped in aluminum foil to avoid
contamination from the soil. A push air pump (PVAS11; Volatile Assay Systems®, Rens-
selaer, NY, USA) was used to carry volatiles from the glass jar to the olfactometer arms.
To avoid any outdoor contamination, air was first purified through a charcoal filter before
entering the jars. Airflow through each of the arms was maintained at 100 mL.min-1.
Two experiments were carried out, during which the effect of O. gratissimum or O.
basilicum on aphid behavior was investigated. In the first experiment, two modalities were
tested: a) a pot with soil alone versus a single O. gratissimum plant and b) a single V. faba
plant versus a single O. gratissimum plant. In the second experiment, O. basilicum was used
in place of O. gratissimum. Aphids were individually introduced into the stem part of the
olfactometer and their position was recorded during three minutes. If an aphid remained
inactive (i.e. did not move from its position) during that time, it was removed from the
experiment. If an aphid crossed the line delimiting the stem area but did not cross the line
marked at the bottom of one of the arms, it was considered as not responding. If an aphid
crossed the line marked on one of the arms before the end of the three minutes, it was
considered as responding (i.e., made a choice). Every 10 aphids, the glass jars and the
olfactometer were cleaned with pure n-hexane (>99.7%; VWR®, Radnor, PA, USA) and
dried at room temperature for five minutes. The odor sources (potting soil and plants)
were replaced and the position of the jars was switched to avoid bias. The choice of aphids
was determined by (a) the first entered zone, (b) the zone where the aphids stayed for the
longest time period, (c) the last entered zone [34] and (d) the number of visits in each
olfactometer arm. Sixty aphids were tested for each modality. All assays were carried
out under laboratory condition (24±1˚C temperature, 45±5% RH, uniform lighting in the
observation chamber) as described by [33].
2.4. Statistical analyses
The observed frequencies related to the choice of aphids in the olfactometer were
analyzed by using a two-sided binomial test. The non-parametric Wilcoxon statistical test
(function: “wilcox.test”) was used to compare the mean time spent by aphids in each arm
of the olfactometer according to the different treatments. The significance level was p<0.05
for all analyses. Statistic tests were performed using R software version 3.3.6 [35].
3. Results
For each comparison, at least 85% of the 60 tested aphids responded (Table 1). Over-
all, aphids significantly directed towards potting soil rather than plants of the two basil
species (Figures 1 and 2). However, a significantly higher number of aphids directed to-
wards V. faba when opposed to O. basilicum (Figure 1), but not when opposed to O. gratis-
simum, although a similar trend was observed (Figure 2). Consistently, the average resi-
dence time of an aphid in the soil pot area or the V. faba plant area was significantly higher
than in the O. basilicum area, whereas it only was the case with O. gratissimum when com-
pared to potting soil (Figure 3).
Proceedings 2021, 68, x FOR PEER REVIEW 4 of 8
Table 1. Responding aphid numbers in dual choice tests including Ocimum plants.
Comparison Responding aphids (%)*
Potting soil versus Ocimum gratissimum 51 (85)
Vicia faba versus Ocimum gratissimum 55 (92)
Potting soil versus Ocimum basilicum 53 (88)
Vicia faba versus Ocimum basilicum 58 (97)
*Responding insects include living individuals present in one of the two side areas of the olfac-
Figure 1. Aphid distribution in dual choice tests including Ocimum basilicum plants in Y olfactom-
eter set-up. ** = p < 0.05, *** = p < 0.001.
Figure 2. Aphid distribution in dual choice tests including Ocimum gratissimum plants in Y olfac-
tometer set-up. ** (p < 0.05), *** (p < 0.001), ns (not significant).
Proceedings 2021, 68, x FOR PEER REVIEW 5 of 8
4. Discussion
Pesticide plants like basil are known to have a repellent effect on crop pests. The re-
pellent activity of O. gratissimum and O. basilicum was tested on A. pisum using a Y-tube
olfactometer. The results of this study showed that basil species tested were effective
against A. pisum. Overall, aphids chose the olfactometer arm connected to a V. faba plant
or a soil pot and stayed there longer. However, the activity varied depending on the Oci-
mum species. Using dual-choice behavioural assays performed in flight tunnels and crop
association design, a repellent effect of O. basilicum and O. gratissimum was reported on
aphids – Aphis craccivora Koch, Aphis fabae Scopoli, Myzus persicae Sulzer – [24]. The repel-
lent effect of O. basilicum was also observed on A. fabae in the laboratory (wind tunnel) and
the field (strip cropping) condition as exemplified by the low infestation level of V. faba
plants associated with basil plant [20]. [32], noted that there were less aphids Brevicoryne
brassicae L. – on cabbage plots intercropped with O. basilicum compared to pure cabbage
plots. Other experiments also demonstrated a good repellent activity of basil species on
other pests such as Agrotis ipsilon H. (Lepidoptera: Noctuidae) [26], Trialeurodes vaporario-
rum W., (Hemiptera: Aleyrodidae) [27], Hellula undalis Fabricius, Spodoptera littoralis B.,
Tuta absoluta M. [28,29]. Essential oils of O. basilicum was found effective on Tetranychus
urticae K., Bemisia tabaci G. or Planococcus ficus S. [36,37].
Figure 3. Duration (mean+SE) spent in each olfactometer arm by Acyrthosiphon pisum aphid includ-
ing Ocimum plants. Ob-Sp (O. basilicum versus soil pot), Ob-Vf (O. basilicum versus V. faba), Og-Sp
(O. gratissimum versus soil pot), Og-Vf (O. gratissimum versus V. faba).
Behavioural evaluation of the A. pisum in the presence of V. faba (host plant of this
aphid) alone as a source of odor would have made it possible to better assess the repellent
effect of basil. However, based on previous reports on the repellent effect of Ocimum spe-
cies on several pests, we can suggest that the two basil species tested in our study have a
repellent activity on A. pisum. Differences in repellency are generally due to the fact that
volatile organic compounds can vary within the same species [38] or between different
plant species of the same family [28]. Thus, the relatively lower repellency level of O. gra-
tissimum on A. pisum compared to O. basilicum could be attributed to a difference in the
organic volatile compounds released by this species and O. basilicum.
Proceedings 2021, 68, x FOR PEER REVIEW 6 of 8
In crop pest management, Ocimum can be used as source of essential oil or associated
with other crops. In the Western context, where vegetables are mainly produced in the
greenhouse, the essential oil of Ocimum can be used as a diffuser in pest management.
However, in developing countries like West African countries, where market gardening
is practiced in open field, the use of essential oil diffusers cannot be generalized among
producers for the moment. In addition to the lack of availability and affordability of es-
sential oil for producers, it is necessary to evaluate the release dynamics of these essential
oils in West African climatic conditions. This will allow to assess the effectiveness of Oci-
mum based essential oils under natural conditions before recommendation of their use for
pest management in open field. Besides, using Ocimum in the intercropping system could
be the most appropriate strategy. Indeed, Ocimum is widely used as a vegetable in this
part of Africa [39] and its integration in crop association systems should not be a problem.
In many cases, crop association appears to be an advantageous agroecological practice for
producers in terms of pest management and crop yield improvement [40]. Also, the pres-
ence of diverse families of entomophagous beneficials on Ocimum crop [41,42] could be
valued in terms of ecosystem services as biological control provided by natural enemies
and pollination for yield improvement [25,43].
5. Conclusions
Ocimum basilicum was the species that induced a stronger repellent effect on A. pisum
compared to O. gratissimum. Further studies are needed to validate these results under
field conditions.
Author Contributions: Conceptualization, B.B.Y.; methodology, B.B.Y.; software, B.B.Y.; valida-
tion, F.F.; formal analysis, B.B.Y.; investigation, B.B.Y.; data curation, B.B.Y.; writing—original
draft preparation, B.B.Y.,T.B.; writing—review and editing, T.B., A.G.C.M, F. A-K and F.F.; super-
vision, F.F.. All authors have read and agreed to the published version of the manuscript.
Acknowledgments: This research was supported by Erasmus Mundus Program of the Dream
ACP project. The authors would like to thank Mathieu Ahenan for reading the manuscript.
Conflicts of Interest: The authors declare no conflict of interest.
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