IJERD – International Journal of Environmental and Rural Development (2020) 11-2

Research article
erd

Combination of Entomopathogenic Fungi with Essential Oilfor Controlling Bean Fly, Ophiomyia phaseoli Tryon(Agromyzidae: Diptera)
MALEE THUNGRABEAB
Agricultural Technology Research Institute, Rajamangala University of Technology Lanna,
Lampang, Thailand

PATCHARIN KRUTMUANG*
Department of Entomology and Plant Pathology, Faculty of Agriculture, ChiangMai University,
ChiangMai, Thailand
Email: patcharink26@gmail.com
Received 6 March 2020

Accepted 12 October 2020(*Corresponding Author)

Abstract The objective of this study was to evaluate the compatibility of entomopathogenic
fungi with repellants to control bean fly, Ophiomyia phaseoli Tryon (Agromyzidae: Diptera).
Nine isolates of entomopathogenic fungi belonging to 3 genera and 5 species were screened
for pathogenicity against bean fly. The results found that Beauveria bassiana isolate Bff andMetarhizium anisopliae isolate Mff caused 100% mortality to both its larval and adult stages.
The repellency of 7 essential oil was also evaluated against adult bean fly in the laboratory.
The results showed significant differences in repellency among essential oil andconcentrations. Essential oil extract from pomelo peeling was the strongest repellant followed
by eucalyptus and sweet basil. However, essential oil extract from camphor was the weakest
repellant. On the other hand, these seven essential oils (pomelo, zingiber, citronella grass,eucalyptus, camphor, kaffir lime and sweet basil) at a concentration of 0.1% had no effect on
the viability of conidia, but at higher concentrations it was found that citronella grass andzingiber reduced viability of B. bassiana. The results on the effect of essential oils on colony
growth showed the same tendency as the one on the viability of conidia.
Keywords bean fly, entomopathogenic fungi, essential oil, repellent

INTRODUCTION
Phaseolus vulgaris L., French bean is a herbaceous annual plant in the Fabaceae family, it grows to
between 20-60 cm high; and twining, climbing vines up to 2-5m long. The stems of bushy types arerather slender, pubescent with many branches (Chávez-Servia et al., 2016). In twinning types, thestems are prostrate for most of their length and rise toward the end. The leaves, borne on long green
petioles, are green or purple in colour and trifoliate. Leaflets are 6-15 cm long and 3-11 cm broad.
The inflorescences are axillary or terminal, 15-35 cm long racemes. The flowers are arranged in pairs
or solitary along the rachis, white to purple and typically papillonaceous. Once pollinated, eachflower gives rise to one pod. Pods are slender, green, yellow, black or purple in colour, sometimes
striped (Caproni et al., 2019) . They can be cylindrical or flat, straight or curved, 1-1.5 cm wide andup to 20 cm in length. The pods may contain 4 to 12 seeds. The seeds are 0.5-2 cm long, kidneyshaped and highly variable in colour depending on the variety: white, red, green, tan, purple, gray orblack.
French bean is a major plant of Royal Project Foundation for the growers of organic vegetablesystem in Thailand. However, the problem of French bean plantation is bean fly, Ophiomyia phaseoli
Tryon (Agromyzidae: Diptera) (Ojwang et al., 2010) .They lay eggs in the leaf tissue or directly on
the stem. Early signs of attack are egg laying punctures on the primary leaves which tend to beconcentrated around the leaf base (Letourneau, 1994). Eggs hatch into small white maggots which
migrate down the stem to the root zone where they pupate into brown puparia. Severe damage isⒸ ISERD

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IJERD – International Journal of Environmental and Rural Development (2020) 11-2

indicated by wilting and dying of seedlings. The attack disrupts nutrient transportation, causing thetap root to die (Ssekandi et al., 2016). The plant attempts to recover by forming adventitious roots
above the damaged area (Ojwang et al., 2010). Young seedlings under stress wilt and die within ashort time. Older and more vigorous plants may tolerate the damage but become stunted and will
have reduced yield. Control strategies for grower to use were as follow: application of botanicalpesticides, natural enemies, sticky trap and also chemical seed dressing with systemic insecticides.
Plutachis sp. Eulophid, Biosteres sp. Pteromalid sp. nematode and Oecophylla smaragdina Fabricius
were effective against bean fly by 52% but bean fly still serious pest of French bean plantation(Jaramillo et al., 2013).
OBJECTIVES
The objectives of this study were: firstly to screen promising entomopathogenic fungi against bean
fly and secondly to evaluate plant-based insect repellants and their compatibility with theentomopathogenic fungi in controlling bean fly.
MATERIALS AND METHODS
Selection of Entomopathogenic Fungi from Collection
Nine isolates of entomopathogenic fungi consisting of 3 different genera and 5 different species wereselected from Agricultural Technology Research Institute, Rajamangala University of Technology
Lanna, Lampang 52000, Thailand. These were screened for their efficacy against bean fly underlaboratory condition.
Selection of Plants as Repellants Against Bean Fly
Selection of plants that have potential to repel bean fly was conducted according to the literature
reviewed that hinted on the plants that have essential oils that repel invertebrates (Showler, 2017 andAbtew et al., 2015). Extraction of essential oil was done using the Steam-distilled method(Mcguinness, 2003). After that, Study on the efficacy of essential oil that repel bean fly at difference
concentration was conducted. Moreover, study on effect of essential oil to germination ofentomopathogenic fungi and study effect of essential oil to growth of entomopathogenic fungi wasalso conducted (Liu, 2012).
Efficacy of Formulation of Entomopathogenic Fungi Combine with Repellant
This was conducted with the aim to produce an efficient formulation of entomopathogenic fungi forcontrolling bean fly. The formulation was produced by mixing 20% of entomopathogenic fungi +1% of Tween 80 at a concentration of 10% + 1% of Glycerin + 5% vegetable oil + 3% Propylene
glycol (Malee, 2015). This was then applied on French bean leaves to assess the spore viability andefficacy of formulation of entomopathogenic fungi against bean fly. The experiments were carriedout in 3 treatments: formulation 1 (Beauveria bassiana), formulation 2 (Metarhizium anisopliae) andcontrol. Suspension was directly sprayed on to the bean fly.
RESULTS AND DISCUSSION
Efficacy of Entomopathogenic Fungi to Infected Bean Fly
Nine isolates of entomopathogenic fungi belonging to M. anisopliae, B. bassiana Isaria
fumosoroseus, Isaria tenuipes and Isaria farinosus were selected for this study. The results forpathogenicities of 9 isolates of these entomopathogenic fungi showed high virulence against larvalⒸ ISERD

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IJERD – International Journal of Environmental and Rural Development (2020) 11-2and adult stage of bean fly. It was observed that B. bassiana isolate Bff and M. anisopliae isolate
Mff caused 100% mortality to both larval and adult stages of bean fly as shown in Table 1. This wasa very impressive result and is in agreement with several studies (Letourneau, 1994). The otheradvantage of these entomopathogenic fungi is that they offer endophytic advantages as well to theplanted when they are inoculated (Mantzoukas and Eliopoulos, 2020).
Table 1 Percent mortality of entomopathogenic fungi to bean fly (larval stages)
Entomopathogenic fungi*
Metarhizium anisopliae

Beauveria bassiana

Isaria fumosoroseus
Isaria tenuipes
Isaria farinosus

Isolates

Percent mortality

Ma.4849
Ma.7965
Mff
Bb.5335
Bb.4591
Bff
Pfu.2507
Pt.6073
If 2549

80.00
93.33
100.0
80.00
80.00
100.0
60.00
46.67
40.00

*Fungus isolates from Agricultural Technology Research Institute, Rajamangala University ofTechnology Lanna, Lampang

Efficacy of Plants as Repellants Against Bean Fly

Repellents (%)

The repellency of 7 essential oil was evaluated against adult bean fly in the laboratory. The results
showed significant differences in the repellency among essential oils in different concentrations.
Essential oil extract from peel of pomelo was the strongest repellent followed by eucalyptus andsweet basil. Essential oil extract from camphor was observed to be the weakest repellent as indicatedin Fig. 1. In addition, the seven essential oils (pomelo, zingiber, citronella grass, eucalyptus,camphor, kaffir lime and sweet basil) a concentration of 0.1% had no effect on the viability ofconidia, but in higher concentrations, it was observed that citronella grass and zingiber reduced
viability of B. bassiana. The results on the effect of essential oils on colony growth showed the sametendency as shown in the viability of conidia (Fig. 2). These results did not deviate much from otherstudies (Liu, 2012). Compatibility of these EPFs is known to be of paramount effect on theireffectiveness and hence cannot be omitted (Neves et al., 2001).

Fig. 1 Percent efficacy of plants as repellents against bean flyⒸ ISERD

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IJERD – International Journal of Environmental and Rural Development (2020) 11-2

0.10% Beaveria

0.10% Metarhizium

1% Beaveria

1% Metarhizium

Inhibition (%)

Inhibition (%)

zingiber

eucalyptus

sweet basil

camphorcitronella grass

kaffir lime

pemeloFig. 2 Percent inhibition of plant repellent to Beauveria spp. and Metarhizium spp.
Efficacy of Formulation of Entomopathogenic Fungi Combine with Repellent
Concerning the potentiality of formulation of entomopathogenic fungi in combination with essential
oils against bean fly, it was observed that when entomopathogenic fungi combine with essential oil
there was increased effectiveness in controlling bean fly. This is an interesting finding as they would
complement without adverse effects. Ali et al (2018) also worked with eucalyptus andentomopathogenic fungi observed remarkable potential in their compatibility and the resultantefficacy.
CONCLUSIONSOut of 9 isolates, 2 isolates i.e., B. bassiana isolate Bff and M. anisopliae isolate Mff showed thehighest mortality against both larval and adult stages of bean fly. Formulation of entomopathogenic
fungi took time to ablate from French bean leave which was very impressive. Essential oil extractfrom peel of pomelo was a stronger repellent followed by eucalyptus and sweet basil. However,essential oil extract from camphor was a weaker repellent. It was also observed that a concentrationof 0.1% had no effect on the viability of conidia, but at higher concentrations it was found thatcitronella grass and zingiber reduced viability of B. bassiana. The results on the effect of essential
oils on colony growth showed the same tendency as the one on the viability of conidia.
Entomopathogenic fungi combined with essential oil had high potential in controlling bean fly.
ACKNOWLEDGEMENTS
This research was funded by Highland Research and Development Institute (Public Organization).
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