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The pathogenicity of entomopathogenic nematodes against Spodopteraexigua

Article in Journal of Engineering and Applied Sciences · December 2017

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 VOL. 12, NO. 24, DECEMBER 2017 ISSN 1819-6608
ARPN Journal of Engineering and Applied Sciences
©2006-2017 Asian Research Publishing Network (ARPN). All rights reserved.

[Link]

THE PATHOGENICITY OF ENTOMOPATHOGENIC NEMATODES

AGAINSTSpodopteraexigua
Dyah Rini Indriyanti, Baharuddin Achmad Fauzi and Yoris Adi Maretta
Biology Department, Faculty of Mathematics and Natural Sciences, Universitas Negeri Semarang,
Jalan Raya Sekaran, Gunungpati Semarang, Indonesia
E-Mail: dyahrini36@[Link]

ABSTRACT
Spodopteraexigua is an insect pest that threat to a wide range of agricultural crops. Entomopatho genicnematodes
are the parasitic nematodes that have the ability to attack the insects pest. The study to analyzed the pathogenicity of the
entomopathogenic nematodes (EPN) from Steinernematidaein controlling the third instar larvae of S. exigua. The study’s
design was completely randomized design (CRD) with six treatment groups and four times of repetition (10 larvae at each
repetition). S. Exigua were taken from the field, andmass breedingwas doneat Laboratoryfor two [Link] was
isolated from organic soil. EPN breeding carried out using a Tenebriomolitor [Link] larvae of S. exigua were
used for Bioassay test. The number of larvae killed by EPN was recorded for 96 [Link] data analysis used probit
analysis from Minitab 1.5 software. The results showed that EPN gave positive response against larvae of S. exigua within
96 hours. The conclusion was LD90 of EPN against the larvae of S. exiguawas 772 IJ/2 ml.

Keywords: pathogenicity, LD90 -96hours, Entomopathogenic Nematodes, Spodopteraexigua.

INTRODUCTION To overcome the negative effect of using chemical

The red onion (Allium cepa) is known as pesticide, it is important to conduct a study on controlling
ingredients to make spices taste in food. One of the main S. exiguapest using an effective and environmentally
production areas on A. cepa in Indonesia is Brebes, friendly biocontrol agent. One of biocontrol agent that can
Central Java. The harvested area is 30954 Ha with be used is Entomopathogenicnematode (EPN). EPN is
production of onion reached 3759742Kwintal and in endoparasiticnematodeespecialy for the insects. EPN used
average was 121.46 Kw/Ha (Brebes District’s Central as biocontrolcome from the family of Steinerematidae and
Bureau of Statistic, 2017). A. cepa from Brebesare well Heterorhabditidae (Boemare et al, 2002; Raucha et al.,
known for their good quality. They are better than A. cepa 2017; Indriyanti et al., 2017).
from other places in Indonesia and even from other Steinernematidae and Heterorhabditidae are
countriessuch as Thailand and China. High economic potentialbiocontrol agents for various pest insects (Ehlers,
value of A. cepa liven up the Farmers interest on 1996). Both of them are effective on contolling pest insect
cultivating A. cepa. However, there werestill many from Lepidoptera, Coleoptera, and Diptera for 24 to 48
problems oncultivatingA. cepa such as the viral diseases hours (Chaerani, 1996) and it is also safe for non target
(Gunaeni et al., 2011), and pest attacks (Basuki, 2014), organisms (Grewal and Richardson, 1993). However, there
including attack from Spodopteraexigua (Marhaen et al. is no information about controlling S. exigua with EPN.
2016; Herwanto et al., 2012). [Link] is aninsect pest So, it is importantt to conducted a study in order to
that usually attacksgrowingplant in Asia, Europe, Africa, determine the effective dosage of EPN (LD90) to control S.
Australia, and America (Agata et al., 2005). S. exigua exigua in Laboratory.
larvae actively attack the A. cepa leaves, especially the
young leaves. The damage can reach 100 percent if not MATERIAL AND METHODS
controlled (Abdi, 2003).
The explotionof S. exigua population at field Mass breedingof S. exigua
drives thefarmers to use chemical pesticide excessively Mass breeding of S. exiguawas conducted at
(Djojosumarto, 2008; Suhartono, 2010). Using chemical Biology Laboratory of Universitas Negeri Semarang. The
pesticide excesssively lead to the distruction of larvae were obtained from onion farmingin Brebes
environment and health problems.S. exigua population in District. They werereared for two generations at
Brebeshas been exposed some pesticides including laboratory. Third instars larvae were used to Bioassay test.
hydrochloride, deltramethrin, methoxy fenozide and
phyraclotose (Moekasan and Basuki, 2007;Wibisono et Isolating and breeding EPN
al., 2007). EPN was isolated from organic soil, it was
Continuous use of pesticides can cause health isolated by the following step: digging the organic soil in
problems, Fikri (2010) found that the there was an arsenic the field as deep as 20 cm, took 250 g of organic soil, and
content in the urine of Farmer in Brebes who sprayed put into container (6 cm in height and 13 cm in diameter).
pesticide everyday, indicating that it was harmful to Tenebriomolitor caterpillar was used to trap the
human being. EPN from soil for 5-7 days. After T. molitor was infected

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 VOL. 12, NO. 24, DECEMBER 2017 ISSN 1819-6608
ARPN Journal of Engineering and Applied Sciences
©2006-2017 Asian Research Publishing Network (ARPN). All rights reserved.

[Link]

by EPN. EPN were reared using T. molitor caterpillar as a P1 = EPN population density 750 JI/2 ml aquadest
host in white trap method. After the process of rearing, P2 = EPN population density 800/2 ml aquadest
EPN were harvested, then used for pathogenicity test P3 = EPN population density 850/2 ml aquadest
(Indriyanti&Muharromah, 2016). The introduction test P4 = EPN population density 900/2 ml aquadest
was conducted to determine the range of [Link] range P5 = EPN population density 950/2 ml aquadest
doses of LD90 for 96 hours EPN was on 800 Jl-1000Jl/2
ml. Data analyzed
The S. exigua mortality data for 96 hours was
EPN pathogenicity test analyzed probit test using Minitab 1.5 program.
Effectiveness test was conducted to determine the
LD90 for 96hours. Research design was Complete Random RESULT AND DISCUSSIONS
Design (CDR), consisting of 6 treatments (P0-P5) by 4 The death of S. Exigualarvae caused by EPN was
times repetition (the number of larva in each repetition characterized by changingin the body color from green to
were 10). There were 24 unit experiments, total larvae dark brown. The body texture of larvae was also became
were 240 (Kamariah et al., 2013). The treatmentstest [Link] use of third instar of S. exigua larvae was
arrangement wereas follow: because it is the most active larvae (Hadi and Soviana,
2000). The result of pathogenicity test is presented on
P0 = EPN population density0 JI (Juvenile infective)/2ml Table-1.
aquadest (Control)

Table-1. S. exigua mortality for 96 hours.

Treatment
Repetition P0 P1 P2 P3 P4 P5
(0 JI/2 ml) (750 JI/2 ml) (800 JI/2 ml) (850 JI/2 ml) (900 JI/2 ml) (950 JI/2 ml)
1 0 7 10 10 10 10
2 0 8 8 10 10 10
3 0 8 10 10 10 10
4 0 10 10 10 10 10
Total 0 33 38 40 40 40
Average 0 8.25 9.5 10 10 10
Percentage 0 82.5 95 100 100 100

Table-1 shows that the rate of death larvae ofS. It was suitable with Wagiman et al. (2001) who stated that
Exigua was diversed based on EPN population density. the superiority of EPN as a biocontrolcan kill insects
More density of EPN given wasresulting in higher rate of through hemolymphquickly (24-48 hours).That ability is
larvae mortality. This result is suitable with Iskandar et al. caused by symbiont bacteria which come out from EPN
(2003) & Kamariah et al. (2013) statement that the more after penetrate into the insect (Kaya, 1993).
density of EPN population speed up the rate of death Steinernematidae family has a symbiosiswith bacteria of
larvae. Xenorhabdusspp. Meanwhile, Heterorhabditidae has
Ninety hours after theapplication, the larvae symbiosis with bacteria of Photorhabdussp (Boemare et
mortality formtreatment P3, P4, and P5 had been reached al., 2002). That bacteria release sometoxins (eksitoxin and
100%. EPN (pathogenicity) effectiveness test for 96 hours endotoxin) like protease, lipase, pectinase. That
showed that values of LD90 on P1 and P2 were 82.5% and combination of bacterial toxins cause the insect die faster
95% respectively. The data of S. exigua mortality was (Dowds, 1998). However, the identification of EPN
counted and EPN population density for each treatment symbiont bacteria was not performed in this research.
was analyzed using probit test (software Minitab 1.5) to The symptoms of S. exigua larval mortality were
determine the value of LD90 and the result obtainedwas the changing in color of body from light green to be dark
772 Jl/2 ml. The result was different with the result of yellow, brownish or blackish, then themushy body texture,
study on [Link] by Uhan (2006). EPN population density production of liquid and no respond of any touch. This
to kill [Link] samples using 95% (pathogenicity) condition indicated that the larvaehad been infected by
effectiveness reached 800 Jl/ml. It is because S. Exigua EPN from Steinernematidae family. It was in accordance
and [Link] are two different species. So, it is possible if with the study of Simoes and Rosa (1996) which showed
the effective EPN population density needed was different. that the larvae that were attacked by Steinernematidae had
Increasing mortality of S. exigua was showedsignificantly, a mushy dark brown body and produced a little liquid.
it indicated that EPN have an ability to kill in a short time.

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ARPN Journal of Engineering and Applied Sciences
©2006-2017 Asian Research Publishing Network (ARPN). All rights reserved.

[Link]

Djojosumarto P. 2008. Pestisida dan Aplikasinya. Jakarta :

Agromedia Pustaka.

Dowds B C. 1998. Bacterial Virulence Mechanisms.

European Cooperation in the Field of Scientific and
Technical [Link]. 819. pp. 9-16.

Ehlers R U. 1996. Current and future use of nematodes in

biocontrol: practice and commercial aspects with regard to
regulatory policy issues. Journal of Biocontrol Science and
Technology. 6(1): 303-316.

Fikri E. 2010. Hubungan Paparan Pestisida Dengan

Figure-1. Larvae of S. exigua after 96 hours of Kandungan Arsen (As) dalam Urin dan Kejadian Anemia.
application EPN. Jurnal Kesehatan Lingkungan Indonesia. 11(1): 29-37

The EPN killing ability was not only determined Grewal P S and Richardson P N. 1993. Effect of
by symbiosis between EPN bacteria simbion but also S. application rates of Steinernema feltiae on biological
exigua ability onself-defence. Ehler (1996) stated that control of the mushroom fly Lyccoriella auripila
EPN killing ability to host insects is not only determined (Diptera: Sciaridae). Journal of Biocontrol Science and
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Gunaeni, N.1, A.W. Wulandari, A.S. Duriat& A. M. 2011.
CONCLUSIONS Insiden Penyakit Virus Tular Umbi pada Tigabelas
Based on the result of the study, it can be Varietas Bawang Merah Asal Jawa Barat dan Jawa
concluded that EPN was effective on controlling S. exigua Tengah. J. Hort. 21(2): 164-172.
pest in A. cepa with the value of LD90for 96 hours was 772
Jl/2ml. Hadi K U dan Soviana S. 2000. Ektoparasit:Pengenalan,
Diagnosis dan Pengendaliannya. Bandung : IPB.
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