THE PRESENCE OF INCLUSION BODIES OF Helicoverpaarmigera NUCLEAR POLYHEDROSIS VIRUS (HANPV) IN THE MIDGUT TISSUE OF FOURTH
INSTARS Spodopteralitura LARVAE
MIA MIRANTI, MUHAMAD RULIAWANSALIM, AND YASMIPURNAMASARIKUNTANA
Department of Biology, Faculty of Mathematics and Natural Sciences, University of Padjadjaran.
Jl. Raya BandungSumedang KM-21 Jatinangor, Indonesia Contact Person : miamiantariksa@yahoo.com
ABSTRACT
The research to determine the presence of inclusion bodies of HaNPV in the midgut tissue of fourth instars Spodopteraliturae larvae has been done. The larval has been infected by 1.13 X 105polihedra/ml concentration of viruses and the presence of inclusion bodies has been conducted every 8 hours from the 1st until 72nd hours after virus infection. The histological slide of midgut tissue of larvae has been stained by Mallory-Azan staining method. The experiment shows that in 8th hours after infection, the inclusion bodies of virus presence in columnar cells of larvae. In 24th hours after infection, inclusion bodies of virus through disgestive tract and appeared in faeces of larvae. In 32nd hours after infection, the inclusion boddies of virus still appeared in regenerative columnar cells until 56th hours after infection. After 64th hours until 72nd hours after infection, the present of polihedra damaged of midgut tissue, made all columnar cells lysis and inclusion bodies form into granul (polyhedral granules). Therefore HaNPV can replicated in Spodopteraliturae larval as an alternative host.
Keywords :Helicoverpaarmigera Nuclear Polyhedrosis Virus (HaNPV),
Spodopteralitura, Inclusion Bodies, Histological midgut structure,
Mallory Azan Staining Method.
INTRODUCTION
Helicoverpaarmigera Nuclear Polyhedrosis Virus (HaNPV) isolated from
cannibalism. Otherwise, in vivo production of HaNPV need another host as an alternative host.
HaNPV can infect another insect larval as Spodopteralitura,
Spodopteraexiguaandcrocidolomiapavonana (Miranti, 2008). The virus is ideal to use
as biological agent to control the population of these insects. Spodopteralitura is an ideal host for HaNPV production because the inclusion bodies can formed perfectly (Miranti and Wardono, 2009). In this research, the presence of inclusion bodies of
HaNPV will observed in histological slide of midgutof S. litura larvae at 1st hour until 72nd hours after larval infection.
MATERIAL AND METHOD
HaNPV collected from Laboratory of Microbiology, Department of Biology,
Faculty of Mathematics and Natural Sciences, University of Padjadjaran,
Bandung-Indonesia is in cadaver of H. armigera larvae. The suspension of virus has been made with Indrayani, et al., (1993) method with modification. Larval fourth instars of S. litura collected from BalaiPenelitianSayuran, Lembang, West Java-Indonesia.
The inclusion bodies of the virus was 1 X 105 Inclusion Bodies/ml concentration. Every 1 ml of virus suspension mixed with 30 gram of kernel sweet corn.
S. litura larval infection within orally method. The histological slide of midgut tissue of
larval body was made every 8 hours from the 1st until the 72nd hour after larval infection, through Mallory-Azan staining method. The inclusion bodies of virus observed with light microscope at 400X.
RESULT
The experiment shows that in the histological slide of midgut tissue, the inclusion bodies of HaNPV can be observed. At 1st hour after larval infection, the inclusion bodies of virus is disappear from the midgut. In the 1st hour after larval infection, the polihedrin of virus is lysis and liberated the virions. The virionscan not be presence in midgut tissue (Flipsen, 1995). They formed a budding virus in midgut cells.
digestive tract and appeared in faeces of larvae. In 32nd hours after infection, the inclusion boddies of virus still appeared in regenerative columnar cells until 56th hours after infection. Hawtin, et al., (1992), and Sudhakar and Mathavan (1999) said that the presence of the inclusion bodies shows that virus can replicated in S. litura perfectly. After 64th hours until 72nd hours after infection, the present of polihedra damaged of midgut tissue, made all columnar cells lysis and inclusion bodies form into granul (polyhedral granules). The midgut cells were lysis and the inclusion bodies liberated to the hemocoel. The midgut cells damaged and the colour of the hemocoel is whitegrey. The presence of the inclusion bodies shows that HaNPV can replicated in
Spodopteraliturae larval as an alternative host.
CONCLUSION
The presence of polyhedral of HaNPV in the body of S. litura larval shows that
S. litura can be used as an alternate host for HaNPV in vivo production.
ACNOWLEDGEMENT
This research was supported in part by HibahBersaing Grant 2008-2010, DirjenDikti, The Ministry of National Education, Republic of Indonesia.
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