Pages from Proceeding Development in Asia and Africa %28Yulia%29
Pra.eedlng, of the
3d l^tetnil1lo^alIeettng lor trw De1t,etopne t of Ip ttt Astd.
altrl
.
$499!-*t*!!4!g!ang) I^!9^esla Decenber 7-9, 2ooe
CSoUoi 0?o\
tOooc4C
Isolatlon ofA Tetravirus from Setothose a rrsignlr Laiwae
Iafected wlth Aa Epldemic Disease In Oil palm plaatatloa
Ia South Sumatra, Indoaesia
Mada Sugltartit, Yulta Pujiaatuti2, Hisanori Balrdol
r. l,aboratory of Applied Molecular Entomolog/, Graduate Schoot of
Agriculture,
Hokkaido Universitt, Japair
2 Department of Plant Pests and Diseases, paculty of Agriculture.
Sriwijaya Universitr-, Irrdonesia
Iltroduction
La.rvae of nettle caterpillai, Sefolhosea asigna, Setora niteiLs, Dan& tima and
Dama Brdd.leqi lLEpidoptera ; Limacodidael, ar.e s],rnpatric and coseasonal; Iimacodid
moths in Southeast Asia and being important defoliator of oil palm arrd coconuts
palms. In severe infestations, SetotLbsea osigna (S.asig4a) larvae consume all
foliage and leave only mid rib of t}te frond, causing sethack oI fruit p.oduction lor a
year. Thus, S. osrgna become an economicaliy impodant defol;ator in oil palm
(Elaeis Wieensis) plantation in norih easteril Surl}atra, Bo.neo aId weitem
Malaysia. Recufing outbreak of S. aslgna, singly, or together wjth heterosF,ecific
limacodids, prcvoked application of broadspectnrm insecricides. However, recently,
the endronmental pollution by agricultural chemical becomes a big social problem in
many countries, and Indonesia is no exception.
Recentiy, natural epidemics among S. dsiqna lar-vae suspected of viral infection
have been occurred in some oil palm 6elds ir] Indonesia. These infectjous diseases
resulted in decrease of pest populatio[ below arl economic tireshold in those
epidemic areas. Suggesting the possibility oI an ellective control of S.astgna by viral
pesticides isolated from epidemic field. However, Iittle is known yei about the
causing agent of ratural epidemic disease among S. osigna observed irr lndonesia
Result and Diacuaaion
S asigna were collected in an epidemic field arrd used
as a starti]:lg material for puri$ing viruses. The existence of RNA with the
sequence homologr at least partly to the RdRp sequence of TaV was
demonstrated by RT-PCR followed by sequence analysis. ihis result suggested
that TaV or Tav-related virus is existed in the infected larvae. purification of
virus particles was then perfo.med by a method described for Dendrolimus
punctatus tetravirus (DpTV) purification. by Fuming yi et a1 (2005). On a sucrose
density gradient of paitialy purified vlal fraction, only one white band was
Diseased larvae of
observed. The vrhite band was collected, precipitated by ultra centrifugation, arrd
exarnined witl electron microscop) .
Negatively stain elec&on microscopy shov/ed tl.at the fraction contained
numerous numbers of non-enveloped, spherical virus-like particles with about
40 nrn in dilmeter. The morphological chatacteristics of the purified particles
resembled thos€ of members of the farrlily Tetrauirid.ae. The cDNAs against RNAS
extracted ftom the puritred viral particles were generated with raadom primers
arrd sequenced usi[g an automatic sequence!. The viral RNA showed a
sirilarity of about 99% in total with that ol Thosea o_signa.vitts (TaV), a member
oI the gel]ns Betatetra.arus previously reported.
74
u;
-E
G!
-ir
Proceedtngs of
Jor t tc DaeloPneftt ol lP tn A,la
Indotesla Deceatbc" 7'9, 2OO9
I'aflpu^g,
dnd Afrlca. B.1tddr
tle 3d Intarndtl6ndl lleeting
lnT
i:3'lre l.
Negattve stain elect.on microscopy oI viral particles purified by sucrc'se
deisity gradient centrifugation' Rod shaped particles itl are tobacco
mosajc virlrses ITMVs) Llsed as:n lndi' rtot
Conclusions
Thosea asigna Vlrus l\'as isolated from S asigna lafl'ae
:=-:ected with an epidemic disease in an oi1 pain plantation in Souih Sumatra'
the possibility that TaV isolated in this study would be a
::--is result
"ogg""i.g
:seful resourci-for a biological contr:ol of S asigna in oil palm plantations-
In conclusion,
Referellces
intwisfle, P.F. 1987. Virus diseases oi limacodidae' In slug and nettle
caterpillar : The Biolog/, taxonomy aild control of the limacodidae of
econ;mic importance on palm in south east asia (pp 213-221].'
Holloway. Walling ford : C.A.B. International'
:lendry, D.A., & D.K. Agrawal. 1994. Tetravirus Encyclopedia of
Virolory.
:iendry, D.A., Johnson, J.E., Rueckert, R.R., Scotti, P'D & Hanzlik, T N'
1995. Family Tetraviridae. In Vims Taxonomy: six Report of the
International Committee on Taxonomy of Viruses lpp' 372-375J'
New York : SPringer verlag
M.F., Gordon, K.H.J., Halzlik, T.N., Kalmatoff, J , Scolii, P D &
?ringle,
.-Wa-rd,
V.K. 1999. A novel capsid expression statery foi Thosea
asigna virus (Tetraviridae). Journal of general Virolory, 80, 18551853. Retrieved from http:/ / vir. sgmjoumals org/
75
Proceedlftgs o! the
rr.q'tlond,l UGet,,^g lot t tc LcoeloP'n'o tof IP ltt A'la
Aftlcc, Banda.r ,,annt tt^g, I^dola.erla Laccmbc,. 7'9' 2OO9
gd I^t
dri,
Rernganum, C., Robertson, J.S., & Tinsley, T-w. 1978 A new group of
RNA r,irrs from insects. Joumal of general Virolos/,. 40' 195-202'
Retriered from http: / /vir.sgmjournals.org/
Yi, F. \'1.. Z:-ang, J. M., Yu, H. Y., Liu, C. F., Wang, J P. and Hu, Y Y'
lO,la Isolation and identification of a new tetravirus from
k-.irclimus Wnctafr)s larvae collected from Yunnal Province,
a -r-,:a Jou;al of generalVirologr,86: 789-796. Retrieved from
.:::
D
rir.sgmjoumals.org/
:E'
-
.=
*(
3T
FjE
tu
:=fi
r-=
-E!E
t=6
:r:
'-r!
ht
iml
76
3d l^tetnil1lo^alIeettng lor trw De1t,etopne t of Ip ttt Astd.
altrl
.
$499!-*t*!!4!g!ang) I^!9^esla Decenber 7-9, 2ooe
CSoUoi 0?o\
tOooc4C
Isolatlon ofA Tetravirus from Setothose a rrsignlr Laiwae
Iafected wlth Aa Epldemic Disease In Oil palm plaatatloa
Ia South Sumatra, Indoaesia
Mada Sugltartit, Yulta Pujiaatuti2, Hisanori Balrdol
r. l,aboratory of Applied Molecular Entomolog/, Graduate Schoot of
Agriculture,
Hokkaido Universitt, Japair
2 Department of Plant Pests and Diseases, paculty of Agriculture.
Sriwijaya Universitr-, Irrdonesia
Iltroduction
La.rvae of nettle caterpillai, Sefolhosea asigna, Setora niteiLs, Dan& tima and
Dama Brdd.leqi lLEpidoptera ; Limacodidael, ar.e s],rnpatric and coseasonal; Iimacodid
moths in Southeast Asia and being important defoliator of oil palm arrd coconuts
palms. In severe infestations, SetotLbsea osigna (S.asig4a) larvae consume all
foliage and leave only mid rib of t}te frond, causing sethack oI fruit p.oduction lor a
year. Thus, S. osrgna become an economicaliy impodant defol;ator in oil palm
(Elaeis Wieensis) plantation in norih easteril Surl}atra, Bo.neo aId weitem
Malaysia. Recufing outbreak of S. aslgna, singly, or together wjth heterosF,ecific
limacodids, prcvoked application of broadspectnrm insecricides. However, recently,
the endronmental pollution by agricultural chemical becomes a big social problem in
many countries, and Indonesia is no exception.
Recentiy, natural epidemics among S. dsiqna lar-vae suspected of viral infection
have been occurred in some oil palm 6elds ir] Indonesia. These infectjous diseases
resulted in decrease of pest populatio[ below arl economic tireshold in those
epidemic areas. Suggesting the possibility oI an ellective control of S.astgna by viral
pesticides isolated from epidemic field. However, Iittle is known yei about the
causing agent of ratural epidemic disease among S. osigna observed irr lndonesia
Result and Diacuaaion
S asigna were collected in an epidemic field arrd used
as a starti]:lg material for puri$ing viruses. The existence of RNA with the
sequence homologr at least partly to the RdRp sequence of TaV was
demonstrated by RT-PCR followed by sequence analysis. ihis result suggested
that TaV or Tav-related virus is existed in the infected larvae. purification of
virus particles was then perfo.med by a method described for Dendrolimus
punctatus tetravirus (DpTV) purification. by Fuming yi et a1 (2005). On a sucrose
density gradient of paitialy purified vlal fraction, only one white band was
Diseased larvae of
observed. The vrhite band was collected, precipitated by ultra centrifugation, arrd
exarnined witl electron microscop) .
Negatively stain elec&on microscopy shov/ed tl.at the fraction contained
numerous numbers of non-enveloped, spherical virus-like particles with about
40 nrn in dilmeter. The morphological chatacteristics of the purified particles
resembled thos€ of members of the farrlily Tetrauirid.ae. The cDNAs against RNAS
extracted ftom the puritred viral particles were generated with raadom primers
arrd sequenced usi[g an automatic sequence!. The viral RNA showed a
sirilarity of about 99% in total with that ol Thosea o_signa.vitts (TaV), a member
oI the gel]ns Betatetra.arus previously reported.
74
u;
-E
G!
-ir
Proceedtngs of
Jor t tc DaeloPneftt ol lP tn A,la
Indotesla Deceatbc" 7'9, 2OO9
I'aflpu^g,
dnd Afrlca. B.1tddr
tle 3d Intarndtl6ndl lleeting
lnT
i:3'lre l.
Negattve stain elect.on microscopy oI viral particles purified by sucrc'se
deisity gradient centrifugation' Rod shaped particles itl are tobacco
mosajc virlrses ITMVs) Llsed as:n lndi' rtot
Conclusions
Thosea asigna Vlrus l\'as isolated from S asigna lafl'ae
:=-:ected with an epidemic disease in an oi1 pain plantation in Souih Sumatra'
the possibility that TaV isolated in this study would be a
::--is result
"ogg""i.g
:seful resourci-for a biological contr:ol of S asigna in oil palm plantations-
In conclusion,
Referellces
intwisfle, P.F. 1987. Virus diseases oi limacodidae' In slug and nettle
caterpillar : The Biolog/, taxonomy aild control of the limacodidae of
econ;mic importance on palm in south east asia (pp 213-221].'
Holloway. Walling ford : C.A.B. International'
:lendry, D.A., & D.K. Agrawal. 1994. Tetravirus Encyclopedia of
Virolory.
:iendry, D.A., Johnson, J.E., Rueckert, R.R., Scotti, P'D & Hanzlik, T N'
1995. Family Tetraviridae. In Vims Taxonomy: six Report of the
International Committee on Taxonomy of Viruses lpp' 372-375J'
New York : SPringer verlag
M.F., Gordon, K.H.J., Halzlik, T.N., Kalmatoff, J , Scolii, P D &
?ringle,
.-Wa-rd,
V.K. 1999. A novel capsid expression statery foi Thosea
asigna virus (Tetraviridae). Journal of general Virolory, 80, 18551853. Retrieved from http:/ / vir. sgmjoumals org/
75
Proceedlftgs o! the
rr.q'tlond,l UGet,,^g lot t tc LcoeloP'n'o tof IP ltt A'la
Aftlcc, Banda.r ,,annt tt^g, I^dola.erla Laccmbc,. 7'9' 2OO9
gd I^t
dri,
Rernganum, C., Robertson, J.S., & Tinsley, T-w. 1978 A new group of
RNA r,irrs from insects. Joumal of general Virolos/,. 40' 195-202'
Retriered from http: / /vir.sgmjournals.org/
Yi, F. \'1.. Z:-ang, J. M., Yu, H. Y., Liu, C. F., Wang, J P. and Hu, Y Y'
lO,la Isolation and identification of a new tetravirus from
k-.irclimus Wnctafr)s larvae collected from Yunnal Province,
a -r-,:a Jou;al of generalVirologr,86: 789-796. Retrieved from
.:::
D
rir.sgmjoumals.org/
:E'
-
.=
*(
3T
FjE
tu
:=fi
r-=
-E!E
t=6
:r:
'-r!
ht
iml
76