PROTEIN TOKSININSEKTISIDAL DARI BAKTERI PATOGEN SERANGGA Photorhabdus luminescens HJ

Alina Akhdiya, Etty Pratiwi, I Made Samudra
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Abstract

Photorhabdus luminescens HJ is an entomopathogenic bacterium that has a high toxicity against Tenebrio molitor larvae.Toxicity assay of crude extra cellular protein precipitated using ammonium sulphate showed that the highest toxin activity was found in 70 % saturation. Purification of the toxin using Hi Prep 16/60 Sephacryl S-200 HR column exhibited one fraction of toxic protein and three fractions of non-toxic protein. Mortality of T. molitor larvae treated with 19.2 nanogram of toxic fraction was up to 80%. Denatured protein analysis using sodium dodecyl sulphate-polyacrylamide gel electrophoresis revealed that the toxic fraction was composed of three proteins, which were 19.5, 42, and 66 kDa respectively. Based on toxin activity bioassay, this toxin type was an injectable toxin and presumably classified as Mcf toxin.

Keywords

Entomopatogen, bakteri patogen, Photorhabdus luminescens, toksisitas protein, larva serangga Tenebrio molitor, bioesei.

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References

Blackburn M, E Golubeva, D Bowen and RH Ffrench-Constan. 1998. A novel insecticidal toxin from Photorhabdus luminescens, toxin complex a (Tea), and its histopatological effects on the midgut of Manduca sexta. Appl. Environ. Microbiol. 64,3036-3041.

Bowen DJ and JC Ensign. 1998. Purification and characterization of a High-molecular-weight insecticidal protein complex produced by the entomopathogenic bacterium Photorhabdus luminescens. Appl. Environ. Microbiol. 64, 3029-3035.

Bowen D, TA Rocheleau, M Blackburn, O Andreev, E Golubeva, R Bhartia and Ffrench-Constant RH. 1988. Insecticidal toxins from the bacterium Photorhabdus luminescens. Science 280,2129-2132.

Brown SE, AT Cao, ER Hines, RJ Akhrust and PD East 2004. A novel secreted protein toxin from the insect pathogenic bacterium Xenorhabdus nematophila. TheJ. Biol. Chem. 279,14595-14601.

Brown SE, AT Cao, P Dobson, ER Hines, RJ Akhrust and PD East 2006. Txp40, a ubiquitous insecticidal toxin protein from Xenorhabdus and Photorhabdus bacteria. Appl. Environt. Microbiol. 72,1653-1662.

Daborn, P J, N Waterfield, CP Silva, CPY Au, S Sharma and RH Ffrench-Constant. 2002. A single Photorhabdus gene, makes caterpillars floppy (mcf), allows Escherichia coli to persist within and kill insects. PNAS. 99, 10742-10747.

Duchaud E, C Rusniok, L Frangeul, C Buchrieser, A Givaudan, S Taourit, S Bocs, C Boursaux-Eude, M Chandler, JF Charles, E Dassa, R Derose, S Derzelle, G Freyssinet, S Gaudriault, C Medigue, A Lanois, K Powell, P Siguier, R Vincent, V Wingate, M Zouine, P Glaser, N Boemare, AA Danchin and F Kunst. 2003. The genome sequence of the enthomopathogenic bacterium Photorhabdus luminescens. Nat. Biotechnol. 21,1307-1313.

Forst S and K Nealson. 1996. Molecular biology of the symbiotic-pathogenic bacteria Xenorhabdus spp. and Photorhabdus spp. Microbiol. Rev. 60, 21-43.

Guo L, RO Fattig III, GL Orr, BW Schafer, JA Strickland, K Sukhapinda, AA Woodsworth and JK Petell. 1999. Photorhabdus luminescens W-14 insecticidal activity consists of at least two similar but distinct proteins. The J. Biological Chemistry 274, 9836-9842.

Hazir S, HK Kaya, SP Stock and N Keskin. 2003. Entomopathogenic nematodes (Steinernematidae and Heterorabtidae) for biological control of soil pests. TurkJ. Biol. 27, 181-202.

Liu D, S Burton, T Glancy, ZS Li, R Hampton, T Meade and DJ Merlo. 2003. Insect resistance conferred by 283-kDaPhotorhabdus luminescens protein TcdA in Arabidobsis thaliana. Nat. Biotechnol. 21,1222-1228.

Miyahara Y. 1977. Rearing method of Spodoptera litura by artificial diet. Annual Report Kyushu Nat. Agric.Expt. Sta., 34-37.

Pratiwi E, A Akhdiya dan IM Samudra. 2005. Eksplorasi Gen Penyandi Toksin Insektisidal dari Bakteri S imbion Nematoda Patogen Serangga. Laporan Hasil Penelitian Balai Besar Bioteknologi dan Sumberdaya Genetik Pertanian Tahun 2005.

Waterfield NR, PJ Daborn, AJ Dowling, G Yang, M Hares and RH Ffrench-Constant. 2003. The insecticidal toxin makes caterpillar floppy 2 (Mcf2) show similarity to HrmA, an avirulence protein from a plant pathogen. FEMS Microbiol. Lett. 229, 265-270.

Waterfield N, SG Kamita, BD Hammock and RH Ffrench-Constant. 2005. The Photorhabdus Pir toxins are similar to a developmentaily regulated insect protein but show no juvenile hormone esterase activity. FEMSMicrobiol. Lett. 229,265-270.

Yang G, AJ Dowling, U Gerike, RH Ffrench-Constant and NR Waterfield. 2006. Photorhabdus virulence cassettes confer injectable insecticidal activity againstthe wax moth. J.Bacteriol. 188,2254-2261.


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