PRODUKSI ANTIBODI POLIKLONAL SUGARCANE STREAK MOSAIC VIRUS

Cahyo Artho Nugroho, Nur Basuki, Arifin Noor Sugiharto

Abstract


Sugarcane Streak Mosaic is an important disease in sugarcane caused by Sugarcane streak mosaic virus (SCSMV). SCSMV attack intensity at the sample location sugarcane crop may reach 62%. The high intensity of the attack on the planting of sugar cane SCSMV make the virus should receive serious attention in order not widespread dissemination. One way to control the spread of the virus is the rapid detection. The rapid and precise mass detection to detect symptoms caused by the virus is by serologic testing. This study aims to produce polyclonal antiserum that can be used to detect SCSMV in sugarcane. Pure virus was analyzed using a spectrophotometer at a wavelength of 260 and 280 nm, a purity virus was 0.997. Pure virus particles injected into rabbits subcutaneously 3 times. Intake of rabbit blood serum performed 5 times and its IgG levels were measured by using a spectrophotometer nanodrop. The resulting antiserum is able to detect the presence of particles SCSMV on leaf tissue of sugarcane with the indication of the color change to yellow in wells ELISA Plate. Results of the process of reading absorbance at a wavelength of 405 nm by using ELISA reader machine showed a positive reaction because it shows the absorbance value is 3x greater than the negative control and the ratio between the sample is infected and a healthy sample or buffer (S/H Ratio)>2. Thus antiserum produced can be used as a detector SCSMV.

Keywords


Polyclonal antiserum, sugarcane streak mosaic virus, indirect ELISA

Full Text:

PDF

References


Agindotan, O., Ahonsi, M.O., Domier L.L., Gray, M.E., and Bradley, C.A. 2010. Protocols application of sequence-independent amplification (SIA) for the identification of RNA viruses in bioenergy crops, Journal of Virological Methods (169): 119–128.

Cooper, H.M and Paterson, Y. 2008. Preparation of polyclonal antisera, production of polyclonal antisera. Current Protocol in Molecular Biology. Immunology. 11.12.1-11.12.10. www.interscience.wiley.com. Published online January 2008.

Damayanti Asmira.,T., Putra .K., dan Juliadi D. 2007. Kajian Sifat bio-ekologi dan biomolekuler virus mosaik bergaris pada tebu di Indonesia. Abstrak Penelitian dan Pengembangan Institut pertanian Bogor. Darmaga Bogor. Abstrak penelitian Litbang. (http://we servebetter.wordpress.com/2008/06/04/Lembaga Penelitian Dan Pengabdian Masyarakat Institut Pertanian Bogor/). Di akses tanggal 06 April 2009.

Damayanti Asmira.T., dan Putra L.K. 2012. Preparasi RNA virus mosaik bergaris dari tanaman tebu menggunakan metode tabung PCR. Jurnal Fitopatologi Indonesia (8) 1:22-27.

Haider, S.M., Afgan S., Riaz H., Tahrir M., Javed A.M., Rashid N., and Iqbal J. 2011. Identification of two sugarcane mosaic virus (scmv) variants from naturally infected sugarcane crop in pakistan. Pak.J.Bot 43(2): 1157-1162,

Hall, J.S., Adams B., Parsons T.J., French R., Lane L.C, and Jensen S.G. 1998 Molecular cloning, sequencing and phylogenetic relationships of a new potyvirus: sugarcane streak mosaic virus, and a reevaluation of the classification of the Potyviridae. Mol Phylogenet Evol. (10):323–332

Hema, M., Sreenivasulu P., and Savithri H.S. 2002. Taxonomic position of sugarcane streak mosaic virus in the family Potyviridae. Arch Virol. (147)

Koenig, R., Fribourg C.E. and. Jones R.A.C. 1979. Symptomological, serological, and electrophoretic diversity of isolates of Andean potato latent virus from different regions of the Andes. Phyton (69): 748–752.

Li W, He Z., Li S., Huang Y., Zhang Z., Jiang D., Wang X., and Luo Z. 2011. Molecular characterization of a new strain of sugarcane streak mosaic virus (SCSMV). Arch Virol. Springer-Verlag 2011. Published Online 17 Sept 2011.

Putra, L.K., dan Damayanti A.T. 2009. Penyakit mosaik pada tebu di Indonesia: survey lapang, deteksi virus, uji penularan, kisaran inang dan ketahanan varietas. Majalah Penelitian Gula 45 (1): 21-38

Ratti C., Budge G., Ward L., Clover G., Autonell C.R., and Henry C. 2004. Detection and relative quantitation of soil-borne cereal mosaic virus (SBCMV) and Polymyxa graminis in winter wheatusing real-time PCR (TaqMan®). Journal of Virological Methods (122) : 95–103.

Sakhuja, A. G., Sears J.L., Alberto Nuñez., and Liu H. 2009. Production of polyclonal antibodies against Pelargonium Zonate Spot Virus Coat Protein expressed in escherichia coli and application for immunodiagnosis. Journal of Virological Methods (160) : 29–37.

Thouvenel, J. C., Givord L., and Pfeiffer P. 1976. Guinea grass mosaic virus, a new member of the potato virus Y group. J Phytopathology 66 : 954-957

Veerisetty, V and Myron K. B. 1978. Purification of Some Legume Carlaviruses. J Phytopathology 68 : 59-64.




DOI: http://dx.doi.org/10.33366/bs.v15i2.367

Article metrics

Abstract views : 193 | views : 296

Refbacks

  • There are currently no refbacks.


Copyright (c) 2017 BUANA SAINS

Plagiarism Check and indexed by:


Universitas Tribhuwana Tunggadewi © 2017