Preparasi DNA Spesies Colletotrichum sp. dan Spesifitas Sistem Fingerprinting RAPD

Jamsari Jamsari

Abstract


DNA preparation from C. capsici dan C. gleosporides is important for PCR based analysis of anthracnose causingpathogens. On the other hand, RAPD as one of DNA-based fingerprinting is hindered by its instability and its nonspecifity problems. For that reason, converting RAPD fragments into other DNA based systems is an alternative toincrease its analysis reliability. The study showed that mycelia grown in 2-days liquid culture is appropriate materialfor DNA preparation especially when combined with Shagai-Maroof protocol as well as Promega Genomic DNAIsolation Kit. The study indicated also that some isolated RAPD-fragments showed its instability character. Thiswas proved by the occurrence of multi different length of fragments after re-amplification of some single RAPDspecific fragments.

Keywords


C. capsici, C. gleosporides, RAPD- analysis

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References


Adetula, O.A. 2006. Genetic diversity of capsicum using Random Am pl if ied P ol ym orphi c DNA s. Af ri can J ournal of Biotechnology 5: 120-122.

Botstein, D., White, R. L., Skolnick, M., Davies, R. W. 1980. Construction of a genetic map in man using restriction fragment lenght polymorphisms. Am. J. Hum. Genet 32: 314- 331.

Cano, J., Guarro,J. and Gene, J. 2004. Molecular and morphological identification of colletotrichum species of clinical interest. Journal of Clinical Microbiology, 42: 2450- 2454

Das, M., Bhattacharya, S. and Pal, A. 2005. Generation and characterization of SCARs by cloning and sequencing of RAPD Products: A strategy for species-specific marker development in bamboo. Annals of Botany 95: 835-841

Dweikat I, Mackerzie S, Levy M, Ohm H. 1993. Pedigree assessment using RAPD-DGGE in cereal crop species. Theor. Appl. Gene 85:497-505.

El-Mezawy, A. 2000. Fine mapping of the bolting gene from sugar beet (Beta vulgaris L.) with molecular markers. Dissertation. Germany: Christian Albrechts Universität zu Kiel.

Guerber, J.C, Liu, B., Correll. J.C. 2003. Characterization of diversity in Colletotrichum acutatum sensu lato by sequence analysis of two gene introns, mtDNA and intron RFLPs, and mating compatibility. Mycologia 95: 872–895.

Hayford, A.E. Petersen, A., Vogensen, F., and Jakobsen, M. 1999. Use of conserved Randomly Amplified Polymorphic DNA (RAPD) Fragm ents and RAP D Pattern f or characterization of Lactobacillus fermentum in Ghanaian fermented maize dough. Appl Environ Microbiol 65: 3213– 3221.

Jamsari, 2003. Construction of high-density genetic and physical maps around the sex gene M of Asparagus officinalis L. Dissertation. Germany: Christian Albrechts Universität zu Kiel.

Jamsari, Darusalam, R., Syahlena, M. Syaputra, R., Darnetty, Putri, N.E. 2007. Seleksi primer RAPD dan studi kekerabatan capsicum sp. Koleksi Dari Sumatera Barat. Aktaagrosia (in Press)

Jiang, C., Lewis, M. E., Sink, K. C. 1997. Combined RAPD and RFLP molecular linkage map of asparagus. Genome 40: 69- 76.

Kim, K.S., and Lee, Y.S. 2001. Selection of RAPD markers for phytophthora infestans and PCR detection of phytophthora 132

MacPherson, J.M., Eckstein, P.E., Scoles, G.J., Gajadhar, A.A. 1993. Variability of the random amplified polymorphic DNA assay among thermal cyclers, and effects of primer and DNA concentration. Mol Cell Probes 7: 293–299.

M eun ier, J. R, Grim on t, P. A. 1993. Fact ors aff ec ti ng reproducibility of random amplified polymorphic DNA fingerprinting. Res. Microbiol 144: 373–379.

Mongkolporn, O., Dokmaihom, Y. 2004. Genetic purity test of F1 hybrid Capsicum using molecular analysis. The Journal of Horticultural Science and Biotechnology 79: 449-451

Noverta, A. 2007. Optimasi Isolasi DNA Tanaman Gambir (Uncaria gambir Roxb) dan Seleksi Primer RAPD (Random Amplified Polymorphic DNA). Skripsi Sarjana Pertanian. Padang: Fakultas Pertanian Unand

Rajput, S.G., Wable, K.J., Sharma, K.M., Kubde, P.D. and Mulay, S.A. 2006. Reproducibility testing of RAPD and SSR markers in Tomato. African Journal of Biotechnology 5: 108- 112.

Reamon-Büttner, S. M., Jung, C. 2000. AFLP-derived STS markers for the identification of sex in Asparagus officinalis L. Theor Appl Genet 100: 432-438.

Roca, M.G. Davide, L.C., Wheals, A.E. 2003. Template Preparation for Rapid PCR in Colletotrichum lindemuthianum. Brazillian Journal of Microbiology 34: 8-12.

Saghai-Maroof, M.A.; Soliman, K.M.; Jorgensen, R.A.; Al lard, R. W. 1984. Ribos om al DNA spacer-length polymorphisms in Barley: Mendelian inheritance, chromosomal location and population dynamics. Proc Natl Acad Sci USA 81: 8014-8018

Schmink, S., Reeves, M.W, Plikaytis, B., and Popovic. T. 2001. Random Amplified Polymorphic DNA assay as a rapid tool in screening for neisseria meningitidis serogroup C isolates of electrophoretic type 24. Journal of Clinical Microbiology 39: 1622–1625

Shaaban, E.A., Abd-El-Aal, S.K.H., Zaied, N.S., and Rizkalla, A.A. 2006. Assessment of genetic variability on some orange accessions uUsing RAPD-DNA markers. Research Journal of Agriculture and Biological Sciences 2: 564-570, 2006

Thomsen, L., and Jensen, A.B. 2002. Application of nested- PCR technique to resting spores from the entomophthora muscae species complex: implications for analyses of hostpathogen population interactions. Mycologia, 94: 794–802.

Vos, P., Hogers, R., Bleeker, M., Reijans, M., van de Lee, T., Hornes, M., Fritjters, A., Pot., J., Peleman, J., Kuiper, M., Zabeau, M. 1995. AFLP: A new technique for DNA fingerprinting. Nucl Acids Res 23: 4407-4414

Williams, J.G.K., Kubelick, A.R., Livak, K.J., Rafalski, J.A., Tingey, S.V. 1990. DNA Polymorphisms Amplified by Arbitrary Primers are Useful as Genetic Markers. Nucleic acids Res. 18:6531-6535

Xu, W.J, Wang, B.W and Cui, K.M. 2004. RAPD and SCAR markers linked to sex determination in eucommia ulmoides oliv. Euphytica 136: 233–238.

Yu, K., K.P. Pauls. 1992. Optimization of the PCR program for RAPD analysis. Nucl. Acids Res. 20: 2606.

Zahner, V., Rabinovitch, L, Suffys, P. and H. Momen. 1999. Genotypic Diversity among Brevibacillus laterosporus Strains. Applied and Environemtal Microbiology 65: 5182– 5185




DOI: http://dx.doi.org/10.31258/jni.11.01.%25p

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