Browse > Article
http://dx.doi.org/10.5423/RPD.2013.19.4.254

Fusaric Acid Production in Fusarium oxysporum Transformants Generated by Restriction Enzyme-Mediated Integration Procedure  

Lee, Theresa (Microbial Safety Team, National Academy of Agricultural Science, Rural Development Administration)
Shin, Jean Young (Microbial Safety Team, National Academy of Agricultural Science, Rural Development Administration)
Son, Seung Wan (Microbial Safety Team, National Academy of Agricultural Science, Rural Development Administration)
Lee, Soohyung (Microbial Safety Team, National Academy of Agricultural Science, Rural Development Administration)
Ryu, Jae-Gee (Microbial Safety Team, National Academy of Agricultural Science, Rural Development Administration)
Publication Information
Research in Plant Disease / v.19, no.4, 2013 , pp. 254-258 More about this Journal
Abstract
Fusaric acid (FA) is a mycotoxin produced by Fusarium species. Its toxicity is relatively low but often associated with other mycotoxins, thus enhancing total toxicity. To date, biosynthetic genes or enzymes for FA have not been identified in F. oxysporum. In order to explore the genetic element(s) for FA biosynthesis, restriction enzyme mediated integration (REMI) procedure as an insertional mutagenesis was employed using FA producing-F. oxysporum strains. Genetic transformation of two F. oxysporum strains by REMI yielded more than 7,100 transformants with efficiency of average 3.2 transformants/${\mu}g$ DNA. To develop a screening system using phytotoxicity of FA, eleven various grains and vegetable seeds were tested for germination in cultures containing FA: Kimchi cabbage seed was selected as the most sensitive host. Screening for FA non-producer of F. oxysporum was done by growing each fungal REMI transformant in Czapek-Dox broth for 3 weeks at $25^{\circ}C$ then observing if the Kimchi cabbage seeds germinated in the culture filtrate. Of more than 5,000 REMI transformants screened, fifty-three made the seeds germinated, indicating that they produced little or fewer FA. Among them, twenty-six were analyzed for FA production by HPLC and two turned out to produce less than 1% of FA produced by a wild type strain. Sequencing of genomic DNA regions (252 bp) flanking the vector insertion site revealed an uncharacterized genomic region homologous (93%) to the F. fujikuroi genome. Further study is necessary to determine if the vector insertion sites in FA-deficient mutants are associated with FA production.
Keywords
Fusaric acid; Fusarium oxysporum; REMI; Transformation;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Namiki, F., Matsunaga, M., Okuda, M., Inoue, I., Nishi, K., Fujita, Y. and Tsuge, T. 2001. Mutation of an arginine biosynthesis gene causes reduced pathogenicity in Fusarium oxysporum f. sp. melonis. Mol. Plant Microbe Interact. 14: 580-584.   DOI   ScienceOn
2 Notz, R., Maurhofer, M., Dubach, H., Haas, D. and Defago, G. 2002. Fusaric acid-producing strains of Fusarium oxysporum alter 2,4-diacetylphloroglucinol biosynthetic gene expression in Pseudomonas fluorescens CHA0 in vitro and in the rhizosphere of wheat. Appl. Environ. Microbiol. 68: 2229-2235.   DOI
3 Smith, T. K. and Sousadias, M. 1993. Fusaric acid content of swine feedstuffs. J. Agric. Food Chem. 41: 2296-2298.   DOI   ScienceOn
4 Srobarova, A., Eged, S., Teixeira da Silva, J., Ritieni, A. and Santini, A. 2009. The use of Bacillus subtilis for screening fusaric acid production by Fusarium spp. Czech J. Food Sci. 27: 203-209.
5 Yabuta, T., Kambe, K. and Hayashi, T. 1937. Biochemistry of the bakanae fungus. I. Fusarinic acid, a new product of the bakanae fungus. J. Agric. Chem. Soc. Jpn. 10: 1059-1068.
6 Bacon, C. W., Porter, J. K., Norred, W. P. and Leslie, J. F. 1996. Production of fusaric acid by Fusarium species. Appl. Environ. Microbiol. 62: 4039-4043.
7 Brown, D. W., Butchko, R. A. E., Busman, M. and Proctor, R. H. 2012. Identification of gene clusters associated with fusaric acid, fusarin, and perithecial pigment production in Fusarium verticillioides. Fungal Genet. Biol. 49: 521-532.   DOI   ScienceOn
8 Desjardins, A. E. 2006. Fusarium mycotoxins: chemistry, genetics, and biology. The American Phytopathological Society, St. Paul, Minnesota, USA. 260 pp.
9 Gaumann, E. 1957. Fusaric acid as a wilt toxin. Phytopathology 47: 342-357.
10 Han, Y.-K., Lee, T., Han, K.-H., Yun, S.-H. and Lee, Y.-W. 2004. Functional analysis of the homoserine O-acetyltransferase gene and its identification as a selectable marker in Gibberella zeae. Curr. Genet. 46: 205-212.   DOI   ScienceOn
11 Linnemannstons, P., Vob, T., Hedden, P., Gaskin, P. and Tudzynski, B. 1999. Deletions in the gibberellins biosynthesis gene cluster of Gibberella fujikuroi by restriction enzymemediated integration and conventional transformationmediated mutagenesis. Appl. Environ. Microbiol. 65: 2558-2564.