The Korean Journal of Mycology (한국균학회지)

The Korean Society of Mycology (한국균학회)
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Protoplast Fusion of Fusarium oxysporum and activity of polygalacturonase

Fusarium of oxysporum의 원형질체 융합과 polygalacturonase의 활성

  • Ha, Kyung-Ran (Department of Biology, College of Natural Sciences, Sang Myung Women's University) ;
  • Min, Byung-Re (Department of Biology, College of Natural Sciences, Sang Myung Women's University)
  • 하경란 (상명여자대학교 생물학과) ;
  • 민병례 (상명여자대학교 생물학과)
  • Published : 1993.09.30
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Abstract

This experiments was designed for development of Fusarium oxysporum strains with enhanced activity of polygalacturonase by means of mutants and protoplasts fusion. Six auxotrophic mutants of F. oxysporum were induced by treatment of MNNG. Protoplasts from mutants were formed from early exoponential mycelium after treatment with driselase(10 mg/ml) using 0.6 M KCl as osmotic stabilizer. Fusion experiments between protoplasts of several auxotrophic mutants were done using polyethyleneglycol 8,000 and $CaCl_2$. The frequency of fusion was $5.0{\times}10^{-3}$ as determined from several experiments. Activity of polygalacturonase was determined by the methods of modified DNS. Consequently, the polygalacturonase activities of mutants and fusant derived F. oxysporum were 1.4 to 3.5 times greater than that of the parental strain, and mutant Fx-2 seemed to be the best strain. Thus, the method we used seemed to be favorable for the improvement of strains.

Fusarium oxysporum을 재료로 하여 polygalacturonase 활성능이 좋은 우량 균주를 개발하기 위하여 돌연변이주와 원형질체융합주를 선별하였다. MNNG를 처리하여 6개의 영양요구성 돌연변이주를 유도하였다. 세포벽 분해효소로는 driselase를 이용하고 삼투안정제로 0.6M KCI을 처리하여 돌연변이체로부터 원형질체를 얻었다. polyethyleneglycol 8,000과 $CaCl_2$를 이용하여 영양요구성 돌연변이체의 원형질체 사이에 융합을 시도하여 1개의 융합주를 얻었으며 융합률은 $5.0{\times}10^{-3}$이었다. polygalacturonase 활성도를 수정된 DNS법을 이용하여 측정하였다. 결과적으로, 선별된 융합주와 돌연변이주의 polygalacturonase 활성은 모균주와 비교하여 약 1.5배에서 3.5배가량 증가하였으며 그 중에서도 mutant Fx-2가 가장 높은 활성도를 나타냈다. 그러므로 이 방법은 우량균주개발을 위한 유용한 방법으로 판단되었다.

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