Isolation of Marine Bacteria Killing Red Tide Microalgae II. Isolation and Algicidal Properties of Pseudomonas sp. LG-2 Possessing Killing Activity for Dinoflagellate, Prorocentrum micans

적조생물 살조세균 탐색 II. 적조생물 Prorocentrum micans 살조세균 Pseudomonas sp. LG-2의 분리와 살조특성

We have isolated a bacterial strain that tends to kill P. micans from the mixed culture of p. minns plus seawater filtrate (poresize, 0.8 $\mu$m) collected at Masan bay in July 1996, in which the mixed culture grown in the f/2 medium. According to the experimental results of the isolated bacterium such as fatty acids analysis, morphological and biochemical characteristic tests, the strain was supposed to be a Pseudomonas and then it was named as Pseudomonas sp. LG-2. The killing effect of Pseudomonas sp. LG-2 against P. micans was proportionally increased with the concentrations of culture filtrate (pore size, 0.8 $\mu$m) is well as with the number of bacterium inoculated. In the mixed culture inoculated with $1.3\times10^6$ cells/ml of Pseudomonas sp. LG-2, the number of P. micans (2,000 cells/ml) was gradually decreased and then killed below 100 cells/ml within 7 days. In addition, the culture filtrate with $30\%$ of final concentration revealed a significant killing effect against P. micans around 3 days after culture. In the relationship between killing effects and growth stage of Pseudomonas sp. LG-2, the culture filtrate at lag phase has little effects on P. micans. In constant, the culture filtrate at mid-log phase showed the killing effect by decreasing P. micans to 112 in number within 5 days. In particular, the culture filtrate at stationary phase showed a significant killing effect against P. micans in which the majority of it was killed after 3 day culture. The species specificity of killing effects of Pseudomonas sp. LG-2 against 5 species of dinoflagellate was only found in P. micans and Scrippsiella trochoidea.

1996년 7월 마산만의 해수를 0.8 $\mu$m filter에 여과한 여과액과 f/2배지에서 배양한 P. micans와의 혼합배양액에서 P. micans를 사멸시키는 해양세균을 분리하였다. 분리균의 형태학적 및 생화학적 검사와 균체지방산을 분석하여 동정한 결과 Pseudomonas 속으로 동정되어 Pseudomonas sp. LG-2로 명명하였으며, 그 살조특성을 조사한 결과는 아래와 같다. Pseudomonas sp. LG-2의 개체수와 배양여과액의 농도가 높을수록 P. micans의 사멸효과가 높게 나타났다. $1.3\times10^5,\;1.3\times10^6$ cells/ml의 농도로 접종한 시험관의 P. micans는 급격히 감소하여 배양 7일 후에는 $10^2$ cells/ml 이하로 사멸되었다. 또한, 배양석과액의 최종농도가 $30\%$일 경우에는 급격히 감소하여 배양 3일 후 거의 사멸하였다. Pseudomonas sp. LG-2의 성장시기에 따른 배양여과액의 사멸효과는 잠복기의 경우 P. micans의 성장에 큰 영향을 미치지 않았으나, 대수증식기 중기의 배양여과액은 접종 5일 후 P. micans의 개체수를 1/2로 감소시켰으며, 정상기의 배양여과액은 접종 후 급격히 감소시켜 배양 3일 후 대부분 사멸시켰다. Alexandrium tamarense, Prorocentrum micans, Scrippsiella trochoidea, Gymnodinium sanguineum, Cochlodinium polykrikoides의 5종의 적조원인 편모조류에 대한 Pseudomonas sp. LG-2의 살조특이성은 P. micans와 S. trochoidea에 살조효과를 나타내었다.