臺灣北部沿岸久置海水中細菌的分離和特性研究

海洋中營養鹽的分布通常是不均勻的,且會依季節與區域變化,但就整體而言,海洋是一廣闊而低營養的環境。為進一步瞭解能夠存活於低營養環境中的海洋細菌,本研究利用採自台灣北部南雅、石城及磅磅子的海水,將之存放超過七年時間之後進行細菌的分離、培養與分析。本研究自三地區水樣共分離出 25 株細菌,根據 16S rRNA gene 的定序及比對,初步鑑定該 25 分離株細菌分別屬於 Stappia(6 株)、Alteromonas(4 株)、Oceanococcus(2 株)、Marinobacter(3 株)、Mesorhizobium(1 株)、Alcanivorax(4 株)、Kangiella(1 株)、Halobacillus (1 株)、Spongiibacter(1 株)、Erythrobacter(1 株)及 Marinobacterium(1 株)各屬。分離株經各種生理、生化特性檢測後,皆無法利用葡萄糖發酵產酸, 均能在 1-5%NaCl 濃度、pH 8-9 及 25-30°C 下生長,均不產生吲哚及硫化氫。 Na1 經 16S rRNA gene 比對,與 Spongiibacter tropicus CL-CB221T 序列相似度為 95.9%,Na1 為好氧且具鈉鹽需求性之革蘭氏陰性細菌,最適合生長於 30-37°C、 1-3%NaCl 及 pH7-8 的培養基中,依初步生理生化特性及序列分析,分離株 Na1 可能為新種細菌。為瞭解實驗菌株在低營養環境中的增殖情況,本研究將分離株 N1C 及 N2B 進行四種濃度的低營養培養,並連日以細胞平板計數法(plate count method)觀察比較其增殖情形,結果顯示二分離株皆能夠在 40 mg/L 濃度碳源下快速生長。

關鍵字

營養鹽；低營養培養；細胞平板計數法； 16s rRNA

並列摘要

The distribution of nutrients in marine environments is usually uneven, and varys between seasons and regions, but on a whole, the ocean is a vast, low-nutrient environment. To further understand the marine bacteria able to survive in low nutritious environments, we isolated, purified and analyzed the bacteria in the aged seawater samples collected from northern Taiwan which have been stored for more than 7 years. We isolated 25 isolates from the samples, and according to the sequences of 16S rRNA gene, they belong to the following genera: Stappia(6 isolates)、Alteromonas(4 isolates)、Oceanococcus(2 isolates)、Marinobacter(3 isolates)、Mesorhizobium(1 isolate)、Alcanivorax(4 isolates)、Kangiella(1 isolate)、Halobacillus(1 isolate)、 Spongiibacter(1 isolate)、Erythrobacter(1 isolate)and Marinobacterium(1 isolate). According to the physiological and biochemical tests, we found that all the isolates are negative for glucose fermentation, indole and H2S production, and can grow under 1-5% NaCl、pH 8-9 and 25-30°C. Based on the 16S rRNA sequence, the similarity between Na1 and Spongiibacter tropicus CL-CB221T is 95.9%. Na1 is a sea salt-needing, aerobic and gram’s negative bacteria, and the optimal range for growth is 30-37°C, 1-3% NaCl and pH7-8. Na1 may be a novel bacterium of genus Spongiibacter according to the cross- matching between the study and reported data. In order to understand the proliferation of the isolates in low-nutrietion environment, we cultivated isolates N1C and N2B in medium, and observed the enumerational change by plate count method. The results showed that the two isolates are able to grow rapidly in a medium with carbon source of 40 mg / L.

並列關鍵字

nutrient ； low-nutrient medium ； plate count method ； 16s rRNA

參考文獻

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國際替代計量

臺灣北部沿岸久置海水中細菌的分離和特性研究

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