Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/22103
標題: 小球藻 Chlorella sp. strain DT 耐乾燥機制的研究. 一. 活細胞染色 與可溶性蛋白質電泳分析
Study of Desiccated-Tolerance Machenism of Chlorella strain DT I. Vital Staining ang Gel Electrophoresis of soluble proteins.
作者: 陳曾二
Chen, Tzeng Err
關鍵字: desiccated-tolerance
耐乾燥機制
vital stain
soluble proteins
viability
活細胞染色
可溶性蛋白質
電泳分析
存活率
出版社: 植物學系
摘要: 一種新分離的小球藻 Chlorella strian DT, 以 40 C 烘乾處理三小時, 並儲存於相對濕度 25% 的乾燥箱中三天後, 重新以液體培養基再培養, 能夠在再培養的第七天重新恢復生長.以活細胞染劑 Evans Blue 測定該 藻在乾燥過程的存活率, 結果分別為:第一小時 99.55 %, 第二小時 99.17 %, 第三小時時為 97.65 % 。對照組小球藻 Chlorella strain 211-8b 的存活率,分別為 89.23 % ,61.39 % 以及 30.37 % 。結果顯 示小球藻 Chlorella strain DT,確實對乾燥有忍耐性,在乾燥的過程中 能保持細胞膜系的完整 。 使用活細胞螢光染劑 Fluorscein diacetate (FDA) 測定 40 C 乾燥三小時過程中,strain DT 藻細胞內能 夠水解該染劑的酵素活性變化,結果分別為 82.97 %, 35.53 % 與 0.34 % 。以 Nitro Blue Tetrazolium (NBT) 測定其在 40 C 乾燥三小 時的過程中,被去氫酵素還原的情形,結果分別為 32.33 % ,16.66% 與 3.35 % 。以附加有分光光譜儀與掃描裝置的光學顯微鏡,掃描經 1 % Triphenyl Tetrazolium Chloride (TTC) 染色的 strain DT 藻細胞 與經 40 C 乾燥三小時過程中 ,活細胞內葉綠素a 、葉綠素b與 zeaxanthin 的變化,發現隨著乾燥時間的加長,藻細胞去氫酵素的活性 會逐漸消失,而經過乾燥處理後,藻細胞內葉綠素a 、葉綠素b與 zea- xanthin 的量均有增加的現象。顯示隨著乾燥時間的延長,小球藻 strain DT 細胞內水解酵素與去氫酵素的活性 ,逐漸的下降至零。使用 不含有清潔劑SDS的還原膠體電泳系統,分析新鮮以及經 40 C 乾燥的 小球藻 strain DT 藻細胞可溶性蛋白質,發現乾燥時間超過兩小時的藻 細胞,在 250 KD 處有一新合成的蛋白質出現,而 60 KD 與 70 KD 處的 蛋白質含量有上升的情形。但是以 40 C 水浴 、60 C 與 70 C 烘乾至含 水量與 40C 烘乾 三小時相同時 ,或是以 40C 烘乾對照組小球藻 strain 211-8b 三小時,均沒有 250 KD 新合成的蛋白質出現,也沒有前 述蛋白質含量增加的情形。這些結果說明了新蛋白質的合成與特定蛋白質 含量的增加,極可能是此一小球藻能忍受乾燥的一項機制。
It was discovered that, a new isolated green alga (Chlorophyceae) Chlorella sp. strain DT, could survive in very desiccated condition. After drying 3 hours at 40 C , it could restore growth 7 days later if rehydrating again in liquid medium. The viability of this alga after desiccated 3 hours at 40C was investigated using three vital stains, Evans Blue 、Fluorescein diacetate (FDA) and Nitro Blue Tetra- zolium (NBT), and showed the rate was 97.65%、0.34% and 3.35% respectively. It means that Chlorella sp. strain DT did keep its membrane intact but lost its enzymes activity when confronted very desiccated condition. And this may be one of the mechanisms that makes it survive under desiccation. A novel soluble protein of Chlorella sp. strain DT was discovered when it was dry at 40 C after 2 hours. The molecular weight of this novel protein was 250 KD. And it was discovered at 40 C drying only, not at 60 C , 70 C or 40 C water bathing. In addition, the protein bands of 60 KD and 70 KD increased after 2 hours drying at 40 C. Since 60 KD and 70 KD proteis are similar to the known heat shock proteins , and therefore , these proteins and novel 250 KD soluble protein may play protecting role when this alga was desiccated.
URI: http://hdl.handle.net/11455/22103
Appears in Collections:生命科學系所

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