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題名 | 以非放射性核酸探針定量土壤中的脫氮菌(2)--土壤中脫氮菌之定量=Quantification of Soil Denitrifier by Non-Isotope Nucleic Acid Probe Part ΙI. Quantification of Soil Denitrifier |
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作者姓名(中文) | 吳秋曄; 梁志欽; | 書刊名 | 健康與照顧科學學刊 |
卷期 | 2:2 2014.08[民103.08] |
頁次 | 頁114-121 |
分類號 | 434.225 |
關鍵詞 | 脫氮菌定量; 核酸探針; Quantification of denitrifier; Nucleic acid probe; |
語文 | 中文(Chinese) |
中文摘要 | 以最大可能計數法測定脫氮菌數需兩周才能得到測定值,因此本研究嘗試建立以DNA探針定量土壤脫氮菌數。首先以來自Alcaligenes eutrophus CCRC 13036 的核酸探針 (781 bp) 與不同菌體濃度的A. eutrophus 進行雜交後,以密度掃描儀掃描定量,顯示菌體濃度在 1.522 ×106 與 3.805×107 cell number ml -1 間,迴歸直線的 R2 值為 0.99。以此核酸探針定量土壤脫氮菌,結果顯示以此探針不能測出壽豐系與龍崗系土壤脫氮菌數,但能測定到二水系每克土壤含有約 3.01 ×106 個 A. eutrophus 等量菌數,此數目約為以最大可能計數法測定的脫氮菌數的 1000 倍。此差異性可能是由於最大可能計數法測定為活菌,其數量決定於培養基的組成,因此測定值會小於實際值。而以核酸探針則無法區別活菌與死菌,所以其值會較實際值高。 |
英文摘要 | Generally, it takes two weeks for determination of denitrifiers by the most probable number (MPN)method. Therefore the quantification method of soil denitrifier by a DNA probe was established in the present study. First we used the 781-bp fragment of Alcaligenes eutrophus CCRC 13036 to detect different concentrations of A. eutrophus cell. The results showed that a linear relationship for the quantitation of A. eutrophus between 1.522×106 and 3.805×107 cell number ml -1 by the densitometric measurement was found. A direct extraction of DNA from Shoufeng, Erhshui and Lungkang soils was applied for the quantification by the probe. The numbers of denitrifiers in Erhshui soil detected by hybidization with the probe was equivalent to 3.01×106 cells of A. eutrophus g-1 soil, however, the denitrifiers in Shoufeng and Lungkang soils could not be determined by the probe. The number of denitrifiers in Erhshui soil was 1000 times of those determined previously by MPN method. MPN is one kind of cultural methods. The community of grown microorganisms was defined by the compositions of the medium used. Therefore the numbers of denitrifiers measured by MPN method may be generally lower than the numbers existed in the soil. Although there was few hybridization with a nondenitrifying bacterium in the early study, the numbers of denitrifiers determined by the probe may be higher than the numbers existed in the soil. |
本系統之摘要資訊系依該期刊論文摘要之資訊為主。