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標題: 固殺草降解性土壤微生物之篩選與鑑定
Screening and identification of glufosinate-degrading bacterium
作者: 蕭巧玲
Hsiao, Chau-Ling
關鍵字: Glufosinate
出版社: 農藝學系所
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摘要: 本研究自中興大學北溝(BG)農場、台中縣霧峰(WF)與南投縣埔里(PL)農地等土壤分別利用固殺草(glufosinate)短期誘導(SI)及長期誘導(LI)耐性菌種後,篩選其中可降解固殺草之微生物。培養之微生物經分離、純化與16S rDNA鑑定所篩選出之PL014、BG007-1、WF004-1 與PL016等菌株,分別判定為Burkholderia sacchari 、Pseudomonas citronellolis、Pseudomonas psychrotolerans與Serratia marcescens。降解測試發現,四菌株於6.28 mM 固殺草處理下,降解能力大小依序為PL016 (S. marcescens)、WF004-1 (P. psychrotolerans)、PL014 (B. sacchari)及BG007-1 (P. citronellolis)。研究結果顯示長期連續性使用固殺草之環境下較易取得分解能力高之菌株,且配合高量固殺草下所篩選之菌株較具降解力。此外,選出之三菌株於滅菌土壤中經培養7天後發現,可顯著降解固殺草,表現其高降解活性,然而於不滅菌土壤中因有其他土壤微生物也參與固殺草降解,使得三菌株降解活性之表現相對減低。本研究利用變性梯度膠體電泳(denaturing gradient gel eletrophoresis)偵測土壤於外加菌種及固殺草後的菌相變化,研究發現經在固殺草施用下,外加特定之菌株於土壤培養後21天,除了P. citronellolis無法偵測外,其餘二菌株皆能於土壤中穩定生長。此外,土壤菌相中之菌群互有消長,出現之菌群多以GC鹼基含量較高的菌群,本研究顯示特定菌種未明顯影響土壤原生菌種之生存。
Glufosinate-degrading bacteria were screened from the soils with or without experience of glufosinate application. After isola- tion and purification of edaphic microbes, coupled with 16S rDNA analysis, 4 strains, i.e., PL014, BG007-1, WF004-1 and Pl016, more tolerant to glufosinate were Burkholderia sacchari, Pseudomonas citronellolis, Pseudomonas psychrotolerans, and Serratia marcescens, respectively. Under the 6.28 mM glufosinate treatment, the degradation abilities of 4 strains were Pl016 > WF004-1> PL014 > BG007-1. Experimental results showed that bacterial strains with higher degradation ability were easily obtained after long-term induction of glufosinate, especially under a screening pressure of high glufosinate. Biodegradation experiment of 3 selected strains in sterilized soils containing glufosinate showed a higher degradation ability 7 days after treatment. However, similar degradation was also found in non-sterilized and non-inoculated soils due to other indigenous microbes. Analysis of denaturing gradient gel electro- phoresis (DGGE) revealed that the propagation of P. citronellolis was apparently inhibited in soils 21 days after inoculation, while that of two other strains were not affected. Besides, the indigenous bacteria flora were more or less altered by selected bacteria.
其他識別: U0005-2706200614010600
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