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標題: | 堆肥中高溫非厭氧性細菌相之研究 The studies of Thermophilic,Non-anaerobic Bacterial Community of Composts |
作者: | 洪新棠 |
出版年 : | 2003 |
學位: | 碩士 |
摘要: | 微生物活動為堆肥品質良□之決定性因素之一。國內目前許多具規模的堆肥生產場,對其使用的微生物組成並不明暸,因此本實驗擬建立一個堆肥中微生物蒐集、鑑定的基礎模式,利用較為精確的判定方式,將台灣本土化菌種蒐集保存起來,希望能發現出各種堆肥中所存在的高溫優勢菌種,用以改善有機堆肥生產的效益。 本實驗利用台糖虎尾堆肥廠的蔗渣堆肥,及果菜市場廢棄物與花卉市場廢棄物堆肥,從中取樣探討其高溫狀態下的微生物相。實驗分三方面同時進行:從堆肥中取樣之後,一方面進行取得菌種之冷凍保存;一方面進行菌株外觀觀察、革蘭氏染色觀察,以及生理代謝測試;另一方面針對16S核醣體核醣核酸基因序列(16S rRNA gene;16S rDNA)進行聚合酵素鏈鎖反應(polymerase chain reaction, PCR),將所得的產物以四種不同的限制核酸內切酵素處理,進行限制內切酵素片段多型性分析(restriction fragment length polymorphism, RFLP),即所謂之 Amplified rDNA restriction analysis (ARDRA),再將所得到之菌株進行分組,更進一步自各組中選擇一代表菌株,作16S rDNA定序,從資料庫中比對出相似菌種,進而瞭解其親緣關係。 所得菌株共412株,比對生理特性及ARDRA之後,可以分為24組,其中最大一組數目為137株。分離菌株多為Gram positive或是Gram variable (96%),全部是桿狀菌,大部份可分解澱粉(65%),且具有還原硝酸鹽之能力(91%)。由ARDRA分析的pattern雖呈現高度的變異,但從16S rDNA定序結果可知大部份分離之菌株為Bacillus licheniformis及其相關菌種,其中少部份為Pseudomonas屬之菌株,此結果與生理特性分析相符合。本實驗發現高溫堆肥中存在的優勢菌種為Bacillus Iicheniformis,及Bacillus相關菌屬。本實驗結果不僅有助於台灣本土堆肥的微生物族群研究,同時也對本土雅肥中微生物族群多樣性的瞭解有所益助。 Microbial activity is one important factor of compost process. To understand microbial community in compost was one of topics interested to people of many large fields of compost. In this study, we tried to establish a basic model in collection and identification of the microorganisms of compost, and tried to find the superior bacterial strains in the hot compost in order to improve the quality of compost. In this experiment, we studied the thermophilic microbial community in the compost composed of bagasse, waste of fruit/food market, and waste of flower market in Huwei compost plant of Taiwan Sugar Corporation in Yunlin County, Taiwan (R.O.C.). The experiments included three parts: preserving the isolated bacteria strains, Gram’s staining, morphology observation and physiological test, and restriction fragment length polymorphism (RFLP) analysis of 16S rRNA gene (16S rDNA). The isolated bacterial strains were grouped based on these analyses, and a phylogenetic tree was constructed on the selected strain from each group. We isolated 412 strains, and divided they into 24 groups. The numbers of the largest group is 137. All of the isolates were rod, and most of them were Gram positive or Gram variable (96%). Most of isolated strains were able to hydrolysis starch (65%) and able to reduce nitrate (91%). The isolates displayed different patterns in amplified rDNA restriction analysis (ARDRA), but most of them belonged to Bacillus spp. and related strains (Bacillus licheniformis and Bacillus sonorensis). Few of isolates were Pseudomonas spp. The morphology and physiological characteristics of isolated strains supported the ARDRA. Our study not only established the local microbial community in Taiwan, but also would improve the benefit of high-quality compost. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/75378 |
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顯示於系所單位: | 植物科學研究所 |
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