Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/28270
標題: 長期施用不同肥料下水稻根圈土壤固氮基因多樣性
Diversity of nifH gene pools in the rhizosphere soil of rice plant after long-term fertilizer application
作者: 朱宏怡
Chu, Horng-I
關鍵字: rhizosphere;根圈;nifH gene;diversity;固氮基因;多樣性
出版社: 土壤環境科學系所
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摘要: 
長期使用化學肥料已知會造成土壤理化學性質改變及微生物族群的改變。生物固氮是土壤中重要的微生物作用,除了幫助植物生長也維持著自然界氮循環的平衡。水稻是世界包括台灣重要的糧食作物,很多協生或受到根分泌物而聚集於水稻根圈的固氮微生物已經被篩選並發表,但在不同施肥管理下關於其根圈固氮微生物組成及影響因子的研究卻很少。因此,本研究之目的在測試水稻田土壤中,固氮微生物族群因長期施用不同肥料而有所變化之假說。利用不需培養的方法研究在長期 (十年) 施用化肥與豬糞堆肥的田間管理及水稻不同生長階段下根圈土壤之固氮微生物固氮基因 (nifH gene) 組成及探討造成固氮基因變化可能的影響因子。結果顯示水稻根圈固氮活性在不同施肥管理下無顯著差異,但在不同生長期下卻有明顯變化。使用限制酵素長度片段多型性分析 (Restriction fragment length polymorphism, RFLP) 結果也顯示,不同生長階段造成不同的nifH基因圖譜,而不同施肥處理間變化不大。因此進一步再利用選殖方式 (cloning) 將所得到之nifH基因片段挑出並定序,發現存在於根圈土壤中的固氮基因在水稻生長期間有部份種類的增減,而長期相同肥料之施用可能造成nifH基因的變異,導致不同施肥處理間沒有發現相同的nifH基因。水稻生長期內,不同施肥處理間固氮基因組成類似,但佔有優勢的固氮基因卻各有不同,顯示施肥處理造成水稻根圈土壤之固氮微生物族群並非完全一致外還有數量上的差別。雖然以RFLP方法無法辨識不同施肥處理間nifH基因圖譜之差異,但以選殖方式將nifH基因定序後顯示不同施肥處理明顯造成土壤中nifH基因組成之變異。此結果支持土壤中固氮微生物族群因長期施用不同肥料而有所變化之假說。本研究亦建立台灣第一筆關於水田土壤中固氮微生物固氮基因的資料庫。並且發現其中存有大量新發現的固氮基因,此大量的未知nifH基因目前雖還不能確立其分類地位,但值得繼續探討。

Long-term fertilizer application are known to cause changes in soil physical, chemical properties and microbial population. Biological nitrogen fixation is an important process by soil microorganisms, not only to help plant growth but also maintain the balance of natural nitrogen cycle. Rice is the most important food crop in the world, including Taiwan. Many of nitrogen-fixing bacteria associated with rice rhizosphere by the accumulation of root exudates have been selected and published, but research about nitrogen-fixing bacteria on the rhizosphere under different fertilization management is rare. The purpose of this study was to test the hypothesis that in paddy soil, nitrogen-fixing bacterium population would be changed considerably after a long-term fertilizer application. Unculturable methods was used to study the composition of the nifH gene in rice rhizosphere at different growth stages after long-term (ten years) chemical fertilizer and swine manure compost application, and the potential factors influencing the population was proposed. The results showed that nitrogen-fixing activities of rice rhizosphere under different fertilizer managements were not significant different, but there were significant changes under different growth stages. Restriction enzyme fragment length polymorphism analysis (RFLP) also showed different nifH gene profiles at different growth stages, but not significantly different among fertilization treatments. Cloning technique was thus used to sequence the nifH gene, The results indicated growth stage, significant change of the rhizospheric nifH pool under long-term applications of fertilizers. Several nifH genes disappeared in some fertilization treatments. nifH gene compositions at different growth stages were similar in different fertilization treatments but varied in from dominancy. Although RFLP method could not distinguish the nifH gene profiles under the different fertilizations, cloned nifH gene sequence revealed significantly different among fertilization treatments. These results support the hypothesis that the nitrogen fixing bacterium population changed considerably after long-term fertilizer application. The study also established the first nifH gene database in Taiwan paddy soil, and found that there are a large number of newly discovered nifH gene unknown.
URI: http://hdl.handle.net/11455/28270
其他識別: U0005-2008201013053700
Appears in Collections:土壤環境科學系

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