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標題: 長期水旱田輪作生態系土壤中不可培養及可培養微生物之功能多樣性與群落變動之研究(I)
Studies on Functional Diversity and Dynamics of Non-Culturable and Culturable Microbial Communities in the Paddy-Upland Crop Rotation Ecosystem (I)
作者: 楊秋忠
關鍵字: 農業化學類, 自然生態保育;基礎研究;Long-term agro-ecosystem;長期農業生態系;不可培養微生物;土壤DNA;變性梯度凝膠電泳;Culturable microorganisms;Non-culturable microorganisms;Soil DNA;Denaturing gradient gel electrophoresis
土壤微生物在農業生態系物質循環及養分釋放中扮演重要角色,除參與各種養分代謝以維持土壤地力外,與農作物產量與品質亦息息相關。由於傳統研究方法的限制,早期僅能對於可在培養基中生長的微生物進行分析。在農業生態系研究中,不可培養之微生物及長期不同田間管理與作物生長對微生物群落影響之研究仍甚為缺乏。台灣位處亞熱帶地區,與歐美溫帶地區不同,值得深入及長期研究。本研究將在已建立九年以玉米與水稻輪作系統研究長期施用化學肥料及不同有機肥料(堆肥、綠肥及泥炭)的農地中,研究土壤中不可培養微生物特性與群落結構之影響。在基因型研究上,本研究將利用土壤總DNA 萃取、菌種專一性引子設計、聚合.連鎖反應、變性梯度凝膠電泳分析、土壤DNA 選殖等技術,進行土壤環境中微生物群落結構分析,並利用功能性基因偵測特定微生物族群之消長情形。另外亦將以傳統培養法自環境中分離菌株,利用選擇性培養基篩選並鑑定可培養部分之微生物族群作為採樣時土壤的比較。分析長期施用不同肥料下土壤之生態功能如固氮活性、硝化作用、脫氮作用、溶磷活性等指標因子與微生物群落結構之相關性。應用分生技術可研究微生物結構受到作物種類、不同肥培管理下所呈現的多樣性,並探討有機質綠肥、豬糞堆肥與泥炭對農田土壤中微生物種類之影響。期能對土壤環境中總體微生物所扮演的角色有更深入之瞭解,有助於農業栽培方法之改進與農業生產品質之提升。

Soil microorganisms play a vital role in the nutrient turnover in natural ecosystem. Soilmicroorganisms are associated not only with crop production but also impact soil fertilitymaintenance. In past, studies on the diversity of natural microbial populations have beenrestricted only to culturable microorganisms, which are far a smaller number than theestimated total bacterial populations. Therefore our knowledge about the dynamics of naturalmicrobial communities and their interactions with plants has been limited, because only aminor fraction of all cells in natural ecosystems are accessible to cultivation techniques.Researches in past on soil fertility and nutrient cycling in Taiwan have made some goodcontributions to our understanding of native soil ecosystem processes and their managementbut, the long-term research is limited and fragmented. Hence, there is a great urgency andneed to understand these ecological functions of microbial communities (culturable andnon-culturable) in long-term agro & soil ecosystem as to gain a better insight to the role ofmicrobial community structure for sustainable soil management.In the current proposal the paddy-upland crop rotation system, which has beenestablished to study the functional diversity and dynamics of culturable microbialcommunities under different organic fertilizer treatments for nine years will be used asexperimental site. Non-culturable microbial genotypic diversity will be studied by the directDNA isolation from soil, genus-specific primers designation, polymerase chain reaction,denaturing gradient gel electrophoresis, soil DNA cloning and sequencing methods. Theculturable microorganisms will also be screened, isolated and identified as a part of totalmicrobial populations. Simultaneously functional diversity of microorganisms such asnitrogen fixation, nitrification, denitrification, ammonia oxidation and phosphatesolubilization will be analyzed phenotypically and genetically as to appreciate their role innutrient cycling and plant community interaction. The present/proposed study willconsequently lead us to gain a better insight into the diverse role played by microorganisms inthe agro-ecosystems, their management and manipulation which, is the key for sustainableagriculture and better crop production.
其他識別: NSC93-2313-B005-049
Appears in Collections:土壤環境科學系

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