請用此 Handle URI 來引用此文件: http://hdl.handle.net/11455/95768
標題: 羽毛分解菌的分離與鑑定及其於芥菜健康管理之應用
Isolation and Identification of Feather-Degrading Bacteria and their Application for Managing Leaf Mustard Health
作者: 林庭緯
Ting-Wei Lin
關鍵字: 羽毛廢棄物
羽毛分解菌
土壤添加物
芥菜黃葉病
Fusarium oxysporum f.sp. rapae
feather wastes
feather-degrading bacteria
organic amendments
Fusarium oxysporum f.sp. rapae
mustard Fusarium wilt
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摘要: 西元2012與2013年,台灣廢棄的羽毛分別高達27,003與27,804公噸,若不妥善處理會有汙染環境之虞。羽毛富含豐富的氮素,其成分係由結構穩定的角蛋白組成,動植物不易直接利用。因此,本研究的主要目的在於篩選與鑑定具有分解羽毛能力的微生物,進而利用分解後的羽毛粉搭配其他有機資材,研製可有效保護芥菜健康的土壤添加物。首先從台南地區的廢棄雞毛中分離出具有分解羽毛能力的細菌共27菌株,其中PDML-0417菌株分解羽毛的能力最佳,分解率可達42%以上,此外有10株地分解率介於26-35%,其餘分離株的分解率則皆小於25%。進一步,利用16S rDNA 序列比對法,將PDML-0417菌株鑑定為Deinococcus ficus Lai et al.。在不同緩衝溶液之1%(w/v)羽毛培養基培養D. ficus PDML-0417後,發現羽毛培養基添加0.1%磷酸鉀緩衝劑(代號FP)可使羽毛分解率顯著提高至80%。將FP培養基作為基礎配方,以單一或不同組合的無機鹽類培養D. ficus PDML-0417,發現皆無法進一步提高羽毛分解率,但若將培養基中的羽毛濃度提高至3、5及7%(w/w),結果顯示以FPK01、FPN01及FPKN01配方培養PDML-0417,其羽毛分解率的下降趨勢皆較FP對照組緩慢。以羽毛粉為基質搭配其他有機資材施用於土壤中,評估其對芥菜生長的影響與防治芥菜黃葉病Fusarium oxysporum Schlecht. f. sp. rapae J. Enya, M. Togawa, T. Takeuchi et T. Arie的效果,結果顯示單獨添加0.25% (w/w)羽毛粉即可顯著促進芥菜植株的鮮重,且使芥菜黃葉病的罹病度從71.1%降低至47.8%;若將羽毛粉搭配蝦蟹殼粉或蓖麻粕則分別可使芥菜鮮重提高13.8%與17.1%,並降低13.3%與3.5%黃葉病的罹病度。進一步,將前述具有促進芥菜生長與防治黃葉病功效的蝦蟹殼粉、蓖麻粕、碳化稻殼及羽毛粉均勻混拌成FSC-35混合物,添加於土壤中,評估其對芥菜生長的影響與防治黃葉病的功效,結果顯示添加1% (w/w)FSC-35混合物可提高芥菜的鮮重與根重分別達159%與198%,且可使芥菜黃葉病的罹病度從95.6%降低至52.2%。在土壤中分別添加FSC-35混合物及其組成分後,每隔6天測量土壤pH值,發現土壤添加FSC-35混合物與蝦蟹殼粉的處理,pH值於第6天時上升至6.0以上,可維持24天均沒有下降的趨勢。進一步,將FSC-35混合物添加於土中,測量微生物相的變化,發現施用後30天內的土壤細菌族群量均較對照組高;惟真菌、放線菌及芥菜黃葉病菌的族群量在添加與未添加的土壤中並無顯著差異。
In 2012 and 2013, the production of feather waste was 27,003 and 27,804 tons respectively in Taiwan. Improper disposal of feather waste causes serious environmental problems. Even though feathers are rich in nitrogen, plants and animals cannot absorb them efficiently because feathers mostly consist of keratin, which is a stable protein. The main purpose of this study was to isolate and identify the feather-degrading bacteria and to evaluate their utilization. Twenty-seven strains of feather-degrading bacteria were obtained from feather wastes. Among them, strain PDML-0417 showed the highest degradation ability (42 %). Ten strains displayed degradation efficiencies between 26-35 %, and the others had less than 25 % degradation rates. According to partial 16s rDNA sequences, PDML-0417 was identified as Deinococcus ficus Lai et al. The ability of PDML-0417 in degradation of feather waste was evaluated by using three different buffers. The results indicated that PDML-0417 cultured in 1% (w/v) feather medium amended with 0.1% (w/v) potassium phosphate buffer (FP medium) enhanced the rates of feather degradation from 42% to 80%. FP medium with PDML-0417 was further formulated with different inorganics individually or in various combinations. The results revealed that none of inorganic salts could further improve the rates of feather degradation when the cultural solution was only amended with 1% (w/v) feather. However, amendment of FPK01, FPN01 or FPKN01 mixture could maintain more than 50% feather-degrading ability of PDML-0417 when the cultural solution included 1-3% (w/v) of feather. Combinations of feather meal with each of eight organic materials were evaluated their efficacy for managing leaf mustard health. In greenhouse trials, amendment of soil with 0.25% (w/w) feather meal alone was effective in increasing 75% of leaf mustard fresh weight and reducing 23.3% of disease severity of leaf mustard Fusarium wilt caused by Fusarium oxysporum f. sp. rapae. Furthermore, feather meal mixed with shrimp and crab shell powder or castor bean pomace was able to further increase fresh weight by 13.8% or 17.1% and reduce disease severity by 13.3 or 3.5% respectively. In advance, feather meal, shrimp and crab shell powder, castor bean pomace, and carbonized rice husk were selected to conduct the formulation of FSC-35 mixture. Amendment of soil with 1% (w/w) FSC-35 mixture was more effective in reducing the disease severity of mustard Fusarium wilt by 43.4% and increasing leaf mustard fresh weight and root weight by 159% and 198% respectively. The pH value of soil was raised to a level more than 6.0 and maintained 6 to 24 days after soil was amended with 1% (w/w) FSC-35 mixture or 0.2% (w/w) shrimp and crab shell powder. The population density of bacteria was significantly increased more than 108 cfu/g soil when soil was amended with 1% (w/v) FSC-35 mixture. However, populations of fungi included leaf mustard Fusarium wilt pathogen and actinomycetes were not markedly changed between soil amendment and non-amendment.
URI: http://hdl.handle.net/11455/95768
文章公開時間: 2020-02-06
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