【作者】 曹广祥；

【导师】 黄为一；

【作者基本信息】 南京农业大学 ， 微生物学， 2003， 硕士

【摘要】 本文分离筛选了两株具有纤维素酶活性的嗜热细菌，两株可降低氨气释放量的耐高温酵母和一株具有多酚氧化酶活性的高温放线菌，建立了人工快速腐熟畜禽粪的菌群，称为NMF菌群，本文研究了该菌群中各菌株的分类地位、发酵条件、植物安全性、对畜禽粪便腐熟的影响和在生物缓释氮肥生产中的作用。 细菌编号B022和B023,酵母编号Y124和Y127，放线菌编号F04。通过形态特征和生理生化特性初步鉴定B022为芽孢杆菌属、B023为芽孢杆菌属，Y124为裂殖酵母属、Y127为汉逊酵母属，F04为高温放线菌属。植物安全性试验没有发现NMF菌剂对水稻、小麦和番茄有致病现象，也没有发现NMF菌群单株发酵液对供试植物病原菌有抑制现象。 通过单因子试验和多因子试验，确定B022的最适发酵培养基是淀粉0.8％、葡萄糖0.2％、黄豆粉0.2％、尿素0.1％、磷酸氢二钾0.01％、温度50℃、起始pH值7.2，B023的最适发酵培养基是淀粉0.8％、葡萄糖0.2％、黄豆粉0.2％、硫酸铵0.1％、磷酸氢二钾0.02％、温度45℃、起始pH值7.5，Y124的最适发酵培养基是葡萄糖0.5％、蔗糖1.5％、酵母膏0.2％、硫酸铵0.1％、磷酸氢二钾0.02％、温度28℃、起始pH值6.5，Y127的最适发酵培养基是葡萄糖1.5％、麦芽糖0.5％、酵母膏0.1％、尿素0.1％、磷酸氢二钾0.02％、温度37℃、起始pH值6.4，F04的最适发酵培养基是淀粉0.8％、葡萄糖0.2％、黄豆粉0.2％、硝酸钠0.1％、磷酸氢二钾0.02％、温度45℃、起始pH值7.8。 奶牛粪便处理腐熟试验发现，接种NMF菌剂的奶牛粪在25天内可以达到腐熟标准，氮磷钾总养分和腐植酸分别比不接菌处理提高18％和45％。NMF菌剂可以通过改变微生物区系、增加纤维素酶活性和多酚氧化酶活性等途径加快腐熟速度，促进腐熟程度。经测定，接种NMF菌剂生产的有机肥完全满足中华人民共和国国家标准GB7959-87无害化要求，对油菜种子发芽的抑制作用基本解除，重金属含量符合商品有机肥质量标准。 奶牛粪便腐熟时添加硫酸铵和尿素发现，接种NMF菌剂时初期pH下降快，有机酸含量和腐植酸含量分别比未接种高54.6％和28.1％，蛋白氮提高了8.1％，速效氮占全氮的22.3％，未接种处理为48.7％。NMF菌剂将速效氮转化为蛋白质或以有机酸铵、腐植酸铵形式固定下来可能是提高氮素缓释效果的主要途径。 人为接种植物病原菌的盆栽试验中，施用NMF菌剂生产的有机肥处理中，小麦纹枯病发病率比不施肥对照和施用堆肥处理分别低54.2％、59.0％；番茄灰霉病发病率比不施肥对照和施用堆肥处理分别低23.7％和17.3％。分析可能是腐熟程度差的堆肥中含有供植物病原菌生长的营养物质，而腐熟程度高的有机肥改变了土壤微生物区系。 有机肥和生物缓释氮肥的甜椒盆栽试验显示，与化学氮肥相比生物缓释氮肥和有机无机复混肥均显著促进甜椒生长，提高土壤有机质的含量，Vc含量提高了25.1％和13.0％，硝酸盐含量降低了59.8％和43.3％。在氮素利用率方面，生物缓释氮肥比有机无机复混肥和化学氮肥分别提高了19.9％和38.8％。更多还原

【Abstract】 The author severed and filtered two thermalphilic bacteria in possession of eellulase, two yeasts depressing NH3 release capacity which can tolerate high temperature , an actinomyceto possessing laccase activity.NMF microbiological fertilizer is constituted by five strains above. Its classification status , fermentation conditions , plant security , effects on decayed animal manure and action on biological slow-release nitrogenous fertilizer are studied in this article.Bacteria were named B022, B023 respectively and yeasts Y124, Y127, actinomyceto F04.Through experiment about their configuration and physiological , biochemical characters,B022, B023 are identified as bacillus spp. And Y124, Y127, F04 are identified as Schijosacchar , Hansenula, Thermoactinomyces respectively. NMF biological fertilizer cannot incur diseases when added in paddy , wheat and tomato fields during plant security experiment.Results show that NMF fermentation liquor cannot inhibit plant pathogenic bacteria offered in this article.Through single-factor and multiplefactor experiment,the optimum fermentation medium of B022 is confirmed as starch 0.8%;glucose 0.2%;soybean powder 0.2%; carbamide 0.1%; potassium phosphate dibasic 0.01%;temperature 50 C; incept pH 7. 2; the optimum medium of B023 is starch 0.8%;glucose 0.2%;soybean powder 0.2%; ammonium sulfate 0.1%; potassium phosphate dibasic 0.02%;temperature 45 C; incept pH 7. 5; the optimum medium of Y124 is glucose 0.5%;sucrose 1.5%;yeast extract 0.2%; ammonium sulfate 0.1%; potassium phosphate dibasic 0.02%;temperature 28 C; incept pH 6. 5; the optimum medium of Y127 is glucose 1.5%;maltose 0.5%;yeast extract 0.1%; carbamide 0.1%;potassium phosphate dibasic 0.02%;temperature 37 C; incept pH 6. 4; the optimum medium of Y124 is glucose 0.5%;sucrose 1.5%;yeast extract 0.2%; ammonium sulfate 0.1%; potassium phosphate dibasic 0.02%;temperature 28 C; incept pH 6. 5; the optimum medium of F04 is starch 0.8%;glucose 0.2%;soybean powder 0.2%;sodium nitrate 0.1%;potassium phosphate dibasic 0.02%;temperature 45 C; incept pH 7.8.When cow dung was disposed, the cow dung inoculated by NMF can achieve standard after 25 days. The general nutrient and humic acid were increased by 18%, 45% respectively.NMF can hasten speed of disposing cow dung by way of alter microbiological fauna, increase laccase activity and cellucase activity.Organi fertilizer produced by NMF can satisfied GB7959-87 and relieve inhibition of germentation of rape.The content of heavy metal accord with quality criterion of commodity organic fertilizer.During disposal of cow dung,chemical nitrogenous fertilizer and NMF were added,at initial stages ,pH descended largely.The content of organic acid and humic acid were increased by 54.6%, 28.1% respectively;protein nitrogen increased by8.1%;valid nitrogen accouted for 22.3% of chief nitrogen ;valid nitrogen accouted for 48.7% of chief nitrogen when treated without inoculation.NMF would transform valid nitrogen into protein or fixed in form of organic acid ammonia and humic acid,which may be the important approach of nitrogen release.During potted plant inoculated by pathogenic microbes of wheat scab and gray mold of tomato,when treated by organic fertilizer produced by NMF,the ratio of wheat scab were reduced by 54.2 % - 59.0 % than comparision without fertilizer and compost treatment respectively ;the ratio of gray mold of tomato were reduced by 23.7% , 17.3% respectively.The reason maybe that there is nutrition for plant pathogenic microbes in compost.The microbes fauna was changed after organic fertilizer disposed.In potted pimiento experiment, after treatment by biological release nitrogenous fertilizer and compound fertilizer constituted by organic and inorganic fertilizer, pimiento grew faster and content of soil organic matter were increased compared with chemical nitrogenous fertilizer addition. Content of Vc were increased by 25.1 %, 13.0% respectively. The content of nitrate salt were decreased by 59.8%, 43.3% respectively. After treated by biological release nitrogen更多还原