高效解磷菌株的筛选及其复合微生物肥料的研制与肥效研究

发布时间：2018-05-08 21:47

本文选题：解磷菌 + Pseudomonas　；参考：《南京农业大学》2015年硕士论文

【摘要】：由于有机肥见效慢、施用面积及用量严重不足和生物菌肥效率低、应用规模小、无区域针对性、市场推广难度大等肥料使用特点,加上长期大量与不合理施用化肥,导致我国土壤结构遭到严重破坏、有机质含量急剧下降、土壤微生物群落结构严重失衡,从而造成农产品产量和质量大幅度下降和肥料利用率逐年降低等一系列突出的问题。同时,大量的农作物秸秆和畜禽粪便等农业废弃物制成的有机肥由于低的肥效和附加值,造成使用面积和使用量严重受阻,导致大量资源浪费和严重的生态环境污染。为了农业可持续发展和农民对便捷高效的耕作模式的追求,复合微生物肥料是未来肥料的发展趋势。复合微生物肥料集微生物的促效增效,有机肥的缓效长效和化肥的速效高效为一体,实现三肥合一,具有营养平衡、改土培肥、增产增收、改善果实品质等特点。本研究利用实验室保存的解淀粉芽孢杆菌Bacillus amyloliquefaciens T-5、NJN-6,以及本研究筛选出的高效解磷菌株——Y2与普通有机肥通过二次发酵生成生物有机肥,再配以一定比例的无机化肥,通过圆盘造粒法研制复合微生物肥料,通过粉状和颗粒复合微生物肥料储存试验,盆栽效果及养分释放试验研究上述肥料的生产实际性。结果表明,本研究首先从马铃薯根际筛选出解磷菌58株,通过定性和定量测定了解磷菌的解磷能力。摇瓶复筛试验结果表明,溶液中速效磷含量升高较大的菌株是Y2和Y40,含量分别达到341.17 μg/mL和300.02 μg/mL,菌株Y40发酵液pH值下降幅度最大,下降到4.04,该两株菌被选择进行进一步研究。在两季解磷菌的盆栽(番茄、茄子、马铃薯、玉米、烟草)促生试验中可以得出添加解磷菌株Y2和Y40的处理对这几种作物均具有较好的促生效果,同时可以得出这两株菌对茄科作物具有较好的促生效果,且功能菌能在植株根际有效定殖。结合实验室及盆栽实验数据最后筛选出菌株Y2为解磷能力较强菌株。经过生理生化结合16SrRNA序列发育分析,该两株菌分别被鉴定为假单胞菌(Pseudomonas frederikbergensi)和乙酸钙不动产杆菌(Acinetobacter calcoaceticus),在复合微生物肥料存放试验中,存放6个月时,分别含T-5、NJN-6菌株的10%和15%养分粒状复合微生物肥料有效活菌数均达相应的国家标准,菌株T-5在其制成的复合微生物肥料中存活能力优于菌株NJN-6,与此同时,10%养分复合微生物肥料中有效活菌数高于15%养分,含水量变化保持在10%-20%之间。在芽孢萌发试验中,复合微生物肥料中功能菌芽孢萌发率均高于60%。盆栽促生试验结果初步表明,颗粒状复合微生物肥料的整体促生效果优于粉状复合微生物肥料。在富含Y2菌株的复合微生物肥料存放试验中,存放6个月时,10%和15%养分粉状及粒状复合微生物肥料中功能菌有效活菌数均高于国家标准,但数量低于生物有机肥中所含活菌数,且10%养分含量复合微生物肥料中活菌数优于15%养分肥料所含活菌数。存储期间,肥料含水量在前3个月时保持在15%-20%之间,后3个月时保持在10%-15%之间。盆栽试验结果表明,分别施用存储6个月后的富含Y2菌株的添加粉状和颗粒状复合微生物肥料处理中,添加10%养分复合微生物肥料处理中番茄长势显著优于添加15%养分复合微生物肥料的处理,且该养分下,颗粒状肥料的促生效果显著优于粉状肥料。
[Abstract]:Due to the slow effect of organic manure, serious shortage of application area and dosage and low biological bacterial fertilizer efficiency, the application of small scale, no regional pertinence, difficult market popularization and so on, combined with long term and irrational application of fertilizer, resulted in serious deterioration of soil structure in China, sharp decline of organic matter content and soil microbial community knot. There is a serious imbalance in structure, which causes a series of outstanding problems, such as the yield and quality of agricultural products and the decrease of the utilization rate of fertilizer year by year. At the same time, the organic manure made from agricultural wastes such as crop straw and livestock waste, caused by low fertilizer efficiency and added value, caused serious obstruction of the area and use, resulting in a large amount of resources. In order to promote the sustainable development of agriculture and the farmers' pursuit of convenient and efficient farming mode, compound microbial fertilizer is the trend of future fertilizer development. Compound microbial fertilizer combines the effect of microorganism to promote efficiency, the slow effect of organic fertilizer and the effective efficiency of chemical fertilizer as a whole, and achieve three fertilizer in one. The characteristics of nutrition balance, soil improvement and fertilization, increase of yield and harvest, improve the quality of fruit, and so on. This study uses the laboratory preserved Bacillus amyloid Bacillus amyloliquefaciens T-5, NJN-6, and the screening of high efficiency phosphate solubilizing bacteria - Y2 and common organic manure by two times of fermentation to produce biological organic manure, and then to a certain proportion of inorganic fertilizer. Chemical fertilizer was prepared by the method of disc granulation. Through the experiment of powdery and particulate compound microbial fertilizer storage, the potted effect and nutrient release test were used to study the practical production of the above fertilizer. The results showed that 58 strains of phosphate solubilizing bacteria were selected from the potato rhizosphere first, and the Phosphorus Solution was determined by qualitative and quantitative determination. The test results of shake flask rescreening showed that the strains with higher content of available phosphorus in the solution were Y2 and Y40, the content reached 341.17 mu g/mL and 300.02 g/mL respectively, the pH value of the strain Y40 fermentation liquid decreased to 4.04, and the two strains were selected for further study. In the two season, the potted plants of the phosphate bacteria (tomato, eggplant, potato and jade) The treatment of Y2 and Y40 with phosphate solubilizing strain can have good growth promoting effect on these crops. At the same time, it can be found that the two strains have good growth promoting effect on the Solanaceae crops, and the functional bacteria can be effectively colonized in the rhizosphere of the plant. The strain Y2 is finally screened out by the laboratory laboratory and the pot experiment data. The two strains were identified as Pseudomonas (Pseudomonas frederikbergensi) and calcium acetate actobacillus (Acinetobacter calcoaceticus) by physiological and biochemical combination of 16SrRNA sequence development. In the storage test of compound microbial fertilizer, 10% and 15 of NJN-6 strains were stored for 6 months, respectively. The number of effective living bacteria of% nutrient granular compound microbial fertilizer reached the corresponding national standard. The surviving ability of strain T-5 in the compound microbial fertilizer made by it was better than that of strain NJN-6. At the same time, the number of effective living bacteria in the 10% nutrient compound microbial fertilizer was higher than that of 15%, and the change of water content was kept between 10%-20%. The germination rate of the spores of functional bacteria in the microbial fertilizer was higher than that of the 60%. pot cultivation test. The results showed that the overall growth promoting effect of the granular compound microbial fertilizer was better than that of the powdery compound microbial fertilizer. In the storage test of the compound microbial fertilizer rich in Y2 strain, the powder and granular compound microorganism were stored for 6 months. The number of active bacteria in functional bacteria in fertilizer is higher than that of the national standard, but the number of active bacteria is lower than that contained in bio organic fertilizer, and the number of living bacteria in the compound microbial fertilizer with 10% nutrient content is better than that of 15% nutrient fertilizer. During the storage, the water content of the fertilizer is kept between 15% -20% in the first 3 months and between the last 3 months. The experiment results showed that in the treatment of adding powder and granular compound microbial fertilizer rich in Y2 strain after 6 months of storage, the growth potential of Tomato in the treatment of 10% nutrient compound microbial fertilizer was significantly better than that of adding 15% nutrient compound microorganism fertilizer, and the growth promoting effect of granular fertilizer was significantly better than that under this nutrient. Powdered fertilizer.

【学位授予单位】：南京农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S144

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