接种菌剂对牛粪堆肥硝化和反硝化细菌群落影响的研究

发布时间：2018-04-26 16:52

本文选题：自然堆肥 + 菌剂堆肥　；参考：《东北农业大学》2015年硕士论文

【摘要】：将具有木质纤维素降解能力的菌剂,接种到以牛粪和水稻秸秆为材料的堆肥中,通过测定温度、p H值、C/N、种子发芽率、NH4+-N、NO3ˉ-N相关指标,考察了接种菌剂堆肥和自然堆肥腐熟情况。采用传统培养与聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)技术相结合的方法,研究了接种菌剂对牛粪堆肥中硝化、反硝化细菌数量及群落结构组成和多样性的影响,为进一步探讨硝化、反硝化细菌在堆肥氮素循环中的作用提供了理论基础。主要研究结果如下:(1)整个堆肥过程共持续45d,两组堆肥都经历了升温、高温、降温和腐熟阶段。菌剂堆肥最高温度为66℃高于自然堆肥的最高温度59℃,高温期分别持续13d和9d,表明接种菌剂可提高堆肥温度,延长高温期,同时p H值、C/N、种子发芽率、NH4+-N和NO3ˉ-N含量变化表明接种菌剂有利于微生物生长,促进堆肥腐熟,达到无害化标准。(2)接种菌剂使堆肥中可培养的硝化细菌数量多于自然堆肥,反硝化细菌数量少于自然堆肥。硝化和反硝化细菌数量与NH4+-N成负相关关系,与NO3ˉ-N成显著正相关关系(P0.05);菌剂堆肥相关系数大于自然堆肥。(3)接种菌剂对amo A基因标记的氨氧化细菌多样性及群落结构组成产生一定影响。和自然堆肥相比,菌剂堆肥中氨氧化细菌种类更加丰富。自然堆肥中亚硝化螺菌属(Nitrosospira)为优势种群,而菌剂堆肥中增加了新的类群,分别为亚硝化弧菌属(Nitrosovibrio)和亚硝化单胞菌属(Nitrosomonas)。(4)接种菌剂对nir S型反硝化细菌群落多样性无明显影响,而使nar G,nap A,nir K和nos Z型发生改变。和自然堆肥相比,nar G多样性指数在堆肥升温阶段较低,降温阶段较高;菌剂堆肥nap A和nos Z种类更加丰富,各时期多样性指数都大于自然堆肥;Nir K型反硝化细菌多样性在菌剂堆肥中减少。(5)接种菌剂对nir S型反硝化群落结构组成无影响,对nar G,nap A,nir K和nos Z影响较大。自然堆肥nar G型主要类群为发硫菌属(Thiothrix),而接种菌剂导致一些新的未培养类群产生,使原优势发硫菌属(Thiothrix)数量和所占比例减小;假单胞菌属(Pseudomonas)和某些未培养细菌是自然堆肥Nap A型反硝化细菌群落的优势菌属,菌剂堆肥除了含有这些类群外还出现了新的反硝化类群,分别属于红细菌属(Rhodobacter)和生丝微菌属(Hyphomicrobium);Nir K在菌剂堆肥后期检测到与红假单胞菌属(Rhodopseudomonas)同源性较高的反硝化细菌,但未检测到自然堆肥降温时期存在的无色杆菌属(Achromobacter)和一些未培养细菌,且使原自然堆肥的慢生根瘤菌属(Bradyrhizobium),根瘤菌属(Rhizobium),中慢生根瘤菌属(Mesorhizobium),副球菌属(Paracoccus)和假单胞菌属(Pseudomonas)表现出消失再出现的变化现象;自然堆肥nos Z型反硝化细菌包括假单胞菌属(Pseudomonas),产碱菌属(Alcaligenes)和无色杆菌属(Achromobacter),其中,假单胞菌属(Pseudomonas)为优势菌群,而菌剂堆肥中检测出自然堆肥所没有的新类群,包括盐单胞菌属(Halomonas),固氮螺菌属(Azospirillum),副球菌属(Paracoccus),草螺菌属(Herbaspirillum)和产碱菌属(Alcaligenes)。
[Abstract]:The microbial agent with lignocellulose degradation ability was inoculated into the composting of cow dung and rice straw. By measuring temperature, P H value, C/N, seed germination rate, NH4+-N, NO3 to -N related indexes, the condition of inoculant compost and natural composting was investigated. The traditional culture and polymerase chain reaction denaturation gradient gel electrophoresis (PC) were used. R-DGGE) the effect of inoculation on nitrification, denitrifying bacteria number, composition and diversity of denitrifying bacteria in cow manure composting was studied, and the theoretical basis for further discussion of nitrification and denitrifying bacteria in the nitrogen cycle of composting was provided. The main results were as follows: (1) the whole composting process lasted for a total of 45d, two The compost of the group experienced the stage of heating, high temperature, cooling and decayed. The highest temperature of the compost was 66 degrees centigrade higher than that of the natural compost at 59 degrees centigrade, and the high temperature period continued 13D and 9D respectively. It showed that the inoculant could increase the temperature of the compost and prolong the high temperature period. At the same time, the P H value, the C/N, the seed germination rate, the NH4+-N and the NO3 -N content showed that the inoculant was favorable. In the growth of microbes, the composting can be promoted to reach the harmless standard. (2) inoculants make the number of culturable nitrifying bacteria more than natural compost, the number of denitrifying bacteria is less than that of the natural compost. The number of nitrification and denitrifying bacteria is negatively correlated with NH4+-N, and the relationship with NO3 -N is significant positive correlation (P0.05); The number was greater than that of natural compost. (3) inoculants had a certain effect on the diversity and community structure of the ammoxidation bacteria marked by amo A gene. Compared with natural compost, the species of ammonia oxidizing bacteria were more abundant in the compost of the bacteria. In natural compost, the genus Nitrosospira was the dominant population, and the new group was added in the microbial composting. Respectively, Nitrosovibrio and Nitrosomonas. (4) inoculants had no obvious effect on the diversity of NIR S type denitrifying bacteria, but NAR G, nap A, NIR K and NOS Z type changed. The variety of nap A and NOS Z was more abundant, and the diversity index of each period was greater than that of natural compost, and the diversity of Nir K denitrifying bacteria decreased in the microbial composting. (5) inoculants had no influence on the structure of NIR S type denitrification community, and the main group of NAR G, nap A, etc. ) and the inoculation of some new uncultivated groups resulted in the production of some new uncultivated groups, which reduced the number and proportion of the original Thiothrix, and the Pseudomonas (Pseudomonas) and some uncultured bacteria were the dominant bacteria of the natural compost Nap A denitrifying bacteria community. Groups, respectively, belong to the genus Rhodobacter and Hyphomicrobium, and Nir K detected the denitrifying bacteria with higher homology with the genus Rhodopseudomonas in the later period of the microbial composting, but not the colorless bacilli (Achromobacter) and some uncultured bacteria in the natural composting period, and the original bacteria were not detected. The slow growing Rhizobium (Bradyrhizobium), the rhizobia (Rhizobium), the middle and slow Rhizobium (Mesorhizobium), the Paracoccus and the Pseudomonas (Pseudomonas) in the natural compost were changed, and the natural compost NOS Z denitrifying bacteria included the Pseudomonas (Pseudomonas) and the alkaloid (Alcaligene). S) and colorless bacilli (Achromobacter), among which Pseudomonas (Pseudomonas) is the dominant group, and new groups of natural compost are detected in microbial composting, including Halomonas, Azospirillum, Paracoccus, Herbaspirillum and Alcaligenes (Alcaligenes).

【学位授予单位】：东北农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S141.4

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