木质纤维素分解复合菌系微生物组成多样性及其在牛粪堆肥化中的应用
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摘要
为了研究高效稳定的木质纤维素降解复合菌系的微生物组成,明确该复合菌系作为接菌剂在牛粪堆肥处理中的应用效果,探索在牛粪堆肥化过程中把添加的无机养分有机化、提高堆肥养分含量的可能性,采用16S rDNA克隆技术分析了复合菌系微生物多样性,通过对比试验研究复合菌系对堆肥发酵进程的影响,研究了添加磷酸二铵、尿素、硫酸钾中氮磷钾元素的有机化率。试验结果如下:
(1)对复合菌系构建克隆文库,通过菌落PCR获得了184个16S rDNA片断,经RFLP筛选获得24个代表性克隆,序列数据库比对结果中有62%的近缘种为已知菌,分别归属于Clostridium属、Brevibacillus属、Ureibacillus属、Proteiniborus属和Rhizobium。其余38%的菌的近缘种属于难培养菌。
(2)以复合菌系为接菌剂进行牛粪堆肥发酵,结果表明添加接菌剂的处理可使堆体提前4天进入高温期,堆体中纤维素、半纤维素、木质素的降解率分别比对照提高了12.8%、3.51%和4.78%,与对照相比,明显降低了堆肥前期的氮损失,总氮损失减少了3.9%。
(3)一次性添加无机养分进行堆肥发酵,与对照(不添加无机养分处理)相比,堆肥中全氮、全磷、全钾含量分别提高了47.5%、115.9%、59.2%,添加的全氮、全磷和全钾有机转化率分别为40.0%、20.34%和19.47%。
通过以上研究明确了复合菌系的微生物组成,复合菌系作为接菌剂应用于堆肥中明显改善堆肥质量,减少养分损失,在牛粪堆肥化过程中一次性添加无机养分可以提高堆肥有机养分和总养分含量。
In order to detecting the microbial diversity of the microbial community with high capacity of lignocelluloses degradation, identifying the effect of microbial community used as inoculant on cattle manure composting, the 16S rDNA clone technique combined with plate isolation was used. Diammonium phosphate, urea, potassium sulfate was added in materials at the beginning of composting for investigating the probability of organification of added chemical nutrient and increasing of the total nutrient during cattle manure composting process with the method of comparative trial. Results were as follows.
One hundred eighty four clones and 24 typical strains were detected by method of restriction fragment length polymorphism. The sixty four percentage of the closest relative among them belonged to Clostridium, Brevibacillus, Ureibacillus, and Rhizobium through DNA sequence aligment. Further more, the rest of thirty six percentage clones belonged to uncultured bacterium clone.
The duration that temperature of compost pile reached 50℃was shorten after the first four days inoculated, and the themophilic phase of temperature of compost pile above 50℃was lengthened by seven days under the condition of inoculated treatment compared with control. The degradation rate of cellulose, hemi-cellulose and lignocelluloses was also increased by 12.8%, 3.51% and 4.78% compared to the control, respectively. The time of nitrogen loss was put off and the lost quantity of nitrogen was decreased by 3.9% under inoculated condition.
The content of total nitrogen, total phosphorus and total potassium was highly increased by adding inorganic nutrient compared to the control treatment that inorganic nutrient was not added, and the increase rate were 47.5%, 115.9% and 59.2% respectively. At the end of the fermentation there were 40.0% of the nitrogen, 20.3% of phosphorus and 19.5% of potassium which had been add into composting were converted to organic nutrient.
The results showed that there were a lot of microbial species consisted in microbial community with high capacity of lignocelluloses degradation. The quality of compost was improved and the nutrient loss was decreased when microbial community was used as inoculum. The organic nutrient and total nutrient content of compost was increased by adding inorganic nutrient.
(1)对复合菌系构建克隆文库,通过菌落PCR获得了184个16S rDNA片断,经RFLP筛选获得24个代表性克隆,序列数据库比对结果中有62%的近缘种为已知菌,分别归属于Clostridium属、Brevibacillus属、Ureibacillus属、Proteiniborus属和Rhizobium。其余38%的菌的近缘种属于难培养菌。
(2)以复合菌系为接菌剂进行牛粪堆肥发酵,结果表明添加接菌剂的处理可使堆体提前4天进入高温期,堆体中纤维素、半纤维素、木质素的降解率分别比对照提高了12.8%、3.51%和4.78%,与对照相比,明显降低了堆肥前期的氮损失,总氮损失减少了3.9%。
(3)一次性添加无机养分进行堆肥发酵,与对照(不添加无机养分处理)相比,堆肥中全氮、全磷、全钾含量分别提高了47.5%、115.9%、59.2%,添加的全氮、全磷和全钾有机转化率分别为40.0%、20.34%和19.47%。
通过以上研究明确了复合菌系的微生物组成,复合菌系作为接菌剂应用于堆肥中明显改善堆肥质量,减少养分损失,在牛粪堆肥化过程中一次性添加无机养分可以提高堆肥有机养分和总养分含量。
In order to detecting the microbial diversity of the microbial community with high capacity of lignocelluloses degradation, identifying the effect of microbial community used as inoculant on cattle manure composting, the 16S rDNA clone technique combined with plate isolation was used. Diammonium phosphate, urea, potassium sulfate was added in materials at the beginning of composting for investigating the probability of organification of added chemical nutrient and increasing of the total nutrient during cattle manure composting process with the method of comparative trial. Results were as follows.
One hundred eighty four clones and 24 typical strains were detected by method of restriction fragment length polymorphism. The sixty four percentage of the closest relative among them belonged to Clostridium, Brevibacillus, Ureibacillus, and Rhizobium through DNA sequence aligment. Further more, the rest of thirty six percentage clones belonged to uncultured bacterium clone.
The duration that temperature of compost pile reached 50℃was shorten after the first four days inoculated, and the themophilic phase of temperature of compost pile above 50℃was lengthened by seven days under the condition of inoculated treatment compared with control. The degradation rate of cellulose, hemi-cellulose and lignocelluloses was also increased by 12.8%, 3.51% and 4.78% compared to the control, respectively. The time of nitrogen loss was put off and the lost quantity of nitrogen was decreased by 3.9% under inoculated condition.
The content of total nitrogen, total phosphorus and total potassium was highly increased by adding inorganic nutrient compared to the control treatment that inorganic nutrient was not added, and the increase rate were 47.5%, 115.9% and 59.2% respectively. At the end of the fermentation there were 40.0% of the nitrogen, 20.3% of phosphorus and 19.5% of potassium which had been add into composting were converted to organic nutrient.
The results showed that there were a lot of microbial species consisted in microbial community with high capacity of lignocelluloses degradation. The quality of compost was improved and the nutrient loss was decreased when microbial community was used as inoculum. The organic nutrient and total nutrient content of compost was increased by adding inorganic nutrient.
引文
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