污泥干化芦苇床中芦苇内生菌群多样性研究

以污泥干化芦苇床中芦苇内生菌的多样性作为主要研究对象,结合16S rDNA高通量测序技术与PCR-DGGE技术,对污泥干化芦苇床和天然湿地中污泥菌群及芦苇内生菌群多样性进行了比较分析。结果表明,污泥干化芦苇床芦苇内生菌的细菌类群包含了20门、30纲、73目、152科、467属、1 167种细菌。其中变形菌门(65.6%)和拟杆菌门(25.5%)占绝对优势,γ-变形菌纲是丰度最高的纲,占总数的43.4%;排前20位的优势菌种相对百分比之和为46.5%,分属于所对应的18个属、12科、10目、6纲、3门。DGGE分析表明,污泥干化芦苇床中的污泥和芦苇内生菌优势菌群的种类与天然湿地相比差异显著;同时由于污泥的影响,芦苇内生菌群种类也发生了较为明显的变化。

On the endophytic bacteria diversity of the reed in the sludge drying reed bed

This study is concerned about the reed endophytic bacteria diversity from the sludge drying reed bed (SDRB).To achieve our study purpose,we have identified and analyzed the great varieties of the reed endophytic bacteria lying between SDRB and natural wetland with 16S rDNA high-throughput sequencing and PCR-based method for denaturing the gradient gel electrophoresis(PCR-DGGE).In our work,we have managed to extract the genomic DNA from the reed endophytic bacteria and the sludge by means of Soil DNA Kit(Omega).16S rDNA high flux sequence has been executed by Misep(Illumina,San Diego,CA) initiated and supplied by Sangon Biotech (Shanghai) Co.,Ltd.The aforementioned comparative analysis has shown that the reed endophytic bacteria of SDBR can be divided and categorized into 20 phylums,30 classes,73 orders,152 families,467 genuses,and 1167 species,wherein Proteobacteria (65.6％) and Bacteroidetes(25.5％) have been found existing in overwhelming majority.And,then,most of bacteria categories belong to γ-Proteobacteria(43.4％) with the top 20 strains(43.4％) being of 18 genuses,12 families,10 orders,6 classes,and 3 phylums.DGGE cluster analysis indicates that Shannon-Wiener indexes(H) of the sludge of 3 SDRB units have statistically found higher than that in the natural wetland,whilst,in contrast,Pielou evenness indexes(E) have been noticed lower than that in the natural wetland.Among the 3 SDRB units,H value of the sludge in Unit 2 (of reed and vent system) has been discovered to be highest whilst E value stands in the middle of the 3 units.On the one hand,H value of the reed endophytic bacteria in Unit 2 has been found the lowest one,but,on the other hand,E value is in the middle among the 3 units.Thus,it can be concluded that the sludge bacteria diversity in SDRB can be said significantly different from that of the natural wetland.At the same time,the variety of the reed endophytic bacteria in SDRB can also been defined and discriminated to be significantly different from that of the natural wetland due to the impact of the different features of the sludge.