摘要
采用Ion Torrent Personal Genome Machine(PGM)测序平台,以16S rRNA基因V4-V5区为标靶,沼泽土壤和草坪土壤为材料,建立土壤微生物16S rDNA检测方法。结果表明,当测序序列数达到15 000条左右,两种土壤样品的Shannon曲线趋于平稳,而且沼泽土壤的多样性指数明显高于草坪土壤。在沼泽土壤和草坪土壤中分别检测到7 615和8 072个OTUs,分属于30门、53纲、93目、130科、182属和29门、52纲、88目、125科、183属。方差分析表明两类土壤存在着丰度差异显著的微生物类群和OTUs。根据样品间相似度分析,所有测试样本可明显分成两组,而且3个重复各自归为一组。可见,该试验所建立的Ion Torrent PGM土壤微生物16S rDNA检测法具有高通量、高灵敏度等优点,能够较为全面和准确的反映土壤微生物群落结构,从而可为土壤微生物群落结构研究提供强有力的检测工具。
We aimed to establish the high throughput sequencing method for soil microbial 16S rDNA using Ion Tor- rent PGM. In this study, the V4-V5 variable region of 16s rDNA was amplified from the marsh soil and meadow soil by PCR and sequenced respectively. The Shannon curves of the two soil samples tended to be stabilized when the se- quencing depth reached 15 000 sequences. The diversity index showed that the bacterial communities in the marsh soil were more abundant than that in the meadow soil. A total of 7 615 and 8 072 OTUs were identified in the marsh soil and meadow soil, respectively, which corresponded to 30 phyla,53 classes,93 orders, 130 families and 182 gen- era in the marsh soil and 29 phyla,52 classes,88 orders, 125 families and 183 genera in the meadow soil. The vari- ance analysis showed that there were significant differences in the abundance of microbial compositions and OTUs be- tween the marsh soil and meadow soil. According to the similarity analysis, all the tested samples could be divided into two groups, and the three repeats were classified into the same group. Therefore, it was suggested that the soilmicrobial 16S rDNA detection method based on Ion Torrent PGM developed in our study could comprehensively and accurately reflect the soil microbial community structure, and provide a powerful tool for the detection of soil microbi- al community structure.
出处
《浙江农业学报》
CSCD
北大核心
2015年第12期2165-2170,共6页
Acta Agriculturae Zhejiangensis
基金
浙江省科技计划项目(2013C37034)
