School of Biological Sciences, The University of Hong Kong, Pokfulam Road, Hong Kong, China.
Appl Microbiol Biotechnol. 2013 Oct;97(19):8741-56. doi: 10.1007/s00253-013-5169-2. Epub 2013 Aug 24.
Repetitive extraction of DNAs from surface sediments of a coastal wetland in Mai Po Nature Reserve (MP) of Hong Kong and surface Baijiang soils from a rice paddy (RP) in Northeast China was conducted to compare the microbial diversity in this study. Community structures of ammonia/ammonium-oxidizing microorganisms in these samples were analyzed by PCR-DGGE technique. The diversity and abundance of ammonia-oxidizing archaea (AOA), ammonia-oxidizing bacteria (AOB), and anaerobic ammonium-oxidizing (anammox) bacteria were also analyzed based on archaeal and bacterial ammonia monooxygenase subunit A encoding (amoA) and anammox bacterial 16S rRNA genes, respectively. DGGE profiles of archaeal and bacterial amoA and anammox bacterial 16S rRNA genes showed a similar pattern among all five repetitively extracted DNA fractions from both MP and RP, except the anammox bacteria in RP, indicating a more diverse anammox community retrieved in the second to the fifth fractions than the first one. Both soil and marine group AOA were detected while soil and coastal group AOB and Scalindua-anammox bacteria were dominant in MP. Soil group AOA and marine group AOB were dominant in RP, while both Scalindua and Kuenenia species were detected in RP. Pearson correlation analysis showed that the abundance of archaeal and bacterial amoA and anammox bacterial 16S rRNA genes was significantly correlated with the DNA concentrations of the five DNA fractions from MP, but not from RP (except the archaeal amoA gene). Results suggest that anammox bacteria diversity may be biased by insufficient DNA extraction of rice paddy soil samples.
本研究从香港米埔自然保护区(MP)的沿海湿地表面沉积物和中国东北稻田的 Baijiang 土壤中重复提取 DNA,以比较这两个地点的微生物多样性。采用 PCR-DGGE 技术分析了这些样品中氨/铵氧化微生物的群落结构。还分别基于古菌氨单加氧酶亚基 A 编码(amoA)和厌氧氨氧化菌 16S rRNA 基因,分析了氨氧化古菌(AOA)、氨氧化细菌(AOB)和厌氧氨氧化(anammox)细菌的多样性和丰度。古菌和细菌 amoA 以及 anammox 细菌 16S rRNA 基因的 DGGE 图谱在 MP 和 RP 的所有五个重复提取的 DNA 分数之间表现出相似的模式,除了 RP 中的 anammox 细菌,表明在第二到第五个分数中回收的 anammox 群落更加多样化。检测到土壤和海洋组 AOA,而土壤和沿海组 AOB 和 Scalindua-anammox 细菌在 MP 中占优势。土壤组 AOA 和海洋组 AOB 在 RP 中占优势,而在 RP 中同时检测到 Scalindua 和 Kuenenia 物种。Pearson 相关分析表明,古菌和细菌 amoA 和 anammox 细菌 16S rRNA 基因的丰度与 MP 中五个 DNA 分数的 DNA 浓度显著相关,但与 RP 中的 DNA 浓度无关(古菌 amoA 基因除外)。结果表明,anammox 细菌的多样性可能受到稻田土壤样品 DNA 提取不足的影响。