Knowledge Resource Center for Ecological Environment in Arid Area
DOI | 10.1002/mbo3.495 |
Capabilities and limitations of DGGE for the analysis of hydrocarbonoclastic prokaryotic communities directly in environmental samples | |
Al-Mailem, Dina M.; Kansour, Mayada K.; Radwan, Samir S. | |
通讯作者 | Radwan, Samir S. |
来源期刊 | MICROBIOLOGYOPEN
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ISSN | 2045-8827 |
出版年 | 2017 |
卷号 | 6期号:5 |
英文摘要 | Prokaryotic communities in pristine and oil-contaminated desert soil, seawater, and hypersaline coastal soil were analyzed using culture-dependent and culture-independent approaches. The former technique was the dilution-plating method. For the latter, total genomic DNA was extracted and the 16S rRNA genes were amplified using a universal bacterial primer pair and primer pairs specific for Actinobacteria, Gammaproteobacteria, and Archaea. The amplicons were resolved using denaturing gradient gel electrophoresis (DGGE) and sequenced, and the sequences were compared to those in GenBank. The plating method offered the advantages of capturing the targeted hydrocarbonoclastic microorganisms, counting them and providing cultures for further study. However, this technique could not capture more than a total of 15 different prokaryotic taxa. Those taxa belonged predominantly to the genera Alcanivorax, Pseudoxanthomonas, Bosea, Halomonas, and Marinobacter. The individual isolates in culture consumed between 19 and 50% of the available crude oil in 10 days. Although the culture-independent approach revealed much more microbial diversity, it was not problem-free. The subdivision primers exhibited satisfactory specificity, but they failed to capture all the available taxa. The universal bacterial primer pair ignored Actinobacteria altogether, although the primer pair specific for Actinobacteria captured many of them, for example, the genera Geodermatophilus, Streptomyces, Mycobacterium, Pontimonas, Rhodococcus, Blastococcus, Kocuria, and many others. Because most researchers worldwide use universal primers for PCR, this finding should be considered critically to avoid misleading interpretations. |
英文关键词 | Archaea bacteria DGGE environmental samples molecular analysis |
类型 | Article |
语种 | 英语 |
国家 | Kuwait |
收录类别 | SCI-E |
WOS记录号 | WOS:000412758000002 |
WOS关键词 | 16S RIBOSOMAL-RNA ; GEL-ELECTROPHORESIS ANALYSIS ; BENCH-SCALE BIOREMEDIATION ; MICROBIAL COMMUNITY ; DESERT SOIL ; OILY SEAWATER ; CRUDE-OIL ; BACTERIA ; PCR ; GENES |
WOS类目 | Microbiology |
WOS研究方向 | Microbiology |
资源类型 | 期刊论文 |
条目标识符 | http://119.78.100.177/qdio/handle/2XILL650/201099 |
作者单位 | Kuwait Univ, Dept Biol Sci, Microbiol Program, Fac Sci, Safat, Kuwait |
推荐引用方式 GB/T 7714 | Al-Mailem, Dina M.,Kansour, Mayada K.,Radwan, Samir S.. Capabilities and limitations of DGGE for the analysis of hydrocarbonoclastic prokaryotic communities directly in environmental samples[J],2017,6(5). |
APA | Al-Mailem, Dina M.,Kansour, Mayada K.,&Radwan, Samir S..(2017).Capabilities and limitations of DGGE for the analysis of hydrocarbonoclastic prokaryotic communities directly in environmental samples.MICROBIOLOGYOPEN,6(5). |
MLA | Al-Mailem, Dina M.,et al."Capabilities and limitations of DGGE for the analysis of hydrocarbonoclastic prokaryotic communities directly in environmental samples".MICROBIOLOGYOPEN 6.5(2017). |
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