Arid
DOI10.1002/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
ISSN2045-8827
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
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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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