柱式法和磁珠法提取原油脱氧核糖核酸

打开文本图片集

中图分类号：Q503.23；TE99 文献标志码：A

Abstract：Direct extractionofdeoxyribonucleic acid（DNA）from petroleum is the most fundamentaland most important step foranalyses of its microbial communityanddiversity.The spincolumn based extraction method（SE）and the magnetisable solid phase extraction method（ME）were used toobtainthetotal DNAof microbial genomeof12 petroleumsamples，respectively.Andtheirdiferenceof microbialdiversityandcommunitywereanalyzedfurther more through Illumina highthroughput sequences. The results show that the DNA mass concentration and purity（ A260/A280 ratio）are 20.17±31.83 ng/μL and （2 1.01±0.63 of SE ，and 11.14±17.27ng/μL and 1.29±0.20 of ME，respectively. SE detects 30 phyla and 507 genera， while ME detects 38 phyla and 677 genera，in which the relative content of unclassified and uncultured genera are 1 .12% （20 and 0.20% （SE）， 1.24% and 1.10% （ME），respectively. Unique genera are detected by both methods （SE：65 genera and ME：235 genera）.SEdetects reservoir microorganismssuch as Geobacter，andME detectssuspected reservoir microorganisms such as Xanthobacter.Both methods have advantagesand disadvantages in petroleum DNA extraction. SE has a higherextraction massconcentrationandbetterresponse tooilreservoir microorganisms.MEhasaslightly strongerabilityto detect microbial diversity in petroleum，but both methods have strong detection bias at the taxonomic level.

Keywords ： DNA extraction;spincolumn based extraction method;magnetisable solid phase extraction method; microbial diversity ；microbial community

油藏中蕴含了丰富的微生物资源[14]，目前研究油藏微生物多样性主要是培养[5-6]和非培养[7-9]方法，但迄今环境中可培养的微生物资源不足1%[10] ，而非培养方法优势在于样本的直接检测更为快速、灵敏，丰富了油藏微生物群落结构和多样性等特征[1-I2]，其首要问题是核酸提取[4，7-8]，原油具有黏稠、浓黑、生物量低和试剂可及性差等特点，这使得提取原油总DNA较为困难[13]，基于传统柱式法（SE）的改进已出现为数不多适用于原油微生物总DNA的提取方法[14-16]，但不同核酸提取方法获取DNA纯度和质量浓度的差异将直接影响测序分析的灵敏度[17-18]和特异性[18-19]。（剩余13130字）