Background:As an important player during food digestion,gut microbiota has attracted much attention in diet adaptation studies in birds.Microbiota extracted from feces has been widely used as a proxy for gut microbiot...Background:As an important player during food digestion,gut microbiota has attracted much attention in diet adaptation studies in birds.Microbiota extracted from feces has been widely used as a proxy for gut microbiota.Although several methods have been developed for microbial DNA extraction,their performances in the bird feces have not been systematacially evaluated yet.Methods:In this study,we applied three DNA extraction methods(Qiagen,MoBio and Bead)to extract DNA from feces of three avian dietary guilds(granivore,omnivore and carnivore),sequenced V4 region of 16S rRNA gene for each extract and evaluated the performances of DNA yield,DNA integrity,microbial composition,cell lysis capacity and alpha diversity for the three methods on each dietary guild.Results:Bead method was the best on the performance of both DNA yield and DNA integrity regardless of dietary guild.In granivore,microbial relative abundance at both species and phylum levels,alpha diversity and cell lysis capacity were comparable among all methods.In omnivore,Qiagen had the best performance on alpha diversity,fol-lowed by Bead and MoBio.There were small variations on microbial relative abundance at both species and phylum levels among different extraction methods.MoBio exhibited the best performance on cell lysis capacity.In carnivore,considerable variations were found on microbial relative abundance at both species and phylum levels.Qiagen had the best performance on alpha diversity,followed by MoBio and Bead.MoBio had the highest cell lysis capacity.Conclusions:DNA yield and integrity have no obvious impact on microbial composition,alpha diversity or cell lysis capacity.The microbiota results(e.g.,microbial composition,cell lysis capacity,alpha diversity)obtained from differ-ent methods are comparable in granivorous avian species but not in omnivorous or carnivorous birds.Either method could be used in granivore microbiota studies.For omnivores and carnivores,we recommend Qiagen method when the research purpose is microbial diversity and MoBio when gram-positive bacteria is the research target.展开更多
从木材识别的角度,对木材DNA提取方法(CTAB、SDS、PTB、DNeasy Plant Mini Kit等)和DNA条形码及DNA指纹图谱等基于DNA的木材树种识别方法进行了综合评述。其中,针对DNA条形码方法,重点阐述了基因组DNA中可用的特征序列来源叶绿体基因(ch...从木材识别的角度,对木材DNA提取方法(CTAB、SDS、PTB、DNeasy Plant Mini Kit等)和DNA条形码及DNA指纹图谱等基于DNA的木材树种识别方法进行了综合评述。其中,针对DNA条形码方法,重点阐述了基因组DNA中可用的特征序列来源叶绿体基因(chloroplast DNA,cpDNA)rbcL、matK、trnH-psbA间隔区序列,核糖体基因(ribosomal DNA,rDNA)ITS序列,以及变异位点的识别和系统进化树的应用等问题;针对DNA指纹图谱方法,重点阐述了(RAPD、ISSR、SSR、SNP)4种DNA分子标记方法在DNA指纹图谱中的研究和应用现状。笔者认为,以DNA特征序列为依据的木材树种识别理论上虽然是可行的,但要应用于实际还需开展更多的研究。展开更多
The 16S rDNA amplicon high-throughput sequencing technique provides a robust and inexpensive approach to detect the gut microbiota of amphibians. Since different experimental protocols generate technical biases in dra...The 16S rDNA amplicon high-throughput sequencing technique provides a robust and inexpensive approach to detect the gut microbiota of amphibians. Since different experimental protocols generate technical biases in drawing the gut microbiota profiles, the integrative analysis of gut microbiota produced by different studies must be performed with circumspection. In this study, we compared the efficacy of two DNA extraction methods （i.e., a phenol-chloroform method and TIANamp Stool DNA Kit） in describing intestinal and fecal bacterial communities of transplanted Asiatic toad （Bufo gargarizans） tadpoles. In terms of the DNA extraction quality （i.e., DNA purity and yield rate） and the consistency in between fecal and intestinal microbiota structures （i.e., a and 13 diversity indices）, the phenol-chloroform method was more robust than this commercial stool kit in profiling gut microbiota of tadpoles with feces.展开更多
基金This study was supported by National Natural Science Foundation of China(No.31930013,31872240)the National Key Program of Research and Development,Ministry of Science and Technology(2016YFC0503200)Youth Innovation Promotion Association of Chinese Academy of Sciences(2020086)to SP.
文摘Background:As an important player during food digestion,gut microbiota has attracted much attention in diet adaptation studies in birds.Microbiota extracted from feces has been widely used as a proxy for gut microbiota.Although several methods have been developed for microbial DNA extraction,their performances in the bird feces have not been systematacially evaluated yet.Methods:In this study,we applied three DNA extraction methods(Qiagen,MoBio and Bead)to extract DNA from feces of three avian dietary guilds(granivore,omnivore and carnivore),sequenced V4 region of 16S rRNA gene for each extract and evaluated the performances of DNA yield,DNA integrity,microbial composition,cell lysis capacity and alpha diversity for the three methods on each dietary guild.Results:Bead method was the best on the performance of both DNA yield and DNA integrity regardless of dietary guild.In granivore,microbial relative abundance at both species and phylum levels,alpha diversity and cell lysis capacity were comparable among all methods.In omnivore,Qiagen had the best performance on alpha diversity,fol-lowed by Bead and MoBio.There were small variations on microbial relative abundance at both species and phylum levels among different extraction methods.MoBio exhibited the best performance on cell lysis capacity.In carnivore,considerable variations were found on microbial relative abundance at both species and phylum levels.Qiagen had the best performance on alpha diversity,followed by MoBio and Bead.MoBio had the highest cell lysis capacity.Conclusions:DNA yield and integrity have no obvious impact on microbial composition,alpha diversity or cell lysis capacity.The microbiota results(e.g.,microbial composition,cell lysis capacity,alpha diversity)obtained from differ-ent methods are comparable in granivorous avian species but not in omnivorous or carnivorous birds.Either method could be used in granivore microbiota studies.For omnivores and carnivores,we recommend Qiagen method when the research purpose is microbial diversity and MoBio when gram-positive bacteria is the research target.
基金supported by the National Natural Science Foundation of China (NSFC 31600104)Key Scientific Research Project of Higher Education in Henan Province (No. 17B180004)+2 种基金National Undergraduate Training Program for Innovation and Entrepreneurship (No. 201610477013)Ph.D. Research Startup Foundation of Xinyang Normal University (No. 0201424)Nanhu Scholars Program for Young Scholars of Xinyang Normal University
文摘The 16S rDNA amplicon high-throughput sequencing technique provides a robust and inexpensive approach to detect the gut microbiota of amphibians. Since different experimental protocols generate technical biases in drawing the gut microbiota profiles, the integrative analysis of gut microbiota produced by different studies must be performed with circumspection. In this study, we compared the efficacy of two DNA extraction methods （i.e., a phenol-chloroform method and TIANamp Stool DNA Kit） in describing intestinal and fecal bacterial communities of transplanted Asiatic toad （Bufo gargarizans） tadpoles. In terms of the DNA extraction quality （i.e., DNA purity and yield rate） and the consistency in between fecal and intestinal microbiota structures （i.e., a and 13 diversity indices）, the phenol-chloroform method was more robust than this commercial stool kit in profiling gut microbiota of tadpoles with feces.
