Standard methods for research on Apis mellifera gut symbionts

被引:96
作者
Engel, Philipp [1 ]
James, Rosalind R. [2 ]
Koga, Ryuichi [1 ,4 ]
Kwong, Waldan K. [1 ]
McFrederick, Quinn S. [3 ]
Moran, Nancy A. [1 ]
机构
[1] Yale Univ, New Haven, CT USA
[2] USDA ARS, Pollinating Insects Res Unit, Logan, UT USA
[3] Univ Texas Austin, Austin, TX 78712 USA
[4] Natl Inst Adv Ind Sci & Technol, Bioprod Res Inst, Tsukuba, Ibaraki 3058566, Japan
基金
美国国家科学基金会;
关键词
COLOSS; BEEBOOK; fluorescent in situ hybridization; gut symbionts; methods; microbial culture; next generation sequencing; honey bee; 16S RIBOSOMAL-RNA; IN-SITU ACCESSIBILITY; WORKER HONEY-BEES; MICROBIAL COMMUNITIES; ESCHERICHIA-COLI; OLIGONUCLEOTIDE PROBES; LACTOBACILLUS-KUNKEEI; DIVERSITY; BACTERIA; IDENTIFICATION;
D O I
10.3896/IBRA.1.52.4.07
中图分类号
Q96 [昆虫学];
学科分类号
摘要
Gut microbes can play an important role in digestion, disease resistance, and the general health of animals, but little is known about the biology of gut symbionts in Apis mellifera. As part of the BEEBOOK series describing honey bee research methods, we provide standard protocols for studying gut symbionts. We describe non-culture-based approaches based on Next Generation Sequencing (NGS), methodology that has greatly improved our ability to identify the microbial communities associated with honey bees. We also describe Fluorescent In Situ Hybridization (FISH) microscopy, which allows a visual examination of the microenvironments where particular microbes occur. Culturing methods are also described, as they allow the researcher to isolate particular bacteria of interest for further study or gene identification, and enable the assignment of particular functions to particular gut community members. We hope these methods will help others advance the state of knowledge regarding bee gut symbionts and the role they play in honey bee health.
引用
收藏
页数:23
相关论文
共 112 条
[1]  
Ahn JH, 2012, J MICROBIOL, V50, P735
[2]   BASIC LOCAL ALIGNMENT SEARCH TOOL [J].
ALTSCHUL, SF ;
GISH, W ;
MILLER, W ;
MYERS, EW ;
LIPMAN, DJ .
JOURNAL OF MOLECULAR BIOLOGY, 1990, 215 (03) :403-410
[3]   Single-cell identification in microbial communities by improved fluorescence in situ hybridization techniques [J].
Amann, Rudolf ;
Fuchs, Bernhard M. .
NATURE REVIEWS MICROBIOLOGY, 2008, 6 (05) :339-348
[4]   Quantifying microbial communities with 454 pyrosequencing: does read abundance count? [J].
Amend, Anthony S. ;
Seifert, Keith A. ;
Bruns, Thomas D. .
MOLECULAR ECOLOGY, 2010, 19 (24) :5555-5565
[5]  
Anderson MJ, 2001, AUSTRAL ECOL, V26, P32, DOI 10.1111/j.1442-9993.2001.01070.pp.x
[6]  
[Anonymous], 2010, J APIPRODUCT AMP API, DOI DOI 10.3896/IBRA.4.02.2.03
[7]  
[Anonymous], 2002, Experimental designs and data analysis for biologists
[8]   Bacterial community structures in honeybee intestines and their response to two insecticidal proteins [J].
Babendreier, Dirk ;
Joller, David ;
Romeis, Joerg ;
Bigler, Franz ;
Widmer, Franco .
FEMS MICROBIOLOGY ECOLOGY, 2007, 59 (03) :600-610
[9]  
Barbosa p, 1974, MANUAL BASIC TECHNIQ
[10]   Is the in situ accessibility of the 16S rRNA of Escherichia coli for Cy3-labeled oligonucleotide probes predicted by a three-dimensional structure model of the 30S ribosomal subunit? [J].
Behrens, S ;
Fuchs, BM ;
Mueller, F ;
Amann, R .
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2003, 69 (08) :4935-4941