Denaturing gradient gel electrophoresis (DGGE) as a tool for identification of marine nematodes

被引:32
作者
Cook, AA
Bhadury, P
Debenham, NJ
Meldal, BHM
Blaxter, ML
Smerdon, GR
Austen, MC
Lambshead, PJD
Rogers, AD
机构
[1] British Antarctic Survey, Nat Environ Res Council, Cambridge CB3 0ET, England
[2] Nat Hist Museum, Dept Zool, Nematode Res Grp, London SW7 5BD, England
[3] Plymouth Marine Lab, Plymouth PL1 3DH, Devon, England
[4] Univ Southampton, Sch Ocean & Earth Sci, Southampton Oceanog Ctr, Southampton SO14 3ZH, Hants, England
[5] Univ Edinburgh, Inst Cell Anim & Populat Biol, Ashworth Labs, Edinburgh EH9 3JT, Midlothian, Scotland
关键词
marine nematodes; 18S ribosomal DNA; denaturing gradient gel electrophoresis;
D O I
10.3354/meps291103
中图分类号
Q14 [生态学(生物生态学)];
学科分类号
071012 ; 0713 ;
摘要
Many phyla of marine invertebrates are difficult to identify using conventional morphological taxonomy. Larvae of a wider set of phyla are also difficult to identify as a result of conservation of morphology between species or because morphological characters are destroyed during sampling and preservation. DNA sequence analysis has the potential for identification of marine organisms to the species level. However, sequence analysis of specimens is time-consuming and impractical when species diversity is very high and densities of individuals huge, as they are in many marine habitats. The effectiveness of the 18S rRNA gene sequences for identification of one speciesrich marine group, the Nematoda, is analysed. Following identification of variable regions of the 18S rRNA gene, primers were designed to amplify a small segment of sequences suitable for denaturing gradient gel electrophoresis (DGGE). The effectiveness of DGGE for identifying individual species is analysed. DGGE analysis of natural communities of nematodes detected less than 2/3 of the species present. This fraction of the community probably represents the abundant species in the original samples. It is concluded that DGGE is not a useful tool for analysis of species richness in marine communities as it fails to detect rare species of which there are usually many in the marine benthic environment. However, DGGE may be a useful method for detecting changes in communities that influence the abundance of the most common taxa.
引用
收藏
页码:103 / 113
页数:11
相关论文
共 51 条
[1]   Limitations of metazoan 18S rRNA sequence data: Implications for reconstructing a phylogeny of the animal kingdom and inferring the reality of the Cambrian explosion [J].
Abouheif, E ;
Zardoya, R ;
Meyer, A .
JOURNAL OF MOLECULAR EVOLUTION, 1998, 47 (04) :394-405
[2]  
[Anonymous], NATURE HUMAN SOC QUE
[3]  
AUSTEN MC, 1986, THESIS U EXETER
[4]  
Avise John C., 1994, pi
[5]   A molecular evolutionary framework for the phylum Nematoda [J].
Blaxter, ML ;
De Ley, P ;
Garey, JR ;
Liu, LX ;
Scheldeman, P ;
Vierstraete, A ;
Vanfleteren, JR ;
Mackey, LY ;
Dorris, M ;
Frisse, LM ;
Vida, JT ;
Thomas, WK .
NATURE, 1998, 392 (6671) :71-75
[6]   Mitochondrial DNA diversity in nematodes [J].
Blouin, MS .
JOURNAL OF HELMINTHOLOGY, 1998, 72 (04) :285-289
[7]  
Chan OC, 2002, FEMS MICROBIOL ECOL, V42, P119, DOI 10.1111/j.1574-6941.2002.tb01001.x
[8]   Sequence differences in the internal transcribed spacers of DNA among four species of hookworm (Ancylostomatoidea: Ancylostoma) [J].
Chilton, NB ;
Gasser, RB .
INTERNATIONAL JOURNAL FOR PARASITOLOGY, 1999, 29 (12) :1971-1977
[9]   The first internal transcribed spacer (ITS-1) of ribosomal DNA as a molecular marker for phylogenetic and population analyses in Crustacea [J].
Chu, KH ;
Li, CP ;
Ho, HY .
MARINE BIOTECHNOLOGY, 2001, 3 (04) :355-361
[10]  
COOK AA, 2001, THESIS U SOUTHAMPTON