Molecular identification of fine roots of trees from the Alps:: reliable and fast DNA extraction and PCR-RFLP analyses of plastid DNA

被引:68
作者
Brunner, I [1 ]
Brodbeck, S [1 ]
Büchler, U [1 ]
Sperisen, C [1 ]
机构
[1] Swiss Fed Res Inst, WSL, CH-8903 Birmensdorf, Switzerland
关键词
cpDNA; fine roots; taxon identification; trees; trnL intron; universal primers;
D O I
10.1046/j.1365-294X.2001.01325.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Fine roots of trees are intensively used as indicators to assess soil alterations, e.g. those owing to atmospheric inputs of acidifying substances, but their identification to species with morphological criteria is difficult. In this study, we established molecular techniques in order to identify fine roots of the 30 most common tree species of the Alps. We developed a protocol for efficient isolation of DNA from fine roots with extraction of DNA in the presence of polyvinylpyrrolidone (PVP) and polyvinylpolypyrrolidone (PVPP). The trnL (UAA) intron of plastid DNA was used as a marker for fine root identification. We amplified and sequenced this intron with plant universal primers. The size of the sequences ranged from 444 to 672 bp. A synoptic key for species identification was designed on the basis of restriction fragment patterns predicted from sequence data. Using the restriction enzyme TaqI as key enzyme, and where necessary HinfI, RsaI and CfoI, 16 taxa, including Picea abies, Larix decidua, Abies alba, and Fagus sylvatica, the dominant tree species of the Alpine region could be identified by agarose gel electrophoresis of restriction fragments. Fourteen taxa could be identified to the genus level, among them Quercus, Salix and Populus species. In a field study, conducted in a 20 x 30 m plot of a mixed forest with five tree species, fine roots of 43 out of 46 samples were identified and their distributions were mapped. These results demonstrate the utility of our DNA extraction method and of the trnL intron for the identification of fine tree roots.
引用
收藏
页码:2079 / 2087
页数:9
相关论文
共 38 条
[1]   Nitrogen saturation in temperate forest ecosystems - Hypotheses revisited [J].
Aber, J ;
McDowell, W ;
Nadelhoffer, K ;
Magill, A ;
Berntson, G ;
Kamakea, M ;
McNulty, S ;
Currie, W ;
Rustad, L ;
Fernandez, I .
BIOSCIENCE, 1998, 48 (11) :921-934
[2]  
Agerer R., 1987, Colour atlas of ectomycorrhizae
[3]  
[Anonymous], 1987, ROOT IDENTIFICATION
[4]   Fine-root parameters as indicators of sustainability of forest ecosystems [J].
Bakker, MR .
FOREST ECOLOGY AND MANAGEMENT, 1999, 122 (1-2) :7-16
[5]   Identification of roots of woody species using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analysis [J].
Bobowski, BR ;
Hole, D ;
Wolf, PG ;
Bryant, L .
MOLECULAR ECOLOGY, 1999, 8 (03) :485-491
[6]  
Chang S. J., 1993, Plant Molecular Biology Reporter, V11, P113, DOI 10.1007/BF02670468
[7]   PHYLOGENETICS OF SEED PLANTS - AN ANALYSIS OF NUCLEOTIDE-SEQUENCES FROM THE PLASTID GENE RBCL [J].
CHASE, MW ;
SOLTIS, DE ;
OLMSTEAD, RG ;
MORGAN, D ;
LES, DH ;
MISHLER, BD ;
DUVALL, MR ;
PRICE, RA ;
HILLS, HG ;
QIU, YL ;
KRON, KA ;
RETTIG, JH ;
CONTI, E ;
PALMER, JD ;
MANHART, JR ;
SYTSMA, KJ ;
MICHAELS, HJ ;
KRESS, WJ ;
KAROL, KG ;
CLARK, WD ;
HEDREN, M ;
GAUT, BS ;
JANSEN, RK ;
KIM, KJ ;
WIMPEE, CF ;
SMITH, JF ;
FURNIER, GR ;
STRAUSS, SH ;
XIANG, QY ;
PLUNKETT, GM ;
SOLTIS, PS ;
SWENSEN, SM ;
WILLIAMS, SE ;
GADEK, PA ;
QUINN, CJ ;
EGUIARTE, LE ;
GOLENBERG, E ;
LEARN, GH ;
GRAHAM, SW ;
BARRETT, SCH ;
DAYANANDAN, S ;
ALBERT, VA .
ANNALS OF THE MISSOURI BOTANICAL GARDEN, 1993, 80 (03) :528-580
[8]   Molecular phylogeny of extant gymnosperms and seed plant evolution: Analysis of nuclear 18S rRNA sequences [J].
Chaw, SM ;
Zharkikh, A ;
Sung, HM ;
Lau, TC ;
Li, WH .
MOLECULAR BIOLOGY AND EVOLUTION, 1997, 14 (01) :56-68
[9]   THE EFFECTS OF NITROGEN ADDITION AND REMOVAL ON NORWAY SPRUCE FINE-ROOT VITALITY AND DISTRIBUTION IN 3 CATCHMENT AREAS AT GARDSJON [J].
CLEMENSSONLINDELL, A ;
PERSSON, H .
FOREST ECOLOGY AND MANAGEMENT, 1995, 71 (1-2) :123-131
[10]   FINE-ROOT VITALITY IN A NORWAY SPRUCE STAND SUBJECTED TO VARIOUS NUTRIENT SUPPLIES [J].
CLEMENSSONLINDELL, A ;
PERSSON, H .
PLANT AND SOIL, 1995, 168 :167-172