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Publication Type J
Authors Blignaut, M; Ellis, AG; Le Roux, JJ
Author Full Name Blignaut, Marguerite; Ellis, Allan G.; Le Roux, Johannes J.
Title Towards a Transferable and Cost-Effective Plant AFLP Protocol
Source PLOS ONE
Language English
Document Type Article
Keywords Plus GENETIC DIVERSITY; POA-ANNUA; POPULATION-GENETICS; MOLECULAR MARKERS; GENOTYPING ERRORS; PATTERNS; AMPLIFICATION; METHYLATION; EVOLUTION; DENSITY
Abstract Amplified fragment length polymorphism (AFLP) is a powerful fingerprinting technique that is widely applied in ecological and population genetic studies. However, its routine use has been limited by high costs associated with the optimization of fluorescently labelled markers, especially for individual study systems. Here we develop a low-cost AFLP protocol that can be easily transferred between distantly related plant taxa. Three fluorescently labelled EcoRI-primers with anchors that target interspecifically conserved genomic regions were used in combination with a single non-labelled primer in our AFLP protocol. The protocol was used to genotype one gymnosperm, two monocot and three eudicot plant genera representing four invasive and four native angiosperm species (Pinus pinaster (Pinaceae), Pennisetum setaceum and Poa annua (Poaceae), Lantana camara (Verbenaceae), Bassia diffusa (Chenopodiaceae), Salvia lanceolata, Salvia africana-lutea, and Salvia africana-caerulea (Lamiaceae)). Highly polymorphic and reproducible genotypic fingerprints (between 37-144 polymorphic loci per species tested) were obtained for all taxa tested. Our single protocol was easily transferred between distantly related taxa. Measures of expected heterozygosity ranged from 0.139 to 0.196 for P. annua and from 0.168 to 0.272 for L. camara which compared well with previously published reports. In addition to ease of transferability of a single AFLP protocol, our protocol reduces costs associated with commercial kits by almost half. The use of highly conserved but abundant anchor sequences reduces the need for laborious screening for usable primers that result in polymorphic fingerprints, and appears to be the main reason for ease of transferability of our protocol between distantly related taxa.
Author Address [Blignaut, Marguerite; Le Roux, Johannes J.] Univ Stellenbosch, Ctr Invas Biol, ZA-7600 Stellenbosch, Western Cape, South Africa; [Ellis, Allan G.] Univ Stellenbosch, Dept Bot & Zool, ZA-7600 Stellenbosch, Western Cape, South Africa
Reprint Address Le Roux, JJ (reprint author), Univ Stellenbosch, Ctr Invas Biol, ZA-7600 Stellenbosch, Western Cape, South Africa.
E-mail Address jleroux@sun.ac.za
Funding Agency and Grant Number South Africa's National Research Foundation (NRF Blue Skies Programme) [BS2008093000013]; DST-NRF Centre of Excellence for Invasion Biology; Working for Water (WfW) Programme through their collaborative project on
Funding Text Funding was provided by South Africa's National Research Foundation (NRF Blue Skies Programme, grant no: BS2008093000013). This work was also partly supported by the DST-NRF Centre of Excellence for Invasion Biology and the Working for Water (WfW) Programme through their collaborative project on
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Cited Reference Count 42
Times Cited 5
Total Times Cited Count (WoS, BCI, and CSCD) 5
Publisher PUBLIC LIBRARY SCIENCE
Publisher City SAN FRANCISCO
Publisher Address 1160 BATTERY STREET, STE 100, SAN FRANCISCO, CA 94111 USA
ISSN 1932-6203
29-Character Source Abbreviation PLOS ONE
ISO Source Abbreviation PLoS One
Publication Date APR 16
Year Published 2013
Volume 8
Issue 4
Article Number e61704
Digital Object Identifier (DOI) 10.1371/journal.pone.0061704
Page Count 7
Web of Science Category Multidisciplinary Sciences
Subject Category Science & Technology - Other Topics
Document Delivery Number 130DR
Unique Article Identifier WOS:000317893400096
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