Biotechnology
| Poster #518 | |
| » | Introduction and Abstract |
| » | Materials and Methods |
| » | Results and Discussion |
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AFLP® Analysis of Lupinus luteus and
L. cosentinii Using Near Infrared Fluorescence
Labeled Primers and LI-COR Automated
DNA Sequencers
J. Qiu1, E. van Santen2, and M.P. Campos-Andrada3
1) LI-COR Inc., Biotechnology Division, 4647 Superior Street, Lincoln, NE 68504, USA.
2) Department of Agronomy and Soils, 202 Funchess Hall, Auburn University, AL 36849, USA.
3) Department of Genetics and Breeding, Estacao Agronomica National (EAN), 2780 Oeiras Portugal.
Abstract
Phylogenetic relationships between Lupinus luteus and L. cosentinii were determined using the AFLP® technique and LI-COR automated genetic analysis system. Genomic DNA of six lupin accessions (three per species) were isolated, purified, quantified, and subjected to AFLP® analysis using infrared dye (IRD) labeled EcoR1 primers. Data were automatically collected during electrophoresis and were analyzed using Gene ImagIR software. A genetic diversity index of 92% between the two species was estimated based on cluster analysis of 276 informative AFLP® fragments ranging from 100 to 700 bp in size. Several polymorphic AFLP® fragments, which differentiate parental lines, were steadily inherited into respective progenies (F3 and/or F10). Thus, it is possible to use these markers for tracking inheritance and perhaps selection history. Our preliminary data suggest that the automated AFLP® technique can be effectively used to characterize lupin germplasm and facilitate marker-assisted breeding programs in plants.
Introduction
Lupinus spp. is a promising protein source competitive to soybean and has played an important role in sustainable agriculture worldwide (Campos-Andrada et al., 1994; Noffsinger and van Santen, 1995). Selection based on certain morphological traits that are affected by significant G x E interactions, however, can be inefficient. The use of molecular markers may provide a better alternative for characterization of germplasm and hence facilitate marker-based breeding programs (Qiu et al., 1995).
Amplified fragment length polymorphism (AFLP®) is a powerful fingerprinting and mapping technique for genome researches (Vos et al., 1995). LI-COR IR2 automated DNA sequencers and associated software have been demonstrated to efficiently generate and analyze complex AFLP® patterns of various genomes (Qiu et al., 1999; Remington et al., 1999). The objectives of this study were to study genetic diversity between L. luteus and L. cosentinii and to examine genetic inheritance of selected AFLP® fragments using defined genetic materials derived from specific crosses.
