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ARS Home » Midwest Area » Madison, Wisconsin » Vegetable Crops Research » Research » Publications at this Location » Publication #95303
Title: CONVERSION OF AN AFLP FRAGMENT LINKED TO THE CARROT Y2 LOCUC TO A SIMPLE, CODOMINANT, PCR-BASED MARKER FORM

Author
item Bradeen, James
item Simon, Philipp

Submitted to: Journal of Theoretical and Applied Genetics
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 9/11/1998
Publication Date: N/A
Citation: N/A

Interpretive Summary: A new method for evaluating plant genetic variation is called AFLP. This is a very powerful tool but requires complex, expensive laboratory equipment and can be difficult to interpret. In this experiment we used AFLP as a starting point and applied several other techniques to result in a technologically simple, inexpensive genetic evaluation system. With this, then, we found an easily evaluated genetic marker located close to a gene of carrots Y2 which results in a root color of yellow, rather than orange. This system will help us understand this Y2 gene and its function better.

Technical Abstract: Recent advances have expanded the potential usefulness of molecular techniques for plant genetic research. The AFLP is a powerful technique, allowing rapid and reliable analysis of multiple potentially polymorphic sites in a single experiment. Because AFLP technology requires no (a priori) knowledge of genome structure or preparation of molecular probes, it is immediately useful for a wide variety of plant species. However, because AFLP markers are dominant, costly, and technologically demanding, the technique has limited application for large scale, locus-specific applications. In carrot, the Y2 locus controls carotene accumulation in the root xylem core. Although carrot is an important source of dietary carotene, little is known about the regulation and biosynthesis of carotenes in carrot. We identified 6 AFLP fragments linked to the Y2 locus through a combination of F2 mapping and bulked segregant analysis. We have developed a procedure for generation simple, codominant, PCR-based markers from dominant AFLP fragments, using a Y2 linked AFLP fragments as a model. Our converted marker requires only a simple PCR followed by standard agarose gel electrophoresis. It is rapid, simple, reliable, comparatively inexpensive, codominant, and non-radioactive. Conversion of AFLP fragments to forms better adapted to large scale, locus-specific applications greatly expands the usefulness of this molecular technique.