Evaluation of inter-simple sequence repeat analysis for mapping in Citrus and extension of the genetic linkage map |
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Authors: | A A Sankar G A Moore |
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Affiliation: | (1) Horticultural Sciences Department, University of Florida, P.O. Box 110690, Gainsville, FL 32611-0690, USA, US;(2) Horticultural Sciences Department, Plant Molecular and Cellular Biology Program, Institute of Food and Agricultural Sciences, University of Florida, P.O. Box 110690, Gainesville, FL 32611-0690, USA e-mail: gam@gnv.ifas.ufl.edu Fax: +1 352-392-5653, US |
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Abstract: | Inter-simple sequence repeat (ISSR) analysis was evaluated for its usefulness in generating markers to extend the genetic
linkage map of Citrus using a backcross population previously mapped with restriction fragment length polymorphism (RFLP), random amplified polymorphic
DNA (RAPD) and isozyme markers. ISSR markers were obtained through the simple technique of PCR followed by analysis on agarose
gels, using simple sequence repeat (SSR) primers. Optimization of reaction conditions was achieved for 50% of the SSR primers
screened, and the primers amplified reproducible polymorphic bands in the parents and progeny of the backcross population.
Mendelian segregation of the polymorphic bands was demonstrated, with an insignificant number of skewed loci. Most of the
SSR primers produced dominant loci; however co-dominance was observed with loci derived from three primers. A new genetic
map was produced by combining the segregation data for the ISSR markers and data for the RFLP, RAPD and isozyme markers from
the previous map and creating genetic linkages among all the markers using JoinMap 2.0 mapping software. The new map has an
improved distribution of markers along the linkage groups with fewer gaps, and marker order showed partial or complete conservation
in the linkage groups. The incorporation of ISSR markers into the genetic linkage map demonstrates that ISSR markers are suitable
for genetic mapping in Citrus.
Received: 3 February 2000 / Accepted: 12 May 2000 |
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Keywords: | Citrus grandis ISSR Marker Microsatellite Poncirus trifoliata |
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