caa a22 4500 475794869 CHVBK 20180406123721.0 cr unu---uuuuu 170329e20000701xx s 000 0 eng 10.1007/s001220051463 doi (NATIONALLICENCE)springer-10.1007/s001220051463 Identification of AFLP fragments linked to seed coat colour in Brassica juncea and conversion to a SCAR marker for rapid selection [Elektronische Daten] [M. S. Negi, M. Devic, M. Delseny, M. Lakshmikumaran] Abstract : A Brassica juncea mapping population was generated and scored for seed coat colour. A combination of bulked segregant analysis and AFLP methodology was employed to identify markers linked to seed coat colour in B. juncea. AFLP analysis using 16 primer combinations revealed seven AFLP markers polymorphic between the parents and the bulks. Individual plants from the segregating population were analysed, and three AFLP markers were identified as being tightly linked to the seed coat colour trait and specific for brown-seeded individuals. Since AFLP markers are not adapted for large-scale application in plant breeding, our objective was to develop a fast, cheap and reliable PCR-based assay. Towards this goal, we employed PCR-walking technology to isolate sequences adjacent to the linked AFLP marker. Based on the sequence information of the cloned flanking sequence of marker AFLP8, primers were designed. Amplification using the locus-specific primers generated bands at 0.5 kb and 1.2 kb with the yellow-seeded parent and a 1.1-kb band with the brown-seeded parent. Thus, the dominant AFLP marker (AFLP8) was converted into a simple codominant SCAR (Sequence Characterized Amplified Region) marker and designated as SCM08. Scoring of this marker in a segregating population easily distinguished yellow- and brown-seeded B. juncea and also differentiated between homozygous (BB) and heterozygous (Bb) brown-seeded individuals. Thus, this marker will be useful for the development of yellow seed B. juncea cultivars and facilitate the map-based cloning of genes responsible for seed coat colour trait. Springer-Verlag Berlin Heidelberg, 2000 Key words Brassica juncea nationallicence Bulked segregant analysis (BSA) nationallicence Seed coat colour nationallicence AFLP nationallicence SCAR nationallicence PCR walking nationallicence Marker conversion nationallicence Negi M. S. Plant Molecular Biology Division, Tata Energy Research Institute, Habitat Place, Lodhi Road, New Delhi 110 003, India e-mail: malaks@teri.res.in, IN aut Devic M. Laboratoire Physiologie et Biologie Moleculaire des Plantes, C.N.R.S. UMR 5545, Universite de Perpignan, 52 Avenue de Villeneuve, 66860 Perpignan cedex, France, FR aut Delseny M. Laboratoire Physiologie et Biologie Moleculaire des Plantes, C.N.R.S. UMR 5545, Universite de Perpignan, 52 Avenue de Villeneuve, 66860 Perpignan cedex, France, FR aut Lakshmikumaran M. Plant Molecular Biology Division, Tata Energy Research Institute, Habitat Place, Lodhi Road, New Delhi 110 003, India e-mail: malaks@teri.res.in, IN aut https://doi.org/10.1007/s001220051463 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s001220051463 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Negi M. S. Plant Molecular Biology Division, Tata Energy Research Institute, Habitat Place, Lodhi Road, New Delhi 110 003, India e-mail: malaks@teri.res.in, IN aut NATIONALLICENCE 700 1- Devic M. Laboratoire Physiologie et Biologie Moleculaire des Plantes, C.N.R.S. UMR 5545, Universite de Perpignan, 52 Avenue de Villeneuve, 66860 Perpignan cedex, France, FR aut NATIONALLICENCE 700 1- Delseny M. Laboratoire Physiologie et Biologie Moleculaire des Plantes, C.N.R.S. UMR 5545, Universite de Perpignan, 52 Avenue de Villeneuve, 66860 Perpignan cedex, France, FR aut NATIONALLICENCE 700 1- Lakshmikumaran M. Plant Molecular Biology Division, Tata Energy Research Institute, Habitat Place, Lodhi Road, New Delhi 110 003, India e-mail: malaks@teri.res.in, IN aut Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer