Screening of Arabidopsis mutants for functional genomic studies
2008
Awan, F.S. (University of Agriculture, Faisalabad (Pakistan). Centre of Agricultural Biochemistry and Biotechnology) | Khan, I.A. (Ludwig-Maximilians-Universitat, Munchen (Germany). Dept. of Biologie) | Khan, A.A. (Ludwig-Maximilians-Universitat, Munchen (Germany). Dept. of Biologie) | Schneider, A. (Ludwig-Maximilians-Universitat, Munchen (Germany). Dept. of Biologie) | Altaf, J. (University of Arid Agriculture, Rawalpindi (Pakistan). Faculty of Sciences) | Ahmad, R. (University of Agriculture, Faisalabad (Pakistan). Dept. of Crop Physiology) | Leister, D.
Eight photosynthetic Arabidopsis mutants were screened for co-segregation of a photosynthetic phenotype with the T-DNA insertion. These mutants were selected from 80 photosynthetic mutants with genetic background of Columbia-0. Two different screening approaches were used to study the T-DNA insertion in the genome of mutant Arabidopsis lines. The sulphonamide sulfadiazine was found to be an effective selective agent and a single copy of sulfonamide resistant gene was found to be completely resistant to the optimal concentration i.e., 5mg mL-1. The maximum number of Arabidopsis mutant plants had confirmed insertions. Some of the plants did not show any amplification with gene specific primer combination, and it was assumed that either they were wild type plants or they had random T-DNA insertion and the insertion was not found in the gene under study but it could be found in any where in the genome. Some mutant plants were morphologically different from the wild type plants e.g., ALP105. These plants grew as small in size and dark green in color. After PCR screening with gene specific and T-DNA border primers all such mutant plants were confirmed as heterozygous T-DNA insertion plants.
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