Institute of Plant Physiology
Sixteen 10-base primers were used to amplify genomic DNA extracted from leaves of three week old plantlets (Table 1).
Table 1. Sequences of 10-base primers
All primers generated from 2 to 17 markers, which varied in size from 200 to 2000 bases.
RAPD analysis did not detect any DNA polymorphism among the individual plants of original inbred A188 (Fig. 1), confirming a high level of inbreeding. At the same time multiplicity qualitative and quantitative differences were found between RAPD profiles A188 and somaclones. One type of the primers, for example QR-1, QR-3, B-2 and B-4, generated an individual spectrum of markers for each somaclone (Fig. 2). All genotypes examined, including A188, could be distinguished on the basis of their RAPD profiles with these primers. RAPD profiles produced by other primers (QR-2, QR-4, B-1, 10, 11, 318 and 340) included both polymorphic and monomorphic bands. The primers of the third type (QR-5, B-3, B-5, B-6 and 450) generated common markers for the somaclones of one group. For example, the RAPD spectrum of clones R11, R14, R27 and R54 produced using the primer QR-5 contained a band of about 870 bases, which was absent in RAPD profiles of A188 and other somaclones. The clones R14, R27 and R54 displayed three specific light amplification products with this primer (Fig. 3). Both groups of somaclones could be distinguished one from the other and from the line A188 with the primer B-5 (Fig. 4). The similarity of RAPD profiles of regenerated plants inside one group seems to be conditioned by common origin .
These results demonstrate that the RAPD technique can be applied for elucidation of genetic polymorphism of regenerated plants.
Figure 1. RAPD profiles of individual plants of inbred A188 generated by primer 11.
Figure 2. Individual spectrum of markers for each somaclone with primer QR-1.
Figure 3. Specific RAPD bands for the somaclones R11, R14, R27 and R54 generated by primer QR-5.
Figure 4. Group-specific RAPD profiles generated by primer B-5.
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