Reproducibility of the differential amplification between leaf and root DNAs in soybean revealed by RAPD markers |
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Authors: | L-F O Chen Hsiao-Ying Kuo Mei-Hwei Chen K-N Lai Shu-Chen Grace Chen |
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Institution: | (1) Institute of Botany, Academia Sinica, Nankang 115 Taipei, Taiwan, Republic of China Fax:+886-2-782-7954 E-mail: ochenlf@gate.sinica.edu.tw, TW |
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Abstract: | Random amplified polymorphic DNA (RAPD) was used to determine whether such markers can be employed for detecting genomic
modification during plant development or under certain stress environments. Pairwise comparisons in RAPD patterns of leaf
and root DNA amplifications were studied for 11 soybean accessions representing different origins. Hydroponic culture was
used for the ease of harvesting roots. From a total of 40 primers screened, it was found that 16 can detect leaf DNA polymorphism,
19 for root DNA polymorphism, while 10 show a greater consistency for detecting polymorphism between leaf and root (L/R) DNAs.
Nevertheless, problems were encountered when the newly synthesized oligo-primers and different thermal cyclers were used to
check the data. Several factors were then tested for their reproducibility. The results indicated that the amplified differences
between root and leaf DNAs are mostly not affected by template DNA concentrations. The addition of DMSO (dimethyl sulphoxide)
or TMAC (tetramethyl-ammonium chloride) also did not mask the L/R differences. However, DNA polymerase and oligo-primers synthesized
from different manufacturers, as well as the thermal cyclers, reacted differently sometimes. Regardless of the general problems
of reproducibility in RAPD patterns, some amplified differences remain between the L/R DNAs. The most distinct patterns involve
differences in the relative intensity of amplified bands. Differential amplification might have occurred during plant leaf
and root development. Southern hybridization of the eluted polymorphic bands against restriction digestion of total genomic
DNA confirms their being homologous to soybean DNA fragments. Polymorphism of these specific L/R differences also exists among
varieties. RAPD should be a useful tool in detecting genomic alterations during plant development or under certain stress
environments, as long as the factors affecting the reproducibility of RAPD patterns can be properly controlled. An additional
cycle of selection would be possible if such a type of polymorphism is proved to be correlated with certain developmental
characters.
Received: 7 October 1996 / Accepted: 20 May 1997 |
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Keywords: | Root Leaf Random amplified polymorphic DNA Polymerase chain reaction Glycine max L Merr |
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