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- Bellini, C., et al.
(författare)
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Stability of a foreign gene in transgenic Nicotiana tabacum plants during a cycle of dedifferentiation-differentiation
- 1992
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Ingår i: Plant Science. - : Elsevier BV. - 0168-9452 .- 1873-2259. ; 82:2, s. 193-200
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Tidskriftsartikel (refereegranskat)abstract
- Protoplasts of Nicotiana tabacum were transformed with the APH(3’)II gene, which confers kanamycin resistance. Plants resistant to kanamycin were differentiated and 3 of them were chosen at random. These were used to study the stability of the foreign gene after a cycle of dedifferentiation, to produce calli, and differentiation, to produce new plants. The effect of the selective pressure was analyzed by performing dedifferentiation and differentiation in the presence or absence of kanamycin. Inbred plants were also produced from the original transformed plants and used as control. Southern blot analysis of DNA extracted from 66 regenerated plants showed in all cases that no detectable alteration occurred both in gene structure and insertion site. Furthermore the specific activity of the APH(3’)II enzyme was shown to be at high level in all regenerated plants regardless of the fact that they were regenerated in the presence or absence of kanamycin. The results described here are experimental evidence that a hybrid forcing gene is rather stable in a heterologous genome even after dedifferentiation of the transformed plants and differentiation in vitro, i.e. in those conditions known to be correlated with extensive somaclonal variation.
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| 6. |
- Santoni, V., et al.
(författare)
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Use of two-dimensional protein pattern analysis for the characterization of Arabidopsis thaliana mutants.
- 1994
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Ingår i: Planta. - 0032-0935 .- 1432-2048. ; 192:4, s. 557-566
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Tidskriftsartikel (refereegranskat)abstract
- Total proteins extracted from wild-type plants of Arabidopsis thaliana Heyhn, an etiolated mutant, a de-etiolated mutant and a mutant affected in cotyledon morphology, were analyzed by two-dimensional gel electrophoresis. Computer analysis of two-dimensional gels allowed the characterization of the mutants by a set of proteins showing a differential expression when compared with the wild-type plant grown under the same conditions. The overlap between comparisons of the different mutants with the wild-type allowed the identification of groups of polypeptides which, since their expression is altered in several mutants, might be involved in certain physiological functions. For example, this approach showed a possible involvement of actin in the elongation process. The simultaneous analysis of the two-dimensional protein patterns of different mutants seems, therefore, to be a promising approach to characterize proteins involved in various physiological functions.
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