Kanamycin-resistant alfalfa has a point mutation in the 16S plastid rRNA.
Plant Cell Rep
; 22(10): 774-9, 2004 May.
Article
en En
| MEDLINE
| ID: mdl-14770263
ABSTRACT
Genes conferring resistance to kanamycin are frequently used to obtain transgenic plants as spontaneous resistance to kanamycin is not known to exist in higher plants. Nevertheless, mutations conferring kanamycin resistance have been identified in Chlamydomonas reinhardtii, raising the question as to why kanamycin-resistant mutants have not been found in higher plants. While attempting plastid transformation of alfalfa, we obtained non-transgenic but kanamycin-resistant somatic embryos following 2 months of culture in the presence of 50 mg l(-1) kanamycin. Sequencing of the plastid DNA region corresponding to the decoding site of the 16S rRNA in ten independent resistant events revealed an A to C transversion at position 1357 of the 16S plastid rDNA, the same site at which an A to G conversion confers kanamycin resistance to C. reinhardtii by reducing the ability of the antibiotic to bind to its target site. All plants derived from the resistant embryos through additional cycles of somatic embryogenesis in the absence of kanamycin retained the mutant phenotype, suggesting that the mutation was homoplastomic. Resistant plants produced 85% less biomass than controls; their leaves were chlorotic during early development and over time slowly turned green. The absence of kanamycin- resistant mutants in higher plants might be explained by the requirement for a regeneration system capable of resulting in homoplastomic individuals, or it may be the result of the detrimental effect of the mutation on the phenotype.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Resistencia a la Kanamicina
/
ARN Ribosómico 16S
/
Mutación Puntual
/
Plastidios
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Medicago sativa
Idioma:
En
Revista:
Plant Cell Rep
Asunto de la revista:
BOTANICA
Año:
2004
Tipo del documento:
Article
País de afiliación:
Italia