Aminoglycoside antibiotics: structure, functions and effects on in vitro plant culture and genetic transformation protocols.
Plant transformation protocols generally involve the use of selectable marker genes for the screening of transgenic material. The bacterial gene nptII, coding for a neomycin phosphotransferase, and the hpt gene, coding for a hygromycin phosphotransferase, are frequently used. These enzymes detoxify aminoglycoside antibiotics by phosphorylation, thereby permitting cell growth in the presence of antibiotics. Nevertheless, the screening for transgenic regenerated shoots is often partial and difficult due to regeneration of escapes and chimeras. These difficulties can be caused, in part, by an incorrect assumption about the mode of action of antibiotics in bacterial and eukaryotic cells and in in vitro tissue culture. The information contained in this review could be useful to establish better selection strategies by taking into account factors such as explant complexity, transformation and selection protocols that allow better accessibility to cells of Agrobacterium and antibiotics, and faster regeneration methods that avoid collateral effects of antibiotics on recovered, putative transgenic shoots.
Padilla
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Grupo de Biotecnología de Frutales, Departamento de Mejora, CEBAS-CSIC, Campus Universitario de Espinardo, Apartado de correos 164, 30100, Murcia, Spain. isabelm.gonzalez.padilla@juntadeandalucia.es
Burgos
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D000617:Aminoglycosides / Q000494:pharmacology*; D003470:Culture Media; D018506:Gene Expression Regulation, Plant; D019868:Kanamycin Kinase / Q000235:genetics; D015394:Molecular Structure; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics; D030821:Plants, Genetically Modified / Q000235:genetics*; D012038:Regeneration; D013329:Structure-Activity Relationship; D046509:Tissue Culture Techniques; D014170:Transformation, Genetic
D000617:Aminoglycosides; D003470:Culture Media; D017853:Phosphotransferases (Alcohol Group Acceptor); C063242:hygromycin-B kinase; D019868:Kanamycin Kinase