Phosphomannose isomerase gene for selection in lettuce (Lactuca sativa L.) transformation.
A positive selection system using phosphomannose isomerase was employed for Agrobacterium tumefaciens mediated transformation of lettuce (Lactuca sativa L. var. 'Achát'). It was shown that the mannose-based selection system works very well with the lettuce genotype used, reaching up to 25% transformation efficiency on the medium with 20 g/L mannose and 20 g/L sucrose. The best transformation efficacy with the commonly-used kanamycin at 100 mg/L as a selection agent was 21%. Southern blot analyses of thirteen chosen mannose-resistant regenerants revealed that some of them have clonal origin, about one-half harbour a single T-DNA copy and one plant contains an incomplete T-DNA segment with only the left part of T-DNA with the pmi gene present in the genomic DNA. The following Northern analysis showed transcriptional activity of the introduced pmi gene in all plants analysed with very high differences in the level of pmi specific mRNA. The results demonstrate that both mannose and kanamycin provide comparable transformation efficiencies in our lettuce genotype. An alternative selection method with mannose as a selection agent is now available for lettuce transgenosis.
Biology Centre of the Academy of Sciences of the Czech Republic, v.v.i., Institute of Plant Molecular Biology, Ceské Budejovice, Czech Republic. briza@umbr.cas.cz
Jindrich
J
Bríza
Nina
N
Růzicková
Hana
H
Niedermeierová
Jana
J
Dusbábková
Josef
J
Vlasák
D016960:Agrobacterium tumefaciens / Q000235:genetics; D018545:Lactuca / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D008359:Mannose-6-Phosphate Isomerase / Q000235:genetics*; D018520:Plant Shoots / Q000201:enzymology / Q000235:genetics / Q000254:growth & development; D030821:Plants, Genetically Modified; D012641:Selection, Genetic; D014170:Transformation, Genetic
D008359:Mannose-6-Phosphate Isomerase