title: Optimization of factors  affecting Agrobacterium-mediated transformation of Micro-Tom tomatoes.
abstract: Micro-Tom  is the smallest known variety of tomatoes. An orthogonal experimental design L(16) (4(5)) was  used to optimize Agrobacterium-mediated transformation of cotyledon  explants of Lycopersicon esculentum cv. Micro-Tom. Four parameters were  investigated to determine their effect on transformation frequency: the  concentration of bacterial suspension, time of dip in bacterial suspension,  co-cultivation time, and concentration of carbenicillin. We also examined the  effect of these parameters on contamination rate, necrosis rate, mortality,  cut-surface browning rate, and undamaged explant rate. Both the bacterial and  carbenicillin concentrations had a significant influence on the rate of  infected explants. The time of co-cultivation also had a significant influence  on the transformation parameters. The optimal transformation protocol  consisted of an Agrobacterium suspension of 0.5 × 10(8) cells/mL  (OD(600) = 0.5) and an infection time of 5 min, one day of  co-cultivation and 500 mg/L carbenicillin. Under these conditions, the  transformation efficiency of the shoots reached 5.1%; the mean transformation  frequency was 3.9% (N = 838).
authors.affiliation: Key Laboratory of Protected Horticulture Ministry of Education, College of Horticulture, Shenyang Agricultural University, Shenyang, PR China.
authors.forename: M
authors.initials: M
authors.lastname: Guo
authors.forename: Y L
authors.initials: YL
authors.lastname: Zhang
authors.forename: Z J
authors.initials: ZJ
authors.lastname: Meng
authors.forename: J
authors.initials: J
authors.lastname: Jiang
mesh_terms: D060054:Agrobacterium / Q000235:genetics / Q000378:metabolism; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D002228:Carbenicillin / Q000494:pharmacology; D018548:Cotyledon / Q000187:drug effects / Q000235:genetics / Q000254:growth & development; D018014:Gene Transfer Techniques; D018551:Solanum lycopersicum / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D010641:Phenotype; D030821:Plants, Genetically Modified / Q000187:drug effects / Q000235:genetics / Q000254:growth & development; D014170:Transformation, Genetic
chemical_list: D000900:Anti-Bacterial Agents; D002228:Carbenicillin