Agrobacterium-mediated transformation of bottle gourd (Lagenaria siceraria Standl.).
We describe a procedure for producing transgenic bottle gourd plants by inoculating cotyledon explants with Agrobacterium tumefaciens strain AGL1 that carries the binary vector pCAMBIA3301 containing a glufosinate ammonium-resistance (bar) gene and the beta-D-glucuronidase (GUS) reporter gene. The most effective bacterial infection was observed when cotyledon explants of 4-day-old seedlings were co-cultivated with Agrobacterium for 6-8 days on co-cultivation medium supplemented with 0.1-0.001 mg/l L-alpha-(2-aminoethoxyvinyl) glycine (AVG). The putatively transformed shoots directly emerged at the proximal end of cotyledon explants after 2-3 weeks of culturing on selection medium containing 2 mg/l DL-phosphinothricin. These shoots were rooted after 3 weeks of culturing on half-strength MS medium containing 0.1 mg/l indole acetic acid and 1 mg/l DL-phosphinothricin. Transgenic plants were obtained at frequencies of 1.9%. Stable integration and transmission of the transgenes in T1 generation plants were confirmed by a histochemical GUS assay, polymerase chain reaction and Southern blot analyses. Genetic segregation analysis of T1 progenies showed that transgenes were inherited in a Mendelian fashion. To our knowledge, this study is the first to show Agrobacterium-mediated transformation in bottle gourd.
Han
J-S
JS
National Horticultural Research Institute, Rural Development Administration, Suwon, 441-440, South Korea.
Kim
C K
CK
Park
S H
SH
Hirschi
K D
KD
Mok
I- G
I
D016960:Agrobacterium tumefaciens / Q000235:genetics*; D000613:Aminobutyrates / Q000494:pharmacology; D015139:Blotting, Southern; D019662:Cucurbitaceae / Q000187:drug effects / Q000235:genetics* / Q000382:microbiology; D005030:Ethylenes / Q000494:pharmacology; D005966:Glucuronidase / Q000235:genetics; D005998:Glycine / Q000031:analogs & derivatives* / Q000494:pharmacology; D018520:Plant Shoots; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction; D012038:Regeneration; D014170:Transformation, Genetic
D000613:Aminobutyrates; D005030:Ethylenes; C003121:phosphinothricin; C036216:ethylene; D005966:Glucuronidase; C088137:aminoethoxyvinylglycine; D005998:Glycine