Title: AGROBACTERIUM-MEDIATED TRANSFORMATION OF BOTTEL GOURD (LAGENARIA SICERARIA STANDL.) Authors
|Han, J - NATL HORT RESEARCH INST|
|Kim, C - SANGJU NATIONAL UNIV|
|Park, S - TEXAS A&M UNIVERSITY|
|Mok, I - NATL HORT RESEARCH INST|
Submitted to: Plant Cell Reports
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: August 2, 2004
Publication Date: March 1, 2005
Citation: Han, J.S., Kim, C.K., Park, S.H., Hirschi, K., Mok, I.G. 2005. Agrobacterium-mediated transformation of bottel gourd (Lagenaria siceraria Standl.). Plant Cell Reports. 23(10-11):692-698. Interpretive Summary: The ability to alter foods through bioengineering requires the ability to move foreign DNA into the crop plant. This process is termed transformation and each crop requires slightly different conditions in order for transformation to occur. Here we describe the transformation protocol for bottle gourd and establish the conditions which optimize the transformation procedure. The technique established here will be used for future studies to improve the nutrient value and growth characteristics of this plant.
Technical Abstract: 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 T(1) generation plants were confirmed by a histochemical GUS assay, polymerase chain reaction and Southern blot analyses. Genetic segregation analysis of T(1) 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.