A multi-needle-assisted transformation of soybean cotyledonary node cells.
A new and simple method for wounding cotyledonary node cells of soybean [Glycine max (L) Merrill] was developed for obtaining a high frequency of transformants. Soybean seeds were germinated for 1 day, and the cotyledonary node cells of half-seeds were wounded mechanically by using a multi-needle consisting of thin 30 fibers. The wounded half-seeds were inoculated with Agrobacterium tumefaciens cells harboring a recombinant DNA that contained the bar and sgfp genes conferring phosphinothricin (PPT)-resistance and green fluorescent protein (GFP) activity, respectively. The inoculated explants were selected on medium containing 5 or 3 mg PPT/l. The transformation efficiency of soybean was up to 12%. Polymerase chain reaction and genomic Southern blot analysis confirmed stable integration of the transgenes in the genome of the PPT-resistant plants. GFP analysis revealed that the transgenes were highly expressed in the plantlets. Adult plants were resistant to 100 mg PPT/l applied on the leaves, demonstrating their herbicide-resistance.
Xue
Ren-Gao
RG
Department of Life and Science, Laiyang Agricultural College, Qingdao 266109, China. xuerengao@163.com
Xie
Hong-Feng
HF
Zhang
Biao
B
D018548:Cotyledon / Q000166:cytology / Q000235:genetics* / Q000382:microbiology; D018014:Gene Transfer Techniques / Q000295:instrumentation*; D053209:Herbicide Resistance / Q000235:genetics; D009339:Needles; D030821:Plants, Genetically Modified; D012231:Rhizobium / Q000235:genetics; D013025:Glycine max / Q000235:genetics*; D014170:Transformation, Genetic