Transformation of sugar beet cell suspension cultures |
| |
Authors: | Snezana?D?Ivic Email author" target="_blank">Ann?C?SmigockiEmail author |
| |
Institution: | (1) Molecular Plant Pathology Laboratory, United States Department of Agriculture, Agricultural Research Service, 20705 Beltsville, MD |
| |
Abstract: | Summary A sugar beet transformation method was developed using particle bombardment of short-term suspension cultures of a breeding
line FC607. Highly embryogenic suspension cultures derived from leaf callus were bombarded with the uidA (gusA) reporter gene under the control of either the osmotin or proteinase inhibitor II gene promoter, and the npt II selectable marker gene. Transient uidA expression was visualized as 500–4000 blue units per 200 mg of bombarded cells 2 d after bombardment. Stably-transformed
calluses were recovered on both kanamycin and paromomycin media. The greatest number of GUS (+) calluses was obtained when
50 or 100 mgl−1 of kanamycin was applied 2 d after transformation for 3–5 wk, followed by either no selection or reduced levels of the antibiotic.
PCR analyses of the GUS (+) callus lines revealed the expected size fragment for uidA and npt II genes. Stable incorporation of the uidA gene into the genome was confirmed by Southern blot analyses. Several transformed embryos were detected by histochemical
β-glucuronidase (GUS) staining. |
| |
Keywords: | Beta vulgaris biolistic embryogenesis npt II particle bombardment uidA |
本文献已被 SpringerLink 等数据库收录! |