Summary
We have used both Escherichia coli cells and Agrobacterium tumefaciens cells as microprojectiles to deliver DNA into suspension-cultured tobacco (Nicotiana tabacum L. line NT1) cells using a helium powered biolistic device. In addition, E. coli cells were used as microprojectiles for the transformation of suspension-cultured maize (Zea mays cv. Black Mexican Sweet) cells. Pretreating the bacterial cells with phenol at a concentration of 1.0%, and combining the bacterial cells with tungsten particles increased the rates of transformation. In N. tabacum, we obtained hundreds of transient transformants per bombardment, but were unable to recover any stable transformants. In Z. mays we obtained thousands of transient transformants and an average of six stable transformants per bombardment. This difference is discussed.
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Abbreviations
- BMS:
-
Black Mexican Sweet
- RPM:
-
revolutions per minute
- uidA:
-
β-glucuronidase gene
- GUS:
-
β-glucuronidase protein
- LB:
-
Luria-Bertani broth
- OD600:
-
optical density at 600 nm
- psi:
-
pounds per square inch
- Apr :
-
ampicillin resistance
- Knr :
-
kanamycinresistance
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Communicated by E. D. Earle
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Rasmussen, J.L., Kikkert, J.R., Roy, M.K. et al. Biolistic transformation of tobacco and maize suspension cells using bacterial cells as microprojectiles. Plant Cell Reports 13, 212–217 (1994). https://doi.org/10.1007/BF00239895
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DOI: https://doi.org/10.1007/BF00239895