Abstract
Leaf pieces of in vitro-cultured plantlets of the wild potato species Solanum brevidens Phil. were cocultivated with Agrobacterium tumefaciens that contained nptII and uidA genes on the disarmed plasmid pBI121. Independent transgenic shoots were regenerated from solidified and liquid medium that contained 50 mg l−1 kanamycin. Two Agrobacterium strains were investigated for transformation efficiency. GV2260, which contained p35SGUSINT, resulted in a 11% transformation frequency, compared with 1% using LBA4404. Transformation rates were 7% in liquid culture and 3% on solidified medium. All kanamycinresistant, putatively transformed plantlets were confirmed positive by histochemical GUS assays. GUS activity in 22 independently transformed plants was quantified by fluorometric assay. Southern analysis of randomly selected transgenic plants showed that each transgenic plant contained at least one copy of the uidA gene.
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Abbreviations
- GUS:
-
ß-glucuronidase
- MS:
-
Murashige-Skoog medium
- BA:
-
6-benzylaminopurine
- 2ip:
-
6-(λ, λ-dimethylallylamino)purine
- IAA:
-
indole-3-acetic acid
- GA3 :
-
gibberellic acid
- npt II :
-
neomycin phosphotransferase II
- NOS:
-
nopaline synthase
- MUG:
-
4-methyl umbelliferyl glucuronide
- MU:
-
7-hydroxy-4-methylcoumarin
- X-gluc:
-
5-bromo-4-chloro-3-indolyl ß-D-glucuronic acid
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Communicated by J. Widholm
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Liu, T.H.A., Stephens, L.C. & Hannapel, D.J. Transformation of Solanum brevidens using Agrobacterium tumefaciens . Plant Cell Reports 15, 196–199 (1995). https://doi.org/10.1007/BF00193719
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DOI: https://doi.org/10.1007/BF00193719