Abstract
Chloroplast expression plasmids pTRBCL-GUS (tobaccorbcL promoter-gusA-tobaccorbcL terminator) and pHHU3004 (spinach ‘x gene’ promoter-gusA-spinachrbcL terminator) and a control nuclear expression plasmid pBI221 (CaMV 35S promoter-gusA-NOS terminator) were introduced separately into cultured cells and tissues of tobacco andArabidopsis thaliana, as well as into cultured cells of the lower land plants liverwort and hornwort by a pneumatic particle gun. The pTRBCL-GUS and pHHU3004 plasmids produced many blue spots in the BY-2 cells and the roots ofArabidopsis thaliana, but not in any of the green cells or tissues. The results suggest that the pTRBCL-GUS and pHHU3004 plasmids are expressed more in proplastids and amyloplasts than in chloroplasts. GUS activities of the BY-2 cells bombarded with pTRBCL-GUS and pHHU3004 were insensitive to α-amanitin treatment (10 and 50 μg/ml), while that of the cells with pBI221 greatly decreased by the same treatment. Hence, it is likely that the pTRBCL-GUS and pHHU3004 plasmids were substantially expressed in the proplastids.
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Abbreviations
- CaMV:
-
cauliflower mosaic virus
- CIM:
-
callusinducing medium
- CRM:
-
callus regeneration medium
- 2,4-D:
-
2,4-dichlorophenoxyacetic acid
- GUS:
-
β-glucuronidase
- LS:
-
Linsmaier and Skoog
- MS:
-
Murashige and Skoog
- MU:
-
4-methylumbelliferone
- MUG:
-
4-methyl umbellifery-β-D-glucuronide
- NAA:
-
1-naphthaleneacetic acid
- NOS:
-
nopaline synthase
- SIM:
-
shoot-inducing medium
- X-Gluc:
-
5-bromo-4-chloro-3-indolyl-β-D-glucuronide
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Seki, M., Shigemoto, N., Sugita, M. et al. Transient expression of β-glucuronidase in plastids of various plant cells and tissues delivered by a pneumatic particle gun. J. Plant Res. 108, 235–240 (1995). https://doi.org/10.1007/BF02344348
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DOI: https://doi.org/10.1007/BF02344348