Document Detail


High-frequency transformation of undeveloped plastids in tobacco suspension cells.
MedLine Citation:
PMID:  15141065     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Although leaf chloroplast transformation technology was developed more than a decade ago, no reports exist of stable transformation of undeveloped plastids or other specialized plastid types, such as proplastids, etioplasts, or amyloplasts. In this work we report development of a dark-grown tobacco suspension cell model system to investigate the transformation potential of undeveloped plastids. Electron microscope analysis confirmed that the suspension cells carry plastids that are significantly smaller (approximately 50-fold less in volume) and have a very different subcellular localization and developmental state than leaf cell chloroplasts. Using antibiotic selection in the light, we demonstrated that both plastid and nuclear transformation of these cell suspensions is efficient and reproducible, with plastid transformation frequency at least equal to that of leaf chloroplast transformation. Homoplasmic plastid transformants are readily obtained in cell colonies, or in regenerated plants, providing a more consistent and versatile model than the leaf transformation system. Because of the uniformity of the cell suspension model, we could further show that growth rate, selection scheme, particle size, and DNA amount influence the frequency of transformation. Our results indicate that the rate-limiting steps for nuclear and plastid transformation are different, and each must be optimized separately. The suspension cell system will be useful as a model for understanding transformation in those plant species that utilize dark-grown embryogenic cultures and for characterizing the steps that lead to homoplasmic plastid transformation.
Authors:
Camri L Langbecker; Guang-Ning Ye; Debra L Broyles; Lisa L Duggan; Charles W Xu; Peter T J Hajdukiewicz; Charles L Armstrong; Jeffrey M Staub
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Plant physiology     Volume:  135     ISSN:  0032-0889     ISO Abbreviation:  Plant Physiol.     Publication Date:  2004 May 
Date Detail:
Created Date:  2004-05-13     Completed Date:  2004-10-14     Revised Date:  2010-09-20    
Medline Journal Info:
Nlm Unique ID:  0401224     Medline TA:  Plant Physiol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  39-46     Citation Subset:  IM    
Affiliation:
Monsanto Company, Chesterfield, Missouri 63017, USA.
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MeSH Terms
Descriptor/Qualifier:
Cell Nucleus / metabolism
Cells, Cultured
Chloroplasts / physiology,  ultrastructure
DNA, Chloroplast / pharmacology
DNA, Plant / pharmacology
Microscopy, Electron
Particle Size
Plastids / physiology*,  ultrastructure
Tobacco / cytology,  genetics*,  physiology
Transformation, Genetic
Chemical
Reg. No./Substance:
0/DNA, Chloroplast; 0/DNA, Plant
Comments/Corrections

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