Document Detail

A novel, two-component system for cell lethality and its use in engineering nuclear male-sterility in plants.
MedLine Citation:
PMID:  12100487     Owner:  NLM     Status:  MEDLINE    
Ablation of cells by the controlled expression of a lethal gene can be used to engineer plant traits such as male sterility and disease resistance. However, it may not be possible to achieve sufficient specificity of expression to prevent secondary effects in non-targeted tissues. In this paper we demonstrate that the extracellular ribonuclease, barnase, can be engineered into two complementary fragments, allowing overlapping promoter specificity to be used to enhance targeting specificity. Using a transient system, we first show that barnase can be split into two inactive peptide fragments, that when co-expressed can complement each other to reconstitute barnase activity. When a luciferase reporter gene was introduced into plant cells along with genes encoding both partial barnase peptides, a substantial reduction in luciferase activity was seen. Cytotoxicity of the reconstituted barnase was demonstrated by crossing together parents constitutively expressing each of the barnase fragments, then assaying their progeny for the presence of both partial barnase genes. None of over 300 tomato seeds planted resulted in a viable progeny that inherited both transgenes. When expression of the partial barnase genes was instead targeted to the tapetum, male sterility resulted. All 13 tomato progeny that inherited both transgenes were male sterile, whereas the three progeny inheriting only the N-terminal barnase gene were male fertile. Finally, we describe how male sterility generated by this type of two-component system can be used in hybrid seed production.
Diane G Burgess; Edward J Ralston; William G Hanson; Matthew Heckert; Minh Ho; Tina Jenq; Joseph M Palys; Keliang Tang; Neal Gutterson
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  The Plant journal : for cell and molecular biology     Volume:  31     ISSN:  0960-7412     ISO Abbreviation:  Plant J.     Publication Date:  2002 Jul 
Date Detail:
Created Date:  2002-07-08     Completed Date:  2002-09-16     Revised Date:  2006-11-07    
Medline Journal Info:
Nlm Unique ID:  9207397     Medline TA:  Plant J     Country:  England    
Other Details:
Languages:  eng     Pagination:  113-25     Citation Subset:  IM    
DNA Plant Technologies, 6701 San Pablo Avenue, Oakland, CA 94608, USA.
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MeSH Terms
Base Sequence
Cell Death
DNA, Plant / genetics
Genes, Plant
Genetic Complementation Test
Genetic Engineering
Lycopersicon esculentum / cytology,  enzymology,  genetics
Models, Molecular
Peptide Fragments / chemistry,  genetics
Plants / cytology,  enzymology*,  genetics*
Plants, Genetically Modified
Reproduction / genetics
Ribonucleases / chemistry,  genetics*
Two-Hybrid System Techniques
Reg. No./Substance:
0/DNA, Plant; 0/Peptide Fragments; EC 3.1.-/Ribonucleases; EC 3.1.4.-/Bacillus amyloliquefaciens ribonuclease

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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