Plant Physiol.
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via CrossRef
Right arrow Citing Articles via Web of Science (88)
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by DeGray, G.
Right arrow Articles by Daniell, H.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by DeGray, G.
Right arrow Articles by Daniell, H.
Agricola
Right arrow Articles by DeGray, G.
Right arrow Articles by Daniell, H.

Plant Physiol, November 2001, Vol. 127, pp. 852-862

Expression of an Antimicrobial Peptide via the Chloroplast Genome to Control Phytopathogenic Bacteria and Fungi

Gerald DeGray, Kanniah Rajasekaran, Franzine Smith, John Sanford, and Henry Daniell*

Department of Molecular Biology and Microbiology and Center for Discovery of Drugs and Diagnostics, 12722 Research Parkway, University of Central Florida, Orlando, Florida 32826-3227 (G.D., H.D.); United States Department of Agriculture, Agricultural Research Service, Southern Regional Research Center, New Orleans, Louisiana 70124-4305 (K.R.); and Sanford Scientific Inc., 877 Marshall Road, Waterloo, New York 13165 (F.S., J.S.)

The antimicrobial peptide MSI-99, an analog of magainin 2, was expressed via the chloroplast genome to obtain high levels of expression in transgenic tobacco (Nicotiana tabacum var. Petit Havana) plants. Polymerase chain reaction products and Southern blots confirmed integration of MSI-99 into the chloroplast genome and achievement of homoplasmy, whereas northern blots confirmed transcription. Contrary to previous predictions, accumulation of MSI-99 in transgenic chloroplasts did not affect normal growth and development of the transgenic plants. This may be due to differences in the lipid composition of plastid membranes compared with the membranes of susceptible target microbes. In vitro assays with protein extracts from T1 and T2 plants confirmed that MSI-99 was expressed at high levels to provide 88% (T1) and 96% (T2) inhibition of growth against Pseudomonas syringae pv tabaci, a major plant pathogen. When germinated in the absence of spectinomycin selection, leaf extracts from T2 generation plants showed 96% inhibition of growth against P. syringae pv tabaci. In addition, leaf extracts from transgenic plants (T1) inhibited the growth of pregerminated spores of three fungal species, Aspergillus flavus, Fusarium moniliforme, and Verticillium dahliae, by more than 95% compared with non-transformed control plant extracts. In planta assays with the bacterial pathogen P. syringae pv tabaci resulted in areas of necrosis around the point of inoculation in control leaves, whereas transformed leaves showed no signs of necrosis, demonstrating high-dose release of the peptide at the site of infection by chloroplast lysis. In planta assays with the fungal pathogen, Colletotrichum destructivum, showed necrotic anthracnose lesions in non-transformed control leaves, whereas transformed leaves showed no lesions. Genetically engineering crop plants for disease resistance via the chloroplast genome instead of the nuclear genome is desirable to achieve high levels of expression and to prevent pollen-mediated escape of transgenes.

* Corresponding author; e-mail daniell{at}mail.ucf.edu; fax 407-384-2062.

© 2001 American Society of Plant Physiologists


This article has been cited by other articles:


Home page
 Tree PhysiolHome page
F.-H. Wu, D.-P. Kan, S.-B. Lee, H. Daniell, Y.-W. Lee, C.-C. Lin, N.-S. Lin, and C.-S. Lin
Complete nucleotide sequence of Dendrocalamus latiflorus and Bambusa oldhamii chloroplast genomes
Tree Physiol, June 1, 2009; 29(6): 847 - 856.
[Abstract] [Full Text] [PDF]

Home page
 Plant Physiol.Home page
S. C. Lee, I. S. Hwang, H. W. Choi, and B. K. Hwang
Involvement of the Pepper Antimicrobial Protein CaAMP1 Gene in Broad Spectrum Disease Resistance
Plant Physiology, October 1, 2008; 148(2): 1004 - 1020.
[Abstract] [Full Text] [PDF]

Home page
 Plant Physiol.Home page
D. Verma and H. Daniell
Chloroplast Vector Systems for Biotechnology Applications
Plant Physiology, December 1, 2007; 145(4): 1129 - 1143.
[Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
H. Daniell
Transgene containment by maternal inheritance: Effective or elusive?
PNAS, April 24, 2007; 104(17): 6879 - 6880.
[Full Text] [PDF]

Home page
 FASEB J.Home page
A. Limaye, V. Koya, M. Samsam, and H. Daniell
Receptor-mediated oral delivery of a bioencapsulated green fluorescent protein expressed in transgenic chloroplasts into the mouse circulatory system
FASEB J, May 1, 2006; 20(7): 959 - 961.
[Abstract] [Full Text] [PDF]

Home page
 Appl. Environ. Microbiol.Home page
M. Donini, C. Lico, S. Baschieri, S. Conti, W. Magliani, L. Polonelli, and E. Benvenuto
Production of an Engineered Killer Peptide in Nicotiana benthamiana by Using a Potato virus X Expression System
Appl. Envir. Microbiol., October 1, 2005; 71(10): 6360 - 6367.
[Abstract] [Full Text] [PDF]

Home page
 Plant Physiol.Home page
O. N. Ruiz and H. Daniell
Engineering Cytoplasmic Male Sterility via the Chloroplast Genome by Expression of {beta}-Ketothiolase
Plant Physiology, July 1, 2005; 138(3): 1232 - 1246.
[Abstract] [Full Text] [PDF]

Home page
 Plant Physiol.Home page
T. Quesada-Vargas, O. N. Ruiz, and H. Daniell
Characterization of Heterologous Multigene Operons in Transgenic Chloroplasts. Transcription, Processing, and Translation
Plant Physiology, July 1, 2005; 138(3): 1746 - 1762.
[Abstract] [Full Text] [PDF]

Home page
 J Exp BotHome page
D. P. Yevtushenko, R. Romero, B. S. Forward, R. E. Hancock, W. W. Kay, and S. Misra
Pathogen-induced expression of a cecropin A-melittin antimicrobial peptide gene confers antifungal resistance in transgenic tobacco
J. Exp. Bot., June 1, 2005; 56(416): 1685 - 1695.
[Abstract] [Full Text] [PDF]

Home page
 Plant Physiol.Home page
P. V. Viitanen, A. L. Devine, M. S. Khan, D. L. Deuel, D. E. Van Dyk, and H. Daniell
Metabolic Engineering of the Chloroplast Genome Using the Echerichia coli ubiC Gene Reveals That Chorismate Is a Readily Abundant Plant Precursor for p-Hydroxybenzoic Acid Biosynthesis
Plant Physiology, December 1, 2004; 136(4): 4048 - 4060.
[Abstract] [Full Text] [PDF]

Home page
 Plant Physiol.Home page
S. Kumar, A. Dhingra, and H. Daniell
Plastid-Expressed Betaine Aldehyde Dehydrogenase Gene in Carrot Cultured Cells, Roots, and Leaves Confers Enhanced Salt Tolerance
Plant Physiology, September 1, 2004; 136(1): 2843 - 2854.
[Abstract] [Full Text] [PDF]

Home page
 MicrobiologyHome page
G. Bell
Response to Hancock
Microbiology, December 1, 2003; 149(12): 3344 - 3345.
[Full Text] [PDF]

Home page
 Plant Physiol.Home page
O. N. Ruiz, H. S. Hussein, N. Terry, and H. Daniell
Phytoremediation of Organomercurial Compounds via Chloroplast Genetic Engineering
Plant Physiology, July 1, 2003; 132(3): 1344 - 1352.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
L. Lin, Y.-G. Liu, X. Xu, and B. Li
Efficient linking and transfer of multiple genes by a multigene assembly and transformation vector system
PNAS, May 13, 2003; 100(10): 5962 - 5967.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
ASPB Publications PLANT PHYSIOLOGY® THE PLANT CELL
Copyright © 2001 by the American Society of Plant Biologists