+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Liposome-mediated delivery of tobacco mosaic virus RNA into petunia protoplast : Improved conditions for liposome-protoplast incubations



Liposome-mediated delivery of tobacco mosaic virus RNA into petunia protoplast : Improved conditions for liposome-protoplast incubations



Plant Molecular Biology 2(1): 5-14



Liposome-mediated delivery of TMV RNA into petunia protoplasts and resulting virus antigen production has been used as an assay for determining incubation conditions which favor increased uptake of vesicle contents by plant cells. Vesicle phospholipid composition, incubation buffer divalent metal ion concentration, the type and concentration of polyalcohol used to stimulate vesicle uptake and the RNA content of the liposome preparation were determined to be critical factors influencing the efficiency of delivery. Manipulation of these parameters resulted in a 50-fold improvement in virus antigen production over that obtained with conditions previously used for liposome-protoplast incubations (Proc Natl Acad Sci 79: 1859-1863, 1982). Virus antigen production could be detected following incubation of protoplasts with <0.5 ng of encapsulated TMV RNA, while at higher concentrations of added liposomes, >80% of the protoplasts could be infected. Comparisons with other techniques used to introduce nucleic acids into plant protoplasts indicated that liposome-mediated delivery was 10-to 1 000-fold more efficient than these other methods. The general use of liposomes to introduce RNA and DNA molecules into plant protoplasts is discussed.

Please choose payment method:






(PDF emailed within 0-6 h: $19.90)

Accession: 054139918

Download citation: RISBibTeXText

PMID: 24318098

DOI: 10.1007/bf00187570


Related references

Liposome-mediated delivery of tobacco mosaic virus RNA into tobacco protoplasts: A sensitive assay for monitoring liposome-protoplast interactions. Proceedings of the National Academy of Sciences of the United States of America 79(6): 1859-1863, 1982

Liposome mediated transformation of tobacco protoplast by delta-endotoxin gene. Bulletin of Faculty of Agriculture, University of Cairo 42(3 Suppl.)): 895-910, 1991

Liposome-mediated infection of plant protoplasts with tobacco mosaic virus RNA. Virology 113(2): 752-760, 1981

Transfection of plant protoplasts with tobacco mosaic virus RNA using electroporation, PEG and cationic liposome - mediated methods. Acta Biochimica et Biophysica Sinica 26(1): 7-13, 1994

Cucumber mosaic virus and tobacco mosaic virus proliferation in transgenic chili peppers and its protoplast. Virologica Sinica 17(3): 243-247, 2002

Liposome-protoplast interactions. Journal of Chemical Technology and Biotechnology 32(1): 347-353, 2007

Effect of protein aggregation state on coat protein-mediated protection against tobacco mosaic virus using a transient protoplast assay. Virology 173(2): 656-663, 1989

Gametosomatic hybridization between egg cell protoplast and mesophyll protoplast of Petunia hybrida. Plant Biotechnology 26(4): 377-383, 2009

The influence of protoplast concentration on retention of tobacco mosaic virus by tobacco protoplasts. Virology 84(2): 544-546, 1978

Interaction of ordinary and tomato tobacco mosaic virus infecting tobacco protoplast. Annals of the Phytopathological Society of Japan 41(1): 86-87, 1975

Tobacco mosaic virus inoculation of tobacco protoplasts in the presence of protoplast fusion agents. Zeitschrift fuer Naturforschung Section C Journal of Biosciences 35(11-12): 1057-1061, 1980

Cell wall and protoplast isoperoxidases in tobacco plants in relation to mechanical injury and infection with tobacco mosaic virus. Plant Physiology 55(4): 611-619, 1975

Liposome-protoplast fusion in Phycomyces blakesleeanus. FEMS microbiology letters Federation of European Microbiological Societies 51(1): 37-40, 1988

The effects of liposome size and surface charge on liposome-mediated delivery of methotrexate-gamma-aspartate to cells in vitro. Biochimica et Biophysica Acta 820(1): 74-84, 1985

The theory and practical applications of liposome-protoplast interactions. Experientia. Supplementum 46: 197-207, 1983