Liposome-mediated delivery of tobacco mosaic virus RNA into tobacco protoplasts: A sensitive assay for monitoring liposome-protoplast interactions

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Liposome-mediated delivery of tobacco mosaic virus RNA into tobacco protoplasts: A sensitive assay for monitoring liposome-protoplast interactions.

Tobacco mosaic virus (TMV) RNA was encapsulated in large, unilamellar phospholipid vesicles (liposomes), and the encapsulated TMV RNA was shown to be infectious when incubated with tobacco protoplasts under appropriate conditions. Maximal virus production in protoplasts was observed after their incubation with TMV RNA entrapped in phosphatidylserine/cholesterol liposomes. Infection was dependen...

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Infection of tobacco mesophyll protoplasts by tobacco mosaic virus.

It was proved that the protoplasts prepared from mesophyll of Nicotiana tabacum are infected by tobacco mosaic virus. The infection occurred when purified tobacco mosaic virus particles were added to a protoplast suspension in the presence of poly-L-ornithine. The virus multiplied in these protoplasts to a level of 10(6) virus particles per infected protoplast during 24 hours of incubation. The...

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High Efficiency Electro-transfection of Tobacco Mesophyll Protoplasts with Tobacco Mosaic Virus RNA

Tobacco mesophyll protoplasts were transfected with tobacco mosaic virus (TMV) RNA in an electric field using a newly devised chamber constructed with gold-coated glass panel electrodes. Up to 95 9/oo of protoplasts subjected to several DC pulses (50 Ixs, 550 to 800 V/cm) while suspended in 0.5 i-mannitol containing 10 ~tg/ml TMV RNA became infected. The treatment did not affect the viability o...

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A simplified method of obtaining tobacco protoplasts for infection with tobacco mosaic virus.

Incubating tobacco leaf tissue, from which the lower epidermis was peeled, overnight with o'3 to o.4 % Macerozyme and o-6 to i-2 % cellulase, depending on leaf condition, produced a good yield of protoplasts that were susceptible to infection by TMV. Fluorescent antibody staining showed that 20 to 8o % ofprotoplasts became infected, and infectivity tests indicated that an average of about 4 × I...

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Protoplasts isolated from tobacco leaves were inoculated with cucumber mosaic virus (CMV) using a method slightly modified from that developed for inoculating tobacco mosaic virus. Multiplication of CMV was demonstrated by staining the protoplasm with fluorescent CMV-antibody, by the electron microscopy of ultrathin sections, and by assaying the infectivity in protoplasts. Seventy to 90% of pro...

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ژورنال

عنوان ژورنال: Proceedings of the National Academy of Sciences

سال: 1982

ISSN: 0027-8424,1091-6490

DOI: 10.1073/pnas.79.6.1859