Abstract
Wheat dwarf virus (WDV) is a geminivirus that infects monocotyledonous plants. To exploit the potential of WDV as a replicative gene vector, we developed a transient replication and expression system based on the transfection of protoplasts derived from Triticum monococcum suspension culture cells. Cloned genomic copies of various WDV isolates as well as mutants constructed in vitro were introduced into the protoplasts and assayed for their ability to replicate. As a result, regions of the WDV genome necessary or dispensable for the viral DNA replication could be defined. In addition, the gene encoding the viral capsid protein was replaced by three different bacterial marker genes, neomycin phosphotransferase, chloramphenicol acetyltransferase, and beta-galactosidase. The beta-galactosidase gene doubled the size of the WDV genome. The replication of the recombinant WDV genomes and the expression of these genes were monitored in suspension culture cells of T. monococcum. The potential of replicative expression vectors based on the WDV genome is discussed.
Full Text
The Full Text of this article is available as a PDF (2.8 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Accotto G. P., Donson J., Mullineaux P. M. Mapping of Digitaria streak virus transcripts reveals different RNA species from the same transcription unit. EMBO J. 1989 Apr;8(4):1033–1039. doi: 10.1002/j.1460-2075.1989.tb03470.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Andersen M. T., Richardson K. A., Harbison S. A., Morris B. A. Nucleotide sequence of the geminivirus chloris striate mosaic virus. Virology. 1988 Jun;164(2):443–449. doi: 10.1016/0042-6822(88)90558-2. [DOI] [PubMed] [Google Scholar]
- Beck E., Ludwig G., Auerswald E. A., Reiss B., Schaller H. Nucleotide sequence and exact localization of the neomycin phosphotransferase gene from transposon Tn5. Gene. 1982 Oct;19(3):327–336. doi: 10.1016/0378-1119(82)90023-3. [DOI] [PubMed] [Google Scholar]
- Briddon R. W., Watts J., Markham P. G., Stanley J. The coat protein of beet curly top virus is essential for infectivity. Virology. 1989 Oct;172(2):628–633. doi: 10.1016/0042-6822(89)90205-5. [DOI] [PubMed] [Google Scholar]
- Donson J., Accotto G. P., Boulton M. I., Mullineaux P. M., Davies J. W. The nucleotide sequence of a geminivirus from Digitaria sanguinalis. Virology. 1987 Nov;161(1):160–169. doi: 10.1016/0042-6822(87)90182-6. [DOI] [PubMed] [Google Scholar]
- Donson J., Gunn H. V., Woolston C. J., Pinner M. S., Boulton M. I., Mullineaux P. M., Davies J. W. Agrobacterium-mediated infectivity of cloned digitaria streak virus DNA. Virology. 1988 Jan;162(1):248–250. doi: 10.1016/0042-6822(88)90416-3. [DOI] [PubMed] [Google Scholar]
- Elmer J. S., Brand L., Sunter G., Gardiner W. E., Bisaro D. M., Rogers S. G. Genetic analysis of the tomato golden mosaic virus. II. The product of the AL1 coding sequence is required for replication. Nucleic Acids Res. 1988 Jul 25;16(14B):7043–7060. doi: 10.1093/nar/16.14.7043. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fromm M., Taylor L. P., Walbot V. Expression of genes transferred into monocot and dicot plant cells by electroporation. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5824–5828. doi: 10.1073/pnas.82.17.5824. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hamilton W. D., Bisaro D. M., Coutts R. H., Buck K. W. Demonstration of the bipartite nature of the genome of a single-stranded DNA plant virus by infection with the cloned DNA components. Nucleic Acids Res. 1983 Nov 11;11(21):7387–7396. doi: 10.1093/nar/11.21.7387. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hanley-Bowdoin L., Elmer J. S., Rogers S. G. Transient expression of heterologous RNAs using tomato golden mosaic virus. Nucleic Acids Res. 1988 Nov 25;16(22):10511–10528. doi: 10.1093/nar/16.22.10511. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hayes R. J., Coutts R. H., Buck K. W. Stability and expression of bacterial genes in replicating geminivirus vectors in plants. Nucleic Acids Res. 1989 Apr 11;17(7):2391–2403. doi: 10.1093/nar/17.7.2391. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Howarth A. J., Caton J., Bossert M., Goodman R. M. Nucleotide sequence of bean golden mosaic virus and a model for gene regulation in geminiviruses. Proc Natl Acad Sci U S A. 1985 Jun;82(11):3572–3576. doi: 10.1073/pnas.82.11.3572. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kalnins A., Otto K., Rüther U., Müller-Hill B. Sequence of the lacZ gene of Escherichia coli. EMBO J. 1983;2(4):593–597. doi: 10.1002/j.1460-2075.1983.tb01468.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Laufs J., Wirtz U., Kammann M., Matzeit V., Schaefer S., Schell J., Czernilofsky A. P., Baker B., Gronenborn B. Wheat dwarf virus Ac/Ds vectors: expression and excision of transposable elements introduced into various cereals by a viral replicon. Proc Natl Acad Sci U S A. 1990 Oct;87(19):7752–7756. doi: 10.1073/pnas.87.19.7752. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lazarowitz S. G. Infectivity and complete nucleotide sequence of the genome of a South African isolate of maize streak virus. Nucleic Acids Res. 1988 Jan 11;16(1):229–249. doi: 10.1093/nar/16.1.229. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lazarowitz S. G., Pinder A. J., Damsteegt V. D., Rogers S. G. Maize streak virus genes essential for systemic spread and symptom development. EMBO J. 1989 Apr;8(4):1023–1032. doi: 10.1002/j.1460-2075.1989.tb03469.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lusky M., Botchan M. R. Transient replication of bovine papilloma virus type 1 plasmids: cis and trans requirements. Proc Natl Acad Sci U S A. 1986 Jun;83(11):3609–3613. doi: 10.1073/pnas.83.11.3609. [DOI] [PMC free article] [PubMed] [Google Scholar]
- MacDowell S. W., Macdonald H., Hamilton W. D., Coutts R. H., Buck K. W. The nucleotide sequence of cloned wheat dwarf virus DNA. EMBO J. 1985 Sep;4(9):2173–2180. doi: 10.1002/j.1460-2075.1985.tb03912.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Morris B., Coates L., Lowe S., Richardson K., Eddy P. Nucleotide sequence of the infectious cloned DNA components of African cassava mosaic virus (Nigerian strain). Nucleic Acids Res. 1990 Jan 11;18(1):197–198. doi: 10.1093/nar/18.1.197. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mullineaux P. M., Donson J., Morris-Krsinich B. A., Boulton M. I., Davies J. W. The nucleotide sequence of maize streak virus DNA. EMBO J. 1984 Dec 20;3(13):3063–3068. doi: 10.1002/j.1460-2075.1984.tb02258.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Peden K. W., Pipas J. M., Pearson-White S., Nathans D. Isolation of mutants of an animal virus in bacteria. Science. 1980 Sep 19;209(4463):1392–1396. doi: 10.1126/science.6251547. [DOI] [PubMed] [Google Scholar]
- Reiss B., Sprengel R., Will H., Schaller H. A new sensitive method for qualitative and quantitative assay of neomycin phosphotransferase in crude cell extracts. Gene. 1984 Oct;30(1-3):211–217. doi: 10.1016/0378-1119(84)90122-7. [DOI] [PubMed] [Google Scholar]
- Rogers S. G., Bisaro D. M., Horsch R. B., Fraley R. T., Hoffmann N. L., Brand L., Elmer J. S., Lloyd A. M. Tomato golden mosaic virus A component DNA replicates autonomously in transgenic plants. Cell. 1986 May 23;45(4):593–600. doi: 10.1016/0092-8674(86)90291-6. [DOI] [PubMed] [Google Scholar]
- Schreier P. H., Seftor E. A., Schell J., Bohnert H. J. The use of nuclear-encoded sequences to direct the light-regulated synthesis and transport of a foreign protein into plant chloroplasts. EMBO J. 1985 Jan;4(1):25–32. doi: 10.1002/j.1460-2075.1985.tb02312.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stanley J., Markham P. G., Callis R. J., Pinner M. S. The nucleotide sequence of an infectious clone of the geminivirus beet curly top virus. EMBO J. 1986 Aug;5(8):1761–1767. doi: 10.1002/j.1460-2075.1986.tb04424.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stanley J., Townsend R. Infectious mutants of cassava latent virus generated in vivo from intact recombinant DNA clones containing single copies of the genome. Nucleic Acids Res. 1986 Aug 11;14(15):5981–5998. doi: 10.1093/nar/14.15.5981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sunter G., Bisaro D. M. Transcription map of the B genome component of tomato golden mosaic virus and comparison with A component transcripts. Virology. 1989 Dec;173(2):647–655. doi: 10.1016/0042-6822(89)90577-1. [DOI] [PubMed] [Google Scholar]
- Sunter G., Gardiner W. E., Bisaro D. M. Identification of tomato golden mosaic virus-specific RNAs in infected plants. Virology. 1989 May;170(1):243–250. doi: 10.1016/0042-6822(89)90372-3. [DOI] [PubMed] [Google Scholar]
- Townsend R., Stanley J., Curson S. J., Short M. N. Major polyadenylated transcripts of cassava latent virus and location of the gene encoding coat protein. EMBO J. 1985 Jan;4(1):33–37. doi: 10.1002/j.1460-2075.1985.tb02313.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Townsend R., Watts J., Stanley J. Synthesis of viral DNA forms in Nicotiana plumbaginifolia protoplasts inoculated with cassava latent virus (CLV); evidence for the independent replication of one component of the CLV genome. Nucleic Acids Res. 1986 Feb 11;14(3):1253–1265. doi: 10.1093/nar/14.3.1253. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Velten J., Schell J. Selection-expression plasmid vectors for use in genetic transformation of higher plants. Nucleic Acids Res. 1985 Oct 11;13(19):6981–6998. doi: 10.1093/nar/13.19.6981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]
- Woolston C. J., Reynolds H. V., Stacey N. J., Mullineaux P. M. Replication of wheat dwarf virus DNA in protoplasts and analysis of coat protein mutants in protoplasts and plants. Nucleic Acids Res. 1989 Aug 11;17(15):6029–6041. doi: 10.1093/nar/17.15.6029. [DOI] [PMC free article] [PubMed] [Google Scholar]
- von Wilcken-Bergmann B., Tils D., Sartorius J., Auerswald E. A., Schröder W., Müller-Hill B. A synthetic operon containing 14 bovine pancreatic trypsin inhibitor genes is expressed in E. coli. EMBO J. 1986 Dec 1;5(12):3219–3225. doi: 10.1002/j.1460-2075.1986.tb04632.x. [DOI] [PMC free article] [PubMed] [Google Scholar]