A plant virus vector for systemic expression of foreign genes in cereals

Summary Inserts bearing the coding sequences of NPT II and β‐glucuronidase (GUS) were placed between the nuclear inclusion b (NIb) and coat protein (CP) domains of the wheat streak mosaic virus (WSMV) polyprotein ORF. The WSMV NIb‐CP junction containing the nuclear inclusion a (NIa) protease cleavag...

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Veröffentlicht in:	The Plant journal : for cell and molecular biology Jg. 23; H. 4; S. 547 - 555
Hauptverfasser:	Choi, Il‐Ryong, Stenger, Drake C., Morris, T. Jack, French, Roy
Format:	Journal Article
Sprache:	Englisch
Veröffentlicht:	Oxford, UK Blackwell Science Ltd 01.08.2000 Blackwell Science
Schlagworte:	Agronomy. Soil science and plant productions Avena sativa beta-glucuronidase Biological and medical sciences Biotechnology Carrier Proteins Carrier Proteins - genetics Carrier Proteins - metabolism cereals coat proteins Edible Grain Edible Grain - genetics Edible Grain - metabolism Edible Grain - virology Fundamental and applied biological sciences. Psychology gene expression Gene Transfer Techniques Genes, Reporter Genetic engineering Genetic engineering applications Genetic technics genetic transformation Genetic Vectors genetics Genetics and breeding of economic plants Glucuronidase Glucuronidase - genetics Glucuronidase - metabolism GUS gene histochemistry Hordeum vulgare infection interspecific variation leaves metabolism Methods. Procedures. Technologies monocotyledonous plants Mosaic Viruses Mosaic Viruses - genetics NIb protein NPT II gene nptII gene open reading frames Plant breeding: fundamental aspects and methodology Plant Roots Plant Roots - metabolism polymerase chain reaction reporter genes Reverse Transcriptase Polymerase Chain Reaction roots seedlings Sodium-Phosphate Cotransporter Proteins Symporters Transgenic animals and transgenic plants Transgenic plants Triticum aestivum virology virulence virus gene expression vector Wheat streak mosaic virus Zea mays Monocotyledones β-Glucuronidase Genetic transformation Zea mays Plant pathogen Enzyme Plant juvenile growth stage Insertion Gene expression Cereal crop Virus Gramineae Glycosidases Angiospermae Cereal Hydrolases Spermatophyta Genetic engineering Transgenic plant O-Glycosidases Vector Triticum aestivum
ISSN:	0960-7412, 1365-313X
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Zusammenfassung:	Summary Inserts bearing the coding sequences of NPT II and β‐glucuronidase (GUS) were placed between the nuclear inclusion b (NIb) and coat protein (CP) domains of the wheat streak mosaic virus (WSMV) polyprotein ORF. The WSMV NIb‐CP junction containing the nuclear inclusion a (NIa) protease cleavage site was duplicated, permitting excision of foreign protein domains from the viral polyprotein. Wheat, barley, oat and maize seedlings supported systemic infection of WSMV bearing NPT II. The NPT II insert was stable for at least 18–30 days post‐inoculation and had little effect on WSMV CP accumulation. Histochemical assays indicated the presence of functional GUS protein in systemically infected wheat and barley plants inoculated with WSMV bearing GUS. The GUS constructs had greatly reduced virulence on both oat and maize. RT–PCR indicated that the GUS insert was subject to deletion, particularly when expressed as a GUS–NIb protein fusion. Both reporter genes were expressed in wheat roots at levels comparable to those observed in leaves. These results clearly demonstrate the utility of WSMV as a transient gene expression vector for grass species, including two important grain crops, wheat and maize. The results further indicate that both host species and the nature of inserted sequences affect the stability and expression of foreign genes delivered by engineered virus genomes.
Bibliographie:	ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 ObjectType-Article-1 ObjectType-Feature-2
ISSN:	0960-7412 1365-313X
DOI:	10.1046/j.1365-313x.2000.00820.x