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6 results on '"MESH: Hordeum"'

1. Methionine Biosynthesis is Essential for Infection in the Rice Blast Fungus Magnaporthe oryzae

Author

Marc-Henri Lebrun, Marie Emmanuelle Saint-Macary, Michel Droux, Océane Frelin, Marie Josèphe Gagey, Roland Beffa, Crystel Barbisan, Microbiologie, adaptation et pathogénie (MAP), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS), Centre de recherche La Dargoire, Bayer Cropscience, Génomique fonctionnelle des champignons pathogènes des plantes (FungiPath), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université Claude Bernard Lyon 1 (UCBL), BIOlogie et GEstion des Risques en agriculture (BIOGER), AgroParisTech-Institut National de la Recherche Agronomique (INRA), Trafic et signalisation membranaires chez les bactéries (MTSB), Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Institut National des Sciences Appliquées (INSA)-Université de Lyon-Institut National des Sciences Appliquées (INSA)-Université de Lyon-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon, Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Institut National de la Recherche Agronomique (INRA)-AgroParisTech, and Bayer Crop Science

Subjects
MESH: Oryza, Homocysteine, amino-acid biosynthesis, lcsh:Medicine, Transsulfuration pathway, MESH: Phenotype, 5-Methyltetrahydrofolate-Homocysteine S-Methyltransferase, chemistry.chemical_compound, Methionine, Gene Expression Regulation, Fungal, Methionine synthase, MESH: Magnaporthe, lcsh:Science, Amino acid synthesis, [SDV.MP.MYC]Life Sciences [q-bio]/Microbiology and Parasitology/Mycology, Sequence Deletion, 2. Zero hunger, chemistry.chemical_classification, Multidisciplinary, biology, lcsh:R, food and beverages, Hordeum, Oryza, MESH: Sequence Deletion, Cystathionine beta synthase, infection, Amino acid, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], [SDV.BV.PEP]Life Sciences [q-bio]/Vegetal Biology/Phytopathology and phytopharmacy, Magnaporthe, Phenotype, chemistry, Biochemistry, MESH: Hordeum, MESH: Methionine, MESH: 5-Methyltetrahydrofolate-Homocysteine S-Methyltransferase, biology.protein, lcsh:Q, methionine, MESH: Gene Expression Regulation, Fungal, Cysteine, Research Article
Abstract

International audience; Methionine is a sulfur amino acid standing at the crossroads of several biosynthetic pathways. In fungi, the last step of methionine biosynthesis is catalyzed by a cobalamine-independent methionine synthase (Met6, EC 2.1.1.14). In the present work, we studied the role of Met6 in the infection process of the rice blast fungus, Magnaporthe oryzae. To this end MET6 null mutants were obtained by targeted gene replacement. On minimum medium, MET6 null mutants were auxotrophic for methionine. Even when grown in presence of excess methionine, these mutants displayed developmental defects, such as reduced mycelium pigmentation, aerial hypha formation and sporulation. They also displayed characteristic metabolic signatures such as increased levels of cysteine, cystathionine, homocysteine, S-adenosylmethionine, S-adenosylhomocysteine while methionine and glutathione levels remained unchanged. These metabolic perturbations were associated with the over-expression of MgCBS1 involved in the reversed transsulfuration pathway that metabolizes homocysteine into cysteine and MgSAM1 and MgSAHH1 involved in the methyl cycle. This suggests a physiological adaptation of M. oryzae to metabolic defects induced by the loss of Met6, in particular an increase in homocysteine levels. Pathogenicity assays showed that MET6 null mutants were non-pathogenic on both barley and rice leaves. These mutants were defective in appressorium-mediated penetration and invasive infectious growth. These pathogenicity defects were rescued by addition of exogenous methionine and S-methylmethionine. These results show that M. oryzae cannot assimilate sufficient methionine from plant tissues and must synthesize this amino acid de novo to fulfill its sulfur amino acid requirement during infection.

Published
2015
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2. Hordeins are expressed in microspore-derived embryos and also during male gametophytic and very early stages of seed development

Author

Amada Pulido, Adela Olmedilla, Beáta Barnabás, Alberto Hernando, Martine Devic, Ferenc Bakos, Enrique Méndez, Laboratoire Génome et développement des plantes (LGDP), and Université de Perpignan Via Domitia (UPVD)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0106 biological sciences, Glutens, Physiology, Blotting, Western, Stamen, Enzyme-Linked Immunosorbent Assay, Plant Science, Biology, 01 natural sciences, Endosperm, [SDV.GEN.GPL]Life Sciences [q-bio]/Genetics/Plants genetics, Tissue Culture Techniques, 03 medical and health sciences, MESH: In Situ Hybridization, Microspore, MESH: Reverse Transcriptase Polymerase Chain Reaction, Botany, MESH: Blotting, Western, MESH: Tissue Culture Techniques, RNA, Messenger, In Situ Hybridization, MESH: RNA, Messenger, Plant Proteins, 030304 developmental biology, 0303 health sciences, MESH: Plant Proteins, Reverse Transcriptase Polymerase Chain Reaction, Embryogenesis, MESH: Enzyme-Linked Immunosorbent Assay, food and beverages, MESH: Immunohistochemistry, Hordeum, Embryo, Immunohistochemistry, Cell biology, MESH: Seeds, Protein body, Hordein, MESH: Hordeum, MESH: Pollen, Seeds, Pollen, Hordeum vulgare, Prolamins, 010606 plant biology & botany
Abstract

Microspore-derived embryos induced by anther or isolated-microspore culture display certain characteristics of zygotic embryos. Furthermore, the expression of certain endosperm genes has been described in these non-zygotic embryos. The expression of hordein genes encoding the main barley endosperm proteins has been studied using a wide range of methods (RT-PCR, in situ hybridization, ELISA sandwich, western blotting immunocytochemistry, and cytochemistry) to ascertain their presence or absence during the induction and first stages of microspore embryogenesis. Due to the very sensitive techniques used it was possible to detect for the first time hordein expression during microspore embryogenesis. Surprisingly, these hordeins were also detected at different stages of male gametophytic development as well as during the very early stages of seed development, when they have not hitherto been detected. The expression and localization of these storage proteins and their corresponding transcripts provide new information about barley microspore embryogenesis and its relationship to zygotic embryogenesis. Although only small quantities of hordeins are accumulated during microspore embryogenesis they seem to be necessary for the initial development of the microspore-derived embryo. This idea is supported by the changes detected in their concentration throughout this process and is in accordance with previously published data concerning the importance of endosperm proteins for embryo development in both microspore culture and in planta.

Published
2006
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3. The fusarium graminearum genome reveals a link between localized polymorphism and pathogen specialization

Author

David R. Nelson, Hans-Werner Mewes, H. Corby Kistler, Weihong Qi, Kerry O'Donnell, John C. Kennell, Scott E. Baker, Liane R. Gale, Igor V. Tetko, Jon K. Magnuson, Gary J. Muehlbauer, Martin Münsterkötter, Martijn Rep, Karen Hilburn, Jiqiang Yao, B. Gillian Turgeon, Thérèse Ouellet, Hadi Quesneville, Li-Jun Ma, Todd J. Ward, Linda J. Harris, Gerhard Adam, Kim E. Hammond-Kosack, Sarah E. Calvo, Scott Kroken, M. Isabel G. Roncero, Kye Yong Seong, Jin-Rong Xu, Gertrud Mannhaupt, Yueh-Long Chang, Antonio Di Pietro, Ulrich Güldener, Christina A. Cuomo, Evan Mauceli, Rudolf Mitterbauer, John F. Antoniw, Rubella S. Goswami, David DeCaprio, Martin Urban, Cees Waalwijk, Jonathan D. Walton, Sante Gnerre, Frances Trail, Thomas K. Baldwin, Bruce W. Birren, Broad Institute of MIT and Harvard (BROAD INSTITUTE), Harvard Medical School [Boston] (HMS)-Massachusetts Institute of Technology (MIT)-Massachusetts General Hospital [Boston], Technische Universität Munchen - Université Technique de Munich [Munich, Allemagne] (TUM), Institute for Bioinformatics (MIPS), GSF National Research Center for Environment and Health, Purdue University [West Lafayette], Michigan State University [East Lansing], Michigan State University System, Cornell University [New York], Universidad de Córdoba [Cordoba], Pacific Northwest National Laboratory (PNNL), University of Amsterdam [Amsterdam] (UvA), Universität für Bodenkultur Wien [Vienne, Autriche] (BOKU), Rothamsted Research, University of Minnesota [Twin Cities] (UMN), University of Minnesota System, Agriculture and Agri-Food [Ottawa] (AAFC), U.S. Department of Agriculture - Agricultural Research Service - Cereal Disease Laboratory (USDA), USDA-ARS : Agricultural Research Service, Saint Louis University (SLU), University of Arizona, University of Tennessee Memphis, The University of Tennessee Health Science Center [Memphis] (UTHSC), USDA ARS, National Center for Agricultural Utilization Research (USDA ARS,), Institut Jacques Monod (IJM (UMR_7592)), Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Institute of Bioorganic Chemistry and Photochemistry, National Ukrainian Academy of Sciences, Institute Bioorganic Chemistry an Photochemistry, Plant Research International (PRI), Wageningen University and Research [Wageningen] (WUR), Molecular Plant Pathology (SILS, FNWI), Technische Universität München [München] (TUM), Cornell University, University of Minnesota [Twin Cities], and Wageningen University and Research Centre [Wageningen] (WUR)

Subjects
localized polymorphism, MESH: Sequence Analysis, DNA, neurospora, Sequence analysis, Molecular Sequence Data, Single-nucleotide polymorphism, Genomics, Gene mutation, Biology, dna, medicine.disease_cause, Genome, Polymorphism, Single Nucleotide, Evolution, Molecular, 03 medical and health sciences, MESH: Plant Diseases, Fusarium, MESH: Polymorphism, Genetic, medicine, Point Mutation, DNA, Fungal, Gene, MESH: Evolution, Molecular, 030304 developmental biology, Plant Diseases, MESH: Point Mutation, Genetics, 0303 health sciences, Mutation, MESH: Fusarium, Multidisciplinary, Polymorphism, Genetic, MESH: Molecular Sequence Data, Biointeracties and Plant Health, 030306 microbiology, Point mutation, MESH: Polymorphism, Single Nucleotide, food and beverages, Hordeum, Sequence Analysis, DNA, [SDV.BIBS]Life Sciences [q-bio]/Quantitative Methods [q-bio.QM], MESH: DNA, Fungal, MESH: Hordeum, MESH: Genome, Fungal, Fusarium graminearum genome, PRI Biointeractions en Plantgezondheid, Genome, Fungal, [INFO.INFO-BI]Computer Science [cs]/Bioinformatics [q-bio.QM]
Abstract

We sequenced and annotated the genome of the filamentous fungus Fusarium graminearum , a major pathogen of cultivated cereals. Very few repetitive sequences were detected, and the process of repeat-induced point mutation, in which duplicated sequences are subject to extensive mutation, may partially account for the reduced repeat content and apparent low number of paralogous (ancestrally duplicated) genes. A second strain of F. graminearum contained more than 10,000 single-nucleotide polymorphisms, which were frequently located near telomeres and within other discrete chromosomal segments. Many highly polymorphic regions contained sets of genes implicated in plant-fungus interactions and were unusually divergent, with higher rates of recombination. These regions of genome innovation may result from selection due to interactions of F. graminearum with its plant hosts.

Published
2007
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4. Effect of dietary fibre of barley variety 'Rihane' on azoxymethane-induced aberrant crypt foci development and on colonic microbiota diversity in rats

Author

Mouledi El Felah, Mohamed Hammami, Ester Pereira, Irène Mangin, Lotfi Achour, Fabien Magne, Philippe Pochart, Antonia Suau, Moncef Mokni, Hichem Ben Salem, Lamia Lahouar, Génomique et évolution des microorganismes, Laboratoire Adaptation et pathogénie des micro-organismes [Grenoble] (LAPM), and Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF)

Subjects
Dietary Fiber, Male, MESH: Hydrogen-Ion Concentration, MESH: Bifidobacterium, 030309 nutrition & dietetics, MESH: Random Allocation, Medicine (miscellaneous), Gastroenterology, MESH: Carcinogens, chemistry.chemical_compound, Random Allocation, Aberrant Crypt Foci, MESH: Animals, Intestinal Mucosa, MESH: Phylogeny, Colonic disease, Phylogeny, 2. Zero hunger, 0303 health sciences, Nutrition and Dietetics, [SDV.BID.EVO]Life Sciences [q-bio]/Biodiversity/Populations and Evolution [q-bio.PE], Hydrogen-Ion Concentration, Gastrointestinal Contents, 3. Good health, MESH: Hordeum, Seeds, MESH: Intestinal Mucosa, Premalignant lesion, MESH: Tunisia, Colorectal Neoplasms, MESH: Azoxymethane, Aberrant crypt foci, medicine.medical_specialty, Tunisia, MESH: Rats, Colon, Azoxymethane, Biology, Body weight, MESH: Aberrant Crypt Foci, Acclimatization, 03 medical and health sciences, MESH: Enterobacteriaceae, Animal science, Enterobacteriaceae, Internal medicine, medicine, Animals, MESH: Colon, Rats, Wistar, Carcinogen, 030304 developmental biology, Dietary fibre, Hordeum, MESH: Rats, Wistar, MESH: Gastrointestinal Contents, MESH: Male, Rats, MESH: Prebiotics, Disease Models, Animal, Prebiotics, chemistry, MESH: Seeds, MESH: Dietary Fiber, Carcinogens, Bifidobacterium, MESH: Disease Models, Animal, MESH: Colorectal Neoplasms
Abstract

Many epidemiological and experimental studies have suggested an important role for dietary fibre (DF) of cereals in the prevention of colon cancer. The objective of the present study was to explain the effects of the DF of barley Rihane (BR) on azoxymethane (AOM)-induced aberrant crypt foci (ACF) and colonic bacterial diversity in rats. Following an acclimatisation period, rats were divided into four groups and fed a control (C) diet or experimental diet containing 30 % of BR. DF content in the experimental diet was twice that of the C diet (total DF was 8·69 % in the C diet and 15·24 % in the BR diet). At 7 and 8 weeks of age, rats received two successive subcutaneous injections of AOM at 20 mg/kg body weight. At 12 weeks after the first injection, ten animals from each group were killed. The BR diet decreased colonic pH (P P P

Published
2012
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5. Diversity and evolution of the Hordeum murinum polyploid complex in Algeria

Author

Malika L. Aïnouche, Spencer Brown, Abdelkader Aïnouche, Olivier Coriton, Rachid Amirouche, Marie-Thérèse Misset, Malika Ourari, Virginie Huteau, Faculté des Lettres et des Sciences humaines - Université de Béjaïa (FLSH), Université Abderrahmane Mira [Béjaïa], Ecosystèmes, biodiversité, évolution [Rennes] (ECOBIO), Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Institut Ecologie et Environnement (INEE), Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)-Observatoire des Sciences de l'Univers de Rennes (OSUR)-Centre National de la Recherche Scientifique (CNRS), Amélioration des Plantes et Biotechnologies Végétales (APBV), Institut National de la Recherche Agronomique (INRA)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-AGROCAMPUS OUEST, Institut des sciences du végétal (ISV), Centre National de la Recherche Scientifique (CNRS), Université des Sciences et de la Technologie Houari Boumediene [Alger] (USTHB), Programme Hubert Curien CMEP-Tassili 08 MDU 724 'Polyploidy, Genome evolution and Biodiversity', Centre National de la Recherche Scientifique (CNRS)-Observatoire des Sciences de l'Univers de Rennes (OSUR)-Institut Ecologie et Environnement (INEE), Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES), Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro), Université de Rennes (UR)-Institut Ecologie et Environnement (INEE), Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)-Observatoire des Sciences de l'Univers de Rennes (OSUR), Université de Rennes (UR)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Rennes 2 (UR2)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Institut national des sciences de l'Univers (INSU - CNRS)-Université de Rennes 2 (UR2)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Recherche Agronomique (INRA)-Université de Rennes (UR)-AGROCAMPUS OUEST, Université des Sciences et de la Technologie Houari Boumediene = University of Sciences and Technology Houari Boumediene [Alger] (USTHB), AGROCAMPUS OUEST-Université de Rennes 1 (UR1), and Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Institut National de la Recherche Agronomique (INRA)

Subjects
0106 biological sciences, MESH: Genome, Plant, Climate, MESH: Flow Cytometry, Evolution moléculaire, MESH: Base Sequence, 01 natural sciences, Genome, MESH: Ploidies, MESH: Chromosomes, Plant, MESH: Genetic Variation, MESH: Phylogeny, In Situ Hybridization, Phylogeny, 0303 health sciences, MESH: DNA, Ribosomal, biology, MESH: Genomics, RNA, Ribosomal, 5S, food and beverages, General Medicine, Polyploid complex, Genomics, Hordeum murinum L, MESH: Climate, Flow Cytometry, Biological Evolution, Meiosis, Phylogeography, MESH: Phylogeography, MESH: Hordeum, Ploidy, MESH: Algeria, Genome, Plant, Biotechnology, DNA, Plant, Mitosis, MESH: Biological Evolution, DNA, Ribosomal, Chromosomes, Plant, 03 medical and health sciences, Cytogenetics, MESH: In Situ Hybridization, Molecular evolution, Hordeum murinum, Botany, Genetics, [SDV.BV]Life Sciences [q-bio]/Vegetal Biology, MESH: RNA, Ribosomal, 5S, POLYPLOIDIE, MESH: DNA, Plant, Molecular Biology, Gene, MESH: Cytogenetics, 030304 developmental biology, Ploidies, Base Sequence, fungi, Genetic Variation, Hordeum, MESH: Mitosis, biology.organism_classification, MESH: Meiosis, Taxon, RNA, Ribosomal, Algeria, FISH–GISH, MESH: RNA, Ribosomal, 010606 plant biology & botany
Abstract

International audience; Population diversity and evolutionary relationships in the Hordeum murinum L. polyploid complex were explored in contrasted bioclimatic conditions from Algeria. A multidisciplinary approach based on morphological, cytogenetic, and molecular data was conducted on a large population sampling. Distribution of diploids (subsp. glaucum) and tetraploids (subsp. leporinum) revealed a strong correlation with a North-South aridity gradient. Most cytotypes exhibit regular meiosis with variable irregularities in some tetraploid populations. Morphological analyses indicate no differentiation among taxa but high variability correlated with bioclimatic parameters. Two and three different nuclear sequences (gene coding for an unspliced genomic protein kinase domain) were isolated in tetraploid and hexaploid cytotypes, respectively, among which one was identical with that found in the diploid subsp. glaucum. The tetraploids (subsp. leporinum and subsp. murinum) do not exhibit additivity for 5S and 45S rDNA loci comparative with the number observed in the related diploid (subsp. glaucum). The subgenomes in the tetraploid taxa could not be differentiated using genomic in situ hybridization (GISH). Results support an allotetraploid origin for subsp. leporinum and subsp. murinum that derives from the diploid subsp. glaucum and another unidentified diploid parent. The hexaploid (subsp. leporinum) has an allohexaploid origin involving the two genomes present in the allotetraploids and another unidentified third diploid progenitor.

Published
2011
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6. BSMV genome mediated expression of a foreign gene in dicot and monocot plant cells

Author

Joshi, Rajiv L, Joshi, R.L., Joshi, V., Ow, D.W., Centre interdisciplinaire de recherche en biologie (CIRB), Labex MemoLife, École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Collège de France (CdF (institution))-Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris), Université Paris sciences et lettres (PSL)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Plant Gene Expression Center [Berkeley], Department of Plant and Microbial Biology [Berkeley], University of California [Berkeley], University of California-University of California-University of California [Berkeley], University of California-University of California, Institut Jacques Monod (IJM (UMR_7592)), Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), and Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Collège de France (CdF)-PSL Research University (PSL)

Subjects
Genes, Viral, Transcription, Genetic, MESH: Oligonucleotide Probes, [SDV]Life Sciences [q-bio], Genetic Vectors, Molecular Sequence Data, MESH: Plants, RNA-dependent RNA polymerase, MESH: Base Sequence, MESH: Mosaic Viruses, Biology, Transfection, General Biochemistry, Genetics and Molecular Biology, Mosaic Viruses, MESH: Genetic Vectors, DNA-directed RNA interference, Transcription (biology), Gene expression, MESH: Coleoptera, Animals, MESH: Animals, MESH: Cloning, Molecular, Cloning, Molecular, Luciferases, Molecular Biology, Gene, MESH: Genes, Viral, MESH: Molecular Sequence Data, Base Sequence, General Immunology and Microbiology, MESH: Transcription, Genetic, MESH: Transfection, General Neuroscience, RNA, Hordeum, Plants, Non-coding RNA, Molecular biology, Coleoptera, RNA silencing, MESH: RNA, Viral, MESH: Hordeum, RNA, Viral, MESH: Luciferases, Oligonucleotide Probes, Research Article
Abstract

International audience; Barley stripe mosaic virus (BSMV) possesses a tripartite genome composed of RNAs alpha, beta and gamma that have been cloned into in vitro transcription vectors from which infectious transcripts can be obtained. The BSMV genome has been engineered here to serve as an expression vector in plant protoplasts. Open reading frame (ORF) b of RNA beta, encoding a non-structural protein, was replaced by the firefly luciferase (luc) reporter gene to yield RNA beta 2-luc. In the presence of both RNAs alpha and gamma, RNA beta 2-luc mediated efficient expression of the luc gene upon transfection into tobacco and maize protoplasts. This expression ranged from 20- to 123-fold higher than the luciferase activity obtained from transfection with a luc gene mRNA. Replication of RNA beta and its derivatives i.e. 'minus' strand synthesis, was confirmed by Northern analysis, indicating that the high level of luc gene expression using RNA beta 2-luc resulted from RNA amplification. ORFa of RNA beta, encoding the coat protein, was also replaced by the luc gene to yield RNA beta 1-luc. Although transfection of RNA beta 1-luc alone produces luciferase efficiently, neither 'minus' strand synthesis nor further increase of luciferase activity was observed in the presence of RNAs alpha and gamma, or RNAs alpha, beta and gamma, suggesting that the deleted sequences within ORFa are cis-acting for replication of RNA beta. Our results demonstrate that a foreign gene can be expressed by replacement of a viral non-structural protein gene that is essential for virus multiplication in plants, leading to a potential strategy for virus 'containment' with use of 'disarmed' plant viral vectors.

Published
1990
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