Abstract: Virus-based expression vectors carrying sequences corresponding to endogenous host genes trigger silencing through a homology-dependent RNA degradation mechanism. Virus-induced gene silencing (VIGS), is useful as a reverse-genetic tool for use in functional genomic programs for loss-of-function transient assays-based screening. Described herein is an approach to enhance the robustness of the VIGS phenotype by increasing the level of dsRNA molecule production.

Abstract: Disclosed herein are novel methods and materials directed to transforming a host cell and expressing exogenous RNA therein. Specifically disclosed are DNA-launching platforms used to introduce a replicating viral segment attached to an exogenous polynucleotide into a cell, whereby the exogenous polynucleotide is expressed in said cell and confers a detectable trait.

Abstract: The invention provides insect viral vectors useful to transfer genes to plants, insects and other hosts.

Abstract: The invention provides novel methods of using viral replicase polypeptides and polynucleotides. Included are methods for modulating endoreduplication and increasing crop yield.

Abstract: The present invention provides nucleic acids, as well as vectors, cells, and plants (including plant parts, seeds, and embryos) containing the nucleic acids. In particular, molecular tools are provided in the form of nucleic acids that encode reverse transcriptases. The invention also features methods for manipulating such nucleic acids. In addition, the invention features methods to introduce nucleic acids containing retroelements or retroelement sequences into cells.

Abstract: The present invention discloses methods for interfering with expression of the genes in plant cells by using replicating recombinant viral vectors. A host plant is infected at one or more locations with a recombinant viral vector. The vector is both an initiator and a target of the RNA-triggered gene silencing in plant cells. The vector upon infection is capable of directing self-replication and producing a transcription product of a nucleic acid segment. The transcription product interferes with the expression of a specific gene in plant cells.

Abstract: The invention relates to &agr;-galactosidase truncated at the carboxy terminus and the production of enzymatically active recombinant human and animal lysosomal enzymes involving construction and expression of recombinant expression constructs comprising coding sequences of human or animal lysosomal enzymes in a plant expression system. The plant expression system provides for post-translational modification and processing to produce a recombinant gene product exhibiting enzymatic activity. The invention is demonstrated by working examples in which transgenic tobacco plants express recombinant expression constructs comprising human glucocerebrosidase nucleotide sequences. The invention is also demonstrated by working examples in which transfected tobacco plants express recombinant viral expression constructs comprising human &agr; galactosidase nucleotide sequences.

Abstract: In general, the invention features a method for selecting a transgenic grapevine or grapevine component having increased resistance to a fanleaf disease, the method including the steps of: (a) transforming a grape plant cell with a grape nepovirus coat protein nucleic acid molecule or fragment thereof which is capable of being expressed in the plant cell; (b) regenerating a transgenic grapevine or grapevine component from the plant cell; and (c) selecting a transgenic grapevine or grapevine component which expresses, at a low level, the nucleic acid molecule or fragment thereof, wherein the low level expression increases the resistance of the transgenic grapevine or grapevine component to fanleaf disease. The invention also relates to an isolated nucleic acid molecule encoding a coat protein or fragment thereof of a ‘Geneva’ isolate of a grapevine nepovirus virus.

Abstract: Disclosed are methods for silencing a target nucleotide sequence (preferably representing one or more endogenous genes, preferably in a systemic fashion) which is present in a first part of the plant, which method comprises transiently introducing into the cytoplasm of a cell in a second part of the plant, which cell comprises a nucleic acid encoding the target sequence and which is remote from said first part of the plant, a nucleic acid construct.

Abstract: In general, the invention features a method for selecting a transgenic grapevine or grapevine component having increased resistance to a grapevine fanleaf disease. The method involves (a) transforming a grape plant cell with a grapevine fanleaf virus coat protein nucleic acid molecule or fragment thereof which is expressed in the plant cell; (b) regenerating a transgenic grapevine or grapevine component from the plant cell; and (c) selecting a transgenic grapevine or grapevine component expressing a grapevine fanleaf virus coat protein or coat protein fragment thereof, where the coat protein or the coat protein fragment thereof is expressed at a level equal to or less than an optical density of about 0.2 at 405 nm as detected by an enzyme-linked immunoassay as compared to a control plant, the selected transgenic grapevine or grapevine component having resistance to the grapevine fanleaf disease.

Abstract: The invention involves combining a peptide toxin effective against insects, including but not limited to thrips, leaf hoppers, and beetles, with a transport peptide capable of facilitating transfer of the peptide toxin from the gut of an insect to the hemocoel. The combination can be effected by a fusion of genetic material encoding the peptide toxin and the transport peptide, such that expression of the genetic material fusion results in synthesis of a fusion protein combining the functions of both the toxin and the transport protein. Ingestion of the fusion protein by the sucking insect transfers the fusion protein into the insect's gut from which it is transferred into the hemocoel due to the functional activity of the transport peptide where the toxin exerts its toxic effect upon the insect. In a preferred embodiment, the invention is effective in control of such sucking insects as aphids, whiteflies and the like, and other vectors that transmit viruses in a circulative manner.

Abstract: The invention described herein discloses a virus-induced resistance that may be transferred from one plant generation to another in which transgenic plants containing a coding sequence, taken from the read-through portion of the replicase portion of the viral genome, are resistant to subsequent disease by the virus. The use of the 54 kDa coding sequence from TMV is described as a specific example of the broader technology. Thus, the invention defines a means for bringing about viral resistance in plants which have been transformed with nucleic acid copies of fragments or segments taken from the replicase portion of the pathogenic virus genome. In addition, the present invention defines transformed plants and their seeds which carry a portion of the viral genome which encodes for a portion of the read-through portion of the replicase genome of the pathogenic virus.

Abstract: This invention relates to an isolated nucleic acid fragment encoding an oxidosqualene cyclase. The invention also relates to the construction of a chimeric gene encoding all or a portion of the isolated polynucleotides of the invention, in sense or antisense orientation, operably linked to a suitable regulatory sequence.

Abstract: The invention describes compositions and methods of use in which an infectious modified Tobacco Mosaic Virus (TMV) virion comprising a coat protein (CP) or a movement protein (MP) gene is replaced with a nuclear inclusion protease (NIa) expression cassette for the expression of a heterologous peptide in a tobacco mosaic virus (TMV) host plant.

Abstract: The present invention relates to DNA constructs which include DNA molecules which affect papaya fruit ripening and DNA molecules which encode papaya ringspot virus coat protein. The present invention further relates to a method of controlling papaya fruit ripening while conferring resistance to Papaya Ringspot Virus by transforming plants with the DNA construct. The present invention also relates to expression systems, host cells, and transgenic plants containing the DNA constructs of the invention.

Abstract: Transgenic plants with increased resistance to geminivirus infection, and nucleic acid constructs useful in producing such plants, are described. The transgenic plants express a mutant AL3/C3 geminivirus protein, which increases resistance to infection by at least one geminivirus, compared to a non-transformed control plant.

Abstract: The invention presents DNA constructs comprising a promoter operably linked to DNA which can be transcribed in a plant cell to an RNA transcript, wherein the RNA transcript comprises plant virus sequence from an RNA virus which confers on the RNA transcript the ability to replicate in the cytoplasm of the plant cell, wherein the transcript lacks all or part of the viral genome not required for replication in the cytoplasm, and further comprises at least one targeting sequence which is foreign to the plant virus sequence and causes post-transcriptional gene silencing of one or more target genes. The invention also presents methods to use the DNA construct to cause post-transcriptional gene silencing of a target gene in a plant.

Abstract: The invention relates to two plant transgene expression systems. The first is comprised of two chromosomally-integrated components that are individually heritable. One component is an inactive replicon, which contains cis-acting viral sequences required for replication and is unable to replicate episomally. The other component is a chimeric transactivating gene comprising a regulated promoter operably-linked to the coding region for a protein that can transactivate replicon replication. Regulated expression of the transactivation protein in plant cells containing the inactive replicon triggers release of free replicon from the integrated inactive replicon and allows episomal replication.

Abstract: The present invention relates to isolated Raspberry Bushy Dwarf Virus (RBDV) nucleic acid sequences which encode RBDV coat and movement proteins or polypeptides and mutant or modified forms thereof. The invention further relates to heterologous nucleic acid constructs, vectors, transformation methods, plant cells and plants comprising such RBDV-encoding nucleic acids and methods for inducing resistance to RBDV by transforming plants with a nucleic acid construct comprising RBDV protein or polypeptide-encoding nucleic acid sequences.

Abstract: The present invention relates to the isolation and identification of nucleic acid sequences encoding the coat protein of papaya ringspot virus in the Kapoho (KA), Keaau (KE), Thailand (TH), Brazil (BR), Jamaica (JA), Mexico (ME), Venezuela (VE), and Oahu (OA) strains, and the uses thereof to impart viral resistance to papaya plants. The present invention also relates to nucleic acid constructs containing individual or multiple papaya ringspot virus coat protein-encoding nucleic acid sequences, and host cells and transgenic plants and seeds containing such constructs. The present invention is also directed to a method of using such constructs to impart to plants resistance to papaya ringspot virus.

Abstract: A strategy for effecting virus resistance in plants causes the transcription in the plant cells of negative RNA strands which are substantially complementary to a target RNA strand. The target RNA strand can be an mRNA transcript created in gene expression, a viral RNA, or other RNA present in the plant cells. The negative RNA strand is complementary to at least a portion of the target RNA strand to inhibit its activity in vivo.

Abstract: The invention involves recombinant, double-stranded DNA that contains a promoter which functions in plant cells to cause the production of RNA sequences of a plant virus, a DNA sequence that causes the production of an RNA sequence encoding the coat protein of said plant virus, and a 3′ non-translated region which functions in plant cells to cause the addition of polyadenylated nucleotides to the 3′ end of said RNA sequence; which double-stranded DNA can be used in a method for genetically transforming plants to produce genetically transformed plant cells and plants that are resistant to virus infection.

Abstract: Retroviral and retroviral-like polynucleotides, and vectors, proteins, and antibodies derived therefrom, that are useful for the introduction of genetic information into soybeans and other plant species.

Abstract: Antifungal peptides which comprise at least six amino acid residues identical to a run of amino acid residues found between position 21 and position 51 of the Rs-AFP2 antifungal protein sequence or of substantially homologous protein sequences. The peptides are useful for combating fungal diseases in agricultural, pharmaceutical or preservative applications.

Abstract: The present invention relates to a recombinant viral nucleic acid selected from a (+) sense, single stranded RNA virus possessing a native subgenomic promoter encoding for a first viral subgenomic promoter, a nucleic acid sequence that codes for a viral coat protein whose transcription is regulated by the first viral subgenomic promoter, a second viral subgenomic promoter and a second nucleic acid sequence whose transcription is regulated by the second viral subgenomic promoter. The first and second viral subgenomic promoters of the recombinant viral nucleic acid do not have homologous sequences relative to each other. The recombinant viral nucleic acid provides the particular adivantage that it systemically transcribes the second nucleic acid in the host. Host organisms encompassed by the present invention include procaryotes and eucaryotes, particularly animals and plants.

Abstract: The invention provides improved methods and means for introducing inhibitory RNA into plant cells. The invention also provides kits comprising viral RNA vectors derived from satellite viruses and corresponding helper viruses for the introduction of inhibitory RNA into plant cells and plants. Further, the invention comprises methods for obtaining an enhanced or improved gene-silencing phenotype.

Abstract: The invention provides methods of suppressing gene silencing and/or stabilizing expression of a coding sequence in a cell comprising expressing a 126 kDa protein and/or the 183 kDa protein of a subgroup sindbis plant virus in the cell. The invention can thus be used to avoid the deleterious effects of gene silencing in a cell. Also provided by the invention are methods of delivering a polypeptide of interest to a limited part of a plant.

Abstract: The invention relates to &agr;-galactosidase truncated at the carboxy terminus and the production of enzymatically active recombinant human and animal lysosomal enzymes involving construction and expression of recombinant expression constructs comprising coding sequences of human or animal lysosomal enzymes in a plant expression system. The plant expression system provides for post-translational modification and processing to produce a recombinant gene product exhibiting enzymatic activity. The invention is demonstrated by working examples in which transgenic tobacco plants express recombinant expression constructs comprising human glucocerebrosidase nucleotide sequences. The invention is also demonstrated by working examples in which transfected tobacco plants express recombinant viral expression constructs comprising human &agr; galactosidase nucleotide sequences.

Abstract: The present invention focuses on the super-expression of foreign genes in transgenic cells by combining within a single cDNA construct and respective RNA transcript, several trans- and cis-acting genetic elements of viral origin which act in concert.

Abstract: This invention is directed to a plus strand RNA viral vector for transformation of a host organism with a foreign RNA, and expression of said foreign RNA. The foreign RNA is inserted into an infective RNA viral segment containing cis-acting viral replication elements, and allowed to infect the host organism. The RNA vector is modified to obtain infectivity by including an intervening sequence between the cap and the 5′ end. The modified RNA is able to tolerate the exogenous RNA segment without disrupting the replication of the modified RNA, in the absence of a trans-acting viral replication element in a single component plant virus host cell.

Abstract: The invention provides novel methods of using viral replicase polypeptides. Included are methods for increasing transformation frequencies, providing a positive growth advantage, and modulating cell division.

Abstract: A gene amplification system based on plant viral genetic elements dramatically increases foreign protein production in plants. A safer and more economical production system for vaccines and antibodies in recombinant plants grown using agricultural practice is described. The high-level expression system uses the replicative process of a plant mastrevirus, exemplified by bean yellow dwarf virus (BeYDV). The expression system is preferably inducible to avoid interference with plant growth and development. Developmental cues, such as fruit ripening, are employed to trigger expression of the foreign protein using a tissue-specific promoter. A single, stably integrated expression cassette for foreign protein is replicated extrachromosomally in ripening fruit, forming hundreds of transcriptionally competent copies. Preferred plant hosts include tomato as a model system and soybean for production of large quantities of protein at high total protein levels.

Abstract: The invention provides a method for imparting resistance in animals to viruses, and in plants to viruses and viroids, that express double-stranded RNA-like structures (dsRNAs). This method enables the binding of pathogenic dsRNAs during the infection process by expression of dsRNA-binding protein in transgenic animal and plant hosts, thus interrupting the infection cycle and inhibiting disease. The presence of a dsRNA-binding protein in a transgenic host renders the transgenic host resistant to the phenotypic symptoms of viral infection and/or decreased pathogen replication. Accordingly, the present invention provides a genetically engineered animal and plant, stably transformed to express a dsRNA-binding protein, such that the transgenic host displays resistance to virus and/or viroid challenge.

Abstract: The present invention relates to isolated Raspberry Bushy Dwarf Virus (RBDV) nucleic acid sequences which encode RBDV coat and movement proteins or polypeptides and mutant or modified forms thereof. The invention further relates to heterologous nucleic acid constructs, vectors, transformation methods, plant cells and plants comprising such RBDV-encoding nucleic acids and methods for inducing resistance to RBDV by transforming plants with a nucleic acid construct comprising RBDV protein or polypeptide-encoding nucleic acid sequences.

Abstract: The present invention relates to foreign peptide sequences fused to recombinant plant viral structural proteins and a method of their production. Fusion proteins are economically synthesized in plants at high levels by biologically contained tobamoviruses. The fusion proteins of the invention have are useful as antigens for inducing the production of antibodies having desired binding properties, e.g., protective antibodies, or for use as vaccine antigens for the induction of protective immunity against the parvovirus. Feline parvovirus epitopes were fused to the N-terminus of the TMV coat protein, expressed in Nicotiana plants, extracted, purified, characterized and administered to animals, resulting in protective immunity.

Abstract: A process is provided of introducing an RNA into a living cell to inhibit gene expression of a target gene in that cell. The process may be practiced ex vivo or in vivo. The RNA has a region with double-stranded structure. Inhibition is sequence-specific in that the nucleotide sequences of the duplex region of the RNA and of a portion of the target gene are identical. The present invention is distinguished from prior art interference in gene expression by antisense or triple-strand methods.

Abstract: Impatiens is a major ornamental bedding and potted plant, and is an important component of the U.S. floral industry. Susceptibility to insect pests and diseases caused by pathogens remains a problem for Impatiens production, even under greenhouse conditions. While chemical treatment can control certain insect pests and disease pathogens, such treatment can also have an adverse effect upon Impatiens. The methods described herein provide a means to genetically engineer transgenic Impatiens that express macromolecules capable of protecting the plant against the insects and pathogens. The production of transgenic plants can also be used to enhance the commercial value of Impatiens by controlling or enhancing native Impatiens characteristics.

Abstract: The present invention relates to an isolated small RNA virus capable of infecting insect species including Heliothis species, and to the nucleotide sequences and proteins encoded thereby. The invention contemplates uses of the virus in controlling insect attack in plants.

Abstract: The present invention provides a method of compiling a plant functional gene profile, a method of changing the phenotype or biochemistry of a plant, a method of determining a change in phenotype or biochemistry of a plant, and a method of determining the presence of a trait in plant. The methods comprise expressing transiently a nucleic acid sequence of a plant into a host plant to affect phenotypic or biochemical changes in the host plant. A viral vector functional genomic screen has been developed to identify nucleotide sequences in transfected plants by systemically knocking out endogenous gene expression in an antisense mechanism. Once the presence of a trait in a plant is identified by phenotypic or biochemical changes in the host plant, the nucleic acid insert in the cDNA clone or in the vector that results in the changes is then sequenced. The present invention exemplifies that genes encoding GTP binding proteins in one plant can silence endogenous gene expression in a different plant.

Abstract: Chimeric expression promoter comprising at least one nucleic acid sequence, derived from a first plant promoter comprising a plant vascular expression promoter region, said plant vascular expression promoter region being replaced with a nucleic acid sequence derived from a second plant promoter and comprising a plant green tissue expression promoter region.

Abstract: The invention relates to the production of gonadotrophins and corresponding receptors in transgenic plants. The invention provides a method to produce a gonadotrophin or its corresponding receptor in a transgenic plant with modified glycosylation machinery, in order to allow for mammalian type of glycosidic side chains of gonadotrophin and its corresponding receptor.

Abstract: The invention provides improved plant transformation methods. In particular the method provides increased transformation freequency, especially in recalcitrant plants. The method comprises stably transforming a target cell with at least one polynucleotide of interest. The target cell has been previously transformed to stimulate growth of the cell and has gone through at least one cell division.

Abstract: The invention describes compositions and methods of use in which an infectious modified Tobacco Mosaic Virus (TMV) virion comprising a coat protein (CP) or a movement protein (MP) gene is replaced with a nuclear inclusion protease (NIa) expression cassette for the expression of a heterologous peptide in a tobacco mosaic virus (TMV) host plant.

Abstract: The present invention provides promoters for driving the expression of heterologous polynucleotide sequences in plant cells. The promoters are derived from strawberry vein banding virus (SVBV), and include variants, derivatives, fragments, and modified versions of the naturally occurring promoter sequences. The present invention also provides expression cassettes comprising the promoters, as well as methods of using the promoters to drive gene expression in plant cells and transgenic plants.

Abstract: The invention relates to genes encoding polyhydroxyalkanoate synthases. Compositions and methods for producing polyhydroxyalkanoate are provided. Such compositions and methods find use in producing biodegradable thermoplastics in host cells and transgenic plants. Isolated nucleotide molecules, isolated polypeptides, expression cassettes and genetically manipulated host cells, plants, plant tissues, plant cells and seeds are also provided.

Abstract: The introduction of DNA episomes into plant cells to reduce or prevent the expression of endogenous plant genes is described. Cabbage leaf curl virus vectors to provide silencing, preferably systemic silencing, of endogenous plant genes in a treated plant are described.

Abstract: A novel method of over expressing genes in plants is provided. This method is based on the RNA amplification properties of plus strand RNA viruses of plants. A chimeric multicistronic gene is constructed containing a plant promoter, viral replication origins, a viral movement protein gene, and one or more foreign genes under control of viral subgenomic promoters. Plants containing one or more of these recombinant RNA transcripts are inoculated with helper virus. In the presence of helper virus recombinant transcripts are replicated producing high levels of foreign gene RNA.

Abstract: The present invention features a multiple component RNA vector system, which consists of RNA virus-derived RNA replicons and helper viruses. The present invention further features a method for producing foreign RNAs, effector RNAs, proteins or peptides in plants using the multiple component RNA vector system. Moreover, the present invention provides a method for stable and systemic production of foreign RNAs, effector RNAs, proteins and peptides using the multiple component RNA vector system.

Abstract: The invention provides novel methods of using viral replicase polypeptides and polynucleotides. Included are methods for increasing transformation frequencies, increasing crop yield, providing a positive growth advantage, modulating cell division, transiently modulating cell division, and for providing a means of positive selection.

Abstract: This invention provides methods for identifying plant cells that exhibit post-transcriptional gene silencing (PTGS) of a chosen gene. The methods involve the use of suppression-sensitive reporter genes that, when introduced into plant cells, are expressed at a lower level in cells that exhibit PTGS than in cells that are not silenced for the particular gene.