Transgenic plants exhibiting heterologous virus resistance

Multicellular living organisms and unmodified parts thereof and – Method of introducing a polynucleotide molecule into or... – The polynucleotide confers pathogen or pest resistance

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435419, 435468, 800301, 800308, C12N 504, C12N 1533, C12N 1582, A01H 100

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060159421

DESCRIPTION:

BRIEF SUMMARY
FIELD OF THE INVENTION

This invention relates to a coat protein gene derived from cucumber mosaic virus strain C. More specifically, the invention relates to the genetic engineering of plants and to a method for conferring viral resistance to a plant using an expression cassette encoding cucumber mosaic virus strain C coat protein.


BACKGROUND OF THE INVENTION

Many agriculturally important crops are susceptible to infection by plant viruses, particularly cucumber mosaic virus, which can seriously damage a crop, reduce its economic value to the grower, and increase its cost to the consumer. Attempts to control or prevent infection of a crop by a plant virus such as cucumber mosaic virus have been made, yet viral pathogens continue to be a significant problem in agriculture.
Scientists have recently developed means to produce virus resistant plants using genetic engineering techniques. Such an approach is advantageous in that the genetic material which provides the protection is incorporated into the genome of the plant itself and can be passed on to its progeny. A host plant is resistant if it possesses the ability to suppress or retard the multiplication of a virus, or the development of pathogenic symptoms. "Resistant" is the opposite of "susceptible," and may be divided into: (1) high, (2) moderate, or (3) low resistance, depending upon its effectiveness. Essentially, a resistant plant shows reduced or no symptom expression, and virus multiplication within it is reduced or negligible. Several different types of host resistance to viruses are recognized. The host may be resistant to: (1) establishment of infection, (2) virus multiplication, or (3) viral movement.
Cucumber mosaic virus (CMV) is a single-stranded (+) RNA plant virus that has a functionally divided genome. The virus genome contains four RNA species designated RNAs 1-4. RNAs 3 and 4 encode the coat protein which is a protein that surrounds the viral RNA and protects the viral RNA from being degraded. Only RNAs 1-3 are required for infectivity because the coat protein, which is encoded by RNA 4, is also encoded by RNA 3.
Several strains of cucumber mosaic virus have been classified using serology, host range, peptide mapping, nucleic acid hybridization, and sequencing analyses. These CMV strains can be divided into two groups, which are designated "WT" (also known as subgroup I) and "S" (also known as subgroup II). The S group consists of at least three members. The WT group is known to contain at least 17 members.
Expression of the coat protein genes from tobacco mosaic virus, alfalfa mosaic virus, cucumber mosaic virus, and potato virus X, among others, in transgenic plants has resulted in plants which are resistant to infection by the respective virus. Heterologous protection can also occur. For example, the expression of coat protein genes from watermelon mosaic virus-2 or zucchini yellow mosaic virus in transgenic tobacco plants has been shown to confer protection against six other potyviruses: bean yellow mosaic virus, potato virus Y, pea mosaic virus, clover yellow vein virus, pepper mottle virus, and tobacco etch virus. However, expression of a preselected coat protein gene does not reliably confer heterologous protection to a plant. For example, transgenic squash plants containing the CMV C coat protein gene, a subgroup I virus, which have been shown to be resistant to the CMV C strain are not protected to the same degree against several highly virulent strains of CMV: CMV V27, CMV V33, and CMV V34 which are all subgroup I viruses.
Thus, a need exists for transgenic plants containing a single viral gene which confers resistance to a plurality of viruses.


SUMMARY OF THE INVENTION

The present invention provides a method of providing heterologous virus resistance to a plant susceptible to infection by two or more viruses by expressing a chimeric recombinant DNA molecule in the cells of the plant which encodes a protein of one class of plant virus, such as a potyvirus protein or cucumovirus protein, i.e., a coat protein or replicase

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