Development of resistance to raspberry bushy dwarf virus

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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C435S320100, C435S419000, C435S468000, C536S023720, C800S301000

Reexamination Certificate

active

06548742

ABSTRACT:

FIELD OF THE INVENTION
The present invention relates to modified raspberry bushy dwarf virus (RBDV) proteins and the nucleic acid sequences which encode them. The invention also relates to RBDV resistant transgenic plants comprising nucleic acid sequences which encode modified RBDV proteins and methods of inducing resistance to RBDV in raspberry plants by transforming them with plant expression vectors comprising such nucleic acid sequences.
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BACKGROUND OF THE INVENTION
Raspberry production in a number of geographic locations, including the Pacific Northwest, Oregon and Washington in the USA and southwestern British Columbia in Canada, has increased dramatically during the past decade. The increase in production has been the result of a move to more mechanical harvesting of raspberries resulting in reduced labor costs, a change in cultivars and an increase in acreage. Prior to 1980, the cultivar, Willamette was the most widely planted raspberry cultivar in this region. Since the early 1980's, the cultivar Meeker has become the cultivar of choice for most growers because of yield, resistance to root rot and the high quality of the berries for the valuable whole frozen and fresh berry markets. Hence, cultivar Meeker has become a suitable replacement for cultfivar Willamette as a processing berry.
The change in cultivars has resulted in the potential for an increased incidence of raspberry bushy dwarf virus (RBDV) since the cultivar Willamette is immune to the type strain of the virus while the cultivar Meeker is susceptible (Daubeny et al., 1982). In 1995, growers in Whatcom Co. (Fraser Valley in Washington) observed increased crumbly fruit in cultivar Meeker plantings, and initial tests showed these plantings had a high incidence of RBDV infection. Such crumbly fruit is not useful for the whole frozen or fresh berry markets. Surveys conducted during 1996-1998 showed that RBDV occurred in most raspberry fields of susceptible cultivars by the second fruiting season in the Fraser Valley (northwest Washington and southwest British Columbia). Also, the infection rate in these fields approached 100% by the fifth cropping season (Martin, 1998).
It is therefore of interest to develop a means to manage RBDV infection in raspberry (
Rubus idaeus
) cultivars such as Meeker that are susceptible to the virus.
SUMMARY OF THE INVENTION
The invention provides RBDV coat and movement protein or polypeptide nucleic acid and amino acid sequences and modified or mutant forms thereof for use in the development of RBDV resistant transgenic plants.
In one aspect, the invention provides an isolated raspberry bushy dwarf virus (RBDV) protein or polypeptide selected from the group consisting of:
(a) a RBDV coat protein or polypeptide with an amino acid sequence having at least 80%, 90% or 95% sequence identity to the sequence presented as SEQ ID NO:2;
(b) a RBDV coat protein or polypeptide with the amino acid sequence presented as SEQ ID NO:2.
(c) a RBDV movement protein or polypeptide with an amino acid sequence having at least 80%, 90% or 95% sequence identity to the sequence presented as SEQ ID NO:4; and
(d) a RBDV movement protein or polypeptide with the amino acid sequence presented as SEQ ID NO:4.
In a related aspect, the invention provides an isolated RBDV polynucleotide selected from the group consisting of:
(a) a polynucleotide with a nucleic acid sequence which encodes or is complementary to a sequence which encodes a RBDV coat protein or polypeptide having at least 80%, 90% or 95% sequence identity to the sequence presented as SEQ ID NO:2;
(b) a polynucleotide with a nucleic acid sequence which encodes or is complementary to a sequence which encodes the RBDV coat protein or polypeptide having the sequence presented as SEQ ID NO:2;
(c) a polynucleotide sequence with the nucleic acid sequence presented as SEQ ID NO:1, which encodes or is complementary to a sequence which encodes a RBDV coat protein or polypeptide;
(d) a polynucleotide with a nucleic acid sequence that hybridizes, under high stringency conditions to the RBDV coat protein-encoding nucleic acid sequence presented as SEQ ID NO:1, or the complement or a fragment thereof;
(e) a polynucleotide comprising a nucleic acid sequence which encodes or is complementary to a sequence which encodes a RBDV movement protein or polypeptide having at least 80%, 90% or 95% sequence identity to the sequence presented as SEQ ID NO:4;
(f) a polynucleotide comprising the nucleic acid sequence which encodes or is complementary to a sequence which encodes a RBDV movement protein or polypeptide having the sequence presented as SEQ ID NO:4;
(g) a polynucleotide sequence with the nucleic acid sequence presented as SEQ ID NO:3, which encodes or is complementary to a sequence which encodes a RBDV movement protein or polypeptide; and
(h) a polynucleotide with a nucleic acid sequence that hybridizes, under high stringency conditions to the RBDV movement protein-encoding nucleic acid sequence presented as SEQ ID NO:3, or the complement or a fragment thereof.
In another aspect, the invention provides an isolated polynucleotide which encodes a raspberry bushy dwarf virus (RBDV) coat or movement protein or pol

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