Multicellular living organisms and unmodified parts thereof and – Plant – seedling – plant seed – or plant part – per se – Higher plant – seedling – plant seed – or plant part
Reexamination Certificate
1998-06-15
2001-05-01
Fox, David T. (Department: 1638)
Multicellular living organisms and unmodified parts thereof and
Plant, seedling, plant seed, or plant part, per se
Higher plant, seedling, plant seed, or plant part
C800S295000, C800S312000, C800S313000, C800S317000, C800S317100, C800S298000, C800S317200, C800S317300, C800S317400, C800S279000, C536S023100, C536S023600, C435S410000, C435S418000, C435S419000, C435S320100, C435S200000, C530S378000
Reexamination Certificate
active
06225531
ABSTRACT:
BACKGROUND OF THE INVENTION
The present invention relates to a glucan elicitor receptor, DNA molecules coding for the glucan elicitor receptor, vectors containing the DNA molecules, plant cells transformed with the DNA molecules, fungus-resistant plants transformed with the DNA molecules and a method for creating such plants. More specifically, the present invention relates to a glucan elicitor receptor derived from a soybean root plasma membrane fraction, DNA molecules coding for the glucan elicitor receptor, vectors containing the DNA molecules, plant cells transformed with the DNA molecules, fungus-resistant plants transformed with the DNA molecules and a method for creating such plants.
It is known that plants synthesize and accumulate an antibiotic agent called phytoalexin in response to infection with pathogens (M. Yoshikawa (1978) Nature 257: 546). Some plant pathogens were found to have the substances that induce them to perform such a resistance reaction (N. T. Keen (1975) Science 187: 74), which are called “elicitors”. The biochemical process from the infection of plants with pathogens to the synthesis and accumulation of phytoalexin is believed to be as follows:
When the mycelium of a pathogen invades a plant cell, glucanase in the plant cell works so as to cleave polysaccharides on the surface of the pathogen mycelial wall, thereby liberating an elicitor. If the elicitor binds to a receptor in the plant cell, a second messenger which plays a role in signal transduction is produced. The signal transduction substance is incorporated in the nucleus of the plant cell and activates the transcription of the genes coding for phytoalexin synthesize enzymes to induce a phytoalexin synthesis. At the same time, the phytoalexin degradation is inhibited. As a result, phytoalexin is efficiently accumulated in the plant cell.
A phytoalexin playing an important role in the resistance of soybean is called glyceollin and its structure has been determined (M. Yoshikawa et al. (1978) Physiol. Plant. Pathol. 12: 73). Elicitor of soybean has a characteristic structure; it has &bgr;-1,6 linked glucan of various lengths as a principal chain from which &bgr;-1,3 linked glucan side chains are branched [J. K. Sharp et al. (1984) J. Biol. Chem. 259: 11321; M. Yoshikawa (1990) Plant Cell Technology 1.2: 695]. A receptor specific for a glucan elicitor derived from a soybean pathogenic mold fungus
Phytophthora megasperma
f. sp.
glycinea
is believed to be a protein which plays an important role in the synthesis and accumulation of the antibiotic agent glyceollin. A method for the purification of the glucan elicitor receptor specific to this elicitor has been disclosed (E. G. Cosio et al., (1990) FEBS 264: 235, E. G. Cosio et al. (1992) Eur. J. Biochem. 204: 1115, T. Frey et al. (1993) Phytochemistry 32; 543). However, the amino acid sequence of a glucan elicitor receptor has not been determined and the gene coding therefor is not yet known. If a gene coding for glucan elicitor receptor is found, it will be possible to create plants having resistance to pathogenic fungi by incorporating the gene into a chromosome of plants and expressing the glucan elicitor receptor in the plants. Thus, improvement of the productivity of agricultural products can be expected.
An object of the present invention is to provide a glucan elictor receptor.
Another object of the present invention is to provide a DNA molecule coding for a glucan elicitor receptor.
A further object of the present invention is to provide a vector containing a DNA molecule coding for a glucan elicitor receptor.
A still further object of the present invention is to provide a plant cell transformed with a DNA molecule coding for a glucan elicitor receptor.
It is an object of the present invention to provide a plant transformed with a DNA molecule coding for a glucan elicitor receptor.
It is another object of the present invention to provide a method for creating a plant transformed with a DNA molecule coding for a glucan elicitor receptor.
SUMMARY OF THE INVENTION
As a result of intensive and extensive researches toward the resolution of the above assignment, the present inventors have succeeded in purifying a soybean root-derived glucan elicitor receptor, cloning a glucan elicitor receptor gene from a soybean cDNA library, transferring this gene into a tobacco plant and expressing it in the plant. Thus, the present invention has been achieved. The present invention provides a glucan elicitor receptor having an amino acid sequence as substantially shown in SEQ ID NO:1. The present invention also provides DNA molecules containing nucleotide sequences coding for a glucan elicitor receptor having an amino acid sequence as substantially shown in SEQ ID NO:1, and fragments thereof. The present invention further provides DNA molecules containing nucleotide sequences coding for a glucan elicitor receptor which are incorporated in plasmid pER23-1, and fragments thereof. The present invention still further provides vectors containing DNA molecules coding for a glucan elicitor receptor and plant cells transformed with DNA molecules coding for a glucan elicitor receptor. Moreover, the present invention provides a method for creating a plant having resistance to pathogenic fungi, comprising incorporating a DNA sequence coding for a glucan elicitor receptor into a chromosome of a plant and expressing the gene in the plant. The present invention also provides a plant having resistance to pathogenic fungi, characterized in that a DNA sequence coding for a glucan elicitor receptor has been transferred into the plant and the gene is expressed in it.
The glucan elicitor receptor of the present invention is useful in the elucidation of resistance to fungi and the development of elicitor derivatives capable of inducing fungal resistance, and it can be used as an antigen for the production of antibodies against glucan elicitor receptors.
The DNA molecules of the present invention which contain nucleotide sequences coding for a glucan elicitor receptor and fragments thereof are useful as materials for establishing techniques for developing fungi-resistant plants. In other words, the DNA molecules of the present invention and fragments thereof may be introduced and expressed in various plants to enhance their fungal resistance.
Antibodies against the glucan elicitor receptor of the present invention, the DNA molecules of the present invention which contain nucleotide sequences coding for a glucan elicitor receptor, their mutants and anti-sense DNAs can be used in the studies of the elicitor-binding site of glucan elicitor receptor and signal transduction.
Further, the information on the amino acid sequence of glucan elicitor receptor and the nucleotide sequence coding therefor can be used in the studies of the elicitor-binding site of glucan elicitor receptor and the signal transduction in which glucan elicitor receptor is involved.
The plant of the present invention has high resistance to fungi.
REFERENCES:
Linthorst et al. Plant Cell. 1989. vol. 1:285-291.*
Carvalho et al. The EMBO J. 1992. vol. 11:2595-2602.*
Takeuchi et al. Plant Physiol. 1990. vol. 93:673-682.*
Spencer et al. Plant Mol Biology. 1992. vol. 18:201-210.*
Yoshikawa et al. Naturwissenschaften. 1993. vol. 80: 417-420. [ref.1].*
Yoshikawa et al. Physiol. Mol. Plant. Path. 1990. vol. 37:367-376. (ref. 2).*
Patent Abstracts of Japan, 6-321995, Nov. 22, 1994.
Frey et al., “Affinity Purification and Characterization of a Binding Protein for a Hepta-&bgr;-Glucoside Phytoalexin Elicitor in Soybean”, Phytochemistry, vol. 32, No. 3, 1993, pp. 543-550.
Schmidt et al., “Specific Binding of a Fungal Glucan Phytoalexin Elicitor to Membrane Fractions from SoybeanGlycine max”, Proc. Natl. Acad. Sci. USA, vol. 84, Jun. 1987, pp. 4117-4121.
Keen, “Gene-for-Gene Complementarity in Plant-Pathogen Interactions”, Annu. Reg. Genet., vol. 24, 1990, pp. 447-463.
Keen et al., &bgr;-1,3-Endoglucanase from Soybean Releases Elicitor-Active Carbohydrates from Fungus Cell Plant Physiol., vol. 71, 1983,
Ishida Isao
Iwamatsu Akihiro
Kakitani Makoto
Umemoto Naoyuki
Yamaoka Naoto
Foley & Lardner
Fox David T.
Kirin Beer Kabushiki Kaisha
Yoshikawa Masahi
Zagmout Ousama
LandOfFree
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