Nitric oxide as an activator of the plant pathogen defense...

Multicellular living organisms and unmodified parts thereof and – Method of introducing a polynucleotide molecule into or...

Reexamination Certificate

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C435S410000, C435S419000, C800S278000, C800S286000, C800S290000, C800S295000, C800S279000, C800S300100, C800S320100

Reexamination Certificate

active

06198020

ABSTRACT:

FIELD OF THE INVENTION
The invention relates to the genetic manipulation of plants, particularly to transforming plants with genes that enhance disease resistance.
BACKGROUND OF THE INVENTION
Disease in plants is caused by biotic and abiotic causes. Biotic causes include fungi, viruses, bacteria, and nematodes. Of these, fungi are the most frequent causative agent of disease on plants. Abiotic causes of disease in plants include extremes of temperature, water, oxygen, soil pH, plus nutrient-element deficiencies and imbalances, excess heavy metals, and air pollution.
A host of cellular processes enables plants to defend themselves from disease caused by pathogenic agents. These processes apparently form an integrated set of resistance mechanisms that is activated by initial infection and then limits further spread of the invading pathogenic microorganism.
Subsequent to recognition of a potentially pathogenic microbe, plants can activate an array of biochemical responses. Generally, the plant responds by inducing several local responses in the cells immediately surrounding the infection site. The most common resistance response observed in both nonhost and race-specific interactions is termed the “hypersensitive response” (HR). In the hypersensitive response, cells contacted by the pathogen, and often neighboring cells, rapidly collapse and dry in a necrotic fleck. Other responses include the deposition of callose, the physical thickening of cell walls by lignification, and the synthesis of various antibiotic small molecules and proteins. Genetic factors in both the host and the pathogen determine the specificity of these local responses, which can be very effective in limiting the spread of infection.
As noted, among the causative agents of infectious disease of crop plants, the phytopathogenic fungi play the dominant role. Phytopathogenic fungi cause devastating epidemics, as well as causing significant annual crop yield losses. All of the approximately 300,000 species of flowering plants are attacked by pathogenic fungi. However, a single plant species can be host to only a few fungal species, and similarly, most fungi usually have a limited host range.
Plant disease outbreaks have resulted in catastrophic crop failures that have triggered famines and caused major social change. Generally, the best strategy for plant disease control is to use resistant cultivars selected or developed by plant breeders for this purpose. However, the potential for serious crop disease epidemics persists today, as evidenced by outbreaks of the Victoria blight of oats and southern corn leaf blight. Accordingly, molecular methods are needed to supplement traditional breeding methods to protect plants from pathogen attack.
SUMMARY OF THE INVENTION
Compositions and methods to aid in protecting a plant from invading pathogenic organisms are provided. The compositions of the invention comprise genes that influence the levels of nitric oxide in plant cells and regulate metal ion transport. Such genes include those encoding nitric oxide synthase (iNOS) as well as natural resistance-associated macrophage proteins (NRAMP) and NRAMP homologues. The compositions and methods of the invention can be used for enhancing resistance to plant pests. The method involves stably transforming a plant with a nucleotide sequence capable of altering the levels of nitric oxide in the plant cell, preferably increasing nitric oxide. The nucleotide sequences will be expressed from a promoter capable of driving expression of a gene in a plant cell. It is recognized that a variety of promoters will be useful in the invention the choice of which will depend in part upon the desired level of expression of the disclosed genes. It is recognized that the levels of expression can be controlled to provide a low level of nitric oxide in the plant to prevent pathogen invasion resulting in levels of immunity in the plant that impart pathogen resistance. Alternatively, the promoters can provide expression at levels to induce cell death. In these latter instances, it is recognized that pathogen inducible promoters will be utilized.
The methods of the invention find use in controlling plant pests, including fungal pathogens, viruses, nematodes, insects, and the like. Transformed plants and seeds, as well as methods for making such plants and seeds are additionally provided.


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