Chemistry: molecular biology and microbiology – Plant cell or cell line – per se ; composition thereof;... – Plant cell or cell line – per se – is pest or herbicide...
Patent
1996-08-09
2000-04-25
Eisenschenk, Chris
Chemistry: molecular biology and microbiology
Plant cell or cell line, per se ; composition thereof;...
Plant cell or cell line, per se, is pest or herbicide...
435419, 4353201, 435183, 530370, 530371, 536 232, 536 237, 536 241, 800278, 800279, 800284, 800287, 800288, 800301, A01H 100, C12N 514, C07H 2104, C07K 14370
Patent
active
060543183
DESCRIPTION:
BRIEF SUMMARY
FIELD OF THE INVENTION.
This invention relates to the production of the transgenic organisms (including for example, plants, microbes and viruses) that express a glucose oxidase gene whose gene product is toxic to economically important pests and diseases of crops. Transgenic plants in accordance with this invention can be sold as improved varieties, whilst transgenic microbes and viruses can be used as biopesticides or as seed coatings or inoculums for soil incorporation.
BACKGROUND OF THE INVENTION.
Production of most crop species is limited by the ravages of pests and diseases. Considerable expense is involved in the protection of crops from these organisms and many current conventional breeding programs are directed at increasing host plant resistance to a variety of invertebrate pests and fungal or viral diseases. Traditional sources of host plant resistance are limited to the same species or species closely related to the crop, but with the advent of genetic engineering novel sources of resistance outside the crop genera are being accessed. Two classic examples are the insecticidal protein genes from Bacillus thuringiensis active against a multitude of insect species (Perlak et al., 1990) and the viral coat protein genes that confer tolerance to a variety of related viral diseases (Powell et al., 1986). Transgenic plants expressing these genes have enormous potential markets once the regulatory hurdles are overcome for their large scale release into agriculture.
New sources of host plant resistance are being sought from a variety of sources, and work leading to the present invention has led to the isolation and characterisation of a glucose oxidase gene from the fungus Talaromyces flavus that has potential for the control of both fungal diseases and arthropod pests when expressed in transgenic crop plants. The particular examples investigated to date are for the control of the cotton pathogen, Verticillium dahliae, the causal agent of verticillium wilt disease, and for the control of Helicoverpa spp. which are economically important Lepidopteran pests of cotton and most summer crops. Use of the glucose oxidase gene for the control of other fungal diseases or pests such as nematodes, mites, aphids, whiteflies, jassids or mirids which are susceptible to hydrogen peroxide produced by glucose oxidase activity is also possible.
Verticillium wilt is a wide-spread disease which affects many different plant species. It is caused by the soil fungus Verticillium dahliae (Leb.), an imperfect fungus first isolated from diseased dahlias in 1913 (Muller, 1928). Isolates of the species vary widely in both morphology and pathogenicity but all produce small, hard black structures called microsclerotia. These structures are composed of melanized cells which store many nutrients and are the means by which V. dahliae survives in the soil.
V. dahliae does not grow saprophytically through the soil, but conidia and microsclerotia germinate in soil if root exudate from desirable plant species are present. The mycelium then invades the plant, entering through the cap of the root region of elongation, root hairs or lower hypercotyl region. In all cases both intercellular and intracellular invasion occurs. In "susceptible" hosts, the fungus successfully penetrates the vascular system of the plant. Here hyphal multiplication occurs and conidiospores are generated which then travel upwards through the xylem thus spreading the fungus rapidly through the vascular system of the plant.
Once spread throughout the vascular system the main effect of the pathogen is to disrupt the passage of water through the stem. This may be attributed to the physical presence of the mycelium of the pathogen, the development of tyloses that block the xylem or to gummosis within the vessels. Further xylem occlusion is caused by high molecular weight polysaccharides produced by the pathogen or cleaved from the plant walls by hydrolytic enzymes. Together this can result in a 40 to 60-fold increase in the resistance of the stem to water flow; thus the
REFERENCES:
Baetselier et al. J Biotech, 1992, vol. 24, pp. 141-148.
Whittington et al. Curr Genet, 1990, vol. 18, pp. 531-536.
Kim et al., Phytopathology, 78: 488-492, 1988 (Exhibit 1).
Cornelissen et al., Plant Physiology, 101: 709-712, 1993 (Exhibit 2).
Peng et al., Phytopathology, 82: 696-699, 1992 (Exhibit 3).
Chen et al., Science, 262, 1883-1886, 1983 (Exhibit 4).
Hain et a;., Nature, 361: 153-156, Jan. 14, 1993 (Exhibit 5).
Huang et al., Scientia Agricultura Sinica, 3: 32-36, 1986 (Exhibit 9).
Perlak et al., 8: 939-943, Oct. 10, 1990 (Exhibit 11).
Wu et al., The Plant Cell 7:1357-1368 (1995) (Exhibit 2).
Hodgkins, Martin et al., "Expression of the Glucose Oxidase Gene from Aspergillus niger in Hansenula polymorpha and its use as a Reporter Gene to Isolate Regulatory Mutations," Yeast, 9: 625-635 (1993).
Kim, K. K. et al., "Production, purification and properties of glucose oxidase from the biocontrol fungus Talaromyces flavus", Can J. Microbiol, 36: 199-205 (1990); and.
Kim, K. K. et al., "Glucose oxidase as the antifungal principle of talaron from Talaromyces flavus", Can J. Microbiol, 36: 760-764 (1990).
Dennis Elizabeth Salisbury
Llewellyn Danny James
Murray Fiona Ruth
Peacock William James
Bui Phuong T.
Commonwealth Scientific and Industrial Research Organization
Eisenschenk Chris
White John P.
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