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
2000-01-12
2002-01-08
McElwain, Elizabeth F. (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
C435S320100, C435S419000, C536S023500, C800S279000, C800S288000
Reexamination Certificate
active
06337432
ABSTRACT:
BACKGROUND OF THE INVENTION
Parasites of plant-eating insects are natural pesticides. One such solitary parasite,
Microplitis croceipes
(a member of the wasp family Braconidae), causes inhibition of growth and permanent developmental arrest of the tobacco budworm, the cotton bollworm (also known as the corn earworm and tomato fruitworm) and the soybean pod worm. The larvae of these insects (Heliothis and Helicoverpa spp.) feed on tobacco, cotton, maize, sorghum, soybeans, sunflower and tomatoes, among other plants. The larvae cause economic losses of over $1 billion annually, primarily in the form of yield reduction and costs related to control. Fitt, 34
Ann. Rev. Entomol
. 17 (1989).
In the past, the United States Department of Agriculture has funded programs designed to increase
Microplitis croceipes
populations in Southern states so as to naturally combat these pests in those areas. These efforts were successful in reducing pest populations. Use of such “biomanagement” techniques has generally been replaced with molecular biology techniques, such as those which allowed engineering of plants which express
Bacillus thuringiensis
toxin.
Certain extra-embryonic cells (teratocytes) from
M. croceipes
have been shown to be involved in impairment of the growth, development and related physiological parameters of Heliothis and Helicoverpa larvae. Although teratocytes do not undergo cell division subsequent to their release into the hemocoel of the host, they do become polyploidal. Injection of one larval equivalent of teratocytes caused characteristic post-wandering, pre-pupation developmental arrest and eventual death associated with parasitization. Zhang and Dahlman, 12 Arch. Insect Biochem. Physiol. 51 (1989). Teratocytes collected from
M. croceipes
eggs hatched in vitro produced responses similar to those collected from parasitized Heliothis larvae. Zhang et al., 43
J. Insect Physiol
. 577 (1997). Furthermore, it has been suggested that when teratocytes are cultured in vitro, they secrete a mixture of proteins (teratocyte secreted proteins or TSP) which, when injected into host larvae, produced responses similar to parasitization. Schepers et al., 44
J. Insect Physiol
. 767 (1998).
In related studies, juvenile hormone esterase and ecdysone titers have been shown to be suppressed by teratocytes to a degree similar to those in parasitized larvae. Zhang, et al. 20
, Arch. Insect Biochem. Physiol
. 231 (1992). Reduced titers of host hemolymph proteins have been observed, particularly in older stages, with a major effect on the 74, 76 and 82 kD storage protein monomers. Zhang et al., 43
J. Insect Physiol
. 577 (1997). Inhibition of storage protein synthesis in the fat body was theorized to be at the level of translation, primarily based on the finding that storage protein mRNA levels did not change, even though protein synthesis declined precipitously after treatment with teratocytes. Dong et al., 32
Arch. Insect Biochem. Physiol
. 237 (1996).
Citation of the above documents is not intended as an admission that any of the foregoing is pertinent prior art. All statements as to the date or representation as to the contents of these documents is based on subjective characterization of information available to the applicant, and does not constitute any admission as to the accuracy of the dates or contents of these documents.
SUMMARY OF THE INVENTION
It is therefore an object of the present invention to provide isolated nucleic acid compounds useful to inhibit insect larval growth and development.
It is a further object to provide isolated amino acid compounds useful to inhibit larval growth and development.
It is yet another object to provide methods to produce isolated amino acid compounds useful to inhibit larval growth and development.
It is yet another object to provide methods to inhibit larval growth and development.
In all of the above embodiments, it is an object to provide methods to reduce crop damage using the compounds and methods herein.
It is an object of the invention to provide a natural pesticide using the compounds and methods herein.
It is an object of the invention to provide vectors, cells and molecular constructs comprising the compounds and methods herein.
It is an object of the invention to provide plants, including plant parts, seeds and embryos comprising the compounds and methods herein.
Definitions:
For the purposes of the present application, the following terms have the following meanings. All other terms have the meaning as generally recognized in the art.
“Allelic variant” is meant to refer to a gene that occurs at essentially the same locus (or loci) as the referent sequence, but which, due to natural variations caused by, for example, mutation or recombination, has a similar but not identical sequence. Allelic variants typically encode proteins having similar activity to that of the protein encoded by the gene to which they are being compared. Allelic variants can also comprise alterations in the 5′ or 3′ untranslated regions of the gene (e.g., in regulatory control regions).
“Fragment” is meant to refer to any nucleic acid or polypeptide subset of the referent compound.
“Inducing agent” means any compound or condition that causes inducement of an inducible promoter, including chemical compounds or environmental conditions, such as drought, wounds, light cycle, etc.
“Maize” and “corn” shall be interchangeable and mean all maize varieties.
“Proteins” means any compounds which comprise amino acids, including peptides, polypeptides, fusion proteins, etc.
“Transform” means delivery of nucleic acid into a cell, including delivery which results in genomic integration and delivery which results in transient localization within the cell membrane.
Moreover, for the purposes of the present invention, the term “a” or “an” entity refers to one or more of that entity; for example, “a protein” or “a nucleic acid molecule” refers to one or more of those compounds or at least one compound. As such, the terms “a” (or “an”), “one or more” and “at least one” can be used interchangeably herein. It is also to be noted that the terms “comprising”, “including”, and “having” can be used interchangeably. Furthermore, a compound “selected from the group consisting of” refers to one or more of the compounds in the list that follows, including mixtures (i.e., combinations) of two or more of the compounds. According to the present invention, an isolated, or biologically pure, protein or nucleic acid molecule is a compound that has been removed from its natural milieu. As such, “isolated” and “biologically pure” do not necessarily reflect the extent to which the compound has been purified. An isolated compound of the present invention can be obtained from its natural source, can be produced using molecular biology techniques or can be produced by chemical synthesis.
BRIEF DESCRIPTION OF THE TABLES
Table 1—Sequence listing of cDNAs, peptides and primers.
Table 2—Responses of Heliothis virescens larvae to feeding on transgenic plants expressing the TSP gene.
Table 3—Responses of Manduca sexta larvae to feeding on transgenic plants expressing the TSP gene.
REFERENCES:
Chen et al, Mutations in Domain I ofBacillus thuringiensis&dgr;-endotoxin CrylAb Reduce the Irreversible Binding of Toxin to Manduca sexta Brush Border Membrane Vesicles (*), Mar. 1995, Journal of Biological Chemistry, vol. 270, No. 11 pp. 6412-6419.*
M. Peferoen, “Progress and prospects for field use of Bt genes in crops”, May 1997, Trends in Biotechnology, vol. 15, pp. 173-177.*
Zhang and Dahlman, 12Arch. Insect Biochem. Physiol. 51 (1989).
Zhang, et al., 43J. Insect Physiol. 577 (1997).
Schepers, et al., 44J. Insect Physiol. 767 (1998).
Zhang, et al., 20Arch. Insect Biochem. Physiol. 231 (1992).
Dong, et al., 32Arch. Insect Biochem. Physiol. 237 (1996).
Dahlman Douglas Lee
Maiti Indu Bhushan
Webb Bruce Allen
Collins Cynthia
McElwain Elizabeth F.
University of Kentucky Research Foundation
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