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
1998-02-23
2001-09-04
Nelson, Amy J. (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, C536S023710
Reexamination Certificate
active
06284949
ABSTRACT:
BACKGROUND OF THE INVENTION
Bacillus thuringiensis
(
B.t.
) is a spore forming soil bacterium which is known for its ability to produce a parasporal crystal protein which is toxic to a wide variety of insects. Most strains are active against Lepidopteran insects (moths and butterflies) and a few are reported to have activity against Dipteran insects (mosquitoes and flies, see Aronson et al. 1986). Toxin genes from a variety of these strains have been cloned and the toxins have been expressed in heterologous hosts (Schnepf et al., 1981; Klier et al., 1982). In recent years,
B.t
. var.
tenebrionis
(
B.t.t
., Krieg et al., 1983; Krieg et al., 1984) and
B.t
. var.
san diego
(
B.t.sd
., Herrnstadt et al., 1986) strains have been identified as having activity against Coleopteran insects. The toxin gene from
B.t.sd
. has been cloned, but the toxin produced in
E. coli
was reported to be a larger size than the toxin from
B.t.sd
. crystals, and activity of this recombinant
B.t.sd
. toxin was implied to be weak.
Insects susceptible to the action of the protein toxin of Coleopteran-type
Bacillus thuringiensis
bacteria include, but are not limited to, Colorado potato beetle (
Leptinotarsa decemlineata
), boll weevil (
Anthonomus grandis
), yellow mealworm (
Tenebrio molitor
), elm leaf beetle (
Pyrrhalta luteola
) and Southern corn rootworm (
Diabrotica undecimpunctata howardi
).
Therefore, the potential for genetically engineered plants which exhibit toxicity or tolerance toward Coleopteran insects was foreseen if such plants could be transformed to express a Coleopteran-type toxin at a insecticidally-effective level. Agronomically important crops which are affected by Coleopteran insects include alfalfa, cotton, maize, potato, rape (canola), rice, tobacco, tomato, sugar beet and sunflower.
BRIEF SUMMARY OF THE INVENTION
Although certain chimeric genes have been expressed in transformed plant cells and plants, such expression is by no means straight forward. Specifically, the expression of Lepidopteran-type
B.t
. toxin proteins has been particularly problematic. It has now been found that the teachings of the art with respect to expression of Lepidopteran-type
B.t
. toxin protein in plants do not extend to Coleopteran-type
B.t
. toxin protein. These findings are directly contrary to the prior teachings which suggested that one would employ the same genetic manipulations to obtain useful expression of such toxins in transformed plants.
In accordance with one aspect of the present invention, there has been provided a method for producing genetically transformed plants which exhibit toxicity toward Coleopteran insects, comprising the steps of:
(a) inserting into the genome of a plant cell susceptible to attack by Coleopteran insects a chimeric gene comprising:
i) a promoter which functions in plant cells to cause production of RNA;
ii) a DNA sequence that causes the production of a RNA sequence encoding a Coleopteran-type toxin protein of
Bacillus thuringiensis
; and
iii) a 3′ non-translated DNA sequence which functions in plant cells to cause the addition of polyadenylate nucleotides to the 3′ end of the RNA sequence;
(b) obtaining transformed plant cells, and
(c) regenerating from the transformed plant cells genetically transformed plants exhibiting resistance to Coleopteran insects.
In accordance with another aspect of the present invention, there has been provided a chimeric plant gene comprising in sequence:
(a) a promoter which functions in plant cells to cause the production of RNA;
(b) a DNA sequence that causes the production of a RNA sequence encoding a Coleopteran-type toxin protein of
Bacillus thuringiensis
; and
(c) a 3′ non-translated region which functions in plant cells to cause the addition of polyadenylate nucleotides to the 3′ end of the RNA sequence.
There has also been provided, in accordance with another aspect of the present invention, bacterial cells, transformed plant cells and plant transformation vectors that contain, respectively, DNA comprised of the above-mentioned elements (a), (b) and (c).
In accordance with yet another aspect of the present invention, a differentiated plant has been provided that comprises transformed plant cells, as described above, which exhibit toxicity to Coleopteran insects. The present invention also contemplates seeds which produce the above-described transformed plants.
REFERENCES:
patent: 4766203 (1988-08-01), Krieg et al.
patent: 4771131 (1988-09-01), Herrnstadt et al.
patent: 5254799 (1993-10-01), DeGreve et al.
patent: 5300629 (1994-04-01), Casteels et al.
patent: 5317096 (1994-05-01), DeGreve et al.
patent: 5338544 (1994-08-01), Donovan
patent: 5495071 (1996-02-01), Fischhoff et al.
patent: 5500365 (1996-03-01), Fischhoff et al.
patent: 5516693 (1996-05-01), Vaeck et al.
patent: 5545565 (1996-08-01), DeGreve et al.
patent: 5760181 (1998-06-01), DeGreve et al.
patent: 5763241 (1998-06-01), Fischhoff et al.
patent: 5767372 (1998-06-01), DeGreve et al.
patent: 5843898 (1998-12-01), DeGreve et al.
patent: 0 142 924 (1985-05-01), None
patent: 0 149 162 (1985-07-01), None
patent: 193 259 (1986-09-01), None
patent: 0 202 739 (1986-11-01), None
patent: 0 206 613 (1986-12-01), None
patent: 0 213 818 (1987-03-01), None
patent: 0 290 395 (1988-11-01), None
patent: 0 292 435 (1988-11-01), None
patent: 0 305 275 (1989-03-01), None
patent: 0 318 143 (1989-05-01), None
patent: 0 359 472 (1990-03-01), None
patent: 0 116 713 (1990-05-01), None
patent: 0 731 170 (1996-09-01), None
patent: WO 84/02913 (1984-08-01), None
patent: WO 86/01536 (1986-03-01), None
Adang, M.J., et al., Applications of aBacillus thuringiensisCrystal Protein for Insect Control, Abstract J20 from Journal of Cellular Biochemistry Supplement 10C-UCLA Symposia on Molecular & Cellular Biology (1986).
Barton, et al.,Bacillus thuringiensis-&-Endotoxin Ecpressin in TransgenicNicotiana tabecumProvides Resistance to Lepidopteran Insects,Plant Physiol., vol. 85, pp. 1103-1109 (1987).
Hernstadt, et al., A new Strain ofBacillus thuringiensiswith Activity against coleopterna Insects, 6062Biotechnology, Apr. No. 4, pp. 305-308 (1986).
Hofte, et al.,Microbiological Reviews, vol. 53(2), pp. 242-255 (Jun. 1989).
Höfte, H., et al., Abstract K86 (no title) from Journal of Cellular Biochemistry Supplement 10C-UCLA Symposia on Molecular & Cellular Biology (1986).
Höfte, H., et al., Structural and Functional Analysis of a Cloned Delta Endotoxin ofBacillus thuringiensisberliner 1715,Eur. J. Bioche., 161: 273-280 (1986).
McPherson, S.A., et al,Biotechnology, vol. 6, pp. 61-66 (1988).
McPherson, S. et al., Abstract H-44,Abstracts of the Annual Meeting of the American Society for Microbiology(VS), vol. 88, No. O, p. 152 (1988).
Whiteley, H.R.,Ann. Rev. Microbiol., vol. 40, pp. 549-576 (1986).
Vaeck, et al., Transgenic plants protected from insect attack,Nature, vol. 328, pp. 30-37 (1987).
Adang, M.J., et al., The reconstruction and expression ofBacillus thuringiensiscryIIIA gene in protoplasts and potato plants,Plant Molecular Biology, 21: 1131-1145 (1993).
Bernhard, Studies on the delta-endotoxin ofBacillus thuringiensisvar. tenebrionis,FEMS Microbiological Letters33, pp. 261-265 (1986).
Klein, T.M., et al., Factors Influencing Gene Delivery into Zea Mays Cells By High-Velocity Microprojectiles,Bio/Technology, vol. 6, pp. 559-563 (1988).
Klier, A., et al., Cloning and expression of the crystal protein genes fromBacillus thuringiensisstrain berliner 1715,EMBO J., 1: 791-799 (1982).
Koziel, M.G., et al., Field Performance of Elite Transgenic Maize Plants Expressing an Insecticidal Protein Derived fromBacillus thuringiensis, Bio/Technology, vol. 11, pp. 194-200 (1993).
Krieg, et al.,Bacillus thuringiensisvar. tenebrionis: ein neuer, gegenüber Larven von Coleopteren wirksamer Pathotyp,Pathotyp. Z. Ang. Ent., 96: 500-508 (1983).
Krieg, et al., Neue Ergebnisse überBacillus thuringiensisvar tenebrionis unter besonderer Berücksichtigung seiner Wirkung and den Kartoffelkäfer (Leptinotarsa decemlineata),Ang. Schadlingshde., Pflanzenschutz., Umweltschutz, 57: 145-1
Fischhoff David A.
Fuchs Roy L.
Lavrik Paul B.
McPherson Sylvia A.
Perlak Frederick J.
Ball T. K.
Howrey Simon Arnold & White , LLP
Monsanto Company
Nelson Amy J.
LandOfFree
Insect-resistant plants comprising a Bacillus thuringiensis... does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Insect-resistant plants comprising a Bacillus thuringiensis..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Insect-resistant plants comprising a Bacillus thuringiensis... will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2455747