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
1999-02-17
2001-08-28
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
C800S279000, C800S288000, C800S320100, C536S023700, C435S418000, C435S419000, C435S320100
Reexamination Certificate
active
06281413
ABSTRACT:
FIELD OF THE INVENTION
The invention relates to novel toxins from
Photorhabdus luminescens,
nucleic acid sequences whose expression results in said toxins, and methods of making and methods of using the toxins and corresponding nucleic acid sequences to control insects.
BACKGROUND OF THE INVENTION
Insect pests are a major cause of crop losses. Solely in the US, about $7.7 billion are lost every year due to infestation by various genera of insects. In addition to losses in field crops, insect pests are also a burden to vegetable and fruit growers, to producers of ornamental flowers, and they are a nuisance to gardeners and home owners.
Insect pests are mainly controlled by intensive applications of chemical insecticides, which are active through inhibition of insect growth, prevention of insect feeding or reproduction, or death of the insects. Good insect control can thus be reached, but these chemicals can sometimes also affect other, beneficial insects. Another problem resulting from the wide use of chemical pesticides is the appearance of resistant insect varieties. This has been partially alleviated by various resistance management strategies, but there is an increasing need for alternative pest control agents. Biological insect control agents, such as
Bacillus thuringiensis
strains expressing insecticidal toxins like d-endotoxins, have also been applied with satisfactory results, offering an alternative or a complement to chemical insecticides. Recently, the genes coding for some of these d-endotoxins have been isolated and their expression in heterologous hosts have been shown to provide another tool for the control of economically important insect pests. In particular, the expression of insecticidal toxins in transgenic plants, such as
Bacillus thuringiensis
d-endotoxins, has provided efficient protection against selected insect pests, and transgenic plants expressing such toxins have been commercialized, allowing farmers to reduce applications of chemical insect control agents. Yet, even in this case, the development of resistance remains a possibility and only a few specific insect pests are controllable. Consequently, there remains a long-felt but unfulfilled need to discover new and effective insect control agents that provide an economic benefit to farmers and that are environmentally acceptable.
SUMMARY OF THE INVENTION
The present invention addresses the need for novel insect control agents. Particularly needed are control agents that are targeted to economically important insect pests and that efficiently control insect strains resistant to existing insect control agents. Furthermore, agents whose application minimizes the burden on the environment are desirable.
In the search of novel insect control agents, certain classes of nematodes from the genera Heterorhabdus and Steinernema are of particular interest because of their insecticidal properties. They kill insect larvae and their offspring feed in the dead larvae. Indeed, the insecticidal activity is due to symbiotic bacteria living in the nematodes. These symbiotic bacteria are Photorhabdus in the case of Heterorhabdus and Xenorhabdus in the case of Steinernema.
The present invention is drawn to nucleic acid sequences isolated from
Photorhabdus luminescens,
and sequences substantially similar thereto, whose expression results in toxins that are highly toxic to economically important insect pests, particularly insect pests that infest plants. The invention is further drawn to the toxins resulting from the expression of the nucleic acid sequences, and to compositions and formulations containing the toxins, which are capable of inhibiting the ability of insect pests to survive, grow or reproduce, or of limiting insect-related damage or loss in crop plants. The invention is further drawn to a method of making the toxins and to methods of using the nucleic acid sequences, for example in microorganisms to control insects or in transgenic plants to confer insect resistance, and to a method of using the toxins, and compositions and formulations comprising the toxins, for example applying the toxins or compositions or formulations to insect-infested areas, or to prophylactically treat insect-susceptible areas or plants to confer protection or resistance to the insects.
The novel toxins are highly active against insects. For example, a number of economically important insect pests, such as the Lepidopterans
Plutella xylostella
(Diamondback Moth),
Trichoplusia ni
(Cabbage Looper),
Ostrinia nubilalis
(European Corn Borer),
Heliothis virescens
(Tobacco Budworm),
Helicoverpa zea
(Corn Earworm),
Manduca sexta
(Tobacco Hornworm),
Spodoptera exigua
(Beet Armyworm), and
Spodoptera frugiperda
(Fall Armyworm), as well as the Coleopterans
Diabrotica virgifera
virgifera (Western Corn Rootworm),
Diabrotica undecimpunctata
howardi (Southern Corn Rootworm), and Leptinotarsa decimlineata (Colorado Potato Beetle) can be controlled by one or more of the toxins. The toxins can be used in multiple insect control strategies, resulting in maximal efficiency with minimal impact on the environment.
According to one aspect, the present invention provides an isolated nucleic acid molecule comprising: (a) a nucleotide sequence substantially similar to a nucleotide sequence selected from the group consisting of: nucleotides 412-1665 of SEQ ID NO:1, nucleotides 1686-2447 of SEQ ID NO:1, nucleotides 2758-3318 of SEQ ID NO:1, nucleotides 3342-4118 of SEQ ID NO:1, nucleotides 4515-9269 of SEQ ID NO:1, nucleotides 15,171-18,035 of SEQ ID NO:11, and nucleotides 31,393-35,838 of SEQ ID NO:11; (b) a nucleotide sequence comprising nucleotides 23,768-31,336 of SEQ ID NO:11; or (c) a nucleotide sequence isocoding with the nucleotide sequence of (a) or (b); wherein expression of the nucleic acid molecule results in at least one toxin that is active against insects.
In one embodiment of this aspect, the nucleotide sequence is isocoding with a nucleotide sequence substantially similar to nucleotides 412-1665 of SEQ ID NO:1, nucleotides 1686-2447 of SEQ ID NO:1, nucleotides 2758-3318 of SEQ ID NO:1, nucleotides 3342-4118 of SEQ ID NO:1, or nucleotides 4515-9269 of SEQ ID NO:1. Preferably, the nucleotide sequence is substantially similar to nucleotides 412-1665 of SEQ ID NO:1, nucleotides 1686-2447 of SEQ ID NO:1, nucleotides 2758-3318 of SEQ ID NO:1, nucleotides 3342-4118 of SEQ ID NO:1, or nucleotides 4515-9269 of SEQ ID NO:1. More preferably, the nucleotide sequence encodes an amino acid sequence selected from the group consisting of SEQ ID NOS:2-6. Most preferably, the nucleotide sequence comprises nucleotides 412-1665 of SEQ ID NO:1, nucleotides 1686-2447 of SEQ ID NO:1, nucleotides 2758-3318 of SEQ ID NO:1, nucleotides 3342-4118 of SEQ ID NO:1, or nucleotides 4515-9269 of SEQ ID NO:1.
In another embodiment of this aspect, the nucleotide sequence is isocoding with a nucleotide sequence substantially similar to nucleotides 15,171-18,035 of SEQ ID NO:11. Preferably, the nucleotide sequence is substantially similar to nucleotides 15,171-18,035 of SEQ ID NO:11. More preferably, the nucleotide sequence encodes the amino acid sequence set forth in SEQ ID NO:12. Most preferably, the nucleotide sequence comprises nucleotides 15,171-18,035 of SEQ ID NO:11.
In still another embodiment of this aspect, the nucleotide sequence is isocoding with a nucleotide sequence substantially similar to nucleotides 31,393-35,838 of SEQ ID NO:11. Preferably, the nucleotide sequence is substantially similar to nucleotides 31,393-35,838 of SEQ ID NO:11. More preferably, the nucleotide sequence encodes the amino acid sequence set forth in SEQ ID NO:14. Most preferably, the nucleotide sequence comprises nucleotides 31,393-35,838 of SEQ ID NO:11.
In yet another embodiment of this aspect, the nucleotide sequence encodes the amino acid sequence set forth in SEQ ID NO:13, and preferably comprises nucleotides 23,768-31,336 of SEQ ID NO:11.
In one embodiment, the nucleotide sequence of the invention comprises the approximately 9.7 kb DNA fragment harbored in
E. co
Anderson Arne Robert
Chen Jeng Shong
Dunn Martha M.
Hart Hope Prim
Kramer Vance Cary
Fox David T.
Holmes Jennifer L.
Kubelik Anne R.
Meigs J. Timothy
Stults Larry W.
LandOfFree
Insecticidal toxins from Photorhabdus luminescens and... does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Insecticidal toxins from Photorhabdus luminescens and..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Insecticidal toxins from Photorhabdus luminescens and... will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2534879