Abstract: The subject invention relates in part to a combination of a cadherin peptide derived from Alphitobius diaperinus and a Cry protein capable of inhibiting an insect. In certain embodiments, the combination is capable of overcoming Cry protein resistance in insects that are resistant to the Cry protein alone.

Abstract: The subject invention relates in part to fragments of non-cadherin Cry toxin binding proteins, wherein the fragments potentiate, or act as synergists with, the insecticidal activity of Cry proteins. In some preferred embodiments, the binding protein (a Cry protein receptor on insect midgut cells) is an aminopeptidase. In preferred embodiments, the fragment comprises a Cry protein binding region.

Abstract: This invention relates in part to modifying BtBooster (BtB) peptides, in part to increase their stability in insect midgut digestive juices. Some preferred embodiments of BtB have removed proteinase cleavage sites resulting in increased stability of the modified BtB in the insect gut, while retaining the ability to enhance B.t. proteins for improved insect control. In some preferred embodiments, the protease-stable BtB is used in combination with B.t. spores and/or crystals comprising a Cry protein. Also reported herein is the significant and increased enhancement of Bt toxins against relatively Bt-tolerant insects (Helicoverpa zea, Spodoptera exigua and Agrotis ipsilon), when used with BtBs. We also describe increased toxin enhancement with cadherin fragments that are stabilized against over-digestion by insect midgut proteinases. We also report enhancement of Bt Cry1F toxin by cadherin fragments.

Abstract: The subject invention relates in part to the surprising and unexpected discovery that insects that are resistant to Bacillus thuringiensis Cry toxins have measurably altered alkaline phosphatase (ALP) activity as compared to insects that are susceptible to Cry toxins. This and other surprising discoveries reported herein have broad implications in areas such as managing and monitoring the development of insect resistance to B.t. toxins. For example, the subject invention provides a simple and fast assay (enzymatic or otherwise) for detecting ALP activity levels and thereby monitoring the development of resistance by insects to crystal protein insect toxins. There was no prior motivation or suggestion to go about resistance monitoring using this simple and easy approach.

Abstract: The subject invention relates in part to the discovery that a fragment from a cadherin of the western corn rootworm enhances Cry3 toxicity to larvae of naturally susceptible species. The subject invention also relates in part to the discovery that a cadherin fragment from a beetle enhances Cry3Aa and Cry3Bb toxicity to coleopteran larvae, particularly those in the family Chrysomelidae. Such cadherin fragments are referred to as Bt Boosters (BtBs). The subject invention can be extended to the use of BtBs with other coleopteran-toxic Cry proteins for controlling a wide range of coleopterans.

Abstract: This invention relates in part to modifying BtBooster (BtB) peptides, in part to increase their stability in insect midgut digestive juices. Some preferred embodiments of BtB have removed proteinase cleavage sites resulting in increased stability of the modified BtB in the insect gut, while retaining the ability to enhance B.t. proteins for improved insect control. In some preferred embodiments, the protease-stable BtB is used in combination with B.t. spores and/or crystals comprising a Cry protein. Also reported herein is the significant and increased enhancement of Bt toxins against relatively Bt-tolerant insects (Helicoverpa zea, Spodoptera exigua and Agrotis ipsilon), when used with BtBs. We also describe increased toxin enhancement with cadherin fragments that are stabilized against over-digestion by insect midgut proteinases. We also report enhancement of Bt Cry1F toxin by cadherin fragments.

Abstract: The subject invention relates in part to the surprising and unexpected discovery that insects that are resistant to Bacillus thuringiensis Cry toxins have measurably altered alkaline phosphatase (ALP) activity as compared to insects that are susceptible to Cry toxins. This and other surprising discoveries reported herein have broad implications in areas such as managing and monitoring the development of insect resistance to B. t. toxins. For example, the subject invention provides a simple and fast assay (enzymatic or otherwise) for detecting ALP activity levels and thereby monitoring the development of resistance by insects to crystal protein insect toxins. There was no prior motivation or suggestion to go about resistance monitoring using this simple and easy approach.

Abstract: The subject invention pertains to the use of peptide fragments of cadherins (including cadherin-like proteins). The subject invention includes a cell (and use thereof) comprising a polynucleotide that expresses the peptide fragment. The subject invention includes methods of feeding the peptides to insects. In preferred embodiments, the peptides are fed to target insects together with one or more insecticidal proteins, preferably (but not limited to) B.t. Cry proteins. When used in this manner, the peptide fragment can not only enhance the apparent toxin activity of the Cry protein against the insect species that was the source of the receptor but also against other insect species. Preferably, the cadherin is a Bacillus thuringiensis (B.t.) insecticidal crystal protein (Cry) toxin receptor. Preferably, the peptide fragment is a binding domain of the receptor. In some preferred embodiments, the peptide is the binding domain nearest to the membrane proximal ectodomain.

Abstract: The subject invention relates in part to the surprising and unexpected discovery that insects that are resistant to Bacillus thuringiensis Cry toxins have measurably altered alkaline phosphatase (ALP) activity as compared to insects that are susceptible to Cry toxins. This and other surprising discoveries reported herein have broad implications in areas such as managing and monitoring the development of insect resistance to B.t. toxins. For example, the subject invention provides a simple and fast assay (enzymatic or otherwise) for detecting ALP activity levels and thereby monitoring the development of resistance by insects to crystal protein insect toxins. There was no prior motivation or suggestion to go about resistance monitoring using this simple and easy approach.

Abstract: The subject invention relates in part to the discovery that BtBooster (BtB) peptides enhance Bt strain and Cry protein toxicity to the lesser mealworm. The subject invention also relates in part to the discovery that Bt tenebrionis producing Cry3Aa crystals and Bt japonensis BuiBui producing Cry8Ca crystals are insecticidal to A. diperinus larvae (darkling beetle or lesser mealworm). The subject invention also relates in part to the discovery that fragments from cadherins of the western corn rootworm and the yellow mealworm enhance the toxicities of Bt tenebrionis and Bt japonensis BuiBui toxicity to coleopteran larvae of the genus Alphitobius. In addition, the subject invention relates in part to the use of cadherin fragments to enhance the toxicities of Cry3Aa, Cry3Bb and Cry8Ca to coleopteran larvae of the genus Alphitobius. The subject invention also relates in part to screening Bt strains and Cry proteins for toxicity to larvae of darkling beetles or lesser mealworm, Alphitobius diaperinus.

Abstract: The subject invention relates in part to the discovery that a fragment from a cadherin of the western corn rootworm enhances Cry3 toxicity to larvae of naturally susceptible species. The subject invention also relates in part to the discovery that a cadherin fragment from a beetle enhances Cry3Aa and Cry3Bb toxicity to coleopteran larvae, particularly those in the family Chrysomelidae. Such cadherin fragments are referred to as Bt Boosters (BtBs). The subject invention can be extended to the use of BtBs with other coleopteran-toxic Cry proteins for controlling a wide range of coleopterans.

Abstract: The subject invention relates in part to a novel protein for binding Bacillus thuringiensis Cry toxins, and fragments of cadherins for enhancing Cry toxicity against dipterans. The subject invention also relates in part to the discovery that fragments of a midgut cadherin from a dipteran insect synergize Cry proteins that are active against dipterans. Thus, the subject invention includes the use of fragments of cadherin ectodomains for controlling dipterans. Such fragments (that bind Crys) can be administered to a dipteran insect for ingestion. In some preferred embodiments, the source cadherin is a dipteran cadherin. Also in some preferred embodiments, the fragment is administered with a Cry protein that is active against a dipteran. Variants of the fragments of naturally occurring cadherins are included within the scope of the subject invention.

Abstract: The subject invention pertains to the use of peptide fragments of cadherins (including cadherin-like proteins). The subject invention includes a cell (and use thereof) comprising a polynucleotide that expresses the peptide fragment. The subject invention includes methods of feeding the peptides to insects. In preferred embodiments, the peptides are fed to target insects together with one or more insecticidal proteins, preferably (but not limited to) B.t. Cry proteins. When used in this manner, the peptide fragment can not only enhance the apparent toxin activity of the Cry protein against the insect species that was the source of the receptor but also against other insect species. Preferably, the cadherin is a Bacillus thuringiensis (B.t.) insecticidal crystal protein (Cry) toxin receptor. Preferably, the peptide fragment is a binding domain of the receptor. In some preferred embodiments, the peptide is the binding domain nearest to the membrane proximal ectodomain.

Abstract: The subject invention pertains to the use of peptide fragments of cadherins (including cadherin-like proteins). The subject invention includes a cell (and use thereof) comprising a polynucleotide that expresses the peptide fragment. The subject invention includes methods of feeding the peptides to insects. In preferred embodiments, the peptides are fed to target insects together with one or more insecticidal proteins, preferably (but not limited to) B.t. Cry proteins. When used in this manner, the peptide fragment can not only enhance the apparent toxin activity of the Cry protein against the insect species that was the source of the receptor but also against other insect species. Preferably, the cadherin is a Bacillus thuringiensis (B.t.) insecticidal crystal protein (Cry) toxin receptor. Preferably, the peptide fragment is a binding domain of the receptor. In some preferred embodiments, the peptide is the binding domain nearest to the membrane proximal ectodomain.

Abstract: The subject invention pertains to the use of peptide fragments of cadherins (including cadherin-like proteins). The subject invention includes a cell (and use thereof) comprising a polynucleotide that expresses the peptide fragment. The subject invention includes methods of feeding the peptides to insects. In preferred embodiments, the peptides are fed to target insects together with one or more insecticidal proteins, preferably (but not limited to) B.t. Cry proteins. When used in this manner, the peptide fragment can not only enhance the apparent toxin activity of the Cry protein against the insect species that was the source of the receptor but also against other insect species. Preferably, the cadherin is a Bacillus thuringiensis (B.t.) insecticidal crystal protein (Cry) toxin receptor. Preferably, the peptide fragment is a binding domain of the receptor. In some preferred embodiments, the peptide is the binding domain nearest to the membrane proximal ectodomain.

Abstract: A method for expressing insecticidal protein structural genes in cotton plant genomes is provided. In the preferred embodiments this invention comprises placing a structural gene for the Bacillus thuringiensis crystal protein under control of a plant or a T-DNA promoter and ahead of a polyadenylation site followed by insertion of said promoter/structural gene combination into a plant genome by utilizing an Agrobacterium tumefaciens Ti plasmid-based transformation system. The modified Ti plasmid is then used to transform recipient plant cells. Also provided are the plants and tissues produced by this method and bacterial strains, plasmids, and vectors useful for execution of this invention.

Abstract: Materials and methods for identifying novel pesticide agents are disclosed herein. Specifically exemplified is a full length aminopeptidase N isolated from Manduca sexta, insect cells expressing APN, and methods of screening pesticide agents using the same. Also disclosed are methods of identifying novel APN inhibitors.

Abstract: A method for expressing insecticidal protein structural genes in plant genomes is provided. In the preferred embodiments this invention comprises placing a structural gene for the Bacillus thuringiensis crystal protein under control of a plant or a T-DNA promoter and ahead of a polyadenylation site followed by insertion of said promoter/structural gene combination into a plant genome by utilizing an Agrobacterium tumefaciens Ti plasmid-based transformation system. The modified Ti plasmid is then used to transform recipient plant cells. Also provided are the plants and tissues produced by this method and bacterial strains, plasmids, and vectors useful for execution of this invention.

Abstract: Disclosed herein are activated Bt toxins expressed in E. coli as a translational fusion with a phage coat protein of filamentous phage. Phage displaying this fusion protein were viable, infectious, and as lethal as pure toxin on a molar basis when fed to insects susceptible to native Bt toxins.

Abstract: Materials and methods for identifying novel pesticide agents are disclosed herein. Specifically exemplified is a full length aminopeptidase N isolated from Manduca sexta, insect cells expressing APN, and methods of screening pesticide agents using the same. Also disclosed are methods of identifying novel APN inhibitors.