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AssigneeUniversity of Georgia Research Foundation, Inc.
InventorsMichael J. Adang, Rebecca McNall, Juan Luis Jurat Fuentes
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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.
A method of screening a lepidopteran for resistance to a Bacillus thuringiensis (B.t) insecticidal Cry1 protein that is toxic to lepidopterans, wherein said lepdopteran is of a species known to develop resistance to said Cry1 protein, wherein said method comprises collecting a lepidopteran from a field of a B.t.crop, obtaining a gut cell membrane sample from said lepidopteran, running said sample on a gel to identify an approximately 68 kDa protein band on said gel, measuring said 68 kDa protein for an amount of alkaline phosphatase enzymatic activity, and comparing said amount to a control level of alkaline phosphatase enzymatic activity determined from a gut cell membrane preparation from a control insect of known susceptibility to said Cry1 protein, wherein resistance to said Cry1 protein by said lepidopteran is indicated if said amount is less than that of a non-resistant insect, wherein said non-resistant insect is of the same genus and species as said lepidopteran.
Dependencies are inferred from the claim text.