US2006216367A1PendingUtilityA1

Insecticidal extract from legume plants and method of preparing the same

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Assignee: CANADA NATURAL RESOURCESPriority: Mar 24, 2005Filed: Mar 24, 2005Published: Sep 28, 2006
Est. expiryMar 24, 2025(expired)· nominal 20-yr term from priority
Inventors:Wesley G. TaylorPaul G. FieldsDaniel H. Sutherland
A01N 65/20A01N 43/90A01N 65/00
46
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Claims

Abstract

The invention may be summarized as follows. The present invention provides insecticides comprised of naturally-occurring compounds, such as, for example, PA1b-related peptides and terpenoid saponins. The present invention also provides for identification and characterization of a synergistic insecticidal effect between peptides and saponins extracted from plants. Furthermore, simplified extraction procedures are provided that avoid the use of a chloroform defatting step or a column chromatography step.

Claims

exact text as granted — not AI-modified
1 . An insecticidal, alcohol-soluble extract of a non-defatted legume seed material, the extract comprising a PA1B-related peptide to saponin ratio ranging from 10:1 to 1:10, the PA1B-related peptide comprising an amino acid sequence of Cys-(Xaa)n-(Xaa)n-Cys-(Xaa)n-Pro-(Xaa)n-(Xaa)n-Pro-Cys-(Xaa)n-(Xaa)n-Cys-(Xaa)n-Cys-(Xaa)n-Pro-(Xaa)n-(Xaa)n-(Xaa)n-Cys (SEQ ID NO:63), where each Xaa is independently selected from any amino acid and each n independently equals 1, 2, 3, 4, or 5.  
     
     
         2 . The extract of  claim 1 , wherein the extract comprises at least one PA1b-related peptide comprising a sequence that is at least 70% identical to an amino acid sequence selected from the group consisting of: SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:8, and SEQ ID NO:9.  
     
     
         3 . The extract of  claim 2 , wherein the PA1b-related peptide comprises a sequence selected from the group consisting of: SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:8, and SEQ ID NO:9.  
     
     
         4 . The extract of  claim 1 , wherein the saponin is a triterpenoid saponin.  
     
     
         5 . The extract of  claim 4 , wherein the triterpenoid saponin is selected from the group consisting of: soyasaponin I, soyasaponin II, soyasaponin III, soyasaponin VI, dehydrosoyasaponin I, echinocystic acid 3-glucoside, glycyrrhizic acid, hederacoside C, beta-escin, and alpha-hederin.  
     
     
         6 . The extract of  claim 1 , wherein the alcohol is selected from the group consisting of methanol, ethanol, n-propanol, isopropanol, n-butanol, tert-butanol, 1-isobutanol, and 2-buten-1-ol.  
     
     
         7 . The extract of  claim 1 , wherein the alcohol is selected from the group consisting of methanol, ethanol, and n-butanol.  
     
     
         8 . The extract of  claim 1 , wherein the alcohol is ethanol.  
     
     
         9 . The extract of  claim 1 , wherein the extract is soluble in an about 50-98% aqueous solution of the alcohol.  
     
     
         10 . The extract of  claim 1 , wherein the extract is soluble in an about 60-95% aqueous solution of the alcohol.  
     
     
         11 . A method of preparing an insecticidal alcohol soluble legume extract comprising: extracting a non-defatted legume seed material with a C(1-4) alcohol.  
     
     
         12 . The method of  claim 11  further comprising precipitating the alcohol soluble extract with an organic acid.  
     
     
         13 . The method of  claim 12 , wherein the organic acid is selected from the group consisting of: benzoic, acetic, trichloroacetic, trifluoroacetic, sorbic, citric, formic, and propionic acid.  
     
     
         14 . The method of  claim 11 , wherein the alcohol is selected from the group consisting of methanol, ethanol, n-propanol, isopropanol, n-butanol, tert-butanol, 1-isobutanol, and 2-buten-1-ol.  
     
     
         15 . The method of  claim 11 , wherein the alcohol is selected from the group consisting of methanol, ethanol, and n-butanol.  
     
     
         16 . The method of  claim 11 , wherein the alcohol is ethanol.  
     
     
         17 . The method of  claim 11 , wherein the alcohol is an about 50-98% aqueous solution of the alcohol.  
     
     
         18 . The method of  claim 11 , wherein the alcohol is an about 60-95% aqueous solution of the alcohol.  
     
     
         19 . The method of  claim 11 , wherein the temperature of the alcohol is increased from room temperature to between 40-85 degree C.  
     
     
         20 . An insecticidal composition comprising an isolated PA1b-related peptide and an isolated saponin, the peptide to saponin ratio ranging from 10:1 to 1:10, the PA1B-related peptide comprising an amino acid sequence of Cys-(Xaa)n-(Xaa)n-Cys-(Xaa)n-Pro-(Xaa)n-(Xaa)n-Pro-Cys-(Xaa)n-(Xaa)n-Cys-(Xaa)n-Cys-(Xaa)n-Pro-(Xaa)n-(Xaa)n-(Xaa)n-Cys (SEQ ID NO:63), where each Xaa is independently selected from any amino acid and each n independently equals 1, 2, 3, 4, or 5.  
     
     
         21 . The insecticidal composition of  claim 20 , wherein the PA1b-related peptide comprises a sequence that is at least 70% identical to an amino acid sequence selected from the group consisting of: SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:8, and SEQ ID NO:9.  
     
     
         22 . The insecticidal composition of  claim 21 , wherein the PA1b-related peptide comprises a sequence selected from the group consisting of: SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:8, and SEQ ID NO:9.  
     
     
         23 . The insecticidal composition of  claim 21 , wherein the saponin is a triterpenoid saponin.  
     
     
         24 . The insecticidal composition of  claim 23 , wherein the triterpenoid saponin is selected from the group consisting of: soyasaponin I, soyasaponin II, soyasaponin III, soyasaponin VI, dehydrosoyasaponin I, echinocystic acid 3-glucoside, glycyrrhizic acid, hederacoside C, beta-escin, and alpha-hederin.  
     
     
         25 . A method of controlling insects comprising exposing the insects to an effective amount of the extract of  claim 1 .  
     
     
         26 . A method of controlling insects comprising exposing the insects to an effective amount of the composition of  claim 20 .  
     
     
         27 . A method of controlling insects comprising exposing the insects to an effective amount of a composition comprising dehydrosoyasaponin I.

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