Compositions and Methods for Controlling Leptinotarsa
First Claim
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1. A method for controlling a Leptinotarsa species infestation of a plant comprising:
- (a) contacting said Leptinotarsa species with a polynucleotide comprising a nucleotide sequence that is complementary to at least 21 contiguous nucleotides of a target gene having a nucleotide sequence selected from the group consisting of;
SEQ ID NO;
730, SEQ ID NO;
807, SEQ ID NOs;
1-725, SEQ ID NOs;
726-729, SEQ ID NOs;
731-806, SEQ ID NOs;
808-830, and SEQ ID NOs;
1087-1094, or an RNA transcribed from said target gene;
or(b) providing in the diet of said Leptinotarsa species a polynucleotide comprising a nucleotide sequence that is complementary to at least 21 contiguous nucleotides of a target gene having a nucleotide sequence selected from the group consisting of;
SEQ ID NO;
730, SEQ ID NO;
807, SEQ ID NOs;
1-725, SEQ ID NOs;
726-729, SEQ ID NOs;
731-806, SEQ ID NOs.;
808-830, and SEQ ID NOs;
1087-1094, or an RNA transcribed from said target gene;
or(c) causing mortality or stunting in larvae of said Leptinotarsa species by providing in the diet of said larvae at least one polynucleotide comprising at least one silencing element comprising 21 contiguous nucleotides that are complementary to a target gene having a nucleotide sequence selected from the group consisting of;
SEQ ID NO;
730, SEQ ID NO;
807, SEQ ID NOs;
1-725, SEQ ID NOs;
726-729, SEQ ID NOs;
731-806, SEQ ID NOs;
808-830, and SEQ ID NOs;
1087-1094, or an RNA transcribed from said target gene;
or(d) topically applying to said plant a composition comprising at least one polynucleotide comprising a nucleotide sequence that is complementary to at least 21 contiguous nucleotides of a target gene having a nucleotide sequence selected from the group consisting of;
SEQ ID NO;
730, SEQ ID NO;
807, SEQ ID NOs;
1-725, SEQ ID NOs;
726-729, SEQ ID NOs;
731-806, SEQ ID NOs;
808-830, and SEQ ID NOs;
1087-1094, or an RNA transcribed from said target gene;
or(e) topically applying to said plant a composition comprising at least one polynucleotide in a manner such that an effective amount of said polynucleotide is ingested by Leptinotarsa species feeding on said plant, said polynucleotide comprising at least 21 contiguous nucleotides that are complementary to a target gene having a nucleotide sequence selected from the group consisting of;
SEQ ID NO;
730, SEQ ID NO;
807, SEQ ID NOs;
1-725, SEQ ID NOs;
726-729, SEQ ID NOs;
731-806, SEQ ID NOs;
808-830, and SEQ ID NOs;
1087-1094, or an RNA transcribed from said target gene;
or(f) expressing in said plant at least one polynucleotide comprising at least one segment that is identical or complementary to at least 21 contiguous nucleotides of a DNA having a sequence selected from the group consisting of;
SEQ ID NO;
730, SEQ ID NO;
807, SEQ ID NOs;
1-725, SEQ ID NOs;
726-729, SEQ ID NOs;
731-806, SEQ ID NOs;
808-830, and SEQ ID NOs;
1087-1094;
or(g) providing to said plant at least one polynucleotide comprising at least one segment that is identical or complementary to at least 21 contiguous nucleotides of a target gene or an RNA transcribed from said target gene, wherein said target gene is selected from the group consisting of;
the genes identified in the Target Gene Sequences Group or an RNA transcribed from said target gene;
or(h) contacting said Leptinotarsa species with an effective amount of a double-stranded RNA, one strand of which is complementary to at least 21 contiguous nucleotides of a gene that encodes a ribosomal protein, wherein RNA interference is induced and mortality occurs;
or(i) contacting said Leptinotarsa species with a polynucleotide comprising at least one segment that is identical or complementary to at least 21 contiguous nucleotides of a target gene selected from the group consisting of;
the genes identified in the Target Gene Sequences Group or an RNA transcribed from said target gene.
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Abstract
Disclosed herein are methods of controlling insect pests, in particular Leptinotarsa spp. which infest crop plants, and methods of providing plants resistant to such pests. Also disclosed are polynucleotides and recombinant DNA molecules and constructs useful in such methods, insecticidal compositions such as topical sprays containing insecticidal double-stranded RNAs, and solanaceous plants with improved resistance to infestation by Leptinotarsa spp. Further disclosed are methods of selecting target genes for RNAi-mediated silencing and control of Leptinotarsa spp.
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Citations
20 Claims
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1. A method for controlling a Leptinotarsa species infestation of a plant comprising:
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(a) contacting said Leptinotarsa species with a polynucleotide comprising a nucleotide sequence that is complementary to at least 21 contiguous nucleotides of a target gene having a nucleotide sequence selected from the group consisting of;
SEQ ID NO;
730, SEQ ID NO;
807, SEQ ID NOs;
1-725, SEQ ID NOs;
726-729, SEQ ID NOs;
731-806, SEQ ID NOs;
808-830, and SEQ ID NOs;
1087-1094, or an RNA transcribed from said target gene;
or(b) providing in the diet of said Leptinotarsa species a polynucleotide comprising a nucleotide sequence that is complementary to at least 21 contiguous nucleotides of a target gene having a nucleotide sequence selected from the group consisting of;
SEQ ID NO;
730, SEQ ID NO;
807, SEQ ID NOs;
1-725, SEQ ID NOs;
726-729, SEQ ID NOs;
731-806, SEQ ID NOs.;
808-830, and SEQ ID NOs;
1087-1094, or an RNA transcribed from said target gene;
or(c) causing mortality or stunting in larvae of said Leptinotarsa species by providing in the diet of said larvae at least one polynucleotide comprising at least one silencing element comprising 21 contiguous nucleotides that are complementary to a target gene having a nucleotide sequence selected from the group consisting of;
SEQ ID NO;
730, SEQ ID NO;
807, SEQ ID NOs;
1-725, SEQ ID NOs;
726-729, SEQ ID NOs;
731-806, SEQ ID NOs;
808-830, and SEQ ID NOs;
1087-1094, or an RNA transcribed from said target gene;
or(d) topically applying to said plant a composition comprising at least one polynucleotide comprising a nucleotide sequence that is complementary to at least 21 contiguous nucleotides of a target gene having a nucleotide sequence selected from the group consisting of;
SEQ ID NO;
730, SEQ ID NO;
807, SEQ ID NOs;
1-725, SEQ ID NOs;
726-729, SEQ ID NOs;
731-806, SEQ ID NOs;
808-830, and SEQ ID NOs;
1087-1094, or an RNA transcribed from said target gene;
or(e) topically applying to said plant a composition comprising at least one polynucleotide in a manner such that an effective amount of said polynucleotide is ingested by Leptinotarsa species feeding on said plant, said polynucleotide comprising at least 21 contiguous nucleotides that are complementary to a target gene having a nucleotide sequence selected from the group consisting of;
SEQ ID NO;
730, SEQ ID NO;
807, SEQ ID NOs;
1-725, SEQ ID NOs;
726-729, SEQ ID NOs;
731-806, SEQ ID NOs;
808-830, and SEQ ID NOs;
1087-1094, or an RNA transcribed from said target gene;
or(f) expressing in said plant at least one polynucleotide comprising at least one segment that is identical or complementary to at least 21 contiguous nucleotides of a DNA having a sequence selected from the group consisting of;
SEQ ID NO;
730, SEQ ID NO;
807, SEQ ID NOs;
1-725, SEQ ID NOs;
726-729, SEQ ID NOs;
731-806, SEQ ID NOs;
808-830, and SEQ ID NOs;
1087-1094;
or(g) providing to said plant at least one polynucleotide comprising at least one segment that is identical or complementary to at least 21 contiguous nucleotides of a target gene or an RNA transcribed from said target gene, wherein said target gene is selected from the group consisting of;
the genes identified in the Target Gene Sequences Group or an RNA transcribed from said target gene;
or(h) contacting said Leptinotarsa species with an effective amount of a double-stranded RNA, one strand of which is complementary to at least 21 contiguous nucleotides of a gene that encodes a ribosomal protein, wherein RNA interference is induced and mortality occurs;
or(i) contacting said Leptinotarsa species with a polynucleotide comprising at least one segment that is identical or complementary to at least 21 contiguous nucleotides of a target gene selected from the group consisting of;
the genes identified in the Target Gene Sequences Group or an RNA transcribed from said target gene. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11)
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12. An insecticidal composition for controlling a Leptinotarsa species, comprising:
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(a) an insecticidally effective amount of a polynucleotide comprising at least 21 contiguous nucleotides that are complementary to a target gene having a nucleotide sequence selected from the group consisting of;
SEQ ID NO;
730, SEQ ID NO;
807, SEQ ID NOs;
1-725, SEQ ID NOs;
726-729, SEQ ID NOs;
731-806, SEQ ID NOs;
808-830, and SEQ ID NOs;
1087-1094, or an RNA transcribed from said target gene;
or(b) an insecticidally effective amount of at least one polynucleotide comprising at least one silencing element that is complementary to at least 21 contiguous nucleotides of a target gene or an RNA transcribed from said target gene, wherein said target gene has a nucleotide sequence selected from the group consisting of;
SEQ ID NO;
730, SEQ ID NO;
807, SEQ ID NOs;
1-725, SEQ ID NOs;
726-729, SEQ ID NOs;
731-806, SEQ ID NOs;
808-830, and SEQ ID NOs;
1087-1094;
or(c) an insecticidally effective amount of at least one RNA comprising at least one segment that is identical or complementary to at least 21 contiguous nucleotides of a target gene having a nucleotide sequence selected from the group consisting of;
SEQ ID NO;
730, SEQ ID NO;
807, SEQ ID NOs;
1-725, SEQ ID NOs;
726-729, SEQ ID NOs;
731-806, SEQ ID NOs;
808-830, and SEQ ID NOs;
1087-1094, or an RNA transcribed from said target gene;
or(d) an RNA molecule that causes mortality or stunting of growth in a Leptinotarsa species when ingested or contacted by said Leptinotarsa species, wherein said RNA molecule comprises at least 21 contiguous nucleotides that are complementary to a target gene having a nucleotide sequence selected from the group consisting of;
SEQ ID NO;
730, SEQ ID NO;
807, SEQ ID NOs;
1-725, SEQ ID NOs;
726-729, SEQ ID NOs;
731-806, SEQ ID NOs;
808-830, and SEQ ID NOs;
1087-1094, or an RNA transcribed from said target gene;
or(e) an insecticidal double-stranded RNA molecule that causes mortality or stunting of growth in a Leptinotarsa species when ingested or contacted by said Leptinotarsa species, wherein at least one strand of said insecticidal double-stranded RNA molecule comprises 21 contiguous nucleotides that are complementary to a target gene or an RNA transcribed from said target gene, wherein said target gene has a sequence selected from the group consisting of;
SEQ ID NO;
730, SEQ ID NO;
807, SEQ ID NOs;
1-725, SEQ ID NOs;
726-729, SEQ ID NOs;
731-806, SEQ ID NOs;
808-830, and SEQ ID NOs;
1087-1094;
or(f) an insecticidally effective amount of at least one double-stranded RNA comprising a sequence selected from the group consisting of;
SEQ ID NOs;
989, 1049, 831, 842, 849, 898, 910, 925, 928, 931, 932, 937, 938, 940, 941, 942, 943, 944, 945, 947, 948, 949, 950, 951, 952, 955, 956, 957, 958, 960, 961, 964, 966, 967, 968, 969, 970, 971, 973, 976, 978, 979, 982, 983, 985, 987, 988, 991, 992, 994, 995, 996, 997, 999, 1006, 1007, 1008, 1009, 1010, 1013, 1018, 1019, 1020, 1022, 1025, 1029, 1030, 1033, 1035, 1036, 1037, 1038, 1039, 1040, 1041, 1042, 1043, 1045, 1046, 1047, 1050, 1053, 1054, 1058, 1060, 1061, 1064, 1065, 1066, 1067, 1068, 1070, 1073, 1074, 1075, 1077, 1078, 1080, 1081, 1082, 1084, 1085, 1095, 1096, 1097, 1098, 1099, 1100, 1101, 1102, 1103, 1104, 1105, 1110, 1111, 1112, 1113, 1114, 1115, 1116, 1117, 1118, 119, 1120, 1121, 1122, 1123, 1124, 1125, and 1126. - View Dependent Claims (13, 14, 15)
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16. A recombinant DNA construct comprising a heterologous promoter operably linked to:
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(a) DNA comprising a nucleotide sequence that is complementary to at least 21 contiguous nucleotides of a target gene having a sequence selected from the group consisting of;
SEQ ID NO;
730, SEQ ID NO;
807, SEQ ID NOs;
1-725, SEQ ID NOs;
726-729, SEQ ID NOs;
731-806, SEQ ID NOs;
808-830, and SEQ ID NOs;
1087-1094, or an RNA transcribed from said target gene;
or(b) a DNA comprising 21 or more contiguous nucleotides having 100% identity to a fragment of equivalent length of a DNA having a sequence selected from the group consisting of;
SEQ ID NO;
730, SEQ ID NO;
807, SEQ ID NOs;
1-725, SEQ ID NOs;
726-729, SEQ ID NOs;
731-806, SEQ ID NOs;
808-830, and SEQ ID NOs;
1087-1094, or the DNA complement thereof;
or(c) DNA encoding at least one silencing element that is complementary to at least 21 contiguous nucleotides of a target gene or an RNA transcribed from said target gene, wherein said target gene has a sequence selected from the group consisting of;
SEQ ID NO;
730, SEQ ID NO;
807, SEQ ID NOs;
1-725, SEQ ID NOs;
726-729, SEQ ID NOs;
731-806, SEQ ID NOs;
880-830, and SEQ ID NOs;
1087-104;
or(d) DNA encoding at least one silencing element comprising at least 21 contiguous nucleotides that are complementary to a target gene selected from the group consisting of;
the genes in the Target Gene Sequences Group or an RNA transcribed from said target gene;
or(e) DNA encoding a RNA comprising at least 21 contiguous nucleotides that are complementary to a nucleotide sequence selected from the group consisting of;
SEQ ID NOs;
989, 1049, 831, 842, 849, 898, 910, 925, 928, 931, 932, 937, 938, 940, 941, 942, 943, 944, 945, 947, 948, 949, 950, 951, 952, 955, 956, 957, 958, 960, 961, 964, 966, 967, 968, 969, 970, 971, 973, 976, 978, 979, 982, 983, 985, 987, 988, 991, 992, 994, 995, 996, 997, 999, 1006, 1007, 1008, 1009, 1010, 1013, 1018, 1019, 1020, 1022, 1025, 1029, 1030, 1033, 1035, 1036, 1037, 1038, 1039, 1040, 1041, 1042, 1043, 1045, 1046, 1047, 1050, 1053, 1054, 1058, 1060, 1061, 1064, 1065, 1066, 1067, 1068, 1070, 1073, 1074, 1075, 1077, 1078, 1080, 1081, 1082, 1084, 1085, 1095, 1096, 1097, 1098, 1099, 1100, 1101, 1102, 1103, 1104, 1105, 1110, 1111, 1112, 1113, 1114, 1115, 1116, 1117, 118, 1119, 1120, 1121, 1122, 1123, 1124, 1125, and 1126, or the complement thereof, or an orthologous nucleotide sequence from a Leptinotarsa species or a Tribolium species, wherein the orthologous nucleotide sequence has at least 95% sequence identity with a nucleotide sequence selected from the group consisting of;
SEQ ID NOs;
989, 1049, 831, 842, 849, 898, 910, 925, 928, 931, 932, 937, 938, 940, 941, 942, 943, 944, 945, 947, 948, 949, 950, 951, 952, 955, 956, 957, 958, 960, 961, 964, 966, 967, 968, 969, 970, 971, 973, 976, 978, 979, 982, 983, 985, 987, 988, 991, 992, 994, 995, 996, 997, 999, 1006, 1007, 1008, 1009, 1010, 1013, 1018, 1019, 1020, 1022, 1025, 1029, 1030, 1033, 1035, 1036, 1037, 1038, 1039, 1040, 1041, 1042, 1043, 1045, 1046, 1047, 1050, 1053, 1054, 1058, 1060, 1061, 1064, 1065, 1066, 1067, 1068, 1070, 1073, 1074, 1075, 1077, 1078, 1080, 1081, 1082, 1084, 1085, 1095, 1096, 1097, 1098, 1099, 1100, 1101, 1102,1103, 1104, 1105, 1110, 111, 1112, 1113, 1114, 1115, 1116, 1117, 1118, 1119, 1120, 1121, 1122, 1123, 1124, 1125, and 1126, wherein the percentage sequence identity is calculated over the same length;
or(f) DNA encoding a RNA comprising at least one double-stranded RNA region, at least one strand of which comprises at least 21 contiguous nucleotides that are complementary to a nucleotide sequence selected from the group consisting of;
SEQ ID NOs;
989, 1049, 831, 842, 849, 898, 910, 925, 928, 931, 932, 937, 938, 940, 941, 942, 943, 944, 945, 947, 948, 949, 950, 951, 952, 955, 956, 957, 958, 960, 961, 964, 966, 967, 968, 969, 970, 971, 973, 976, 978, 979, 982, 983, 985, 987, 988, 991, 992, 994, 995, 996, 997, 999, 1006, 1007, 1008, 1009, 1010, 1013, 1018, 1019, 1020, 1022, 1025, 1029, 1030, 1033, 1035, 1036, 1037, 1038, 1039, 1040, 1041, 1042, 1043, 1045, 1046, 1047, 1050, 1053, 1054, 1058, 1060, 1061, 1064, 1065, 1066, 1067, 1068, 1070, 1073, 1074, 1075, 1077, 1078, 1080, 1081, 1082, 1084, 1085, 1095, 1096, 1097, 1098, 1099, 1100, 1101, 1102, 1103, 1104, 1105, 1110, 1111, 1112, 1113, 1114, 1115, 1116, 1117, 1118, 1119, 1120, 1121, 1122, 1123, 1124, 1125, and 1126, or the complement thereof, or an orthologous nucleotide sequence from a Leptinotarsa species or a Tribolium species, wherein the orthologous nucleotide sequence has at least 95% sequence identity with a nucleotide sequence selected from the group consisting of SEQ ID NOs;
989, 1049, 831, 842, 849, 898, 910, 925, 928, 931, 932, 937, 938, 940, 941, 942, 943, 944, 945, 947, 948, 949, 950, 951, 952, 955, 956, 957, 958, 960, 961, 964, 966, 967, 968, 969, 970, 971, 973, 976, 978, 979, 982, 983, 985, 987, 988, 991, 992, 994, 995, 996, 997, 999, 1006, 1007, 1008, 1009, 1010, 1013, 1018, 1019, 1020, 1022, 1025, 1029, 1030, 1033, 1035, 1036, 1037, 1038, 1039, 1040, 1041 1042, 1043, 1045, 1046, 1047, 1050, 1053, 1054, 105, 1060, 1061, 1064, 1065, 1066, 1067, 1068, 1070, 1073, 1074, 1075, 1077, 1078, 1080, 1081, 1082, 1084, 1085, 1095, 95, 1096, 1097, 1098, 1099, 1100, 1101, 1102, 1103, 1104, 1105, 1110, 1111, 1112, 1113, 1114, 1115, 1116, 1117, 1118, 1119, 1120, 1121, 1122, 1123, 1124, 1125, and 1126, wherein the percentage sequence identity is calculated over the same length;
or(g) DNA encoding RNA comprising a nucleotide sequence selected from the group consisting of;
SEQ ID NOs;
989, 1049, 831, 842, 849, 898, 910, 925, 928, 931, 932, 937, 938, 940, 941, 942, 943, 944, 945, 947, 948, 949, 950, 951, 952, 955, 956, 97, 958, 960, 961, 964, 966, 967, 968, 969, 970, 971, 973, 976, 978, 979, 982, 983, 985, 987, 988, 991, 992, 994, 995, 996, 997, 999, 1006, 1007, 1008, 1009, 1010, 1013, 1018, 1019, 1020, 1022, 1025, 1029, 1030, 1033, 1035, 1036, 1037, 1038, 1039, 1040, 1041, 1042, 1043, 1045, 1046, 1047, 1050, 1053, 1054, 1058, 1060, 1061, 1064, 1065, 1066, 1067, 1068, 1070, 1073, 1074, 1075, 1077, 1078, 1080, 1081, 1082, 1084, 1085, 1095, 1096, 1097, 1098, 1099, 1100, 1101, 1102, 1103, 1104, 1105, 1110, 1111, 1112, 1113, 1114, 1115, 1116, 1117, 111, 1119, 1120, 1121, 1122, 1123, 1124, 1125, and 1126, or the complement thereof. - View Dependent Claims (17, 18, 19, 20)
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Specification