Biological controls of coleopteran pests
AssigneeSyngenta Participations AG
InventorsKevin Donohue, Renshui Liu, Jeng Shong Chen
Life of this patent
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Abstract
Disclosed are double stranded RNA molecules that are toxic to coleopteran insects. In particular, dsRNA molecules that capable of interfering with pest IAP genes and that are toxic to the target pest are provided. Further, methods of making and using the interfering RNA, for example in transgenic plants to confer protection from insect damage are disclosed.
Claims37
A transgenic plant, or part thereof, comprising a nucleic acid molecule that is or is capable of producing a double stranded (dsRNA) molecule comprising a sense strand and an antisense strand, wherein:
- a) at least 20 consecutive nucleotides of the antisense strand is complementary to a portion of a mRNA polynucleotide transcribable from a Diabrotica insect IAP gene, wherein the mRNA polynucleotide comprises at least 20 consecutive nucleotides of SEQ ID NO:
- 3, SEQ ID NO:
- 12, SEQ ID NO:
- SEQ ID NO:
- 15, or SEQ ID NO:
- 18;
- or
- b) at least 200 consecutive nucleotides of the antisense strand is at least 90% complementary to a portion of a mRNA polynucleotide transcribable from a Diabrotica insect IAP gene, wherein the mRNA polynucleotide comprises at least 200 consecutive nucleotides of SEQ ID NO:
- 3, SEQ ID NO:
- 12, SEQ ID NO:
- SEQ ID NO:
- 15, or SEQ ID NO:
- 18,
- wherein the dsRNA molecule is toxic to a Diabrotica insect.
A method of controlling a Diabrotica insect comprising contacting the Diabrotica insect with a nucleic acid molecule that is or is capable of producing an interfering RNA for inhibiting expression of an IAP gene in the Diabrotica insect thereby controlling the Diabrotica insect.
A method of reducing an adult Diabrotica insect population on a transgenic plant expressing a Cry protein, a hybrid Cry protein or modified Cry protein comprising expressing in the transgenic plant a nucleic acid molecule that is or is capable of producing an interfering RNA capable of inhibiting expression of an IAP gene in an adult Diabrotica insect thereby reducing the adult Diabrotica insect population.
A method of reducing resistance development in a Diabrotica insect population to an interfering RNA capable of inhibiting expression of an IAP gene in a Diabrotica insect, the method comprising expressing in a transgenic plant fed upon by the Diabrotica insect population an interfering RNA that is capable of inhibiting expression of an IAP gene in a larval and adult Diabrotica insect, thereby reducing resistance development in the Diabrotica insect population compared to a Diabrotica insect population exposed to an interfering RNA capable of inhibiting expression of an IAP gene in a larval or adult Diabrotica insect.
A method of reducing the level of a target RNA transcribed from a IAP gene in a Diabrotica insect comprising contacting the Diabrotica insect with a composition comprising a nucleic acid construct that is or is capable of producing a dsRNA molecule comprising a sense strand and an antisense strand, wherein:
- a) at least 20 consecutive nucleotides of the antisense strand is complementary to a portion of a mRNA polynucleotide transcribable from a Diabrotica insect IAP gene wherein the mRNA polynucleotide comprises at least 20 consecutive nucleotides of SEQ ID NO:
- 3, SEQ ID NO:
- 12, SEQ ID NO:
- 15, or SEQ ID NO:
- 18, or
- b) at least 200 consecutive nucleotides of the antisense strand is at least 90$ complementary to a portion of a mRNA polynucleotide transcribable from a Diabrotica insect IAP gene, wherein the mRNA polynucleotide comprises at least 200 consecutive nucleotides of SEQ ID NO:
- 3, SEQ ID NO:
- 12, SEQ ID NO:
- SEQ ID NO:
- 15, or SEQ ID NO:
- 18,
- wherein the dsRNA molecule reduces the level of the target RNA in a cell of the Diabrotica insect.
A method of conferring Diabrotica insect tolerance to a plant, or part thereof, comprising introducing into the plant, or part thereof, a nucleic acid molecule that is or is capable of producing a dsRNA molecule comprising a sense strand and an antisense strand, wherein:
- a) at least 20 consecutive nucleotides of the antisense strand is complementary to a portion of a mRNA polynucleotide transcribable from a Diabrotica insect IAP gene wherein the mRNA polynucleotide comprises at least 20 consecutive nucleotides of SEQ ID NO:
- 3, SEQ ID NO:
- 12, SEQ ID NO:
- 15, or SEQ ID NO:
- 18, or
- b) at least 200 consecutive nucleotides of the antisense strand is at least 90% complementary to a portion of a mRNA polynucleotide transcribable from a Diabrotica insect IAP gene, wherein the mRNA polynucleotide comprises at least 200 consecutive nucleotides of SEQ ID NO:
- 3, SEQ ID NO:
- 12, SEQ ID NO:
- SEQ ID NO:
- 15, or SEQ ID NO:
- 18,
- thereby conferring tolerance of the plant or part thereof to the Diabrotica insect.
A method of reducing root damage to a plant fed upon by a Diabrotica insect, comprising introducing into cells of the plant a nucleic acid molecule that is or is capable of producing a dsRNA molecule comprising a sense strand and an antisense strand, wherein:
- a) at least 20 consecutive nucleotides of the antisense strand is complementary to a portion of a mRNA polynucleotide transcribable from a Diabrotica insect IAP gene wherein the mRNA polynucleotide comprises at least 20 consecutive nucleotides of SEQ ID NO:
- 3, SEQ ID NO:
- 12, SEQ ID NO:
- 15, or SEQ ID NO:
- 18, or
- b) at least 200 consecutive nucleotides of the antisense strand is at least 90% complementary to a portion of a mRNA polynucleotide transcribable from a Diabrotica insect IAP gene, wherein the mRNA polynucleotide comprises at least 200 consecutive nucleotides of SEQ ID NO:
- 3, SEQ ID NO:
- 12, SEQ ID NO:
- SEQ ID NO:
- 15, or SEQ ID NO:
- 18,
- thereby reducing root damage to the plant.
A method of producing a transgenic plant cell having toxicity to a Diabrotica insect, comprising introducing into a plant cell a nucleic acid molecule that is or is capable of producing a dsRNA molecule comprising a sense strand and an antisense strand, wherein:
- a) at least 20 consecutive nucleotides of the antisense strand is complementary to a portion of a mRNA polynucleotide transcribable from a Diabrotica insect IAP gene wherein the mRNA polynucleotide comprises at least 20 consecutive nucleotides of SEQ ID NO:
- 3, SEQ ID NO:
- 12, SEQ ID NO:
- 15, or SEQ ID NO:
- 18, or
- b) at least 200 consecutive nucleotides of the antisense strand is at least 90% complementary to a portion of a mRNA polynucleotide transcribable from a Diabrotica insect IAP gene, wherein the mRNA polynucleotide comprises at least 200 consecutive nucleotides of SEQ ID NO:
- 3, SEQ ID NO:
- 12, SEQ ID NO:
- SEQ ID NO:
- 15, or SEQ ID NO:
- 18,
- thereby producing the transgenic plant cell having toxicity to the Diabrotica insect compared to a control plant cell.
A method of producing a transgenic plant having enhanced tolerance to Diabrotica insect feeding damage, comprising introducing into a plant a nucleic acid molecule that is or is capable of producing a dsRNA molecule comprising a sense strand and an antisense strand, wherein:
- a) at least 20 consecutive nucleotides of the antisense strand is complementary to a portion of a mRNA polynucleotide transcribable from a Diabrotica insect IAP gene wherein the mRNA polynucleotide comprises at least 20 consecutive nucleotides of SEQ ID NO:
- 3, SEQ ID NO:
- 12, SEQ ID NO:
- 15, or SEQ ID NO:
- 18, or
- b) at least 200 consecutive nucleotides of the antisense strand is at least 90% complementary to a portion of a mRNA polynucleotide transcribable from a Diabrotica insect IAP gene, wherein the mRNA polynucleotide comprises at least 200 consecutive nucleotides of SEQ ID NO:
- 3, SEQ ID NO:
- 12, SEQ ID NO:
- SEQ ID NO:
- 15, or SEQ ID NO:
- 18,
- thereby producing a transgenic plant having enhanced tolerance to Diabrotica insect feeding damage compared to a control plant.
Dependencies are inferred from the claim text.