The disclosure provides nucleic acids, and variants and fragments thereof, obtained from strains of Bacillus thuringiensis encoding polypeptides having pesticidal activity against insect pests, including Lepidoptera. Particular embodiments of the disclosure provide isolated nucleic acids encoding pesticidal proteins, pesticidal compositions, DNA constructs, and transformed microorganisms and plants comprising a nucleic acid of the embodiments. These compositions find use in methods for controlling pests, especially plant pests.

Disclosed are double stranded RNA (dsRNA) molecules that are toxic to coleopteran and/or hemipteran insect pests. In particular, interfering RNA molecules capable of interfering with pest insect target genes and that are toxic to the target insect pest are provided. Further, methods of making and using the interfering RNA, for example in transgenic plants or as the active ingredient in an insecticidal composition, to confer protection from insect damage are disclosed.

Disclosed are double stranded RNA (dsRNA) molecules that are toxic to coleopteran and/or hemipteran insect pests. In particular, interfering RNA molecules capable of interfering with pest insect target genes and that are toxic to the target insect pest are provided. Further, methods of making and using the interfering RNA, for example in transgenic plants or as the active ingredient in an insecticidal composition, to confer protection from insect damage are disclosed.

Insect control nanobodies are provided. Accordingly there is provided a nanobody which specifically binds to an insect polypeptide selected from the group consisting of: a polypeptide comprising a chitin binding domain (CBD), V-ATPase subunit c, trehalase, cytochrome p450 monooxygenase, chitin deacetylase, chitin synthase and NPC1 sterol transporter, wherein binding of the nanobody to the insect polypeptide confers an insect control activity to the nanobody. Also provided are polynucleotides encoding the nanobody, host cells expressing the nanobody and methods of using it.

Compositions for providing stability to RNA may include a primary surfactant, and a metal-ion sequestrant. The primary surfactant may be a nonionic surfactant. The composition may be in a soluble liquid concentrate form and may be sufficient to provide shelf stability to RNA for one year at room temperature. Compositions for delivering RNA to a pest via exogenous, foliar application of the composition to a plant may comprise RNA; a primary surfactant; and a metal-ion sequestrant.

Compositions for providing stability to RNA may include a primary surfactant, and a metal-ion sequestrant. The primary surfactant may be a nonionic surfactant. The composition may be in a soluble liquid concentrate form and may be sufficient to provide shelf stability to RNA for one year at room temperature. Compositions for delivering RNA to a pest via exogenous, foliar application of the composition to a plant may comprise RNA; a primary surfactant; and a metal-ion sequestrant.

Provided are an isolated polynucleotide and a method for controlling insect invasion. The isolated polynucleotide is a plurality of target sequences for controlling target gene c35112 of a coleopteran pest, Monolepta hieroglyphica, comprising: a) a polynucleotide sequence shown as SEQ ID NO: 1; or (b) a polynucleotide sequence having at least 15 or 17 or 19 or 21 contiguous nucleotides of SEQ ID NO: 1, a double-stranded RNA comprising at least one strand complementary to the polynucleotide sequence being capable of inhibiting the growth of coleopteran pests after being ingested by the coleopteran pests; or (c) any one of polynucleotide sequences shown as SEQ ID NO: 3 to SEQ ID NO: 20; or (d) a polynucleotide sequence that hybridizes under stringent conditions to a polynucleotide sequence as defined in (a), (b) or (c).

Provided are an isolated polynucleotide and a method for controlling insect invasion. The isolated polynucleotide is a plurality of target sequences for controlling target gene c35112 of a coleopteran pest, Monolepta hieroglyphica, comprising: a) a polynucleotide sequence shown as SEQ ID NO: 1; or (b) a polynucleotide sequence having at least 15 or 17 or 19 or 21 contiguous nucleotides of SEQ ID NO: 1, a double-stranded RNA comprising at least one strand complementary to the polynucleotide sequence being capable of inhibiting the growth of coleopteran pests after being ingested by the coleopteran pests; or (c) any one of polynucleotide sequences shown as SEQ ID NO: 3 to SEQ ID NO: 20; or (d) a polynucleotide sequence that hybridizes under stringent conditions to a polynucleotide sequence as defined in (a), (b) or (c).

Nucleic acid delivery vehicles for delivering nucleic acid, e.g., for RNAi to cells that are typically refractory to RNAi by using extracellular vesicles (EVs) from cultured beetle cells as delivery vehicles. Instead of using high levels of long dsRNA and transfection reagents to accomplish suppression of an mRNA target in cells that don't respond to treatment with naked dsRNA, this approach applies the dsRNA to cultured beetle cells, collects nucleic-acid loaded EVs from the culture, then treats our target cells with the intracellularly loaded EVs, which results in significant enhancement of the RNAi response and greater suppression of transcript levels.

Nucleic acid delivery vehicles for delivering nucleic acid, e.g., for RNAi to cells that are typically refractory to RNAi by using extracellular vesicles (EVs) from cultured beetle cells as delivery vehicles. Instead of using high levels of long dsRNA and transfection reagents to accomplish suppression of an mRNA target in cells that don't respond to treatment with naked dsRNA, this approach applies the dsRNA to cultured beetle cells, collects nucleic-acid loaded EVs from the culture, then treats our target cells with the intracellularly loaded EVs, which results in significant enhancement of the RNAi response and greater suppression of transcript levels.