Plant vectors, compositions and uses relating thereto
The present disclosure relates to a single stranded RNA vector suitable for introducing a therapeutic agent, such as a peptide, a protein or a small RNA, into a host plant. The vector does not encode for any movement protein or coat protein, but is capable of capable of systemic and phloem-limited movement and replication within the host plant.
1 . A plus sense single stranded ribonucleic acid (RNA) vector consisting essentially of SEQ ID NO:1 with one or more heterologous segment(s) inserted therein.
2 . The RNA vector of claim 1 wherein the heterologous segment(s) is located between an ORF encoding an RdRp and a 3′ CITE of SEQ ID NO:1.
3 . The RNA vector of claim 2 , wherein said 3′ CITE comprises the nucleic acid sequence(s) of SEQ ID NO: 4 and/or SEQ ID NO: 5.
4 . The RNA vector of claim 2 , wherein said 3′ CITE comprises the nucleic acid sequence of SEQ ID NO: 3.
5 . The RNA vector of claim 1 , which is functionally stable for replication, movement and/or translation within the host plant for at least one month after infection thereof.
6 . The RNA vector of claim 1 , wherein said heterologous segment(s) comprises a polynucleotide that encodes at least one polypeptide selected from the group consisting of a reporter molecule, a peptide, and a protein.
7 . The RNA vector of claim 6 , wherein said polypeptide is an insecticide, an antibacterial, an antiviral, or an antifungal.
8 . The RNA vector of claim 7 , wherein said antibacterial is an enzybiotic.
9 . The RNA vector of claim 7 , wherein said antibacterial targets a bacterium Candidatus Liberibacter species.
10 . The RNA vector of claim 9 , wherein said Candidatus Liberibacter species is Candidatus Liberibacter asiaticus (CLas).
11 . The RNA vector of claim 1 , wherein said heterologous segment(s) comprises a small non-coding RNA molecule and/or an RNA interfering molecule.
12 . The RNA vector of claim 11 , wherein said small non-coding RNA molecule and/or said RNA interfering molecule targets an insect vector, a virus, or a fungus.
13 . The RNA vector of claim 12 , wherein said small non-coding RNA molecule and/or said RNA interfering molecule targets a nucleic acid of said insect vector, said virus, or said fungus.
14 . The RNA vector of claim 12 , wherein said targeted virus is selected from the group consisting of Citrus vein enation virus (CVEV) and Citrus tristeza virus (CTV).
15 . The RNA vector of claim 1 , wherein said heterologous segment(s) comprises a polynucleotide that encodes for a protein or peptide that alters a phenotypic trait.
16 . The RNA vector of claim 15 , wherein said phenotypic trait is selected from the group consisting of pesticide tolerance, herbicide tolerance, insect resistance, reduced callose production, increased growth rate, and dwarfism.
17 . A host plant comprising the RNA vector of claim 1 , wherein said host plant is a whole plant, a plant organ, a plant tissue, or a plant cell.
18 . The host plant of claim 17 , wherein said host plant is in a genus selected from the group consisting of citrus, vitis, ficus and olea.
19 . The host plant of claim 18 , wherein said host plant is a citrus tree or a citrus tree graft.
20 . A method for introducing a heterologous segment(s) into a host plant comprising introducing into said host plant the RNA vector of claim 1 .
21 . The method of claim 20 , wherein said introducing step comprises grafting a plant organ or plant tissue of a plant that comprises the RNA vector to a plant organ or plant tissue of another plant that does not comprise the RNA vector prior to said introduction.
22 . The method of claim 20 , wherein the RNA vector systemically infects the host plant.
23 . The method of claim 20 , wherein said host plant is in a genus selected from the group consisting of citrus, vitis, ficus and olea.
24 . The RNA vector of claim 1 , wherein the ORF that encodes a RNA-dependent RNA polymerase (RdRp) comprises conserved polynucleotide sequences having the nucleic acid sequence of: SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13and SEQ ID NO:14.
25 . The RNA vector of claim 24 , wherein said ORF further comprises conserved polynucleotide sequences having the nucleic acid sequence of: SEQ ID NO:15 and SEQ ID NO:16.
26 . The RNA vector of claim 1 , comprising conserved polynucleotide sequences having the nucleic acid sequence of: SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO:15 and/or SEQ ID NO:16.
27 . The RNA vector of claim 1 , comprising conserved polynucleotide sequences having the nucleic acid sequence of: SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO:15 and SEQ ID NO:16.
28 . The RNA vector of claim 1 , wherein the heterologous segment(s) is inserted between positions corresponding to positions 2158 and 2468 of SEQ ID NO:1.