Isolated nucleic acid molecules from the genome of citrus leprosis virus and uses thereof
The present disclosure embraces nucleic acid molecules found in the genome of the Citrus Leprosis Virus (CiLV), which is associated to Citrus Leprosis (CiL) disease. The cloned CiLV nucleic acid molecules can be used as probes or can be used to design oligonucleotide primers for detecting CiLV in biological samples, particularly leaves, roots and other tissues or organs of plants, such as plants from the genera Citrus and Poncirus . The cloned CiLV nucleic acid molecules are expressed in cells to provide immunogenic proteins for raising antibodies against CiLV, which can then be used to detect CiLV in biological samples. It also comprises the nucleic acid molecules represented in SEQ ID Nos. 5 and 8, in whole or part, as well as transgenic plants, such as monocots and dicots, containing the CiLV nucleic acid molecules, in any kind of combination, so that expression increases resistance to CiL disease.
1. A recombinant expression vector comprising an isolated cDNA molecule which encodes a full length citrus leprosis virus protein, the complementary sequence of which hybridizes, under stringent conditions, over the full length sequence set forth in SEQ ID NO: 5, operably linked to a heterologous promoter, wherein the isolated cDNA molecule encodes a protein having the amino acid sequence set forth in SEQ ID NO: 7.
2. The recombinant expression vector of claim 1 , wherein the isolated cDNA molecule comprises the isolated nucleic acid molecule, and at least one other nucleic acid molecule.
3. An isolated, recombinant cell, transformed or transfected with the recombinant expression vector of claim 1 .
4. The isolated, recombinant cell of claim 3 , wherein said cell is a eukaryotic cell.
5. The isolated, recombinant cell of claim 4 , wherein said eukaryotic cell is a plant cell.
6. The isolated recombinant cell of claim 5 , wherein said plant cell is a Citrus cell or Poncirus cell.
7. The isolated recombinant cell of claim 5 , wherein said plant cell is a dicot cell.
8. A transgenic plant or plant part comprising the recombinant expression vector of claim 1 .
9. The transgenic plant of claim 8 , wherein said plant is selected from a Citrus plant, Poncirus plant, or other plant used in a citrus breeding program.
10. The transgenic plant of claim 8 , wherein said plant is a dicot.
11. The transgenic plant part of claim 8 , wherein said plant part is a leaf, flower, stem, root, tuber, fruit, pollen, or seed.
12. The transgenic plant of claim 10 , wherein said plant is a citrus plant.
13. A method for determining if a plant is susceptible to citrus leprosis, comprising contacting a sample of said plant with the recombinant expression vector, and determining hybridization of said recombinant expression vector to a target as a determination of susceptibility to citrus leprosis.
14. The method of claim 13 , comprising polymerase chain reaction.
15. A recombinant expression vector comprising an isolated cDNA molecule encoding a citrus leprosis virus protein, wherein said nucleic acid molecule has the full length sequence set forth in SEQ ID NO: 5, operably linked to a heterologous promoter.
16. A recombinant expression vector comprising an isolated cDNA molecule which encodes a citrus leprosis virus protein, having the amino acid sequence recited in SEQ ID NO: 7, operably linked to a heterologous promoter.
17. An isolated, recombinant cell comprising a recombinant expression vector of claim 15 .
18. The isolated, recombinant cell of claim 17 , wherein said cell is a plant cell.
19. The isolated, recombinant cell of claim 17 , wherein said cell is a plant cell is from a Citrus plant, Poncirus plant, or other plant used in a citrus breeding program.
20. The recombinant expression vector of claim 1 , wherein the cDNA molecule encodes a protein having the amino acid sequence set forth in SEQ ID NO: 6.