Plants modified with mini-chromosomes
The invention is generally related to methods of generating plants transformed with novel autonomous mini-chromosomes. Mini-chromosomes with novel compositions and structures are used to transform plants cells which are in turn used to generate the plant. Methods for generating the plant include methods for delivering the mini-chromosome into plant cell to transform the cell, methods for selecting the transformed cell, and methods for isolating plants transformed with the mini-chromosome. Plants generated in the present invention contain novel genes introduced into their genome by integration into existing chromosomes.
1. A plant comprising at least one functional, stable, and autonomous mini-chromosome, wherein the mini-chromosome comprises a plant centromere, wherein the centromere (i) comprises a polynucleotide comprising at least 5 repeated nucleotide sequences, each comprising a nucleic acid sequence having at least 95% sequence identity with SEQ ID NO:24, and (ii) confers an ability to segregate the mini-chromosome to daughter cells through cell division.
2. The plant according to claim 1 , wherein the mini-chromosome is 1000 kilobases or less in length.
3. The plant according to claim 1 , wherein the mini-chromosome comprises a site for site-specific recombination.
4. The plant according to claim 1 , wherein the mini-chromosome comprises a centromeric nucleic acid insert derived from a crop plant centromere.
5. The plant according to claim 1 , wherein the mini-chromosome comprises a centromeric nucleic acid insert that comprises artificially synthesized repeated nucleotide sequences.
6. The plant according to claim 1 , wherein the mini-chromosome is derived from a donor clone or a centromere clone and has substitutions, deletions, insertions, duplications or arrangements of one or more nucleotides in the mini-chromosome compared to the nucleotide sequence of the donor clone or centromere clone.
7. The plant of claim 1 , wherein the mini-chromosome is obtained by passage of the mini-chromosome through one or more hosts.
8. The plant according to claim 1 , wherein the mini-chromosome comprises one or more exogenous nucleic acids.
9. The plant according to claim 8 , wherein at least one exogenous nucleic acid is operably linked to a heterologous regulatory sequence functional in plant cells.
10. The plant according to claim 1 , wherein the mini-chromosome comprises an exogenous nucleic acid that confers herbicide resistance, insect resistance, disease resistance, or stress resistance on the plant.
11. The plant according to claim 1 , wherein the centromere of the mini-chromosome comprises n copies of a repeated nucleotide sequence, wherein n is less than 1000.
12. The plant according claim 1 , wherein the mini-chromosome comprises a telomere.
13. The plant according to claim 1 , wherein the mini-chromosome is circular.
14. The plant according to claim 1 , wherein the plant is a monocotyledon.
15. The plant according to claim 1 , wherein the plant is a dicotyledone.
16. A part of the plant according to claim 1 , wherein the plant part comprises the minichromosome.
17. A meiocyte, gamete, ovule, pollen, or endosperm of the plant according to claim 1 , wherein the meiocyte, gamete, ovule, pollen or endosperm comprises the minichromosome.
18. A seed, embryo or propagule of the plant according to claim 1 , wherein the seed, embryo or propagule comprises the minichromosome.
19. A progeny of the plant according to claim 1 , wherein the progeny comprises the minichromosome.
20. The progeny of claim 19 wherein the progeny is the result of self-breeding.
21. The progeny of claim 19 wherein the progeny is the result of cross-breeding.
22. The progeny of claim 19 wherein the progeny is the result of apomyxis.
23. The progeny of claim 19 wherein the progeny is the result of clonal propagation.
24. A method of using a plant according to claim 1 to produce a food product comprising the steps of growing the plant, and harvesting or processing the plant.
25. A method of using a plant according to claim 1 , to produce a recombinant protein comprising the step of growing a plant comprising a mini-chromosome that comprises an exogenous nucleic acid encoding the recombinant protein.
26. A method of using a plant according to claim 1 to produce a chemical product comprising the step of growing a plant comprising a mini-chromosome that comprises an exogenous nucleic acid encoding an enzyme involved in synthesis of the chemical product.