Plant Centromere Compositions

Abstract

The present invention provides for the nucleic acid sequences of plant centromeres. This will permit construction of stably inherited recombinant DNA constructs and minichromosomes which can serve as vectors for the construction of transgenic plant and animal cells.

1 - 127 . (canceled) 
     
     
         128 . An isolated minichromosome comprising a centromere from sugarcane genomic DNA and at least one structural gene,
 wherein the centromere (i) confers the ability to segregate to daughter cells, and (ii) comprises at least five repeated nucleotide sequences, and wherein the repeated nucleotide sequence is about 75 base pairs to about 210 base pairs.   
     
     
         129 . The isolated minichromosome of  claim 128 , wherein the minichromosome further comprises a site for site-specific recombination. 
     
     
         130 . The isolated minichromosome of  claim 128 , wherein at least one structural gene is operably linked to a heterologous regulatory sequence functional in plant cells. 
     
     
         131 . The isolated minichromosome of  claim 131 , wherein at least one structural gene is operably linked to a plant promoter. 
     
     
         132 . The minichromosome of  claim 128 , wherein the structural gene is selected from the group consisting of a herbicide resistance gene, a nitrogen fixation gene, an insect resistance gene, a disease resistance gene, a plant stress-induced gene, a nutrient utilization gene, a gene that affects plant pigmentation, a gene that encodes an antisense or ribozyme molecule, a gene encoding a secretable antigen, a toxin gene, a receptor gene, a ligand gene, a seed storage gene, a hormone gene, an enzyme gene, an interleukin gene, a clotting factor gene, a cytokine gene, an antibody gene, and a growth factor gene. 
     
     
         133 . The minichromosome of  claim 132 , wherein the disease resistance gene confers resistance to a virus, bacteria, fungi or nematode. 
     
     
         134 . The minichromosome of  claim 132 , wherein the enzyme gene is selected from the group consisting of a gene that encodes an enzyme involved in metabolizing biochemical wastes for use in bioremediation, a gene that encodes an enzyme for modifying pathways that produce secondary plant metabolites, a gene that encodes an enzyme that produces a pharmaceutical, a gene that encodes an enzyme that improves the nutritional content of a plant, a gene that encodes an enzyme involved in vitamin synthesis, a gene that encodes an enzyme involved in carbohydrate or starch synthesis, a gene that encodes an enzyme involved in mineral accumulation or availability, a gene that encodes a phytase, a gene that encodes an enzyme involved in fatty acid or oil synthesis, a gene that encodes an enzyme involved in synthesis of chemicals or plastics, a gene that encodes an enzyme involved in synthesis of a fuel and a gene that encodes enzyme involved in synthesis of a fragrance. 
     
     
         135 . The minichromosome of  claim 128 , wherein the structural gene encodes a protein conferring resistance to drought, heat, chilling, freezing, excessive moisture, or salt stress. 
     
     
         136 . The minichromosome of  claim 128 , wherein the structural gene is a glyphosate resistance gene. 
     
     
         137 . The minichromosome of  claim 128 , wherein the structural gene is a  Bacillus thuringiensis  toxin gene. 
     
     
         138 . The minichromosome of  claim 128 , wherein the centromere comprises n copies of the repeated nucleotide sequence, wherein n is less than 1000. 
     
     
         139 . The minichromosome of  claim 128 , wherein the centromere comprises n copies of the repeated nucleotide sequence, wherein n is at least 15. 
     
     
         140 . The minichromosome of  claim 128 , wherein the centromere comprises n copies of the repeated nucleotide sequence, wherein n is at least 50. 
     
     
         141 . The minichromosome of  claim 128 , wherein the minichromosome is circular. 
     
     
         142 . The minichromosome of  claim 128  comprising a plurality of structural genes associated with at least two different agronomic traits. 
     
     
         143 . The minichromosome of  claim 142 , wherein the minichromosome comprises at least three structural genes. 
     
     
         144 . The minichromosome of  claim 143  wherein the minichromosome comprises at least five structural genes. 
     
     
         145 . The minichromosome of  claim 144  wherein the minichromosome comprises at least ten structural genes. 
     
     
         146 . The minichromosome of  claim 128 , wherein segregation efficiency of the minichromosome through meiotic or mitotic division in sugarcane cells is at least 60%. 
     
     
         147 . The minichromosome of  claim 146 , wherein segregation efficiency of the minichromosome through meiotic or mitotic division in sugarcane cells is at least 80%. 
     
     
         148 . The minichromosome of  claim 146 , wherein segregation efficiency of the minichromosome through meiotic or mitotic division in sugarcane cells is at least 90%. 
     
     
         149 . The minichromosome of  claim 146 , wherein segregation efficiency of the minichromosome through meiotic or mitotic division in sugarcane cells is at least 95%. 
     
     
         150 . A sugarcane plant cell comprising a minichromosome of  claim 128  that (i) is not integrated into the plant cell genome and (ii) confers an altered phenotype on the plant cell associated with at least one structural gene within the minichromosome. 
     
     
         151 . Plant tissue comprising the sugarcane plant cell of  claim 150 . 
     
     
         152 . A sugarcane plant comprising the sugarcane plant cell of  claim 150 . 
     
     
         153 . A sugarcane plant part comprising the sugarcane plant cell of  claim 150 . 
     
     
         154 . A sugarcane seed comprising the sugarcane plant cell of  claim 150 .