Genetically engineered duckweed

Abstract

Methods and compositions for the efficient transformation of duckweed are provided. Preferably, the methods involve transformation by either ballistic bombardment or Agrobacterium. In this manner, any gene or nucleic acid of interest can be introduced and expressed in duckweed plants. Transformed duckweed plants, cells, tissues are also provided. Transformed duckweed plant tissue culture and methods of producing recombinant proteins and peptides from transformed duckweed plants are also disclosed.

That which is claimed is:  
     
         1 . A stably transformed duckweed plant comprising a heterologous nucleic acid of interest incorporated in its genome.  
     
     
         2 . The stably transformed duckweed plant according to  claim 1 , wherein said duckweed plant comprises fewer than 5 copies of said heterologous nucleic acid of interest.  
     
     
         3 . The stably transformed duckweed plant according to  claim 1 , wherein said duckweed plant is selected from the group consisting of the genus Spirodela, genus Wolffia, genus Wolfiella, and genus Lemna.  
     
     
         4 . The stably transformed duckweed plant according to  claim 3 , wherein said duckweed plant is selected from the genus Lemna.  
     
     
         5 . The stably transformed duckweed plant according to  claim 4 , wherein said duckweed plant is selected from the group consisting of a species of  Lemna minor,  a species of  Lemna miniscula,  and a species of  Lemna gibba.    
     
     
         6 . The stably transformed duckweed plant according to  claim 5 , wherein said duckweed plant is from a species of  Lemna minor.    
     
     
         7 . The stably transformed duckweed plant according to  claim 1 , wherein said nucleic acid comprises at least one expression cassette comprising a gene which confers resistance to a selection agent.  
     
     
         8 . The stably transformed duckweed plant according to  claim 7 , wherein said gene which confers resistance to a selection agent is selected from the group consisting of neo, bar, pat, ALS, HPH, HYG, EPSP and Hml.  
     
     
         9 . The stably transformed duckweed plant according to  claim 1 , wherein said nucleic acid comprises two genes of interest.  
     
     
         10 . The stably transformed duckweed plant according to  claim 1 , wherein said nucleic acid encodes a protein or peptide selected from the group consisting of insulin, growth hormone, α-interferon, β-glucocerebrosidase, retinoblastoma protein, p53 protein, angiostatin, leptin, and serum albumin.  
     
     
         11 . The stably transformed duckweed plant according to  claim 1 , wherein said nucleic acid encodes at least one protein or peptide subunit of a multimeric protein selected from the group consisting of hemoglobin, collagen, P450 oxidase, and a monoclonal antibody.  
     
     
         12 . The stably transformed duckweed plant according to  claim 1 , wherein said nucleic acid encodes a secreted protein or peptide.  
     
     
         13 . A stably transformed duckweed plant tissue comprising a heterologous nucleic acid of interest incorporated in its genome.  
     
     
         14 . The stably transformed duckweed plant tissue according to  claim 13 , wherein said plant tissue is meristematic tissue.  
     
     
         15 . The stably transformed duckweed plant tissue according to  claim 13 , wherein said plant tissue is frond tissue.  
     
     
         16 . The stably transformed duckweed plant tissue according to  claim 13 , wherein said plant tissue is callus tissue.  
     
     
         17 . The stably transformed duckweed plant tissue according to  claim 16 , wherein said plant tissue is Type I callus tissue.  
     
     
         18 . A duckweed tissue culture comprising the stably transformed duckweed plant tissue of  claim 13 .  
     
     
         19 . A stably transformed duckweed cell comprising a heterologous nucleic acid of interest incorporated in its genome.  
     
     
         20 . A stably transformed duckweed plant comprising a chimeric nucleic acid of interest incorporated in its genome, wherein said chimeric nucleic acid comprises a coding sequence operably linked to a transcription initiation region that is heterologous to said coding sequence.  
     
     
         21 . The stably transformed duckweed plant according to  claim 20 , wherein said chimeric nucleic acid comprises a duckweed coding sequence operably linked to a transcription initiation region that is heterologous to said coding sequence.  
     
     
         22 . The stably transformed duckweed plant accordingly to  claim 20 , wherein said chimeric nucleic acid is flanked by T-DNA border sequences.  
     
     
         23 . The stably transformed duckweed plant according to  claim 20 , wherein said duckweed plant comprises fewer than 5 copies of said chimeric nucleic acid.  
     
     
         24 . The stably transformed duckweed plant according to  claim 20 , wherein said duckweed plant is selected from the group consisting of the genus Spirodela, genus Wolffia, genus Wolfiella, and genus Lemna.  
     
     
         25 . The stably transformed duckweed plant according to  claim 24 , wherein said duckweed plant is selected from the genus Lemna.  
     
     
         26 . The stably transformed duckweed plant according to  claim 25 , wherein said duckweed plant is selected from the group consisting of a species of  Lemna minor,  a species of  Lemna miniscula,  and a species of  Lemna gibba.    
     
     
         27 . The stably transformed duckweed plant according to  claim 26 , wherein said duckweed plant is from a species of  Lemna minor.    
     
     
         28 . The stably transformed duckweed plant according to  claim 20 , wherein said chimeric nucleic acid of interest comprises at least one expression cassette comprising a gene which confers resistance to a selection agent.  
     
     
         29 . The stably transformed duckweed plant according to  claim 28 , wherein said gene which confers resistance to a selection agent is selected from the group consisting of neo, bar, pat, ALS, HPH, HYG, EPSP and Hml.  
     
     
         30 . The stably transformed duckweed plant according to  claim 20 , wherein said chimeric nucleic acid comprises two genes of interest.  
     
     
         31 . The stably transformed duckweed plant according to  claim 20 , wherein said chimeric nucleic acid encodes a protein or peptide selected from the group consisting of insulin, growth hormone, α-interferon, β-glucocerebrosidase, retinoblastoma protein, p53 protein, angiostatin, leptin, and serum albumin.  
     
     
         32 . The stably transformed duckweed plant according to  claim 20 , wherein said chimeric nucleic acid encodes at least one protein or peptide subunit of a multimeric protein selected from the group consisting of hemoglobin, collagen, P450 oxidase, and a monoclonal antibody.  
     
     
         33 . The stably transformed duckweed plant according to  claim 20 , wherein said chimeric nucleic acid encodes a secreted protein or peptide.  
     
     
         34 . A stably transformed duckweed plant tissue comprising a chimeric nucleic acid of interest incorporated in its genome, wherein said chimeric nucleic acid comprises a coding sequence operably linked to a transcription initiation region that is heterologous to said coding sequence.  
     
     
         35 . The stably transformed duckweed plant tissue according to  claim 34 , wherein said plant tissue is meristematic tissue.  
     
     
         36 . The stably transformed duckweed plant tissue according to  claim 34 , wherein said plant tissue is frond tissue.  
     
     
         37 . The stably transformed duckweed plant tissue according to  claim 34 , wherein said plant tissue is callus tissue.  
     
     
         38 . The stably transformed duckweed plant tissue according to  claim 37 , wherein said plant tissue is Type I callus tissue.  
     
     
         39 . A duckweed tissue culture comprising the stably transformed duckweed plant tissue of  claim 34 .  
     
     
         40 . A stably transformed duckweed cell comprising a chimeric nucleic acid of interest incorporated in its genome, wherein said chimeric nucleic acid comprises a coding sequence operably linked to a transcription initiation region that is heterologous to said coding sequence.