Biological Containment System

Abstract

The invention relates to materials and methods useful for controlling the unwanted spread of transgenic traits. The methods involve an apomictic plant containing a transgene for a desired trait and a transgene causing seed infertility. The methods also involve one or more transcription activators that activates expression of both transgenes carried by the apomictic plant. The transcription activator(s) activates expression of both transgenes in the female. Seeds that are formed on such plants are infertile.

1 . A method for making infertile seed, said method comprising:
 a) permitting seed development to occur on a plurality of monocotyledonous first plants that have been pollinated by a plurality of monocotyledonous second plants, wherein said first plants are genetically or cytoplasmically male-sterile, wherein said male-sterility is not conferred via a transgene, and wherein said first plants comprise first and second nucleic acids, said first nucleic acid comprising a first transcription activator recognition site and a first promoter, said first recognition site and said first promoter operably linked to a sequence to be transcribed, said second nucleic acid comprising a second transcription activator recognition site and a second promoter, said second recognition site and said second promoter operably linked to a coding sequence that results in seed infertility, wherein said second plants are male-fertile and comprise at least one activator nucleic acid comprising at least one coding sequence for a transcription activator that binds to at least one of said recognition sites, each said at least one transcription activator coding sequence having a promoter operably linked thereto, and wherein said seeds are infertile.   
     
     
         2 . The method of  claim 1 , wherein said sequence that results in seed infertility is selected from the group consisting of SEQ ID NOs: 95, 1, and 103; 51, 5, 107, 133, 168, and 215; 54, 18, 154, 179, and 211; 58, 47, 114, 118, 122, and 129; 66, 26, 29, 126, and 188; 8, 16, 136, 150, 158, 175, and 223; 73, 22, 32, and 146; 76, 11, 160, 164, and 196; 80, 38, 138, and 204; 84, 36, 142, 183, 192, 208, and 219; 87, 41, 110, 171, 185, and 200; and 91 and 94. 
     
     
         3 . The method of  claim 1 , wherein said first and second promoters and said promoter operably linked to said at least one transcription activator coding sequence are selected from a CaMV35 S minimal promoter and a rice or maize ubiquitin minimal promoter. 
     
     
         4 . The method of  claim 1 , wherein said transcriptional activator is a chimeric polypeptide comprising a yeast HAP1 DNA binding domain and a herpes simplex VP16 transcription activator domain. 
     
     
         5 . The method of  claim 1 , wherein said first and second transcription activator recognition sites comprise from 1 to 5 copies of a UAS Hap1  upstream activator sequence element. 
     
     
         6 . An article of manufacture comprising:
 a) a container;   b) a first type of seeds within said container, wherein said seeds are monocot seeds, said first type of seeds comprising at least one first nucleic acid comprising: i) a first transcription activator recognition site and a first promoter, said first recognition site and said first promoter operably linked to a sequence to be transcribed; and ii) a second transcription activator recognition site and a second promoter, said second recognition site and said second promoter operably linked to a sequence causing seed infertility, and wherein plants grown from said first type of seeds are genetically or cytoplasmically male-sterile, wherein the male-sterility is not conferred via a transgene; and,   c) a second type of seeds within said container, wherein said seeds are monocot seeds, said second type of seeds comprising at least one activator nucleic acid encoding at least one transcription activator that binds to at least one of said recognition sites, each said at least one transcription activator having a promoter operably linked thereto, wherein plants grown from said second type of seeds are male-fertile.   
     
     
         7 . The article of  claim 6 , wherein said sequence causing seed infertility is selected from the group consisting of SEQ ID NOs: 95, 1, and 103; 51, 5, 107, 133, 168, and 215; 54, 18, 154, 179, and 211; 58, 47, 114, 118, 122, and 129; 66, 26, 29, 126, and 188; 8, 16, 136, 150, 158, 175, and 223; 73, 22, 32, and 146; 76, 11, 160, 164, and 196; 80, 38, 138, and 204; 84, 36, 142, 183, 192, 208, and 219; 87, 41, 110, 171, 185, and 200; and 91 and 94. 
     
     
         8 . The article of  claim 6 , wherein said first and second promoters and said promoter operably linked to said at least one transcription activator coding sequence are selected from a CaMV35 S minimal promoter and a rice or maize ubiquitin minimal promoter. 
     
     
         9 . The article of  claim 6 , wherein said transcriptional activator is a chimeric polypeptide comprising a yeast HAP1 DNA binding domain and a herpes simplex VP16 transcription activator domain. 
     
     
         10 . The article of  claim 6 , wherein said first and second transcription activator recognition sites comprise from 1 to 5 copies of a UAS Hap1  upstream activator sequence element. 
     
     
         11 . A monocotyledonous plant comprising:
 a) a first nucleic acid comprising a first transcription activator recognition site and a first promoter, said first recognition site and said first promoter operably linked to a sequence to be transcribed; and,   b) a second nucleic acid comprising a second transcription activator recognition site and a second promoter, said second recognition site and said second promoter operably linked to a sequence causing seed infertility.   
     
     
         12 . The plant of  claim 11 , wherein said sequence causing seed infertility is selected from the group consisting of SEQ ID NOs: 95, 1, and 103; 51, 5, 107, 133, 168, and 215; 54, 18, 154, 179, and 211; 58, 47, 114, 118, 122, and 129; 66, 26, 29, 126, and 188; 8, 16, 136, 150, 158, 175, and 223; 73, 22, 32, and 146; 76, 11, 160, 164, and 196; 80, 38, 138, and 204; 84, 36, 142, 183, 192, 208, and 219; 87, 41, 110, 171, 185, and 200; and 91 and 94. 
     
     
         13 . The plant of  claim 11 , wherein said first and second promoters and said promoter operably linked to said at least one transcription activator coding sequence are selected from a CaMV35 S minimal promoter and a rice or maize ubiquitin minimal promoter. 
     
     
         14 . The plant of  claim 11 , wherein said transcriptional activator is a chimeric polypeptide comprising a yeast HAP1 DNA binding domain and a herpes simplex VP16 transcription activator domain. 
     
     
         15 . The plant of  claim 11 , wherein said first and second transcription activator recognition sites comprise from 1 to 5 copies of a UAS Hap1  upstream activator sequence element.