Inducible promoter sequences for regulated expression and methods of use

Abstract

The plant promoter of a CBSU-Anther_Subtraction library (CAS1) gene encoding a mannitol dehydrogenase, and fragments thereof, and their use in promoting the expression of one or more heterologous nucleic acid fragments in an inducible manner in plants are described. These promoter fragments are also useful in creating recombinant DNA constructs comprising nucleic acid sequences encoding a desired gene product operably linked to such promoter fragments which can be utilized to transform plants and bring the expression of the gene product under external chemical and/or heat control in monocotyledonous and dicotyledonous plants.

1 . A recombinant DNA construct comprising:
 a) a nucleotide sequence comprising any one of the sequences set forth in SEQ ID NO:9 or SEQ ID NO:10, or a full-length complement thereof;   b) a nucleotide sequence comprising a functional fragment of SEQ ID NO:10, or a full-length complement thereof;   c) a nucleotide sequence comprising a sequence having at least 85% sequence identity, based on the BLASTN method of alignment, when compared to the nucleotide sequence of (a) or (b);   d) a nucleotide sequence which hybridizes to SEQ ID NO:9 under highly stringent conditions of a wash of 0.1 SSC, 0.1% (w/v) SDS at 65° C.;   e) a nucleotide sequence comprising all or a fragment of a 1.7 kb 5′ non-coding sequence of a mannitol dehydrogenase; or,   f) a derivative of one of the nucleotide sequences indicated in (a), (b), (c), (d) or (e) obtained by substitution, addition and/or deletion of one or more nucleotides; and,   wherein said nucleotide sequence is an inducible promoter.   
     
     
         2 . The recombinant DNA construct of  claim 1 , wherein the nucleotide sequence of c) has at least 90% identity, based on the BLASTN method of alignment, when compared to the sequence set forth in SEQ ID NO:1. 
     
     
         3 - 4 . (canceled) 
     
     
         5 . The recombinant DNA construct of  claim 1  wherein said inducible promoter is induced by a chemical or stress treatment. 
     
     
         6 . The recombinant DNA construct of  claim 1  wherein said inducible promoter is induced by a safener or heat treatment. 
     
     
         7 . (canceled) 
     
     
         8 . The recombinant DNA construct of  claim 6 , wherein said heat treatment comprises a temperature greater than 26° C. 
     
     
         9 - 11 . (canceled) 
     
     
         12 . A vector comprising the recombinant DNA construct of  claim 1 . 
     
     
         13 . A cell comprising the recombinant DNA construct of  claim 1 . 
     
     
         14 . The cell of  claim 13 , wherein the cell is a plant cell. 
     
     
         15 . (canceled) 
     
     
         16 . A transgenic plant having stably incorporated into its genome the recombinant DNA construct of  claim 1 . 
     
     
         17 - 20 . (canceled) 
     
     
         21 . Transgenic seed produced by the transgenic plant of  claim 16 . 
     
     
         22 . A plant stably transformed with a recombinant expression construct comprising a plant promoter and a heterologous nucleic acid fragment operably linked to said promoter, wherein said promoter is an inducible promoter and capable of controlling expression of said heterologous nucleic acid fragment in a plant cell, and further wherein said promoter comprises a fragment of SEQ ID NO:10. 
     
     
         23 . A method of expressing a coding sequence or a functional RNA in a plant cell comprising:
 a) introducing the recombinant DNA construct of  claim 1  into a plant cell, wherein the at least one heterologous sequence comprises a coding sequence or a functional RNA;   b) growing the plant cell of step a);   c) induction of the inducible promoter by chemical or stress treatment on the plant cell of b); and,   d) selecting a plant cell displaying expression of the coding sequence or the functional RNA of the recombinant DNA construct.   
     
     
         24 - 29 . (canceled) 
     
     
         30 . A method for altering expression of at least one heterologous nucleic acid fragment in a plant comprising:
 (a) transforming a plant cell with the recombinant expression construct of  claim 1 ;   (b) induction of the inducible promoter by chemical or stress treatment on the cell of (a)   (c) growing fertile mature plants from the transformed plant cell of step (a); and,   (d) selecting plants containing the transformed plant cell wherein the expression of the heterologous nucleic acid fragment is increased or decreased.   
     
     
         31 . A method of transgenically altering a marketable plant trait, comprising:
 a) introducing a recombinant DNA construct of  claim 1  into a plant;   b) induction of the inducible promoter by chemical or stress treatment on the plant of (a);   c) growing a fertile, mature plant resulting from step b); and   d) selecting a plant expressing the at least one heterologous nucleotide sequence in at least one plant tissue based on the altered marketable trait.   
     
     
         32 . (canceled) 
     
     
         33 . A recombinant DNA construct comprising:
 a) a nucleotide sequence comprising all or a functional fragment of SEQ ID NO: 19 or SEQ ID NO: 22;   b) a nucleotide sequence comprising a full-length complement of the nucleotide sequence (a); or,   c) a nucleotide sequence comprising a sequence having at least 90% sequence identity, based on the BLASTN method of alignment, when compared to the nucleotide sequence of (a) or (b); and,   wherein said nucleotide sequence is a promoter.