Promoter, promoter control elements and combinations and uses thereof

Abstract

The present invention is directed to nitrogen responsive promoter sequences and promoter control elements, polynucleotide constructs comprising the nitrogen responsive promoters and control elements and methods of identifying the nitrogen responsive promoters, control elements, or fragments thereof. The invention further relates to the use of the present nitrogen responsive promoters or promoter control elements to modulate transcript levels.

1 . An isolated nitrogen responsive promoter capable of modulating transcription comprising a nucleic acid molecule having at least 85% sequence identity to any one of SEQ ID NOs: 1-17, or a complement thereof. 
     
     
         2 . The isolated promoter of  claim 1 , wherein said nucleic acid comprises a sequence corresponding to any one of SEQ ID NOs: 1-17 having at least one of the corresponding optional promoter fragments identified in Table 1 deleted therefrom. 
     
     
         3 . A vector construct comprising:
 a) a nitrogen responsive promoter capable of modulating transcription comprising a first nucleic acid molecule having at least 80% sequence identity to any one of SEQ ID NOs: 1-17; and   b) a second nucleic acid molecule having to be transcribed, wherein said first and second nucleic acid molecules are heterologous to each other and are operatively linked together.   
     
     
         4 . The vector construct according to  claim 3 , wherein said nucleic acid comprises a sequence according to any one of SEQ ID NOs: 1-17 with at least one of the corresponding optional promoter fragments identified in Table 1 deleted therefrom. 
     
     
         5 . A host cell comprising an isolated nitrogen responsive promoter according to  claim 1 , wherein said nucleic acid molecule is flanked by exogenous sequence. 
     
     
         6 . A host cell comprising a vector construct of  claim 3 . 
     
     
         7 . A method of modulating transcription by combining, in an environment suitable for transcription:
 a) a nitrogen responsive promoter capable of modulating transcription comprising a first nucleic acid molecule having at least 80% sequence identity to a sequence according to any one of SEQ ID NOs: 1-17; and   b) a second molecule to be transcribed;   wherein the first and second nucleic acid molecules are heterologous to each other and operatively linked together.   
     
     
         8 . The method according to  claim 7 , wherein said first nucleic acid molecule is inserted into a plant cell and said plant cell is regenerated into a plant. 
     
     
         9 . A plant comprising a vector construct according to  claim 3 . 
     
     
         10 . A method of introducing an isolated nucleic acid into a host cell comprising:
 a) providing an isolated nucleic acid molecule according to  claim 1 ; and   b) contacting said isolated nucleic acid with said host cell under conditions that permit insertion of said nucleic acid into said host cell.   
     
     
         11 . A method of transforming a host cell that comprises contacting a host cell with a vector construct according to  claim 3 . 
     
     
         12 . A method for detecting a nucleic acid in a sample which comprises:
 a) providing an isolated nucleic acid molecule according to  claim 1 ;   b) contacting said isolated nucleic acid molecule with a sample under conditions which permit a comparison of the sequence of said isolated nucleic acid molecule with the sequence of DNA in said sample; and   c) analyzing the result of said comparison.   
     
     
         13 . A plant, plant cell, plant material or seed of a plant which comprises a nucleic acid molecule according to  claim 1  which is exogenous or heterologous to said plant or plant cell. 
     
     
         14 . A plant, plant cell, plant material or seed of a plant which comprises a vector construct according to  claim 3 . 
     
     
         15 . A plant that has been regenerated from a plant cell or seed according to  claim 13  or  14 . 
     
     
         16 . A plant, plant cell, plant material or seed of a plant which comprises a nucleic acid molecule according to  claim 1 , wherein said plant has improved nitrogen responsiveness characteristics as compared to a wild-type plant cultivated under the same conditions. 
     
     
         17 . A method for increasing nitrogen responsiveness in a plant comprising transforming a plant with a nucleic acid sequence according to  claim 1 . 
     
     
         18 . A plant having a gene construct comprising a nucleic acid encoding a nitrogen responsive promoter operatively linked to a coding sequence so that the coding sequence is ectopically overexpressed in the plant in response to abnormal nitrogen conditions, and the plant exhibits:
 i) faster rate of growth,   ii) greater fresh or dry weight at maturation,   iii) greater fruit or seed yield,   iv) higher tolerance to abnormal nitrogen conditions,   v) greater germination rate under abnormal nitrogen conditions,   viii) reduced nitrogen needs,   ix), greater tolerance to excess nitrogen, or   x) improved performance   than a progenitor plant when the plant and the progenitor plant are cultivated under identical environmental conditions, wherein the nitrogen responsive promoter is promoter sequence according to  claim 1 .   
     
     
         19 . A crop plant having a gene construct comprising a nucleic acid encoding a nitrogen responsive promoter operatively linked to a coding sequence so that the coding sequence is ectopically overexpressed in the crop plant in response to sub-optimal nitrogen conditions, and the crop plant exhibits:
 i) faster rate of growth,   ii) greater fresh or dry weight at maturation,   iii) greater fruit or seed yield,   iv) higher tolerance to sub-optimal nitrogen conditions,   v) greater germination rate under sub-optimal nitrogen conditions,   viii) reduced nitrogen needs, or   ix), greater tolerance to excess nitrogen, or   x) improved performance   than a progenitor plant when the crop plant and the progenitor plant are cultivated under identical environmental conditions, wherein the nitrogen responsive promoter is a promoter sequence according to  claim 1 .   
     
     
         20 . A plant having a gene construct comprising a nucleic acid encoding a nitrogen responsive promoter operatively linked to a coding sequence so that the coding sequence is ectopically overexpressed in the plant under normal nitrogen conditions, and the plant exhibits:
 i) faster rate of growth,   ii) greater fresh or dry weight at maturation,   iii) greater fruit or seed yield,   iv) higher tolerance to normal nitrogen conditions,   v) greater germination rate under normal nitrogen conditions,   viii) reduced nitrogen needs,   ix) greater tolerance to excess nitrogen, or   x) improved performance   than a progenitor plant when the transgenic plant and the progenitor plant are cultivated under identical environmental conditions, wherein the nitrogen responsive promoter is a promoter sequence according to  claim 1 .