Expression Cassettes For Seed-Preferential Expression In Plants

Abstract

The present invention relates to expression cassettes comprising transcription regulating sequences with seed-preferential or seed-specific expression profiles in plants obtainable from Arabidopsis thaliana genes At4g12910, At1g66250, At4g00820, At2g36640, At2g34200, At3g11180, At4g00360, At2g48030, At2g38590, At1g23000, At3g15510, At3g50870, At4g00220, or At4g26320.

1 . An expression cassette for seed-specific or seed-preferential transcription in plants comprising
 i) at least one transcription regulating nucleotide sequence of a plant gene, wherein said plant gene is described by the GenBank  Arabidopsis thaliana  genome loci At4g00820, or a functional equivalent thereof, and functionally linked thereto   ii) at least one nucleic acid sequence which is heterologous in relation to said transcription regulating nucleotide sequence.   
     
     
         2 . The expression cassette of  claim 1 , wherein the transcription regulating nucleotide sequence is selected from the group of sequences consisting of
 i) the sequence described by SEQ ID NO: 23, 24, 25, 26, 27, or 28,   ii) a fragment of at least 50 consecutive bases of the sequence under i) which has substantially the same promoter activity as the corresponding transcription regulating nucleotide sequence described by SEQ ID NO: 23, 24, 25, 26, 27, or 28,   iii) a nucleotide sequence having substantial similarity with a sequence identity of at least 40% to the transcription regulating nucleotide sequence described by SEQ ID NO: 23, 24, 25, 26, 27, or 28,   iv) a nucleotide sequence capable of hybridizing under conditions equivalent to hybridization in 7% sodium dodecyl sulfate (SDS), 0.5 M NaPO4, 1 mM EDTA at 50° C. with washing in 2×SSC, 0.1% SDS at 50° C. to a transcription regulating nucleotide sequence described by SEQ ID NO: 23, 24, 25, 26, 27, or 28, or the complement thereof,   v) a nucleotide sequence capable of hybridizing under conditions equivalent to hybridization in 7% sodium dodecyl sulfate (SDS), 0.5 M NaPO4, 1 mM EDTA at 50° C. with washing in 2×SSC, 0.1% SDS at 50° C. to a nucleic acid comprising 50 to 200 or more consecutive nucleotides of a transcription regulating nucleotide sequence described by SEQ ID NO: 23, 24, 25, 26, 27, or 28, or the complement thereof, and   vi) a nucleotide sequence which is the complement or reverse complement of any of the previously mentioned nucleotide sequences under i) to v).   
     
     
         3 . The expression cassette of  claim 1 , wherein the functional equivalent of the transcription regulating nucleotide sequence is obtained or obtainable from plant genomic DNA from a gene encoding a polypeptide which has at least 70% amino acid sequence identity to the polypeptide sequence of SEQ ID NO: 30. 
     
     
         4 . The expression cassette of  claim 1 , wherein expression of the nucleic acid sequence results in expression of a protein, or expression of an antisense RNA, a sense RNA, or a double-stranded RNA. 
     
     
         5 . The expression cassette of  claim 1 , wherein expression of the nucleic acid sequence confers to the plant an agronomically valuable trait. 
     
     
         6 . A vector comprising the expression cassette of  claim 1 . 
     
     
         7 . A transgenic host cell or non-human organism comprising the expression cassette of  claim 1  or a vector comprising said expression cassette. 
     
     
         8 . A transgenic plant comprising the expression cassette of  claim 1 , or a vector comprising said expression cassette, or a transgenic cell comprising said expression cassette or said vector. 
     
     
         9 . A method for producing a transgenic plant cell, comprising transforming a plant cell with the expression cassette of  claim 1 . 
     
     
         10 . A method for producing a transgenic plant, comprising transforming a plant cell with the expression cassette of  claim 1 , and generating from the plant cell the transgenic plant. 
     
     
         11 . A method for identifying and/or isolating a sequence with seed-specific or seed-preferential transcription regulating activity comprising utilizing
 i) a nucleic acid sequence encoding the amino acid sequence as described by SEQ ID NO: 30, or a part of at least 15 bases thereof or   ii) the nucleic acid sequence of SEQ ID NO: 29, or a part of at least 15 bases thereof.   
     
     
         12 . The method of  claim 11 , wherein said identification and/or isolation is realized by polymerase chain reaction, hybridization, or database screening. 
     
     
         13 . A method for providing a transgenic expression cassette for seed-specific or seed-preferential transcription in plants comprising:
 I. isolating a seed-specific or seed-preferential transcription regulating nucleotide sequence utilizing at least one nucleic acid sequence or a part thereof, wherein said sequence is encoding the polypeptide described by SEQ ID NO: 30, or a part of at least 15 bases thereof, and   II. functionally linking said seed-specific or seed-preferential transcription regulating nucleotide sequence to another nucleotide sequence of interest, which is heterologous in relation to said seed-specific or seed-preferential transcription regulating nucleotide sequence.   
     
     
         14 . An expression cassette for seed-specific or seed-preferential transcription in plants comprising
 i) at least one transcription regulating nucleotide sequence of a plant gene, and   ii) at least one nucleic acid sequence which is operably linked to and heterologous in relation to said transcription regulating nucleotide sequence;   wherein the transcription regulating nucleotide sequence comprises:   a) at least one transcription regulating nucleotide sequence of the plant gene encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 30,   b) the nucleotide sequence of SEQ ID NO: 23, 24, 25, 26, 27, or 28,   c) a nucleotide sequence having at least 70% identity to the nucleotide sequence of SEQ ID NO: 23, 24, 25, 26, 27, or 28, or   d) a fragment of SEQ ID NO: 23, 24, 25, 26, 27, or 28,   wherein the transcription regulating nucleotide sequence has seed-specific or seed-preferential expression activity.   
     
     
         15 . The expression cassette of  claim 14 , wherein expression of the nucleic acid sequence results in expression of a protein, or expression of an antisense RNA, a sense RNA, or a double-stranded RNA. 
     
     
         16 . The expression cassette of  claim 14 , wherein expression of the nucleic acid sequence confers to the plant an agronomically valuable trait. 
     
     
         17 . A vector comprising the expression cassette of  claim 14 . 
     
     
         18 . A transgenic host cell or non-human organism comprising the expression cassette of  claim 14  or a vector comprising said expression cassette. 
     
     
         19 . A transgenic plant comprising the expression cassette of  claim 14 , or a vector comprising said expression cassette, or a transgenic cell comprising said expression cassette or said vector. 
     
     
         20 . A method for producing a transgenic plant or plant cell, comprising transforming a plant cell with the expression cassette of  claim 14 .