Expression cassettes for vascular tissue-preferential expression in plants

Abstract

The present invention relates to expression cassettes comprising transcription regulating sequences with vascular tissue-preferential or vascular tissue-specific expression profiles in plants obtainable from Arabidopsis thaliana genes At4g00140, At5g45350, At2g39830, At1g68430, or At5g67280.

1 . An expression cassette for regulating vascular tissue-preferential or vascular tissue-specific expression in plants comprising
 i) at least one transcription regulating nucleotide sequence of a plant gene, said plant gene selected from the group of genes described by the  Arabidopsis thaliana  genome loci At5g45350, At2g39830, At1g68430, and At5g67280, 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 nucleotide sequence described by SEQ ID NOs: 8, 9, 10, 11, 12, 13, 14, 15, 18, 19, 20, 21, 22, 23, 24, 25, 28, 29, 30, 31, 32, 33, 34, 35, 38, 39, 40, 41, 42, 43, 44, or 45,   ii) a fragment of at least 50 consecutive bases of a sequence under 1) which has substantially the same promoter activity as the corresponding transcription regulating nucleotide sequence described by SEQ ID NO: 8, 0, 10, 11, 12, 13, 14, 15, 18, 19, 20, 21, 22, 23, 24, 25, 28, 29, 30, 31, 32, 33, 34, 35, 38, 39, 40, 41, 42, 43, 44, or 45:   iii) a nucleotide sequence having substantial similarity with a sequence identity of at least 40% to a transcription regulating nucleotide sequence described by SEQ ID NO: 8, 9, 10, 11, 12, 13, 14, 15, 18, 19, 20, 21, 22, 23, 24, 25, 28, 29, 30, 31, 32, 33, 34, 35, 38, 39, 40, 41, 42, 43, 44, or 45;   iv) a nucleotide sequence capable of hybridizing under conditions equivalent to hybridization in 7% sodium dodecyl sulfate (SDS), 0.5 M NaPO 4 , 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: 8, 9, 10, 11, 12, 13, 14, 15, 18, 19, 20, 21, 22, 23, 24, 25, 28, 29, 30, 31, 32, 33, 34, 35, 38, 30, 40, 41, 42, 43, 44, or 45, 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 NaPO 4 , 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: 8, 9, 10, 11, 12, 13, 14, 15, 18, 19, 20, 21, 22, 23, 24, 25, 28, 29, 30, 31, 32, 33, 34, 35, 38, 39, 40, 41, 42, 43, 44, or 45, 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 a polypeptide comprising SEQ ID NO: 17, 27, 37, or 47. 
     
     
         4 . The expression cassette of  claim 1 , wherein expression of the nucleic acid sequence results in expression of a protein, or expression of a antisense RNA, sense or 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 the expression cassette. 
     
     
         8 . A transgenic plant comprising the expression cassette of  claim 1 , a vector comprising the expression cassette, a cell comprising the expression cassette, or a cell comprising the vector. 
     
     
         9 . A method for identifying and/or isolating a sequence with vascular tissue-preferential or vascular tissue-specific transcription regulating activity comprising utilizing a nucleic acid sequence encoding a amino acid sequence as described by SEQ ID NO: 17, 27, 37, or 47, or a part of at least 15 bases thereof for identifying and/or isolating a sequence with vascular tissue-preferential or vascular tissue-specific transcription regulating activity. 
     
     
         10 . The method of  claim 9 , wherein the nucleic acid sequences is described by SEQ ID NO: 16, 26, 36, or 46, or a part of at least 15 bases thereof. 
     
     
         11 . The method of  claim 9 , wherein said identification and/or isolation is realized by a method selected from polymerase chain reaction, hybridization, or database screening. 
     
     
         12 . A method for providing a transgenic expression cassette for vascular tissue-preferential or vascular tissue-specific expression comprising the steps of:
 I. isolating of a vascular tissue-preferential or vascular tissue-specific transcription regulating nucleotide sequence utilizing at least one nucleic acid sequence or a part thereof wherein said sequence is encoding a polypeptide comprising SEQ ID NO: 17, 27, 37, or 47, or a part of at least 15 bases thereof, and   II. functionally linking said vascular tissue-preferential or vascular tissue-specific transcription regulating nucleotide sequence to another nucleotide sequence of interest, which is heterologous in relation to said vascular tissue-preferential or vascular tissue-specific transcription regulating nucleotide sequence.