Blueberry red ringspot virus, sequences, promoters, and uses thereof

Abstract

A nucleic acid sequence of the blueberry red ringspot virus is disclosed. Also disclosed are putative promoter regions of the sequence and promoter regions capable of directing transgene expression in plants, including tissue-specific expression. Also disclosed are expression vectors, transformed plant cells and plants containing a blueberry red ringspot virus promoter and an encoded product for expression. Methods for diagnosis of blueberry red ringspot virus infection are also provided.

1 - 5 . (canceled) 
     
     
         6 . A nucleic acid probe or primer of at least 10 consecutive nucleotides of the sequence as set forth in SEQ ID NO: 1 or the complement thereof, wherein said probe or primer specifically hybridizes under stringent conditions to a blueberry red ringspot virus nucleic acid sequence. 
     
     
         7 . The nucleic acid probe or primer of  claim 6  which specifically hybridizes under stringent conditions to a promoter region comprising BRRV promoter region A as set forth in SEQ ID NO: 2, BRRV promoter region B as set forth in SEQ ID NO: 3, BRRV promoter region C as set forth in SEQ ID NO: 4, BRRV promoter region D as set forth in SEQ ID NO: 5, BRRV promoter region E as set forth in SEQ ID NO: 6, BRRV promoter region F as set forth in SEQ ID NO: 7, BRRV promoter region G as set forth in SEQ ID NO: 8, or a BRRV promoter region as set forth in SEQ ID NO: 9. 
     
     
         8 . A recombinant nucleic acid comprising a promoter from the blueberry red ringspot virus operably linked to a DNA sequence encoding a polypeptide or an RNA. 
     
     
         9 . The recombinant nucleic acid of  claim 8  wherein the promoter comprises BRRV promoter region A as set forth in SEQ ID NO: 2, BRRV promoter region B set forth in SEQ ID NO: 3, BRRV promoter region C set forth in SEQ ID NO: 4, BRRV promoter region D set forth in SEQ ID NO: 5, BRRV promoter region E set forth in SEQ ID NO: 6, BRRV promoter region F set forth in SEQ ID NO: 7, BRRV promoter region G as set forth in SEQ ID NO: 8, or a BRRV promoter region as set forth in SEQ ID NO: 9. 
     
     
         10 . The recombinant nucleic acid of  claim 9  wherein the promoter region is operably linked to a gene encoding a polypeptide or an RNA. 
     
     
         11 . A transgenic plant cell comprising the recombinant nucleic acid of  claim 10 . 
     
     
         12 . A transgenic dicotyledonous plant comprising the plant cell of  claim 11 . 
     
     
         13 . A transgenic monocotyledonous plant comprising the plant cell of  claim 11 . 
     
     
         14 . The transgenic plant of  claim 11 , wherein the plant is of the species  Arabidopsis thaliana.    
     
     
         15 . A method for transforming a plant cell comprising transforming a plant cell with a recombinant DNA construct comprising a blueberry red ringspot virus promoter and a DNA sequence which encodes a polypeptide or an RNA; wherein the promoter regulates the transcription of the DNA sequence. 
     
     
         16 . A method for diagnosing the presence of blueberry red ringspot virus in a host plant comprising (a) incubating a nucleic acid sample obtained from a plant suspected of containing the virus with a nucleic acid probe of  claim 6  under conditions in which the probe can hybridize with any blueberry red ringspot virus nucleic acid present in the sample and (b) detecting the presence of any hybridization complex formed. 
     
     
         17 . A method for diagnosing the presence of blueberry red ringspot virus in a host plant comprising (a) providing two oligonucleotides which are primers for a polymerase chain reaction method and which flank a target sequence which lies within a blueberry red ringspot virus nucleic acid as shown in  FIG. 2  or the complement thereof; (b) incubating the oligonucleotides with a nucleic acid sample obtained from a plant suspected of containing the virus; (c) amplifying the target DNA sequence if it is present in the nucleic acid sample by the polymerase chain reaction method and (d) detecting the presence of the any amplified target DNA sequence. 
     
     
         18 . A recombinant nucleic acid comprising a sequence having promoter activity in a plant cell and which hybridizes under high stringency conditions to a fragment of a blueberry red ringspot virus nucleic acid which has promoter activity in the plant cell, or the complement thereof. 
     
     
         19 . The recombinant nucleic acid of  claim 18  comprising BRRV promoter region A as set forth in SEQ ID NO: 2, BRRV promoter region B as set forth in SEQ ID NO: 3, BRRV promoter region C as set forth in SEQ ID NO: 4, BRRV promoter region D as set forth in SEQ ID NO: 5, BRRV promoter region E as set forth in SEQ ID NO: 6, or BRRV promoter region F as set forth in SEQ ID NO: 7. 
     
     
         20 . The recombinant nucleic acid of  claim 19  wherein the plant cell is a dicotyledonous plant cell. 
     
     
         21 . The recombinant nucleic acid of  claim 19  wherein the plant cell is a monocotyledonous plant cell. 
     
     
         22 . The recombinant nucleic acid of  claim 18  wherein the sequence comprises a consensus promoter sequence as set forth in SEQ ID NO: 10.