Novel plant genes

Abstract

The present invention relates to a DNA which encodes a polypeptide having flavonoid-3′,5′-hydroxylase activity, a recombinant DNA containing said DNA, and a plant having a pigment pattern which the plant does not originally have and which is acquired by transformation with said recombinant DNA.

1 ) A DNA encoding a polypeptide which has flavonoid-3′,5′-hydroxylase activity and which is represented by the amino acid sequence shown by SEQ ID NO: 1, 63 or 64, or a DNA which hybridizes with said DNA.  
     
     
         2 ) The DNA according to claim ( 1 ), wherein a part of the nucleotide sequence of said DNA is deleted or replaced by another nucleotide sequence.  
     
     
         3 ) A recombinant DNA composed of a vector DNA and the DNA of claim ( 1 ) or claim ( 2 ) which is inserted in the vector DNA.  
     
     
         4 ) A plant or plant cell which carries the recombinant DNA according to claim ( 3 ).  
     
     
         5 ) The plant or plant cell according to claim ( 4 ), wherein said plant belongs to the genus Rosa, Nicotiana, Petunia, or Dianthus.  
     
     
         6 ) A DNA which hybridizes with a DNA represented by the nucleotide sequence shown by SEQ ID NO: 1, 63 or 64 in 2×SSC (0.3 M sodium chloride, 0.03 M sodium citrate, pH 7.0) at 50° C.  
     
     
         7 ) A method for producing a plant, which comprises: introducing a recombinant DNA composed of a vector DNA fragment and a DNA fragment which encodes a polypeptide having flavonoid-3′,5′-hydroxylase activity into a plant; breeding a plant which can express a pigment based on the genetic information of the DNA encoding said polypeptide; and harvesting said plant thus obtained.  
     
     
         8 ) The method for producing a plant according to claim ( 7 ), wherein said DNA fragment which encodes the polypeptide having flavonoid-3′,5′-hydroxylase activity is the DNA of claim ( 1 ) or claim ( 2 ).  
     
     
         9 ) A DNA which has the nucleotide sequence shown by any of SEQ ID NO: 2 to 29.  
     
     
         10 ) A DNA which has a sequence comprising a sequence identical with the eight-nucleotide sequence from the 3′-terminus in the sequence of the DNA of claim ( 9 ).  
     
     
         11 ) A method for amplifying and isolating a gene fragment which encodes the amino acid sequence of the heme-binding region of cytochrome P450 enzyme, by polymerase chain reaction (PCR) using the DNA of claim ( 9 ) or claim ( 10 ) as primers.