Rice Pollen-Preferential Promoters And Uses Thereof

Abstract

The present invention relates to rice genomic promoter sequences which promote transcription preferentially in microspores and/or pollen of plants. Also provided are chimeric genes comprising these promoter sequences, and plant transformation vectors comprising these chimeric genes. The present invention also discloses plant cells, plant tissues, plants, seeds and grains comprising these chimeric genes. The invention further discloses methods for expressing foreign nucleic acid sequences preferentially in pollen and for producing plants with modified pollen fertility.

1 . An isolated pollen-preferential promoter region comprising:
 (a) the nucleotide sequence of SEQ ID No 7 from the nucleotide at position 16 to the nucleotide at position 1126, or parts thereof having promoter activity;   (b) the nucleotide sequence of SEQ ID No 7 from the nucleotide at position 16 to the nucleotide at position 940, or parts thereof having promoter activity;   (c) the nucleotide sequence of SEQ ID No 7 from the nucleotide at position 231 to the nucleotide at position 932, or parts thereof having promoter activity; or   (d) a nucleotide sequence having at least 90%, or at least 95%, or 96%, 97%, 98%, 99% sequence identity with any of said nucleotide sequence mentioned under (a), (b), or (c).   
     
     
         2 - 5 . (canceled) 
     
     
         6 . A pollen-preferential chimeric gene comprising the following operably linked DNA fragments:
 (a) the pollen-preferential promoter region of  claim 1 ; and   (b) a heterologous DNA region encoding a biologically active RNA, protein, or polypeptide.   
     
     
         7 . The chimeric gene of  claim 6 , wherein said DNA region encodes a protein or polypeptide, which when produced in a plant cell, significantly disturbs its metabolism, functioning and/or development. 
     
     
         8 . (canceled) 
     
     
         9 . The chimeric gene of  claim 6 , wherein said DNA region encodes an antisense RNA, a sense RNA, or a double-stranded RNA, which when produced in a plant cell, inhibits or reduces the expression of a target gene of interest in said plant cell, compared to the expression of said gene in a plant cell not comprising said DNA region. 
     
     
         10 . (canceled) 
     
     
         11 . The chimeric gene of  claim 6 , wherein said DNA region encodes a ribozyme, which when produced in a plant cell, specifically cleaves a target gene sequence in said plant cell. 
     
     
         12 - 15 . (canceled) 
     
     
         16 . A plant transformation vector comprising the pollen-preferential chimeric gene of  claim 6 . 
     
     
         17 . The plant transformation vector of  claim 16 , which further comprises a second chimeric gene comprising
 (a) a second heterologous DNA region encoding a biologically active RNA, protein or polypeptide, which when produced together with the gene product of said pollen-preferential chimeric gene, counteracts, prevents or inhibits the activity of said gene product; and   (b) a plant expressible promoter, which directs expression of said second heterologous DNA region at least in cells other than microspore and/or pollen cells.   
     
     
         18 . A plant cell, a plant tissue, a plant, a plant seed, or a plant grain comprising the pollen-preferential chimeric gene of  claim 6 . 
     
     
         19 . The plant cell, plant tissue, plant, plant seed, or plant grain of  claim 18 , which further comprises a second chimeric gene comprising
 (a) a second heterologous DNA region encoding a biologically active RNA, protein or polypeptide, which when produced together with the gene product of said pollen-preferential chimeric gene, counteracts, prevents or inhibits the activity of said gene product; and   (b) a plant expressible promoter, which directs expression of said second heterologous DNA region at least in cells other than microspore and/or pollen cells.   
     
     
         20 - 21 . (canceled) 
     
     
         22 . The plant cell, plant tissue, plant, or plant seed of  claim 18 , which is a monocotyledonous plant cell, plant tissue, plant, or plant seed. 
     
     
         23 . The plant cell, plant tissue, plant, plant seed, or plant grain of  claim 18 , which is a cereal plant cell, plant tissue, plant, or plant seed. 
     
     
         24 . The plant cell, plant tissue, plant, plant seed, or plant grain of  claim 18 , which is a rice, corn, or wheat plant cell, plant tissue, plant, or plant seed. 
     
     
         25 . A method for expressing a biologically active RNA, protein, or polypeptide preferentially in microspore and/or pollen cells of a plant, comprising the steps of:
 (a) providing a plant with the pollen-preferential chimeric gene of  claim 6 ; and   (b) growing said plant.   
     
     
         26 . The method of  claim 25 , wherein said plant is provided with said pollen-preferential chimeric gene by transforming a cell of a plant with said chimeric gene and by regenerating said plant from said transformed cell. 
     
     
         27 . The method of  claim 26 , wherein said plant cell is transformed with the plant transformation vector comprising the pollen-preferential chimeric gene comprising the following operably linked DNA fragments:
 (a) the pollen-preferential promoter region comprising
 (i) the nucleotide sequence of SEQ ID No 7 from the nucleotide at position 16 to the nucleotide at position 1126, or parts thereof having promoter activity; 
 (ii) the nucleotide sequence of SEQ ID No 7 from the nucleotide at position 16 to the nucleotide at position 940, or parts thereof having promoter activity; 
 (iii) the nucleotide sequence of SEQ ID No 7 from the nucleotide at position 231 to the nucleotide at position 932, or parts thereof having promoter activity; or 
 (iv) a nucleotide sequence having at least 90%, or at least 95%, or 96%, 97%, 98%, 99% sequence identity with any of said nucleotide sequence mentioned under (i), (ii), or (iii); and 
   (b) a heterologous DNA region encoding a biologically active RNA, protein, or polypeptide.   
     
     
         28 . The method of  claim 25 , which further comprises the step of providing said plant with a second chimeric gene comprising
 (a) a second heterologous DNA region encoding a biologically active RNA, protein or polypeptide, which when produced together with the gene product of said pollen-preferential chimeric gene, counteracts, prevents or inhibits the activity of said gene product; and   (b) a plant expressible promoter, which directs expression of said second heterologous DNA region at least in cells other than microspore and/or pollen cells.   
     
     
         29 . The method of  claim 28 , wherein said plant is provided with said second chimeric gene by transforming a cell of a plant with said second chimeric gene and by regenerating said plant from said transformed cell. 
     
     
         30 . The method of  claim 29 , wherein the plant cell is transformed with the plant transformation vector comprising the pollen-preferential chimeric gene comprising the following operably linked DNA fragments:
 (a) the pollen-preferential promoter region comprising
 (i) the nucleotide sequence of SEQ ID No 7 from the nucleotide at position 16 to the nucleotide at position 1126, or parts thereof having promoter activity; 
 (ii) the nucleotide sequence of SEQ ID No 7 from the nucleotide at position 16 to the nucleotide at position 940, or parts thereof having promoter activity; 
 (iii) the nucleotide sequence of SEQ ID No 7 from the nucleotide at position 231 to the nucleotide at position 932, or parts thereof having promoter activity; or 
 (iv) a nucleotide sequence having at least 90%, or at least 95%, or 96%, 97%, 98%, 99% sequence identity with any of said nucleotide sequence mentioned under (i), (ii), or (iii); and 
   (b) a heterologous DNA region encoding a biologically active RNA, protein, or polypeptide and   further comprises a second chimeric gene comprising   (c) a second heterologous DNA region encoding a biologically active RNA, protein or polypeptide, which when produced together with the gene product of said pollen-preferential chimeric gene, counteracts, prevents or inhibits the activity of said gene product; and   (d) a plant expressible promoter, which directs expression of said second heterologous DNA region at least in cells other than microspore and/or pollen cells.   
     
     
         31 - 34 . (canceled) 
     
     
         35 . A method for producing a plant with modified pollen-fertility properties, which comprises the steps of:
 (a) providing a plant with the pollen-preferential chimeric gene of  claim 7 ; and   (b) growing said plant.   
     
     
         36 . The method of  claim 35 , which further comprises the step of providing said plant with a second chimeric gene comprising
 (a) a second heterologous DNA region encoding a biologically active RNA, protein or polypeptide, which when produced together with the gene product of said pollen-preferential chimeric gene, counteracts, prevents or inhibits the activity of said gene product; and   (b) a plant expressible promoter, which directs expression of said second heterologous DNA region at least in cells other than microspore and/or pollen cells.   
     
     
         37 - 72 . (canceled)