Compositions affecting programmed cell death and their use in the modification of plant development

Abstract

Novel isolated polynucleotides associated with programmed cell death and various plant developmental mechanisms are provided, together with genetic constructs comprising such sequences. Methods for the modulation of the content, structure and metabolism of plants, and particularly for the modulation of PCD and various plant developmental mechanisms in plants, are also disclosed, the methods comprising incorporating one or more of the polynucleotides or genetic constructs of the present invention into the genome of a plant.

We claim:  
     
         1 . An isolated polynucleotide comprising a nucleotide sequence selected from the group consisting of: SEQ ID NOS: 1-45, 90-140, 192-195, 200, 204 and 206-248.  
     
     
         2 . An isolated polynucleotide comprising a nucleotide sequence selected from the group consisting of: 
 (a) complements of the sequences recited in SEQ ID NOS: 1-45, 90-140, 192-195, 200, 204 and 206-248;    (b) reverse complements of the sequences recited in SEQ ID NOS: 1-45, 90-140; 192-195, 200, 204 and 206-248; and    (c) reverse sequences of the sequences recited in SEQ ID NOS: 1-45, 90-140, 192-195, 200, 204 and 206-248.    
     
     
         3 . An isolated polynucleotide comprising a nucleotide sequence selected from the group consisting of: 
 (a) sequences having at least 75% identity to a sequence of SEQ ID NOS: 1-45, 90-140, 192-195, 200, 204 and 206-248;    (b) sequences having at least 90% identity to a sequence of SEQ ID NOS: 1-45, 90-140, 192-195, 200, 204 and 206-248;    (c) sequences having at least 95% identity to a sequence of SEQ ID NOS: 1-45, 90-140, 192-195, 200, 204 and 206-248; and    (d) sequences that hybridize to a sequence of SEQ ID NOS: 1-45, 90-140, 192-195, 200, 204 and 206-248 under stringent hybridization conditions.    
     
     
         4 . The isolated polynucleotide of  claim 3 , wherein the polynucleotide encodes a polypeptide having activity in a plant cell death pathway.  
     
     
         5 . An isolated polynucleotide comprising a nucleotide sequence selected from the group consisting of: 
 (a) sequences that are 200-mers of a sequence of SEQ ID NOS: 1-45, 90-140, 192-195, 200, 204 and 206-248; and    (b) sequences that are 100-mers of a sequence of SEQ ID NOS: 1-45, 90-140, 192-195, 200, 204 and 206-248.    
     
     
         6 . An isolated oligonucleotide probe or primer comprising at least 10 contiguous resides complementary to 10 contiguous residues of a nucleotide sequence of SEQ ID NO: 1-45, 90-140, 192-195, 200, 204 and 206-248.  
     
     
         7 . A genetic construct comprising at least one polynucleotide according to any one of claims  1 - 3 .  
     
     
         8 . A transgenic plant cell comprising at least one genetic construct according to  claim 7 .  
     
     
         9 . A genetic construct comprising, in the 5′-3′ direction: 
 (a) a gene promoter sequence;  
 (b) a polynucleotide sequence comprising at least one of the following: (1) a sequence comprising a coding region of a polynucleotide of any one of claims  1 - 3 ; and (2) a sequence comprising a coding region of a polynucleotide of any one of claims  1 - 3 ; and  
 (c) a gene termination sequence.  
 
     
     
         10 . The construct of  claim 9 , wherein the polynucleotide sequence is in a sense orientation.  
     
     
         11 . The construct of  claim 9 , wherein the polynucleotide sequence is in an antisense orientation.  
     
     
         12 . The construct of  claim 9 , wherein the gene promoter sequence and gene termination sequences are functional in a plant host.  
     
     
         13 . A transgenic plant cell comprising a construct of  claim 9 .  
     
     
         14 . A plant comprising a transgenic plant cell according to  claim 13 , or fruit or seeds or progeny thereof.  
     
     
         15 . The plant of  claim 14 , wherein the plant is a woody plant.  
     
     
         16 . The plant of  claim 15 , wherein the plant is selected from the group consisting of eucalyptus and pine species.  
     
     
         17 . A method for modulating a plant cell death pathway in a plant, comprising stably incorporating into the genome of the plant a genetic construct of  claim 9 .  
     
     
         18 . The method of  claim 17 , wherein the genetic construct incorporated into the genome of the plant provides a plant cell death pathway that is not present in a native form of the plant.  
     
     
         19 . A method for producing a plant having an altered cell death pathway, comprising: 
 (a) transforming a plant cell with a genetic construct of  claim 9  to provide a transgenic plant cell; and    (b) cultivating the transgenic plant cell under conditions conducive to regeneration and mature plant growth.    
     
     
         20 . An isolated polypeptide encoded by a polynucleotide of any one of claims  1 - 3 .  
     
     
         21 . An isolated polypeptide comprising an amino acid sequence selected from the group consisting of: SEQ ID NOS: 46-89, 141-191, 196-199, 201, 205 and 249-290.  
     
     
         22 . An isolated polypeptide comprising an amino acid sequence selected from the group consisting of: 
 (a) sequences having at least 75% identity to a sequence of SEQ ID NOS: 46-89, 141-191, 196-199, 201, 205 and 249-290;    (b) sequences having at least 90% identity to a sequence of SEQ ID NOS: 46-89, 141-191, 196-199, 201, 205 and 249-290; and    (c) sequences having at least 95% identity to a sequence of SEQ ID NOS: 46-89, 141-191, 196-199, 201, 205 and 249-290,    wherein the polypeptide is active in a plant cell death pathway.    
     
     
         23 . A method for modifying the activity of a polypeptide involved in a plant cell death pathway in a plant, comprising introducing into cells of the plant RNA corresponding to a polynucleotide of any one of claims  1 - 3 , thereby inhibiting expression of a polypeptide encoded by the polynucleotide.  
     
     
         24 . A method for modifying the activity of a polypeptide involved in a plant cell death pathway in a plant, comprising introducing into cells of the plant double stranded RNA corresponding to a polynucleotide of any one of claims  1 - 3 , thereby inhibiting expression of a polypeptide encoded by the polynucleotide.