Method for the alteration of plants using cle polypeptides/peptides

Abstract

The present invention relates to altering the biomass and/or structure of a plant, in order to maximise its potential as a source of feedstock or increase its potential as a feedstock for the paper industry. CLE41 and/or CLE42 are used to manipulate growth and structure of the vascular tissue of the plant. The present invention also provides plants in which the levels of CLE41 and/or CLE42 are increased compared to those of a native plant grown under identical conditions, and parts of such plants. Also provided are methods for using such plants or plant parts in the production of plant derived products such as paper or biofuels.

1 . A method of manipulating plant growth and/or structure, comprising:
 providing polypeptide selected from the group consisting of:   i) a CLE41 polypeptide;   ii) a CLE42 polypeptide;   iii) a polypeptide comprising an amino acid sequence that is at least 70% identical to the amino acid sequence of amino acids 124 to 137 of the consensus sequence of  FIG. 10 ;   iv) a polypeptide comprising an amino acid sequence that is at least 70% identical to the amino acid sequence of CLE41 of  FIG. 13A  or CLE42 of  FIG. 14A ; and   v) a polypeptide encoded by a nucleic acid molecule that is at least 70% identical to the nucleotide sequence of CLE41 of  FIG. 13B  or CLE42 of  FIG. 14B ; and   using the polypeptide in the manipulation of plant growth and/or structure.   
     
     
         2 . A method of manipulating plant growth and/or structure, comprising:
 providing a nucleic acid molecule selected from the group consisting of:   i) a nucleic acid molecule that encodes a CLE41 polypeptide;   ii) a nucleic acid molecule that encodes a CLE42 polypeptide;   iii) a nucleic acid molecule that encodes a polypeptide comprising an amino acid sequence that is at least 70% identical to the amino acid sequence of amino acids 124 to 137 of the consensus sequence of  FIG. 10 ;   iv) a nucleic acid molecule which is at least 70% identical to the nucleotide sequence of CLE41 of  FIG. 13B  or CLE42 of  FIG. 14B ; and   v) a nucleic acid molecule that is hybridizes under stringent conditions to the nucleotide sequence i) or ii); and   using the nucleic acid molecule in the manipulation of plant growth and/or structure.   
     
     
         3 . The method according to  claim 1  further comprising using the polypeptide in combination with a nucleic acid molecule selected from the group consisting of:
 i) a nucleic acid molecule that encodes a CLE41 receptor; 
 ii) a nucleic acid molecule that encodes a CLE42 receptor; 
 iii) a nucleic acid molecule that encodes a polypeptide comprising an amino acid sequence that is at least 70% identical to the consensus sequence of  FIG. 12 , or a functional equivalent thereof; 
 iv) a nucleic acid molecule that is at least 70% identical to the nucleotide sequence of  FIG. 15B  or  15 C; 
 v) a nucleic acid molecule that is at least 70% identical to a nucleic acid molecule of i) or ii); and 
 vi) a nucleic acid molecule that is hybridizes under stringent conditions to the nucleotide sequence i) or ii). 
 
     
     
         4 . The method according to  claim 1  further comprising using the polypeptide in combination with a second polypeptide selected from the group consisting of:
 i) a CLE41 receptor; 
 ii) a CLE42 receptor; 
 iii) a polypeptide comprising an amino acid sequence that is at least 70% identical to the consensus sequence of  FIG. 12 , or a functional equivalent thereof; 
 iv) a polypeptide comprising an amino acid sequence that is at least 70% identical to the PXY sequence of  FIG. 15A ; 
 v) a polypeptide sequence comprising an amino acid sequence which is at least 70% identical to a sequence encoding i) or ii); and 
 vi) a polypeptide comprising an amino acid sequence encoded by a nucleic acid molecule that is at least 70% identical to the PXY nucleotide sequence of  FIG. 15B  or  15 C. 
 
     
     
         5 . The method according to  claim 3  wherein said CLE41 or CLE42 receptor is PXY or a functional equivalent thereof. 
     
     
         6 . A method of manipulating the growth and/or structure of a plant, comprising modulating the level of CLE41 and/or CLE42 or a functional equivalent thereof, in the plant. 
     
     
         7 . A method according to  claim 6  wherein the levels of CLE41 and/or CLE42 are modulated by introducing into a cell of the plant:
 i) a CLE41 polypeptide; 
 ii) a CLE42 polypeptide; 
 iii) a polypeptide comprising an amino acid sequence that is at least 70% identical to the amino acid sequence of amino acids 124 to 137 of the consensus sequence of  FIG. 10 ; 
 iv) a polypeptide comprising an amino acid sequence that is at least 70% identical to the amino acid sequence of CLE41 of  FIG. 13A  or CLE42 of  FIG. 14A ; or 
 v) a polypeptide encoded by a nucleic acid molecule that is at least 70% identical to the nucleotide sequence of CLE41 of  FIG. 13B  or CLE42 of  FIG. 14B . 
 
     
     
         8 . A method according to  claim 6  wherein the levels of CLE41 and/or CLE42 are modulated by introducing into a cell of the plant:
 i) a nucleic acid molecule that encodes a CLE41 polypeptide; 
 ii) a nucleic acid molecule that encodes a CLE42 polypeptide; 
 iii) a nucleic acid molecule that encodes a polypeptide comprising an amino acid sequence that is at least 70% identical to the amino acid sequence of amino acids 124 to 137 of the consensus sequence of  FIG. 10 ; 
 iv) a nucleic acid molecule which is at least 70% identical to the nucleotide sequence of CLE41 of  FIG. 13B  or CLE42 of  FIG. 14B ; or 
 v) a nucleic acid molecule that is hybridizes under stringent conditions to the nucleotide sequence i) or ii). 
 
     
     
         9 . A method according to  claim 6  wherein the levels of levels of CLE41 and/or CLE42 or a functional equivalent thereof are upregulated. 
     
     
         10 . A method according to  claim 6  further comprising introducing into a cell of the plant:
 i) a nucleic acid molecule that encodes a CLE41 receptor; 
 ii) a nucleic acid molecule that encodes a CLE42 receptor; 
 iii) a nucleic acid molecule that encodes a polypeptide comprising an amino acid sequence that is at least 70% identical to the consensus sequence of  FIG. 12 , or a functional equivalent thereof; 
 iv) a nucleic acid molecule that is at least 70% identical to the nucleotide sequence of  FIG. 15B  or  15 C; 
 v) a nucleic acid molecule that is at least 70% identical to a nucleic acid molecule of i) or ii); or 
 vi) a nucleic acid molecule that is hybridizes under stringent conditions to the nucleotide sequence i) or ii). 
 
     
     
         11 . A method according to  claim 6  further comprising introducing into a cell of the plant:
 i) a CLE41 receptor; 
 ii) a CLE42 receptor; 
 iii) a polypeptide comprising an amino acid sequence that is at least 70% identical to the consensus sequence of  FIG. 12 , or a functional equivalent thereof; 
 iv) a polypeptide comprising an amino acid sequence that is at least 70% identical to the PXY sequence of  FIG. 15A ; 
 v) a polypeptide sequence comprising an amino acid sequence which is at least 70% identical to a sequence encoding i) or ii); or 
 vi) a polypeptide comprising an amino acid sequence encoded by a nucleic acid molecule that is at least 70% identical to the PXY nucleotide sequence of  FIG. 15B  or  15 C. 
 
     
     
         12 . A method according to  claim 11  wherein said CLE41 and/or CLE42 receptor is PXY or a functional equivalent thereof. 
     
     
         13 . A plant cell manipulated to express:
 i) a CLE41 polypeptide;   ii) a CLE42 polypeptide;   iii) a polypeptide comprising an amino acid sequence that is at least 70% identical to the amino acid sequence of amino acids 124 to 137 of the consensus sequence of  FIG. 10 ;   iv) a polypeptide comprising an amino acid sequence that is at least 70% identical to the amino acid sequence of CLE41 of  FIG. 13A  or CLE42 of  FIG. 14A ; or   v) a polypeptide encoded by a nucleic acid molecule that is at least 70% identical to the nucleotide sequence of CLE41 of  FIG. 13B  or CLE42 of  FIG. 14B ;   optionally in combination with expression of a receptor for CLE41 and/or CLE42.   
     
     
         14 . A plant cell manipulated to express
 i) a nucleic acid molecule that encodes a CLE41 polypeptide;   ii) a nucleic acid molecule that encodes a CLE42 polypeptide;   iii) a nucleic acid molecule that encodes a polypeptide comprising an amino acid sequence that is at least 70% identical to the amino acid sequence of amino acids 124 to 137 of the consensus sequence of  FIG. 10 ;   iv) a nucleic acid molecule which is at least 70% identical to the nucleotide sequence of CLE41 of  FIG. 13B  or CLE42 of  FIG. 14B ; or   v) a nucleic acid molecule that is hybridizes under stringent conditions to the nucleotide sequence i) or ii).   
     
     
         15 . A plant cell according to  claim 13  further manipulated to express a nucleic acid molecule selected from the group consisting of:
 i) a nucleic acid molecule that encodes a CLE41 receptor; 
 ii) a nucleic acid molecule that encodes a CLE42 receptor; 
 iii) a nucleic acid molecule that encodes a polypeptide comprising an amino acid sequence that is at least 70% identical to the consensus sequence of  FIG. 12 , or a functional equivalent thereof; 
 iv) a nucleic acid molecule that is at least 70% identical to the nucleotide sequence of  FIG. 15B  or  15 C; 
 v) a nucleic acid molecule that is at least 70% identical to a nucleic acid molecule of i) or ii); and 
 vi) a nucleic acid molecule that is hybridizes under stringent conditions to the nucleotide sequence i) or ii). 
 
     
     
         16 . A plant cell according to  claim 13  further manipulated to express a polypeptide selected from the group consisting of:
 i) a CLE41 receptor; 
 ii) a CLE42 receptor; 
 iii) a polypeptide comprising an amino acid sequence that is at least 70% identical to the consensus sequence of  FIG. 12 , or a functional equivalent thereof; 
 iv) a polypeptide comprising an amino acid sequence that is at least 70% identical to the PXY sequence of  FIG. 15A ; 
 v) a polypeptide sequence comprising an amino acid sequence which is at least 70% identical to a sequence encoding i) or ii); and 
 vi) a polypeptide comprising an amino acid sequence encoded by a nucleic acid molecule that is at least 70% identical to the PXY nucleotide sequence of  FIG. 15B  or  15 C. 
 
     
     
         17 . A plant comprising a cell according to  claim 13 . 
     
     
         18 . A method of producing a plant-derived product, comprising using a cell or plant according to  claim 13  in the production of the plant-derived product. 
     
     
         19 . A method of manipulating the growth and/or structure of a plant, comprising the steps of:
 i) providing a cell/seed according to  claim 13 ;   ii) regenerating said cell/seed into a plant; and optionally   iii) monitoring the levels of CLE41 and/or CLE42 or a receptor thereof, and and/or PXY or functional equivalents thereof in said regenerated plant.   
     
     
         20 . An expression construct comprising a first nucleic acid sequence selected from the group consisting of:
 i) a nucleic acid molecule that encodes a CLE41 polypeptide;   ii) a nucleic acid molecule that encodes a CLE42 polypeptide;   iii) a nucleic acid molecule that encodes a polypeptide comprising an amino acid sequence that is at least 70% identical to the amino acid sequence of amino acids 124 to 137 of the consensus sequence of  FIG. 10 ;   iv) a nucleic acid molecule which is at least 70% identical to the nucleotide sequence of CLE41 of  FIG. 13B  or CLE42 of  FIG. 14B ; and   v) a nucleic acid molecule that is hybridizes under stringent conditions to the nucleotide sequence i) or ii);   and optionally a second nucleotide sequence encoding a regulatory sequence capable of expressing the first nucleotide sequence specifically in or adjacent to the vascular tissue of a plant.   
     
     
         21 . An expression cassette according to  claim 20  further comprising:
 i) a nucleic acid molecule that encodes a CLE41 receptor; 
 ii) a nucleic acid molecule that encodes a CLE42 receptor; 
 iii) a nucleic acid molecule that encodes a polypeptide comprising an amino acid sequence that is at least 70% identical to the consensus sequence of  FIG. 12 , or a functional equivalent thereof; 
 iv) a nucleic acid molecule that is at least 70% identical to the nucleotide sequence of  FIG. 15B  or  15 C; 
 v) a nucleic acid molecule that is at least 70% identical to a nucleic acid molecule of i) or ii); or 
 vi) a nucleic acid molecule that is hybridizes under stringent conditions to the nucleotide sequence i) or ii) 
 
     
     
         22 . A host cell or organism comprising an expression construct of  claim 20 . 
     
     
         23 . A transgenic plant seed comprising a cell according to  claim 13 . 
     
     
         24 . A plant derived product produced by a method according to  claim 1 . 
     
     
         25 . A method of producing a plant-derived product comprising:
 a) manipulating the growth and/or structure of a plant produced according to  claim 1 ;   b) growing the plant until it reaches a pre-determined lateral size; and optionally   c) harvesting the plant derived product of the plant.   
     
     
         26 . A method of altering the mechanical properties of a plant or plant derived product comprising:
 a) manipulating the growth and/or structure of a plant according to  claim 1 ;   b) growing the plant until it reaches a pre-determined size; and optionally   c) harvesting a plant derived product of the plant.   
     
     
         27 . The method according to  claim 2  further comprising using the nucleic acid molecule in combination with a second nucleic acid molecule selected from the group consisting of:
 i) a nucleic acid molecule that encodes a CLE41 receptor; 
 ii) a nucleic acid molecule that encodes a CLE42 receptor; 
 iii) a nucleic acid molecule that encodes a polypeptide comprising an amino acid sequence that is at least 70% identical to the consensus sequence of  FIG. 12 , or a functional equivalent thereof; 
 iv) a nucleic acid molecule that is at least 70% identical to the nucleotide sequence of  FIG. 15B  or  15 C; 
 v) a nucleic acid molecule that is at least 70% identical to a nucleic acid molecule of i) or ii); and 
 vi) a nucleic acid molecule that is hybridizes under stringent conditions to the nucleotide sequence i) or ii). 
 
     
     
         28 . The method according to  claim 2  further comprising using the nucleic acid molecule in combination with a polypeptide selected from the group consisting of:
 i) a CLE41 receptor; 
 ii) a CLE42 receptor; 
 iii) a polypeptide comprising an amino acid sequence that is at least 70% identical to the consensus sequence of  FIG. 12 , or a functional equivalent thereof; 
 iv) a polypeptide comprising an amino acid sequence that is at least 70% identical to the PXY sequence of  FIG. 15A ; 
 v) a polypeptide sequence comprising an amino acid sequence which is at least 70% identical to a sequence encoding i) or ii); and 
 vi) a polypeptide comprising an amino acid sequence encoded by a nucleic acid molecule that is at least 70% identical to the PXY nucleotide sequence of  FIG. 15B  or  15 C. 
 
     
     
         29 . The method according to  claim 4  wherein said CLE41 or CLE42 receptor is PXY or a functional equivalent thereof. 
     
     
         30 . A plant cell according to  claim 14  further manipulated to express a nucleic acid molecule selected from the group consisting of:
 i) a nucleic acid molecule that encodes a CLE41 receptor; 
 ii) a nucleic acid molecule that encodes a CLE42 receptor; 
 iii) a nucleic acid molecule that encodes a polypeptide comprising an amino acid sequence that is at least 70% identical to the consensus sequence of  FIG. 12 , or a functional equivalent thereof; 
 iv) a nucleic acid molecule that is at least 70% identical to the nucleotide sequence of  FIG. 15B  or  15 C; 
 v) a nucleic acid molecule that is at least 70% identical to a nucleic acid molecule of i) or ii); and 
 vi) a nucleic acid molecule that is hybridizes under stringent conditions to the nucleotide sequence i) or ii). 
 
     
     
         31 . A plant cell according to  claim 14  further manipulated to express a polypeptide selected from the group consisting of:
 i) a CLE41 receptor; 
 ii) a CLE42 receptor; 
 iii) a polypeptide comprising an amino acid sequence that is at least 70% identical to the consensus sequence of  FIG. 12 , or a functional equivalent thereof 
 iv) a polypeptide comprising an amino acid sequence that is at least 70% identical to the PXY sequence of  FIG. 15A ; 
 v) a polypeptide sequence comprising an amino acid sequence which is at least 70% identical to a sequence encoding i) or ii); and 
 vi) a polypeptide comprising an amino acid sequence encoded by a nucleic acid molecule that is at least 70% identical to the PXY nucleotide sequence of  FIG. 15B  or  15 C. 
 
     
     
         32 . A plant comprising a cell according to  claim 14 . 
     
     
         33 . A method of producing a plant-derived product, comprising using a cell or plant according to  claim 14  in the production of the plant-derived product. 
     
     
         34 . The method of  claim 33 , wherein the plant-derived product is selected from the group consisting of biomass, fibres, forage, biocomposites, biopolymers, wood, biofuel and paper. 
     
     
         35 . A method of manipulating the growth and/or structure of a plant, comprising the steps of:
 i) providing a cell/seed according to  claim 14 ;   ii) regenerating said cell/seed into a plant; and optionally   iii) monitoring the levels of CLE41 and/or CLE42 or a receptor thereof, and/or PXY or functional equivalents thereof in said regenerated plant.   
     
     
         36 . A transgenic plant seed comprising a cell according to  claim 14 . 
     
     
         37 . A plant derived product produced by a method according to  claim 2 . 
     
     
         38 . A method of producing a plant-derived product comprising:
 a) manipulating the growth and/or structure of a plant produced according to  claim 2 ;   b) growing the plant until it reaches a pre-determined lateral size; and optionally   c) harvesting the plant derived product of the plant.   
     
     
         39 . A method of altering the mechanical properties of a plant or plant derived product comprising:
 a) manipulating the growth and/or structure of a plant according to  claim 2 ;   b) growing the plant until it reaches a pre-determined size; and optionally   c) harvesting a plant derived product of the plant.   
     
     
         40 . A plant derived product produced by a method according to  claim 6 . 
     
     
         41 . A method of producing a plant-derived product comprising:
 a) manipulating the growth and/or structure of a plant produced according to  claim 6 ;   b) growing the plant until it reaches a pre-determined lateral size; and optionally   c) harvesting the plant derived product of the plant.   
     
     
         42 . A method of altering the mechanical properties of a plant or plant derived product comprising:
 a) manipulating the growth and/or structure of a plant according to  claim 6 ;   b) growing the plant until it reaches a pre-determined size; and optionally   c) harvesting a plant derived product of the plant.   
     
     
         43 . The method of  claim 18 , wherein the plant-derived product is selected from the group consisting of biomass, fibres, forage, biocomposites, biopolymers, wood, biofuel and paper.