Method for producing transgenic plants with modified 5-aminolevulinic acid biosynthesis, and method for identifying 5-aminolevulinic acid synthesis effectors

Abstract

The invention relates to a method for producing transgenic plants by means of modified 5-aminolevulinic acid biosynthesis, transgenic plant cells, transgenic plant parts, transgenic seeds, and transgenic reproduction material. The invention is characterised in that one or several nucleic acid molecules coding for a protein with a 5-aminolevulinic acid synthase function (ALAS), selected from a group of feedback regulated ALAS, animal ALAS and bacterial ALAS, an active fragment thereof or an antisense or complementary sequence thereof, are integrated into the plant genome in a stable form. Effectors of natural 5-aminolevulinic acid biosynthesis in plants are found using a method for determining effector efficacy of a test substance in relation to a GSAAT, by: a) determining the enzymatic activity of the GSAAT in the absence of a test substance b) determining the enzymatic activity of the GSAAT in the presence of a test substance; and c) by comparing the enzymatic activities determined under a) and b).

What is claimed is:  
     
         1 . A method for producing transgenic plants, transgenic plant cells, transgenic plant parts, transgenic plant seeds, and transgenic propagation material, wherein one or more nucleic acid molecules encoding a protein with the function of a 5-aminolevulinic acid synthase (ALAS) selected from the group of the feedback-regulated ALASs, animal ALASs and bacterial ALASs, an active fragment thereof or an antisense or complementary sequence thereof are stably integrated into the plant genome.  
     
     
         2 . The method as claimed in  claim 1 , wherein one or more nucleic acid molecules encoding a protein with the function of an ALAS selected from the group of the feedback-regulated ALASs, animal ALASs and bacterial ALASs, an active fragment thereof or an antisense or complementary sequence thereof are stably integrated into the plant plastome by plastid transformation.  
     
     
         3 . The use of at least one nucleic acid molecule encoding a protein with the function of an ALAS selected from the group of the feedback-regulated ALASs, animal ALASs and bacterial, an active fragment thereof or an antisense or complementary sequence thereof for producing transgenic plant cells or plants.  
     
     
         4 . The use of at least one nucleic acid molecule encoding a protein with the function of an ALAS selected from the group of the feedback-regulated ALASs, animal ALASs and bacterial ALASs, or an active fragment thereof for expressing a protein with the function of an ALAS or an active fragment thereof in transgenic plant cells or plants.  
     
     
         5 . The use of at least one nucleic acid molecule encoding a protein with the function of an ALAS selected from the group of the feedback-regulated ALASs, animal ALASs and bacterial ALASs, or an active fragment thereof or an antisense or complementary sequence thereof for producing transgenic plant cells or plants whose expression of a protein with the function of a GSAAT or an active fragment thereof is suppressed or inhibited.  
     
     
         6 . A non-naturally occurring chimeric gene, at least containing a promoter which is suitable for expressing, in plants, an ALAS selected from the group of the feedback-regulated ALASs, animal ALASs and bacterial ALASs and which is functionally linked to a DNA molecule encoding a protein with the function of an ALAS or an active fragment thereof or an antisense or complementary sequence thereof.  
     
     
         7 . A transgenic plant, a transgenic plant cell, a transgenic plant part, a transgenic plant seed or transgenic propagation material containing a DNA molecule encoding a protein with the function of an ALAS selected from the group of the feedback-regulated ALASs, animal ALASs and bacterial ALASs or an active fragment thereof or antisense or complementary sequence thereof.  
     
     
         8 . A method for producing transgenic plants, transgenic plant cells, transgenic plant parts, transgenic plant seeds, and transgenic propagation material where one or more nucleic acid molecules encoding a protein with the function of a 5-aminolevulinic acid synthase (ALAS), an active fragment thereof or an antisense or complementary sequence thereof, are stably integrated into the plant genome, wherein the plant is a monocotyledonous plant.  
     
     
         9 . The method as claimed in  claim 8 , wherein one or more nucleic acid molecules encoding a protein with the function of an ALAS, an active fragment thereof or an antisense or complementary sequence thereof are stably integrated into the plant plastome by plastid transformation.  
     
     
         10 . The use of at least one nucleic acid molecule encoding a protein with the function of an ALAS or an active fragment thereof or an antisense or complementary sequence thereof for the production of transgenic plant cells or plants, wherein the plant is a monocotyledonous plant.  
     
     
         11 . The use of at least one nucleic acid molecule encoding a protein with the function of an ALAS or an active fragment thereof for expressing a protein with the function of an ALAS or an active fragment thereof in transgenic plant cells or plants, wherein the plant is a monocotyledonous plant.  
     
     
         12 . The use of at least one nucleic acid molecule encoding a protein with the function of an ALAS or an active fragment thereof or an antisense or complementary sequence thereof for producing transgenic plant cells or plants whose expression of a protein with the function of a GSAAT or an active fragment thereof is suppressed or inhibited, wherein the plant is a monocotyledonous plant.  
     
     
         13 . A non-naturally occurring chimeric gene, at least containing a promoter which is suitable for expressing an ALAS in plants and which is functionally linked to a DNA molecule encoding a protein with the function of an ALAS or an active fragment thereof or an antisense or complementary sequence thereof, wherein the plant is a monocotyledonous plant.  
     
     
         14 . A transgenic plant, a transgenic plant cell, a transgenic plant part, a transgenic plant seed or transgenic propagation material containing a DNA molecule encoding a protein with the function of an ALAS or an active fragment thereof or an antisense or complementary sequence thereof, wherein the plant is a monocotyledonous plant.  
     
     
         15 . The method as claimed in  claim 1 ,  2 ,  8  and/or  9 , wherein said nucleic acid molecules are under the control of a feedback-regulated promoter.  
     
     
         16 . The use of a feedback-regulated promoter in the method as claimed in one or more of claims  1 ,  2 ,  8 ,  9  and  15 .  
     
     
         17 . A recombinant vector containing a chimeric gene as claimed in  claim 6  or  13 .  
     
     
         18 . A transgenic plant cell which is transformed and/or genetically modified with a chimeric gene as claimed in  claim 6  or  13  or a vector as claimed in  claim 17 .  
     
     
         19 . A plant, plant cell, plant part, part seed or propagation material as claimed in any of claims  7 ,  14  or  18  which can be produced by a method as claimed in one or more of claims  1 ,  2 ,  8 ,  9  or  15 .  
     
     
         20 . The plant as claimed in  claim 19 , which is a useful plant or ornamental.  
     
     
         21 . The plant as claimed in  claim 20 , which is a maize plant.  
     
     
         22 . A method for determining the effector efficiency of a test substance in relation to a GSAAT activity, wherein 
 a) the enzymatic GSAAT activity is determined in the absence of a test substance;    b) the enzymatic GSAAT activity is determined in the presence of a test substance; and    c) the enzymatic activities determined under a) and b) are compared.    
     
     
         23 . The method as claimed in  claim 22 , which is applied in an automated test system for testing the effector efficacy of test substances.  
     
     
         24 . A method for finding herbicidal active substances, wherein the test substance is tested in the method as claimed in  claim 22 .  
     
     
         25 . The method as claimed in  claim 24 , wherein the test substance is additionally subjected to a Lemna test.  
     
     
         26 . The use of GSAAT effectors as herbicide.  
     
     
         27 . The use as herbicide of a compound which, in a test method for effector properties in relation to GSAAT as claimed in  claim 22 , has in at least a molar concentration in the range of 10 −4  to 10 −7  and an at least 30% inhibitor action.  
     
     
         28 . The use as claimed in  claim 27 , wherein the compound is also active in the Lemna test.  
     
     
         29 . A method for controlling undesired vegetation in crops of useful plants, wherein a herbicidal composition is applied to the location of the useful plant, the useful plant or its propagation material, which comprises a compound which in the test method as claimed in  claim 22  in at least a molar concentration in the range of 10 −4  to 10 −7  molar an at least 30% inhibitor action, as herbicide, an at least 30% inhibitor action in relation to GSAAT.  
     
     
         30 . The method as claimed in  claim 29 , wherein the compound is also active in the Lemna test.  
     
     
         31 . The method as claimed in  claim 29  and/or  30 , wherein the useful plant is a transgenic plant as claimed in  claim 7  or  14 .  
     
     
         32 . A method for controlling undesired vegetation in crops of useful plants, wherein a useful plant as claimed in one or more of claims  7 ,  14 ,  19 ,  20  and/or  21  is employed and a herbicidal composition is applied to the location of this plant, the plant itself or its propagation material whose mechanism of action is based at least in part on inhibition of plant GSAAT.