Increased fungal resistance in plants via modulation of a wall-associated kinase or benzoxazinoids

A method for selecting a maize plant having Helminthosporium turcicum resistance, wherein the Helminthosporium turcicum resistance is regulated by at least one wall-associated kinase, the method comprising:

• (i) providing at least one maize plant cell, tissue, organ, or whole plant comprising at least one gene encoding a wall-associated kinase in the genome of said maize plant cell, tissue, organ, or whole plant;
• (ii) determining a reduced synthesis of at least one benzoxazinoid in response to a Helminthosporium turcicum pathogen infection in at least one maize plant cell, tissue, organ, or whole plant,
• wherein said plant cell, tissue, organ, or whole plant exhibit an amount of at least one benzoxazinoid which is reduced by at least 25% as compared to a corresponding control plant cell, control tissue, control organ, or control whole plant of the same genotype, but lacking the reduced expression level of the at least one molecule within the signaling pathway from the at least one wall-associated kinase to the synthesis of at least one benzoxazinoid or within the synthesis pathway of at least one benzoxazinoid,
• wherein the synthesis of the at least one benzoxazinoid is regulated by the at least one wall-associated kinase and reduced synthesis is caused by a reduced expression level of at least one molecule within a signalling pathway from the at least one wall-associated kinase to the synthesis of at least one benzoxazinoid or within a synthesis pathway of at least one benzoxazinoid in said maize plant cell, tissue, organ, or whole plant, the reduced expression level being relative to the expression level of the at least one molecule within the signalling pathway in the control plant cell, control tissue, control organ, or control whole plant,
• wherein the at least one wall-associated kinase:
• a) is encoded by a nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO:
• 1,
• b) is encoded by a nucleic acid molecule comprising the nucleotide sequence having at least 95% sequence identity to the nucleotide sequence of SEQ ID NO:
• 1,
• c) is encoded by a nucleic acid molecule comprising the nucleotide sequence coding for the amino acid sequence of SEQ ID NO:
• 2,
• d) is encoded by a nucleic acid molecule comprising the nucleotide sequence coding for an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:
• 2,
• e) comprising the amino acid sequence of SEQ ID NO:
• 2, or
• f) comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO:
• 2,
• wherein the benzoxazinoid whose synthesis is regulated by the at least one wall-associated kinase, is selected from at least one of 7,8-dimethoxy-2,4-dihydroxy-1,4(2H)-benzoxazin-3-one (DIM2BOA)-glucoside, 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA), DIMBOA-glucoside, 2-O-Glucosyl-7-methoxy-1,4(2H)-benzoxazin-3-one (HMBOA)-glucoside, and any combination thereof, and
• wherein the at least one molecule within the signalling pathway from the at least one wall-associated kinase to the synthesis of at least one benzoxazinoid or within the synthesis pathway of at least one benzoxazinoid is selected from the group consisting of:
• (i) the nucleic acid sequences of any one of SEQ ID NOs:
• 10, 12, and 16, or
• (ii) a nucleic acid sequence having identity of at least 98% to the nucleic acid sequence of any one of SEQ ID NOs:
• 10, 12, and 16, over the full length of the coding regions, or
• (iii) the amino acid sequences of SEQ ID NOs:
• 11, 13, and 17, or
• (iv) an amino acid sequence having identity of at least 98% to the amino acid sequence of anyone of SEQ ID NOs:
• 11, 13, and 17, over the full length of the sequence.

A method for identifying at least one gene involved in increased pathogen resistance in a plant cell, tissue, organ, whole plant, or plant material the method comprising:

• (i) determining the genotype of at least one plant cell, tissue, organ, whole plant, or plant material with respect to the presence of at least one gene encoding a wall-associated kinase in the genome of said plant cell, tissue, organ, whole plant or plant material;
• (ii) optionally determining the benzoxazinoid signature of the at least one plant cell, tissue, organ, whole plant, or plant material of step (i);
• (iii) exposing the at least one plant cell, tissue, organ, whole plant, or plant material of step (i) or (ii) to a stimulus, optionally wherein the stimulus is correlated with the benzoxazinoid signature in the at least one plant cell, tissue, organ, whole plant, or plant material;
• (iv) performing an analysis of at least one analyte obtained from the at least one plant cell, tissue, organ, whole plant, or plant material of step (i) or (ii) after exposition to the stimulus;
• (v) determining at least one gene being regulated upon exposition to a stimulus according to step (iii) in at least one cell of the at least one plant cell, tissue, organ, whole plant, or plant material as derivable from the analysis of at least one analyte as defined in step (iv),
• (vi) subjecting the at least one gene as determined in step (v) to a functional characterization;
• and
• (vii) providing at least one gene involved in increased pathogen resistance in a plant cell, tissue, organ, whole plant, or plant material.

A method of increasing pathogen resistance in a plant cell, tissue, organ, whole plant, or plant material, the method comprising:

• (i) providing at least one plant cell, tissue, organ, whole plant or plant material;
• (ii) (a) treating the at least one plant cell, tissue, organ, whole plant or plant material according to step (i) with a substance neutralizing an effect of at least one benzoxazinoid, and/or
• (ii) (b) treating the at least one plant cell, tissue, organ, whole plant or plant material according to step (i) with a substance activating the signalling pathway downstream of at least one wall-associated kinase;
• and/or
• (ii) (c) treating the at least one plant cell, tissue, organ, whole plant or plant material according to step (i) with a substance modulating or modifying the activity of at least one promoter or at least one regulatory sequence of at least one gene of the at least one plant cell, tissue, organ, whole plant or plant material of step (i), wherein said promoter or said regulatory sequence is involved in the regulation of transcription of at least on gene involved in the signalling pathway of, or downstream of at least one wall-associated kinase or involved in the synthesis pathway of at least one benzoxazinoid;
• (ii) (d) treating the at least one plant cell, tissue, organ, whole plant or plant material according to step (i) with a substance inhibiting the synthesis of at least one benzoxazinoid;
• and
• (iii) reducing an amount of at least one benzoxazinoid and thereby increasing pathogen resistance in at least one plant cell, tissue, organ, whole plant, or plant material.