Generation of hapoloid plants based on knl2

Abstract

Non-transgenic and transgenic plants, preferably crop plants, comprising at least one mutation of the KINTEOCHORE NULL2 (KNL2) protein, especially a mutation causing a substitution of an amino acid within the KNL2 protein, preferably within the C-terminal region of the KNL2 protein, which preferably have the biological activity of a haploid inducer. Further are methods of generating plants and haploid and double haploid plants obtainable by crossing non-transgenic and transgenic plants with wildtype plants as well as methods of facilitating cytoplasm exchange.

What is claimed is: 
     
         1 . A plant, wherein the plant comprises a nucleotide sequence encoding a KINETOCHORE NULL2 (KNL2) protein comprising a CENPC-k motif, wherein the nucleotide sequence comprises at least one mutation in the CENPC-k motif encoding sequence, and wherein the plant is a crop or agricultural plant or ornamental flower or ornamental crop. 
     
     
         2 . The plant according to  claim 1 , wherein the at least one mutation is a deletion, addition or substitution of at least one nucleotide in the nucleotide sequence for the CENPC-k motif. 
     
     
         3 . The plant according to  claim 1 , wherein the plant has biological activity of a haploid inducer. 
     
     
         4 . The plant according to  claim 1 , wherein the plant is sunflower, peanut, corn, potato, sweet potato, bean, pea, chicory, lettuce, endive, cabbage, cauliflower, broccoli, turnip, radish, spinach, onion, garlic, eggplant, celery, carrot, squash, pumpkin, zucchini, cucumber, apple, pear, melon, strawberry, grape, raspberry, pineapple, soybean, Cannabis, Humulus (hop), tomato, sorghum, sugar cane, in particular maize, wheat, barley, sorghum, rye, oats, turf and forage grasses, millet, rice and sugar cane. 
     
     
         5 . The plant according to  claim 1 , wherein the plant expresses a KNL2 protein having at least one amino acid addition, amino acid deletion and/or amino acid substitution in the CENPC-k motif. 
     
     
         6 . The plant according to  claim 1 , wherein the KNL2 protein comprises an amino acid sequence according to one of SEQ ID No. 23 to SEQ ID No. 123 or SEQ ID No. 164 to SEQ ID No. 274. 
     
     
         7 . The plant according to  claim 1 , wherein the amino acid arginine at position 10 of SEQ ID No. 4 to SEQ ID No. 22 is substituted, preferably substituted for alanine and/or wherein the amino acid tryptophan at position 19 of SEQ ID No. 4 to SEQ ID No. 22 is substituted, preferably substituted for arginine or wherein the amino acid arginine at position 8 or 7 or 6 of SEQ ID No. 127 to SEQ ID No. 163 is substituted, preferably substituted for alanine and/or wherein the amino acid tryptophan at position 17 or 16 or 15 of SEQ ID No. 4 to SEQ ID No. 22 is substituted, preferably substituted for arginine. 
     
     
         8 . The plant according to  claim 1 , wherein the wildtype CENPC-k motif comprises an amino acid sequence according to one of SEQ ID No. 124 to SEQ ID No. 126. 
     
     
         9 . The plant according to  claim 1 , wherein the non-mutated CENPC-k motif comprises an amino acid sequence according to table 1 or table 2 or table 3. 
     
     
         10 . A part of the plant according to  claim 1 , where in the part is a shoot vegetative organ, root, flower or floral organ, seed, fruit, ovule, embryo, plant tissue or cell. 
     
     
         11 . A haploid plant obtainable by crossing the plant of  claim 1  with a plant expressing wildtype KNL2 protein. 
     
     
         12 . A double haploid plant obtainable by converting the haploid plant according to  claim 11  into a double haploid plant, preferably via colchicine treatment. 
     
     
         13 . A haploid plant obtainable by crossing in a first step the plant according to any of  claim 1  with a plant comprising a nucleotide sequence encoding a centromer histone H3 (CENH3) protein comprising a CATD domain, wherein the nucleotide sequence comprises at least one mutation causing in the CATD domain an amino acid substitution which confers the biological activity of a haploid inducer, and crossing in a second step a plant obtained in the first step with a plant expressing wildtype KNL2 protein and wildtype CENH3 protein. 
     
     
         14 . A double haploid plant obtainable by converting the haploid plant according to  claim 13  into a double haploid plant, preferably via colchicine treatment. 
     
     
         15 . A plant having biological activity of a haploid inducer and comprising a nucleotide sequence encoding a KINETOCHORE NULL2 (KNL2) protein, wherein the nucleotide sequence comprises at least one mutation. 
     
     
         16 . A nucleotide sequence encoding at least the C-terminal part of KNL2 protein comprising at least one mutation causing an amino acid substitution, deletion or addition, preferably in the CENPC-k motif. 
     
     
         17 . A plant cell or host cell comprising the nucleotide sequence of  claim 16  or an according vector as a transgene.