Haploid inducers

Abstract

The present invention relates to the provision of technical means such as nucleic acids which, after transcription or after expression in a plant, are suitable for mediating the property of a haploid inductor or for increasing the induction capability of a haploid inductor, as well as methods and uses for the production and identification of non-transgenic and transgenic plant haploid inductors, as well as the improvement of existing plant haploid inductors.

1 - 18 . (canceled) 
     
     
         19 . A method for production of a plant which is suitable for use as a haploid inductor, the method comprising:
 mutagenizing plant cells;   regenerating plants from the mutagenized plant cells; and   identifying the regenerated plant which has at least one mutation in an endogenous DNA sequence which is identical to an isolated nucleic acid comprising a nucleotide sequence, which is   (i) selected from the group consisting of SEQ ID Nos: 8, 9, 10 and 11,   (ii) complementary to the sequence selected from the group consisting of SEQ ID Nos: 8, 9, 10 and 11,   (iii) is at least 80% identical to the sequence selected from the group consisting of SEQ ID Nos: 8, 9, 10 and 11,   (iv) hybridizes with a sequence at least 80% identical to the sequence selected from the group consisting of SEQ ID Nos: 8, 9, 10 and 11 under stringent conditions,   or has at least one mutation in a regulatory sequence of the endogenous DNA sequence, which mutation produces a change in expression rate of the endogenous DNA sequence in the identified plant, in comparison to a non-mutagenized wild-type plant, or a change in the activity or stability of a protein or polypeptide encoded by the endogenous DNA sequence in the identified plant, in comparison to a non-mutagenized wild-type plant,   wherein the at least one mutation causes a property of a haploid inducer to be mediated or the induction capability of a haploid inducer to be increased in the identified plant.   
     
     
         20 . The method of  claim 19 , wherein the change in the expression rate of the endogenous DNA sequence in the identified plant is detected in a pollen or a tissue of a pollen of the identified plant. 
     
     
         21 . The method of  claim 19 , wherein the change in the activity or stability of the protein or the polypeptide encoded by the endogenous DNA sequence is detected in a pollen or a tissue of a pollen of the identified plant. 
     
     
         22 . The method of  claim 19 , wherein the plant comprises the at least one mutation in the regulatory sequence of the endogenous DNA sequence. 
     
     
         23 . The method of  claim 22 , wherein the at least one mutation is the deletion of a nucleotide or the addition of a nucleotide. 
     
     
         24 . The method of  claim 22 , wherein the regulatory sequence of the endogenous DNA sequence is a promoter. 
     
     
         25 . The method of  claim 24 , wherein the promoter comprises a modification in a cis-regulatory element. 
     
     
         26 . The method of  claim 25 , wherein the activity of the promoter is altered as compared to the activity of a promoter without the modification in the cis-regulatory element. 
     
     
         27 . The method of  claim 26 , wherein the activity of the promoter is increased as compared to the activity of a promoter without the modification in the cis-regulatory element. 
     
     
         28 . The method of  claim 26 , wherein the activity of the promoter is reduced as compared to the activity of a promoter without the modification in the cis-regulatory element. 
     
     
         29 . The method of  claim 19 , wherein the at least one mutation is an exchange, addition or deletion of at least one nucleobase in the encoding region of the endogenous DNA sequence, and wherein the exchange, addition or deletion of the at least one nucleobase leads to an amino acid exchange in the encoded protein and produces an alteration in the activity or stability of the protein, in comparison to the wild-type protein. 
     
     
         30 . The method of  claim 19 , wherein the identifying step comprises contacting a sample from the plant with a molecular marker, wherein the molecular marker is a DNA primer or a pair of DNA primers. 
     
     
         31 . The method of  claim 30 , wherein a sample from the plant is contacted with the DNA primer or the pair of DNA primers in a polymerase chain reaction (PCR). 
     
     
         32 . The method of  claim 31 , wherein the PCR is reverse transcription PCR (RT-PCR). 
     
     
         33 . The method of  claim 29 , wherein the molecular marker demonstrates the presence or absence of the at least one mutation. 
     
     
         34 . The method of  claim 33 , wherein the molecular markers are KASPar or TaqMan markers. 
     
     
         35 . The method of  claim 29 , wherein the sample is a pollen sample. 
     
     
         36 . The method of  claim 31 , wherein the DNA primer or the pair of DNA primers comprises a sequence for detection of the at least one mutation in a gene encoding for phospholipase. 
     
     
         37 . The method of  claim 31 , wherein the DNA primer or the pair of DNA primers are specific for a transposon or a target gene. 
     
     
         38 . The method of  claim 31 , wherein the at least one mutation in the endogenous DNA sequence is a point mutation. 
     
     
         39 . A plant or plant part produced according to the method of  claim 19 . 
     
     
         40 . A descendant of the plant according to  claim 39 .