Methods for Preparing Mutant Plants

A method for identifying a mutant microorganism of a predefined species carrying one or more mutation(s) in nucleotide(s) of interest [NOI(s)], in a predetermined target sequence, said method comprising the steps of:

• I. Providing Np microorganisms of said species, wherein Np is an integer of at least 5000;
• II. Subjecting said microorganisms to a step of mutagenesis, leading to a low rate of mutagenesis, wherein said mutagenesis leads to a survival rate of at least 30%, thereby generating a pool of microorganisms representing a plurality of genotypes;
• III. Dividing said mutagenized microorganisms into physically separated sub-pools;
• IV. Subjecting each sub-pool to a step of reproduction so that each sub-pool in theory comprises more than one microorganisms of each genotype,
• V. Preparing DNA samples, such as cDNA samples prepared from mRNA samples or gDNA samples, from each sub-pool;
• VI. Identifying sub-pool(s) comprising DNA comprising the mutation(s) using sensitive detection means;
• and
• VII. Identifying microorganisms within said sub-pool comprising said mutation.