The invention relates to a method for producing a chromosome substitution library, comprising: a) producing a population of DH lines from an at least partially heterozygous starting organism b) identifying a first DH line in the population that has at least one non- recombined chromosome pair c) identifying a matching second DH line in the population that has at least one non-recombined chromosome pair that is the same as the non-recombined chromosome pair of the first DH line, and at least one non-recombined chromosome pair that is different from the non-recombined chromosome pair of the first DH line d) selecting the first DH line and the second DH line as a pair of parents, the progeny of which has at least three non-recombined chromosomes, at least two of which form a pair e) repeating steps b), c) and d) one or more times on the DH population of the heterozygous starting organism, or repeating steps a)-d) on a DH population produced from a progeny plant of a pair of parents selected in step d), until preferably all chromosomes from the selected DH lines are non-recombined and preferably all possible combinations of the parental chromosomes are selected, to assemble a library of chromosome substitution lines. The invention further relates to the libraries thus obtained.Linvention concerne un procédé de fabrication dune bibliothèque de substitution de chromosome qui comprend : a) la génération dune population de lignées HD à partir dun organisme de départ au moins partiellement hétérozygote b) lidentification dune première lignée HD dans la population qui possède au moins une paire de chromosomes non recombinés c) lidentification dune seconde lignée HD dans la population qui possède au moins une paire de chromosomes non recombinés qui est la même que la paire de chromosomes non recombinés de la première lignée HD, et dau moins une paire de chromosomes non recombinés qui est différente de la paire de chromosomes non recombinés de la première lignée HD d) la sélect
