Construction of a high density linkage map of Lentinula edodes by the gene mapping.

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We have previously constructed the genetic linkage map of the shiitake mushroom, Lentinula edodes, based on the segregation of 203 AFLP (Amplified Fragment Length Polymorphism) markers (Terashima et al., 2002). The aim of this study is to improve our previous linkage map in terms of the marker density and the accuracy of the linkage relationships among markers by the development of gene markers and the mapping of them. The results were as follows ; (1) Eighty-five gene (cDNA) markers which are amplified with the PCR primers designed from the cDNA sequence of L. edodes were developed. (2) The 29 cDNA markers were assigned into 7 linkage groups of the previous AFLP-based linkage map which consists of 11 linkage groups. The map re-constructed has a total length of 1907.7 centimorgans (cM) with an average interval between markers of 9.2 cM and is comprised of 11 linkage groups as follows ; LG I (Linkage group I), 344.8cM, 42 markers ; LG II, 257.1cM, 40 markers ; LG III, 248.6cM, 28 markers ; LG IV, 216.1cM, 25 markers ; LG V, 207.1cM, 26 markers ; LG VI, 174.8cM, 18 markers ; LG VII, 168.4cM, 19 markers ; LG VIII, 129.4cM, 19 markers ; LG IX, 97.9 cM, 6 markers ; LG X, 44.9 cM, 7 markers ; LG XI, 18.6 cM, 4 markers. (3) About 50 % of cDNA-sequences of 277 (the average size, about 1.2 kb) showed significant homology to the sequences registered in the DNA Data Bank of Japan (DDBJ) with less than 10^<-10> of expected value (E-value) probability. The improved linkage map in this study would contribute to the developments of the research on molecular breeding of L. edodes, because the cDNA markers are able to use as an anchor marker for combining maps from different populations.