Scar Markers Used In Constructing Genetic Linkage Map Of Volvariella Volvacea

In this study, three kinds of random primer molecular markers (RAPD, ISSR, SRAP) were used to transformed into SCAR markers which were more stable and specificity. At the same time, 112 single-spore group of volvariella volvacea were used to to Preliminary construct genetic linkage map of SCAR markers.With the strain PY of volvariella volvacea as the research material, 26-regenerated strains were got by the mononuclear protoplast technology. Further, 11 strains of these were proved to be monokaryotic strains by the biological characteristics test and the nuclear staining experiment of mycelium.These 11 strains and 5strains of single-spore isolates of the strain PY producing were paired with each other 22. Then, 2 groups of steady hybrid strains were got by purification and separation test, biology test and the identification of molecular biology tests. These 2 hybrid strains were named PYy14-PYy8 and PYd15-PYd21. Finally, the strain PYd15-PYd21 was used as the original strain which were immediately produced and Collected 112 single-spore isolates to construct the genetic linkage map.Hybrid strain PYd15-PYd21, strain PYd15, strain PYd21 as the templates and random primers(RAPD,ISSR,SRAP) as the primes, amplified by PCR, specific DNA bands were recycled, transformed, Sequenced, designed of primers and finally turned into special, steady SCAR markers. Conclusions were as follows: I There were 116 polymorphic DNA fragments were Screened from 2000 RAPD primers; 30 polymorphic DNA fragments from 153 pairs of SRAP primers (17 upstream primers, 9 downstream primers) and 9 fragments from 20 ISSR primers. II 39 RAPD markers, 4 SRAP markers, 3 ISSR markers were transformed into SCAR markers and the success rate each was 33.6%, 13.3%, 33.3%. What's more, there were 4 Co-dominant markers in the 39 SCAR markers of RAPD and the rate was 10.3%.MAPMAKER3.0, 48 SCAR markers and 112 single-spore strains of group were used to get Linkage analysis and construct the genetic linkage map. Conclusions as follows: I 26 SCAR markers distributed in seven linkage groups, but 22 markers are not linked with each other. II Each linkage group had 2-8 markers and the total length: 535.44 cM, Linkage group length range: 20.8cM ~ 177.1cM, the average distance 20.59cM between two adjacent markers.