The Original Study On Molecular Genetics Structure Of Mating-type Locus In Lentinula Eodes

Lentinula edodes is an important cultivated mushroom in China. The development of Lentinus edodes production promotes more studies on it. Lentinula edodes was heterothallic and carry tetrapolar incompatibility factors: A and B factors, which controlled their mating. polymorphism of Mating type gene is the basic theory of Heterothallic mushroom for crossbreeding, such as Lentinula edodes. Now still fewer research at Molecular level on mating-type genes and mating type loci about Lentinula edodes and other species.Our laboratory access the whole genome sequence information for two different mating types of 135 strain by use the 454 high-throughput sequencing technology for 6 times coverage of the whole genome sequence of 135 monokaryons. Based on this work, this study Comprehensive research for mating type locus on molecular genetics structure about ten monokaryons of Lentinula edodes, by doing this we access the sequence existence on mating type locus about this ten strains and also analysis it by using Bioinformatics. The results of this study are as follows:We obtained the sequence on A mating type locus of the ten strains, it’s about 10-12kb in length. Every A mating type locus contain a pair of homeodomain proteins respectively, by cDNA amplification we established the coding region of each homeodomain protein contains homology domain, N-terminal sequence, the role of the dimerization motif, nuclear localization signal, the charge of mating-type protein and other structure or motif which is closely related to its function. Polymorphism analysis showed that there are 4 kinds of different A mating type genes in ten monokaryons strains.We obtained the sequence on B mating type locus of the ten strains, it’s about 50-70kb in length. There are in each strain, the pheromone receptor genes all have transmembrane seven times on structure characteristics, it have a high same rate and similar rates with the pheromone receptor sequences of Coprinopsis cinerea and Schizophyllum, they can be determined as homologous sequences. Pheromone precursor gene only can be found around two of the seven seven pheromone receptor genes and all have characteristics of CaaX motif. Polymorphism analysis showed that there are 3 kinds of different B mating type genes in ten monokaryons strains.According to the 4 different pheromone receptor gene DNA sequences of 135P1 and 135P32, we design 4 pairs of spcific primers, select about 120 proginies of Lentinula edodes strain 135 spore monokaryons for PCR amplification and restructure analysis, the result showed recombination is cann’t be found.This is the first report about the complete sequence of mating type loci was obtained from Lentinula edodes strain and This study firstly reveals the molecular organization of the B mating type locus of Lentinula edodes.