Morphological And B Mating-type Locus Anylysis Of Lentinus Squarrosulus

Edible fungi as a healthy food,which is low in fat content and rich in dietary fiber and protein,are very popular among consumers.Lentinus squarrosulus with big market potential is one kind of recently developed fungus strains.The main findings of this study are as follows:1.From the cultivating result of 8 wild strains of Lentinus squarrosulus.We get the growing condition of this kind of mushroom.The temperature is 3-4℃higher than that of Lentinula edodes.Thus,other conditions are similar to the growing conditions of Lentinula edodes.Temperature changing and clour changing into dark are needed for fruitbody growing.We get fruitbodies from only 6 strains.There are 3 kinds of the fruitbodies.2.ITS region of 8 strains and Lentinula edodes 135 strain were amplified by PCR, sequenced and compared.The result indicated that there are 3groups between the ITS regions of Lentinus squarrosulus.This was equal to the result of cultivating and proved that studying ITS region can help us know the differences between different strains.3.By degenerate PCR of two dikaryon strains 0826,QL7-6,which were morphological diversity.We obtained two pheromone receptor genes.It was identified that the two pheromone receptors shared the homology.4.There are incompatibility factors:A and B factors,which control their mating,in tetrapolar heterothallic mushroom.By means of degenerate PCR and gene walking,we cloned the pheromone receptor gene on B mating-type loci.We obtained one pheromone receptor gene LS-rcb1-B1 from the spore monokaryon strain NO.2 of L.squarrosulus strain NO.7-4; some sequences of pheromone receptor gene LS-rcb2-B2 from the spore monokaryon strain NO.5 of L.squarrosulus strain NO.7-4.Based the DNA sequences of pheromone receptor gene from B mating type factor in L.squarrosulus,we designed two groups of specific primers:rcb1-B1-F,rcb1-B1-R;rob-B2-1,rcb1-B2-R,to amplify the fragment of pheromone receptor from the protoplast monokaryon strains and spore monokaryon strains.We obtained the pheromone receptor fragment from monokaryon strains with the mating type factor B1 using specific primers rcb1-B1-F rcb1-B1-R;also obtained the pheromone receptor fragment from monokaryon strains with the mating type factor B2 using specific primers rcb-B2-F rcb1-B2-R.5.The pheromone receptor gene encoded by LS-rcb1-B1 were seven transmembrane proteins following the analysis using the TMpred software.Phylogenetic analysis on amino acids of pheromone receptor gene LS-rcb1-B1 as well as other 15 kinds of basidiomycetes was done by using the NJ method in PHYLIP software package.The result showed that the Lentinus squarrosulus shared the same cluster with the Lentinula edodes.