| This article studied the Pleurotus geesteranus, Pleurotus pulmonariu, Pleurotus cornucopiae, Pleurotus cystidious,from these aspects of cultivation experiment, sexual compatibility tests and cloning their mating factors, these stains were preserved in Fujian germplasm resources preservation center . We examined their sexual compatibility. By amplifying the partial fragments of A and B mating locus ,we wanted to find one method to clone their full length,furtherly studied the structures and fuctions of them.。the main result were summarized as fellows:1.Cultivation experiments By the cultivation experiments,we found that they had similar morphological characteristics between Pleurotus geesteranus and Pleurotus pulmonariu,the difference of morphological characteristics was obvious among the strains of Pleurotus cornucopiae Pleurotus cystidious were divided into two groups:one was similar to Pleurotus cornucopiae,the other to Pleurotus pulmonariu. we found that three strains were not belong to Pleurotus genus simply through their morphological characteristics,pl.g0016 was a Lentinus edodes. pl.co0007,pl.co0008 and pl.co0029 were the strains of Hypsizigus marmoreus.,pl.g0003 and pl.g0009 were not the Pleurotus geesteranus, pl.p0006 and pl.p0009 were also not Pleurotus pulmonariu,because their morphological characteristics were similar to Pleurotus cornucopiae.2.Identification of mating factos Edible fungi of pleurotus genus belonged to the typical tetrapolar heterothallic mating system,Identification of mating factors was important to breeding and strain identification。65 strains were classified to two groups: Pleurotus pulmonariu and Pleurotus cornucopiae,the results of study showed that the two were absolutely reproductive segregation,they may had a distant genetic relationship,42 putative strains of Pleurotus pulmonariu were classified into 3 classes by mating reactions, 4 A mating-type factors and 3 B mating-type factors were obtained ; 23 putative strains of Pleurotus cornucopiae were classified into 10 classes by mating reactions,and 15 A mating-type factors and 14 B mating-type factors were got。3.Cloning the MIP gene It had been shown that the gene encoding a mitochondrial intermediate peptidase (MIP) was tightly linked (<1 kbp) to the A mating-type locus in two model mushroom species. In this study we got a MIP nucleotide fragments of Pleurotus about 470bp by Degenerate PCR.It had been found that the pair of MIP-2F,MIP-3R primers had better amplified effect.By sequence alignment analysis,we found that the majority of MIP sequences had highly homology with the known fungi MIP genes,they all belonged to Peptidase-M13-superfamily ,But they were not highly conserved gene,some fragments had lowerly conservative,According to the reasults of MIP gene sequencing,nested primers were designed to amplify the downsteam flanked fragments by TAIL-PCR.But the sequenceing results indicated that they were non-specific strips,the feadibility needed furtherly reseach.4.Cloning the pheromone receptor gene of B mating-type factor Degenerate PCR of B mating-type pheromone receptor homologues from homokaryotic strains of Pleurotus was accomplished using primers br1-F and br1-1R targeting amino acids 26–212 of S. commune receptor BAR1.By amino acid sequences alignment ,it proved that they were the members of STE3- receptor superfamily.The results of SOSUI analysis on line had shown that part of fragments included transmembrane protein.But at the same time,it also had been found that there were 4 groups of almostly same fragments of the pheromone receptor gene.Perhaps because the separated DNA fragments were located in the highly conservative region. |
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