| This study did the cultivation experiment, antagonistic test and esterase isozyme analysis of24Ganoderma lucidum strains in order to provide effective analysis and verification method for G. lucidum identification. We studied mating types of G. lucidum population, found some complex alleles,-explicited the polarity and hybrid affinity, and. constructed the fertile group for hybrid breeding and cultivars identification in future. The main results were summarized as follows:1. Cultivation experiments:Through the cultivation tests, we found that No.31and43stains grew more quickly than others, with the average speed of0.788cm/d and0.785cm/d. However, about the average yield per bag. No.36stain was the highest up to89.91g/bag. the second was No.19stain with85.81g/bag. Combinated consideration with morphology and color of G. lucidum. No.11.17and18strains existed large genetic differences with others.Considering both agronomic characteristics and production, we recommend No.05.15.24.25.54stains to be the main cultivated species.2. Antagonistic test:276tests were set in antagonistic test. There were111tests antagonizing obviously and106tests antagonizing less obviously which accounted for78.62%of the antagonistic tests. The results showed that24G. lucidum strains were riched in genetic affinity. There were55tests antagonizing weakly,4tests without antagonism which accounted for21.38%. The results showed that close genetic distance was existed between some strains. Moreover, No.11and54strains antagonized strongly against others in the antagonistic tests with antagonistic line and/or obvious precipitated pigment at mycelia intersecting point. The results revealed that the genetic distances between NO.11/54strains and others were far.3. Esterase isozyme analysis:Esterase isoenzyme patterns of24G. lucidum strains were determined by using vertical polyacrylamide gel electrophoresis.28different types of esterase bands were determined with Rf between0.0508and0.9153. The4bands with Rfof0.5111.0.5631,0.6911.0.7552were the common enzymes of 24tested strains. The genetic similarity coefficients of the24tested strains ranged from0.536to0.964. The genetic similarity coefficients between No.12+2and No.40, No.25and No.36were the highest while between No.11and No.19, No.11and No.50+2were the lowest. All24strains could be divided into four large groups by the cluster analysis with the genetic similarity coefficient of0.6922. Large group I could be divided into four small groups and large group II could be divided into three small groups at genetic similarity coefficient of0.7321.4. Mating type analysis:-Four mating types of haploids in24tested strains were determined by using improved OWE-SOJ technique. We found that two types of colonies appeared, the combanations that formed fluffy colony were named as the A≠B=mating interaction, others that formed the barrage colony were named as the A≠B≠mating interaction. The two colonies could be distinguished clearly from each other. Besides, pseudoclamp connections could be observed in the combined tests with fluffy colony. In terms of the total mating types, to some extent, four mating types showed distorted segregation among monokaryotic mycelia. All24tested strains could be divided into7large groups according to the mating types determination results.There were four alleles in A factor, four alleles in B factor and one mixed allele in A factor. The tested result of mating type analysis was highly consistent with the result from the analysis of esterase isozyme. Therefore, mating type analysis is a very important supplementary method for strain identification and genetic diversity analysis. |
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