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对收集的糙皮侧耳Pleurotus ostreatus19个栽培菌株在贫营养的半PDA培养基中配对进行拮抗线实验,考察其相互间体细胞不亲和性。结果表明,此19个菌株间出现拮抗线的比率是100%,形成的拮抗线类型可分为菌丝集结型(A型)、隔离带型(B型)和暗线型(D型)三类,出现的比例分别是24.9%、23.7%和51.4%。显微观察暗线型拮抗线区域有菌丝自融形成的溶菌沟,另2种拮抗线中没有菌丝自溶。以拮抗线类型分别作为变量进行NTSYS-PC聚类分析,从相似性系数0.48处截断可将其分为3大类,第一类包括802和黑平王等9个菌株;第二类包括苏引6号和江都71等6个菌株;第三类包括夏王40和早秋高丰等4个菌株。应用13条随机引物扩增此19株糙皮侧耳菌株总DNA,共扩增出917条不同分子量的DNA条带。RAPD聚类分析表明,这些菌株的遗传相似性高达85%以上。从相似性系数0.86处截断,也可将这些菌株分为3大类,第一类包括了802和黑平王等17个菌株;第二类仅包括推广一号和夏王40两个菌株;第三类只有831一个菌株。两种方法的聚类分析结果没有相关性。因此,拮抗反应可以作为评价糙皮侧耳栽培菌株之间遗传多样性的方法之一,但与基因组DNA指纹多样性分析结果并不完全吻合,更不能代替分子指纹分析。
Nineteen cultivated strains of Pleurotus ostreatus harvested from Pleurotus ostreatus collected in antagonistic line experiments were paired in oligotrophic half-PDA medium to examine the somatic cell incompatibility with each other. The results showed that the antagonistic lines appeared in 100% of the 19 strains, and the antagonistic lines formed were classified into three types: hyphae (A type), barrier type (B type) and dark line type (D type) , With the proportions appearing 24.9%, 23.7% and 51.4% respectively. Microscopic observation of the dark line of antagonistic line with mycelium melt Rongdiogou formation, the other two antagonistic lines without hyphae autolysis. NTSYS-PC cluster analysis was performed using antagonistic line types as variables respectively. The truncated similarity coefficient of 0.48 could be divided into three categories: the first one consisted of 9 isolates including 802 and Heian Wang; 6 and Jiangdu 71 strains such as six strains; the third category, including Xiawang 40 and early autumn Gao Feng other 4 strains. Thirteen primers were used to amplify the total DNA of nine strains of Pleurotus ear Pleurotus, 917 DNA bands of different molecular weights were amplified. RAPD cluster analysis showed that the genetic similarity of these strains was as high as 85% or more. From the similarity coefficient of 0.86 at the cut-off, these strains can also be divided into three categories, the first category includes 802 and black flat king and other 17 strains; the second category includes only the promotion of the first two and Xia Wang 40 strains; There are only 831 strains in the three categories. There is no correlation between the clustering results of the two methods. Therefore, the antagonistic reaction can be used as one of the methods to evaluate the genetic diversity between Pleurotus ostreatus strains, but it is not completely consistent with the results of genomic DNA fingerprinting analysis, and can not replace the molecular fingerprinting analysis.