中国栽培糙皮侧耳品种体细胞不亲和性实验与RAPD分析结果的比较

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。

Comparison between somatic incompatibility and RAPD analysis of cultivated Pleurotus ostreatus in China

Nineteen China cultivated strains of Pleurotus ostreatus were antagonistically paired on poor nutrient half-PDA plates. The rate of barrage formation was 100% among these strains. The barrages could be identified into three types: raised aggregate of hyphae (Type A), clear zone (Type B), and dark line (Type D). The formation rate of the three barrage types was 24.9%, 23.7% and 51.4%, respectively. The bacteriolytic channel formed from hyphal self-digestion was found in the area of dark line barrage by microscopic observation, while no hyphal self-digestion was found in the other two barrage types. NTSYS-PC clustering analysis using barrage type as a variable indicated that the 19 strains of Pleurotus ostreatus were divided into three groups cut in similar coefficient at 0.48. First group included 9 strains, e.g. 802 and Hei Pin Wang; second group 6 ones, e.g. Su Yin 6 and Jiang Du 71; third group 4 ones, e.g. Xia Wang 40 and Zao Qiu Gao Feng. 917 RAPD markers were yielded by using 13 random primers to amplify the total DNA of the 19 strains. RAPD analysis showed that these strains had high identity over 85%. These strains were also divided into three groups cut in similar coefficient at 0.86. First group included 17 strains, e.g. 802 and Hei Pin Wang; second group included 2 ones, Tuiguang 1 and Xia Wang 40; third group included only one, 831. There was no correlation between the results of the two clustering analysis. The results indicated that barrage test was a useful method for analyzing population structure and genetic diversity of intraspecies fungi, but it could not replace the other DNA finger methods for genotype or relatedness identification.