六株野生羊肚菌的分子鉴定及高活力菌株筛选
摘要： 为筛选活力强、能够稳定出菇的优质羊肚菌菌株，本研究采用单基因及多基因联合的方法对收集的六株野生羊肚菌进行分子生物学鉴定，通过分析液体培养条件下菌丝体的过氧化氢酶、多酚氧化酶、过氧化物酶等抗氧化酶活力和丙二醛（MDA）含量，利用隶属函数综合判定筛选出一株适合栽培的优质菌株。结果表明，收集的六株野生羊肚菌均属于六妹羊肚菌（Morchella sextelata），黑色羊肚菌（Elata Clade）类群。羊肚菌菌丝形态特征均由白色变为微黄，菌丝排列紧密，菌丝末端较钝，菌丝粗细不均且部分表面均分布有球状体，菌丝生长速度在0.40-0.45mm/d之间。交配型基因结果显示LM1、LM3和LM4三个菌株含有MAT1-1、MAT1-2两种交配型基因，能进行有性生殖。尤其，菌株LM4的PPO、POD、CAT、TTC-脱氢酶酶活力均显著高于其他二株菌，且MDA含量最低。结合隶属函数分析， LM4菌株的平均隶属函数为0.9647，是一株活力较强的优质菌，可以作为栽培试验菌株。
关键词: 羊肚菌, 分子鉴定, 交配型基因, 生物量, 抗氧化酶活力
Abstract: In order to screen high-quality Morchella esculenta strains with stable production capacity, the single-gene and multi-gene combination methods were used to identify six wild morel strains collected in this study. Moreover, the activity of antioxidant enzymes such as catalase, polyphenol oxidase, peroxidase, and the content of malondialdehyde (MDA) of mycelia in liquid culture conditions were analyzed. The expected strain suitable for cultivation was screened out by using the subordinate function method. The results showed that all the six wild morels collected belonged to Morchella sextelata, and the black morel (Elata Clade) group. The morphological characteristics of mycelium for Morchella esculenta changed from white to yellowish, the mycelium arranged closely, the end of mycelium blunt, the thickness of mycelium was uneven and a part of surfaces were distributed with globules, and the growth rate of mycelium was in the range of 0.40-0.45mm/d. The results of mating type genes showed that three strains LM1, LM3, and LM4 contained two mating type genes, MAT1-1 and MAT1-2, which were capable of sexual reproduction. In particular, strain LM4 had significantly higher PPO, POD, CAT, and TTC-dehydrogenase enzyme activities than the others two strains, accompanied by the lowest MDA content. Combined with the affiliation function analysis, the average affiliation function of strain LM4 was 0.9647, which is a high-quality Morchella esculenta with high vigor and can be used as a cultivation test strain.
Key words: Morchella esculenta, strain identification, mating type genes, biomass, antioxidant enzyme activity
中图分类号:
Q93
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