Liu Zongying, Wang Qinglong, Cui Jian, Wang Lili, Xiong Lili, Wang Wei, Li Diqiang, Liu Na, Wu Yiran, Mao Canquan
laboratory of Molecular Evolution and Applied Biology, School of Life Science and Engineering, Southwest Jiaotong University, Chengdu, China.
Institute of Forest Ecology, Environment and Protection, Chinese Academy of Forestry, Beijing, China.
Int J Med Mushrooms. 2015;17(3):219-29. doi: 10.1615/intjmedmushrooms.v17.i3.20.
Hericium erinaceus possesses multiple medicinal values. To date, however, there have been few studies of the systemic screening of H. erinaceus strains, and the neuroprotective effects of H. erinaceus prepared from homogenized, fresh fruiting bodies are not fully understood. In this study, 4 random primers were selected and used in random amplified polymorphic DNA (RAPD) polymerase chain reaction (PCR) to screen and evaluate the genetic diversity of 19 commercial strains of H. erinaceus from different localities in China. A total of 66 bands were obtained, and the percentage of polymorphic loci reached 80.30%. Five dendrograms were constructed based on RAPD by Jaccard cluster and within-group linkage analysis. Primer S20 as well as all 4 primers had great potential as specific primers for RAPD-PCR molecular identification and differentiation of H. erinaceus strains. Based on the results of submerged culture and fruiting body cultivation, strains HT-N, HT-J1, HT-C, and HT-M were identified as superior among the 19 H. erinaceus strains. Further study showed that the oral preparation of homogenized, fresh fruiting bodies of H. erinaceus could attenuate the Aβ25-35-triggered damage in PC12 cells by significantly increasing cell viability and by decreasing the release of lactate dehydrogenase. In conclusion, RAPD-PCR combined with liquid and solid cultures can be used well in the screening and identification of H. erinaceus strains, and products prepared from homogenized, fresh fruiting bodies of H. erinaceus had neuroprotective effects on PC12 cells.
猴头菇具有多种药用价值。然而，迄今为止，对猴头菇菌株进行系统筛选的研究较少，且对由新鲜子实体匀浆制备的猴头菇的神经保护作用尚未完全了解。在本研究中，选择了4条随机引物用于随机扩增多态性DNA（RAPD）聚合酶链反应（PCR），以筛选和评估来自中国不同地区的19种市售猴头菇菌株的遗传多样性。共获得66条带，多态性位点百分比达到80.30%。基于RAPD数据，通过Jaccard聚类和组内连锁分析构建了5个系统发育树。引物S20以及所有4条引物在用于猴头菇菌株的RAPD-PCR分子鉴定和区分方面具有很大潜力。根据液体培养和子实体栽培结果，菌株HT-N、HT-J1、HT-C和HT-M被鉴定为19种猴头菇菌株中的优良菌株。进一步研究表明，口服新鲜猴头子实体匀浆制剂可通过显著提高细胞活力和降低乳酸脱氢酶释放来减轻Aβ25-35诱导的PC12细胞损伤。总之，RAPD-PCR结合液体和固体培养可很好地用于猴头菇菌株的筛选和鉴定，且新鲜猴头子实体匀浆制备的产品对PC12细胞具有神经保护作用。
