羊肚菌子实体组织细胞的低温冻存与复苏萌发现象的初步观察

A Preliminary Observation of Cryopreservation and Resuscitation Germination of Fruiting Body Cells of Morchella

为保存新鲜羊肚菌子实体组织细胞并观察其复苏后的细胞萌发情况及菌丝早期萌发形态;对采集的野生新鲜羊肚菌子实体进行处理、超低温冷冻保藏,对超低温保藏后的组织细胞快速解冻后进行培养板内4℃萌发培养,倒置显微镜下观察并记录羊肚菌子实体组织细胞萌发现象,挑取非孢子萌发的、无污染的、符合羊肚菌菌丝形态的培养孔内菌丝进行扩大培养,通过羊肚菌通用抗原的ELISA检测及18 s序列测序比对鉴定分离物;结果显示分离得到M10H3、M10H4两株分离物是羊肚菌子实体构成细胞复苏萌发的羊肚菌菌丝。

In order to store the tissue of fresh Morchella and observe the germination and early morphology of hyphae after resuscitation,the feuiting bodies of wild fresh morels were cryopreserved in negative 84°C fridge.Then repidly re-suscitated the cryopreserved tissue and put it into the cell culture plate with 96pores.Subsequently,to placed the resus-citation tissue piate at 4°surrounding.observeing the single cell sprouting of the fruiting body tissue by inverted micro-scope,and picked the non-spore germinated,non-contaminated mycelium in the form of the mycelial mycelium to cul-ture.Though the ELISA assay for the common antigen of Morchella and an 18 s sequence sequencing alignment to identify isolates.The results showed that the two strains M10H3 and M10H4 were the mycelia hyphae of the germina-tion of the Morchella fruit body cells.