基于二代测序技术的MNP标记鉴别刺芹侧耳菌株

Identification of Pleurotus eryngii Strains by MNP Makers Based on Next-Generation Sequencing

以31个刺芹侧耳(Pleurotus eryngii)菌株及福建省主栽品种‘Ple 0100’为材料,经二代测序与标记序列筛选,构建32个菌株的MNP标记数据库,结果表明,32个菌株之间的遗传相似度为1.79%~99.60%,其中DUS16与DUS17的遗传相似度为99.60%,DUS02与DUS23、DUS30的遗传相似度均为99.01%,DUS23与DUS30的遗传相似度为99.20%。拮抗实验结果表明,遗传相似度高的菌株之间无拮抗现象,可能是极近似品种或相同品种。对3家刺芹侧耳生产企业的子实体、子实体组织分离物、废菌渣、菇脚进行MNP检测,发现这些样品与‘Ple 0100’的遗传相似度均为100.00%,表明利用MNP标记进行品种鉴别的材料不限于菌丝体。对5个自交菌株及其亲本进行MNP标记检测,结果表明,自交菌株之间遗传相似度为26.84%~61.43%,与亲本之间的遗传相似度为28.43%~78.33%,表明刺芹侧耳减数分裂过程中存在广泛的染色体重组和同源染色体交换。对国内52家刺芹侧耳生产企业的56份鲜菇样品的组织分离物进行MNP检测,发现与‘Ple 0100’的相似度均为100.00%,表明国内刺芹侧耳栽培品种高度一致。研究结果表明基于二代测序技术的MNP标记技术可用于刺芹侧耳菌株间遗传相似度分析及新菌株鉴别。

Using Illumina sequencing and MNP marker screening,31 strains of Pleurotus eryngii and the main cultivated variety‘Ple 0100’in Fujian Province were used to construct a database containing 503 MNP markers for genetic similarity(GS)analysis and variety identification.The results showed that the GS of the 32 strains ranged between 1.79%and 99.60%,among which the GS of DUS 16 and DUS 17 was 99.60%,that between DUS02 and DUS 23 or DUS 30 was 99.01%,and that between DUS 23 and DUS 30 was 99.20%.There was no antagonistic reaction between strains with a high GS,and they may be very similar or the same.MNP markers were used to analyze fruiting bodies,fruiting body tissue isolates,cultivation residue,fruiting body base samples from three P.eryngii production factories,and the results showed that these samples were 100%similar to‘Ple 0100’,suggesting that MNP markers can be used for variety identification of samples other than mycelium.Five inbred strains and their parents were detected by MNP markers.The results showed that the GS between the inbred strains ranged between 26.84%and 61.43%,and that between the inbred strains and their parents ranged between 28.43%and 78.33%,suggesting that there were extensive chromosome recombination and homologous chromosome exchange during the meiosis of P.eryngii.Tissue isolates of 56 commercial mushrooms from 52 P.eryngii factories in China were detected by MNP markers,and their similarity with‘Ple 0100’was 100.00%,indicating that the cultivated varieties of P.eryngii in China were highly consistent.The MNP molecular marker technique in combination with next-generation sequencing can be used to analyze the genetic similarity between P.eryngii strains and identify new strains.