Sichuan Key Laboratory of Quality and Innovation of Traditional Chinese Medicine, Institute of Fungus Medicinal Materials, Sichuan Academy of Chinese Medicine Science, Chengdu, 610041 Sichuan Province, People's Republic of China.
Int J Med Mushrooms. 2022;24(4):43-52. doi: 10.1615/IntJMedMushrooms.2022043141.
This study aimed to obtain a set of specific inter simple sequence repeat (ISSR) primers and establish a stable and accurate intraspecific identification method for Ganoderma lingzhi. A total of 117 G. lingzhi strains were identified using internal transcribed spacer sequences from 147 strains determined as G. lingzhi via simple morphological identification. Based on the sequences obtained, specific ISSR primers for G. lingzhi were screened and validated, and 15 specific ISSR primers showed polymorphic banding pattern with clear band resolution. Subsequently, ISSR PCRs of the 15 specific primers were performed for the 117 G. lingzhi strains. As expected, DNA analysis of the ISSR markers could distinguish G. lingzhi strains, with similarity coefficients ranging from 0.11 to 0.89. Thus, the 15 specific ISSR primers can be used for intraspecific identification and polymorphism analysis of G. lingzhi.
本研究旨在获得一套特定的内部简单重复序列(ISSR)引物,并为灵芝建立一种稳定、准确的种内鉴定方法。通过对 147 株经简单形态鉴定为灵芝的菌株进行内部转录间隔区序列分析,共鉴定出 117 株灵芝菌株。基于获得的序列,筛选并验证了灵芝特异性 ISSR 引物,15 个特异性 ISSR 引物显示出多态性带型,带型清晰可辨。随后,对 117 株灵芝菌株进行了 15 个特异性引物的 ISSR-PCR。结果表明,ISSR 标记的 DNA 分析可以区分灵芝菌株,相似系数范围为 0.11 至 0.89。因此,这 15 个特异性 ISSR 引物可用于灵芝的种内鉴定和多态性分析。