基于流式细胞技术的灵芝基因组大小估测

Estimation of the genome size of Ganoderma lucidum based on a flow cytometric analysis

以药典规定的灵芝正品来源灵芝Ganoderma lucidum作为研究对象,利用已完成全基因组测序的黑曲霉Aspergillus niger作为内标,通过机械破碎菌丝体的方法获得合适浓度的细胞核悬液,碘化丙啶荧光染色后成功应用流式细胞术进行基因组大小估测。经过优化材料培养、样品制备、上机分析等实验条件,估测得出灵芝基因组大小(48.98±0.60)Mb,为灵芝基因组学研究提供重要数据。该方法简捷稳定,在蕈菌范围内,首次得到了全基因组测序数据与光学图谱结果验证,为蕈菌基因组学研究提供重要技术平台。

The genome size of Ganoderma lueidum was estimated by analyzing the PI-stained nucleus samples prepared by mechanical disintegration from fungal mycelium pellet. Through optimizing the parameters of fungal cultivation, sample preparation and flow cytometry （FCM） setups, the genome size of G lucidum was estimated to be （48.98＋0.60）Mb by using the genome data of Aspergillus niger as the internal standard. This work provided genome size information for G. lucidum, and was important for future research on genomic study. The estimated value was verified by next-generation sequencing and optical mapping approaches, suggesting a feasible technology platform for genomic research on macrofungi.