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【目的】寡肽转运蛋白(Oligopeptide transporters,OPTs)通过细胞质膜,将细胞外环境中的物质转入细胞内,从而参与各种生理活动。植物中OPTs蛋白参与金属运输和分配,从而促进植物对金属的利用和适应,但真菌中OPTs是否参与这一过程的相关报道较少。探究OPTs是否在灵芝对金属的适应过程中发挥作用。【方法】以灵芝荣保1号为材料,利用平板实验分析不同浓度的Cu2+、Fe2+和Se4+对菌丝体生长和生物量的影响,并采用实时荧光定量PCR分析不同金属离子胁迫下,两个培养阶段(15 d和30 d)灵芝寡肽转运蛋白基因(OPTs)的转录表达水平。【结果】Cu2+、Fe2+和Se4+三个金属离子在实验使用的浓度范围内抑制灵芝菌丝体的生长量和生物量,且在相同处理条件下的生长量与生物量的变化趋势相同。荧光定量PCR结果显示,除未检测到转录本的3个基因(OPT7、OPT8和OPT9),其余10个基因均表现出了差异性表达模式。在培养前期(15 d),有6个寡肽转运蛋白基因(OPT1、OPT3-6和OPT10)的相对表达量在Cu2+的胁迫下发生上调,而在Fe2+和Se4+的胁迫下,所有的OPTs基因的表达量均被下调。在培养后期(30 d),Se4+胁迫下的OPTs基因的表达量仍旧被下调,而Cu2+和Fe2+的胁迫下表达量均显著上调。【结论】金属能够在不同的程度上影响灵芝的生长量和生物量,同时寡肽转运蛋白基因对这些金属胁迫在转录水平上做出了响应,表明寡肽转运蛋白基因可能在灵芝对金属的适应过程中发挥作用。
【OBJECTIVE】 Oligopeptide transporters (OPTs) are involved in various physiological activities by transferring substances in the extracellular environment into the cell through the plasma membrane. Plant OPTs protein involved in metal transport and distribution, thereby promoting the use of plants and metal adaptation, but the fungi in the OPTs involved in the process of less relevant reports. Explore if OPTs play a role in the adaptation of ganoderma to metals. 【Method】 Using Ganoderma lucidum Rongbao 1 as material, the effects of different concentrations of Cu2 +, Fe2 + and Se4 + on the mycelial growth and biomass of the mycelia were analyzed by using the plate experiment. Real-time fluorescence quantitative PCR was used to analyze the effects of two metal ions The transcriptional level of OPTs in the culture stage (15 d and 30 d). 【Result】 The results showed that three metal ions, Cu2 +, Fe2 + and Se4 +, inhibited the growth and biomass of G. ganoderma lucidum at the concentrations used in the experiment. The growth and biomass of G. ganoderma under the same treatment showed the same trend. Fluorescence quantitative PCR results showed that the other 10 genes showed a differential expression pattern except for the three transcripts (OPT7, OPT8 and OPT9). At the early stage of culture (15 d), the relative expression levels of six oligopeptide transporter genes (OPT1, OPT3-6 and OPT10) were up-regulated under the stress of Cu2 +, while under the stress of Fe2 + and Se4 +, all OPTs genes The expression levels were down-regulated. At late stage of culture (30 d), the expression level of OPTs gene was still down-regulated under Se4 + stress, while the expression of OPTs gene was up-regulated under the stress of Cu2 + and Fe2 +. 【Conclusion】 The metal can affect the growth and biomass of Ganoderma lucidum to different extents, and the oligopeptide transporter gene responded to these metal stresses at the transcriptional level, indicating that the oligopeptide transporter gene may play an important role in the growth of Ganoderma lucidum on metal Adapt to play a role in the process.