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金属胁迫下灵芝寡肽转运蛋白基因家族的转录表达
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四川省教育厅重点项目(No. 15ZA0013)


Transcriptional analysis of oligopeptide transporter gene family under metal stress of Ganoderma lucidum
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    摘要:

    【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。

    Abstract:

    [Objective] Substrates in the environment can be transported into the cell by oligopeptide transporters (OPTs), and then contribute to various biological processes. OPTs proteins are involved in metal transports and distributions in plants, which can promote the plants to use and adapt various metals. However, if OPTs from fungi participates in this process is still unknown. To explore if OPTs play a role in the adaptation process of Ganoderma lucidum to metals. [Methods] Mycelium growth and biomass of G. lucidum (No. 1 Rongbao) were tested in the presence of different concentrations of Cu2+, Se4+ and Fe2+. The specific or differential expression profiles of OPTs were analyzed by real-time quantitative PCR (RT-PCR) from the samples cultured for 15 d and 30 d. [Results] Growth and biomass were inhibited under all metal concentrations of Cu2+, Fe2+ and Se4+. The results of RT-PCR showed that, except three genes (OPT7, OPT8 and OPT9), whose transcripts were not detected, the other 10 OPT genes have differential expression profiles. At the early culture stage (15 d), the relative expression levels of 6 OPT genes (OPT1, OPT3-6 and OPT10) were up-regulated under Cu2+ stress, but the expression levels of all OPTs gene were down-regulated under Fe2+ and Se4+ treatments. At the later culture stage (30 d), the relative expression levels of all OPTs gene were down-regulated under Se4+ stress, but the expression levels were significantly up-regulated under Cu2+ and Fe2+ treatments. [Conclusion] These results indicate that these metals can affect the growth and biomass of G. lucidum, while the oligopeptide transporter genes responded to metal stress at transcriptional levels, suggesting that OPT genes may function in the metal adaption processes in G. lucidum.

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何茂兰,沈柯宇,秦澎,向泉桔. 金属胁迫下灵芝寡肽转运蛋白基因家族的转录表达[J]. 微生物学通报, 2017, 44(9): 2120-2127

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  • 在线发布日期: 2017-09-05
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