过量草酸抑制广叶绣球菌降解松木的分子机制研究

Sparassis latifolia is a kind of edible and medicinal fungi with great economic and scientific value. However, its wild strain is difficult to tame and grow slowly. Up to now, no solution has been found. Previous studies showed that there was a large amount of oxalic acid secretion in the process of growing, and it was proved that excessive oxalate had a significant inhibitory effect on the degradation of wood. However, the research on the mechanism of this inhibition is not yet clear. We have obtained experimental materials and whole-genome sequence of S. latifolia in the early stage. Based on these data, the applicant intends to compare the transcriptome and metabonomics information of S. latifolia under different concentrations of oxalic acid, and screen out the obstacle factors and key genes inhibited by excessive oxalic acid; CRISPR/Cas9 technology will be used to verify the function of the key genes inhibited, to explore the molecular mechanism of this inhibition effect. It lays a foundation for the selection of high quality and high yield strains of S. latifolia, and it also good for which breeding.

广叶绣球菌是一类极具经济和科研价值的食药用菌，但在驯化栽培中存在难以解决的吃料缓慢问题。申请人前期研究发现，广叶绣球菌在菌丝生长过程中能分泌大量草酸，过量草酸对广叶绣球菌降解松木有显著的抑制作用。但目前关于这一抑制作用的机制研究尚不明确。申请人拟以前期获得的广叶绣球菌代表性试验材料及已完成的全基因组测序数据为基础，①通过比较分析不同草酸环境中的广叶绣球菌菌丝的转录组、代谢组信息，筛选过量草酸抑制广叶绣球菌降解松木的障碍因子及受抑制的关键基因；②利用CRISPR/Cas9技术对关键基因的功能进行验证，探究这一抑制作用发生的分子机制。这一研究结果可为选育广叶绣球菌优质高产菌株和制定有效选育手段奠定基础。

广叶绣球菌是一类极具经济和科研价值的食药用菌，但在驯化栽培中存在难以解决的吃料缓慢问题。该项目研究表明，广叶绣球菌在菌丝生长过程中分泌大量草酸，过量草酸致使松木栽培基质pH环境下降，与降解相关的木质纤维素降解酶活性均显著降低；不同草酸环境中的广叶绣球菌菌丝的转录组分析进一步表明，过量草酸影响了广叶绣球菌TCA循环，使其菌丝代谢出现呼吸障碍，其能量传递及各种氨基酸合成均受到抑制。联合代谢组分析发现，过量草酸抑制了广叶绣球菌对松木中烯烃、烷烃等非环状芳香类物质的降解，但促进了对多环芳香烃类物质的降解，同时也促进了自身特有的香气成分及药理活性成分的合成。经验证，松树中多环芳香烃类物质可诱导广叶绣球菌产生大量草酸，从而将其分解并合成特殊的萜类化合物，据此可判定，过量草酸导致广叶绣球菌降解松木进程受阻的障碍因子是松木中的多环芳香烃类化合物，生产中可将松木栽培基质中的该类物质有针对性的进行消除，进而加快其吃料速度，缩短其栽培周期，推动绣球菌产业发展。同时这一研究结果也将为广叶绣球菌优质高产菌株选育指标提供理论依据。

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