为了解平菇生长发育过程中子实体形成的代谢物基础,采用超高效液相色谱-电喷雾串联四级杆质谱(UPLC-ESI-MS/MS)技术结合多变量统计分析方法对发菌完成期(MM)、原基期(MP)及子实体分化期(MF)的平菇菌丝体进行代谢组学分析。结果表明,主成...为了解平菇生长发育过程中子实体形成的代谢物基础,采用超高效液相色谱-电喷雾串联四级杆质谱(UPLC-ESI-MS/MS)技术结合多变量统计分析方法对发菌完成期(MM)、原基期(MP)及子实体分化期(MF)的平菇菌丝体进行代谢组学分析。结果表明,主成分(PCA)模型分析结果显示3个时期平菇菌丝体中的代谢产物具有明显差异。通过正交偏最小二乘判别分析(OPLS‐DA),以VIP(varible importance in the projection)>1和差异倍数值(fold change)≥2或≤0.5为条件对MM vs MP、MM vs MF和MP vs MF中的差异代谢物进行比较分析,分别获得139个、147个和67个差异代谢物,变化倍数最大的物质包括氨基酸及其衍生物、脂质、生物碱、有机酸等,说明这些差异代谢物对平菇子实体发育具有重要影响。KEGG分析表明,苯丙氨酸代谢、色氨酸代谢、嘌呤代谢等20条代谢通路表现活跃。在子实体发育过程中,脂质、有机酸、核苷酸及其衍生物、氨基酸及其衍生物之间明显相关。以上研究结果为平菇子实体发育机制和标准化栽培提供了理论依据。展开更多
Blocking the development of edible mushrooms will affect the production cycle and yield of fruiting bodies.Phenylalanine ammonia lyase(PAL,EC 4.3.1.24.)is an enzyme that catalyzes the deamination of phenylalanine to f...Blocking the development of edible mushrooms will affect the production cycle and yield of fruiting bodies.Phenylalanine ammonia lyase(PAL,EC 4.3.1.24.)is an enzyme that catalyzes the deamination of phenylalanine to form trans-cinnamic acid.Previous studies have shown that a decrease in pal1 gene transcription delays fruiting body development in Pleurotus ostreatus.Herein,we used wild type(WT)and RNA interference(RNAi)strains to study the molecular regulation of pal1 by RNA sequencing and Agrobacterium-mediated genetic transformation.Our results showed that interference with the pal1 gene resulted in reductions in the total PAL enzyme activity and the total phenol content,as well as an increase in the intracellular H_(2)O_(2)content.RNA-Seq data demonstrated that the significantly enriched KEGG terms were mainly related to the peroxisome pathway,MAPK signaling pathway-yeast and three other pathways,and the catalase(CAT)gene cat1 is also involved in multiple pathways that were enriched above.Exogenous H_(2)O_(2)significantly enhanced the transcription of the cat1 gene and elevated total CAT enzymatic activity.Moreover,the levels of cat1 gene transcription and the total CAT enzymatic activity in the RNAi-pal1 strains gradually become closer to those in the WT strain through the removal of H_(2)O_(2),which indicated that pal1 regulated the expression of cat1 by affecting the intracellular H_(2)O_(2)content.Finally,the overexpression of the cat1 gene in P.ostreatus caused growth retardation,especially during the process of primordia formation.In conclusion,this study demonstrated that PAL1 affects cat1 gene expression through the signaling molecule H_(2)O_(2)and regulates the development of P.ostreatus.The findings of this study enhance our understanding of the molecular developmental mechanism of edible mushrooms.展开更多
文摘为了解平菇生长发育过程中子实体形成的代谢物基础,采用超高效液相色谱-电喷雾串联四级杆质谱(UPLC-ESI-MS/MS)技术结合多变量统计分析方法对发菌完成期(MM)、原基期(MP)及子实体分化期(MF)的平菇菌丝体进行代谢组学分析。结果表明,主成分(PCA)模型分析结果显示3个时期平菇菌丝体中的代谢产物具有明显差异。通过正交偏最小二乘判别分析(OPLS‐DA),以VIP(varible importance in the projection)>1和差异倍数值(fold change)≥2或≤0.5为条件对MM vs MP、MM vs MF和MP vs MF中的差异代谢物进行比较分析,分别获得139个、147个和67个差异代谢物,变化倍数最大的物质包括氨基酸及其衍生物、脂质、生物碱、有机酸等,说明这些差异代谢物对平菇子实体发育具有重要影响。KEGG分析表明,苯丙氨酸代谢、色氨酸代谢、嘌呤代谢等20条代谢通路表现活跃。在子实体发育过程中,脂质、有机酸、核苷酸及其衍生物、氨基酸及其衍生物之间明显相关。以上研究结果为平菇子实体发育机制和标准化栽培提供了理论依据。
基金supported by the National Key R&D Program of China(2022YFD1200600)the National Natural Science Foundation of China(32002110)the earmarked fund for China Agriculture Research System(CARS-20)。
文摘Blocking the development of edible mushrooms will affect the production cycle and yield of fruiting bodies.Phenylalanine ammonia lyase(PAL,EC 4.3.1.24.)is an enzyme that catalyzes the deamination of phenylalanine to form trans-cinnamic acid.Previous studies have shown that a decrease in pal1 gene transcription delays fruiting body development in Pleurotus ostreatus.Herein,we used wild type(WT)and RNA interference(RNAi)strains to study the molecular regulation of pal1 by RNA sequencing and Agrobacterium-mediated genetic transformation.Our results showed that interference with the pal1 gene resulted in reductions in the total PAL enzyme activity and the total phenol content,as well as an increase in the intracellular H_(2)O_(2)content.RNA-Seq data demonstrated that the significantly enriched KEGG terms were mainly related to the peroxisome pathway,MAPK signaling pathway-yeast and three other pathways,and the catalase(CAT)gene cat1 is also involved in multiple pathways that were enriched above.Exogenous H_(2)O_(2)significantly enhanced the transcription of the cat1 gene and elevated total CAT enzymatic activity.Moreover,the levels of cat1 gene transcription and the total CAT enzymatic activity in the RNAi-pal1 strains gradually become closer to those in the WT strain through the removal of H_(2)O_(2),which indicated that pal1 regulated the expression of cat1 by affecting the intracellular H_(2)O_(2)content.Finally,the overexpression of the cat1 gene in P.ostreatus caused growth retardation,especially during the process of primordia formation.In conclusion,this study demonstrated that PAL1 affects cat1 gene expression through the signaling molecule H_(2)O_(2)and regulates the development of P.ostreatus.The findings of this study enhance our understanding of the molecular developmental mechanism of edible mushrooms.
