蛋白组学结合代谢组学研究工业大麻响应低钾胁迫的分子机制

Industrial hemp is a significant green fiber crop,which has huge value in the devolopment of industrial and medicinal. As the potassium has a significant impact on the yield and quality of industrial hemp, Studying the molecular mechanism of industrial hemp in low potassium environment helps to improve its industrial development potential. At present, there is no research reports about the molecular mechanisms of industrial hemp in response to low potassium stress, the combination of proteomics and metabolomics can help us gain key proteins and metabolins associated with potassium absorpion and tolerance in a quick and high throughput manner.This study uses efficient potassium absorption variety"Bamahuoma " and low potassium absorption variety"Jinma #1" as experimental materials, using solution culture, with a low potassium stress（0.4mmol/L K+）to deal with normal potassium levels（2mmol/L K+）for comparison.According to the sampling period which is confirmed by physiological, biochemical and agronomic character indexes, we have a research on proteomics and metabonomics separetely.Through referencing genome data of hemp and making correlation analysis of proteomics and metabolomics, we hope that we can find about 30 kinds of key proteins and 10 kinds of metabolins ,indentified 2～3 key metabolic pathways of industrial hemp in response to low potassium stress as expected.Finally, we verify these important proteins and metabolins by qRT-PCR and MRM technology separetely, and on this basis, we build translation and metabolism regulatory network of industrial hemp in response to low potassium stress , which provides the molecular basis for industrial hemp of efficient potassium absorption in response to low potassium stress .

工业大麻为重要的绿色纤维作物，极具工业、药用开发价值。钾对工业大麻产量和品质有显著影响，研究工业大麻在低钾环境下的分子机制有助于提高其开发潜力。目前尚未见大麻响应低钾胁迫的分子机制研究报道，蛋白组学与代谢组学结合可快速、高通量、直接地获取与钾吸收、耐性相关的关键蛋白和代谢产物。本研究以钾高效型品种“巴马火麻”，低效型品种“晋麻1号”为实验材料，采用溶液培养，以低钾胁迫(0.4mmol/LK+)为处理，正常钾水平(2mmol/LK+)为对照。根据生理生化及农艺性状确定的取样时期，分别进行蛋白组和代谢组学研究。参考大麻基因组数据，进行蛋白组和代谢组学关联分析，预期挖掘关键蛋白约30种，代谢产物约10种，鉴定出工业大麻响应低钾胁迫关键代谢通路2-3条。最后对关键蛋白和代谢物进行qRT-PCR和MRM技术验证，以此为基础构建工业大麻响应低钾胁迫的翻译、代谢调控网络，为工业大麻响应低钾奠定分子基础。

工业大麻为重要的绿色纤维作物，极具工业、药用开发价值。钾对工业大麻产量和品质有显著影响，研究工业大麻在低钾环境下的分子机制有助于提高其开发潜力。.1.本项目完成工业大麻响应低钾胁迫的生理生化机制，工业大麻可能通过增加茎粗、根冠比、降低蒸腾速率、提高SOD活性、增加可溶性蛋白含量等适应低钾胁迫。.2.通过SWATH蛋白质组学技术，完成耐低钾籽用大麻品种（BM）和低钾敏感型纤用品种（Y1）快速生长期叶片和茎差异蛋白组学，通过LC-ESI-MS/MS质谱鉴定得到BM和Y1差异蛋白，茎：288和353，叶：226和256。分析两个品种的KEGG，Y1叶片、BM叶片、Y1茎和BM茎的差异蛋白分别显著富集在15、3、11和4个pathway，获得与大麻低钾适应相关的关键蛋白。通过比较分析差异蛋白，发现耐低钾品种BM主要通过以下机制适应低钾胁迫：茎叶皆表现出叶绿素合成下降，降低光合作用；茎杆提高淀粉代谢，提高抗氧化能力，提高木质素代谢，促进细胞膜合成和细胞分裂，促进乙烯合成；叶片调节气孔关闭，加强光呼吸和光能传递，蛋白合成代谢旺盛。.3.代谢组学显示工业大麻茎叶中代谢物丰富，代谢物的成分依次为有机酸和衍生物，有机氧化合物，有机杂环化合物，脂类和类脂分子等。代谢组学和蛋白组学联合分析，大麻钾高效吸收的关键代谢通路有蛋白质合成代谢及淀粉和蔗糖代谢。. 研究结果可为未来工业大麻高效吸收钾素的分子研究提供重要的参考蛋白和通路信息。为最终揭示工业大麻低钾适应的机制，以及培育钾高效品种打下基础。