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为探索苹果矮化砧木抗寒的生理和分子机制,筛选抗寒相关基因,分析了苹果高抗寒矮化砧木‘71-3-150’在冷胁迫(4℃)0、2、6、12和24 h时的膜伤害、活性氧代谢、渗透调节物质积累等生理指标及转录组的变化。结果表明:‘71-3-150’在冷胁迫0~2 h即表现出抗氧化和渗透调节反应,6 h后SOD、CAT等活性氧清除酶活性及主要渗调物质维持在较高水平趋于稳定,膜伤害加重趋势缓解,说明其具有快速响应冷胁迫的生理机制,0~6 h是其冷适应的关键时期;转录组分析表明,0 h vs 2 h、0 h vs6 h、0 h vs 12 h和0 h vs 24 h等比较组分别含有491、1 115、828和1 047个上调差异表达基因及537、688、708和1 016个下调差异表达基因,其中4个组共有的115个上调基因和136个下调基因表现为6种表达模式;筛选出28个‘71-3-150’冷适应过程中存在显著差异表达的功能基因与转录因子基因,调控低温信号感知与传导的基因在0~2 h出现显著性上调表达,参与渗透调节、活性氧代谢、膜和蛋白保护等过程的基因随低温处理呈现多样性表达模式,表明该砧木在冷适应过程中存在快速和多样的低温信号感知、转导和响应的分子机制。
In order to explore the physiological and molecular mechanisms of cold-tolerant apple dwarfing stock, we screened the genes related to cold-resistance, analyzed the effects of ’71 -3-150 ’ And 24 h when the membrane damage, reactive oxygen metabolism, osmotic adjustment material accumulation and other physiological indicators and transcriptome changes. The results showed that ’71-3-150 ’showed antioxidant and osmoregulation reaction at 0-2 h after cold stress. After 6 h, the activities of active oxygen scavenging enzymes such as SOD and CAT and their main infiltration substances maintained at higher levels The trend of aggravation and membrane damage aggravating tended to be alleviated, indicating that it has a physiological mechanism of rapidly responding to cold stress and 0 to 6 h is the critical period of its cold acclimation. Transcriptome analysis showed that 0 h vs 2 h, 0 h vs 6 h, 0 h 491, 115828 and 1 047 up-regulated differentially expressed genes and 537,688,708 and 1 016 down-regulated differentially expressed genes in 12 h vs 0 h vs 24 h groups, respectively Six up-regulated genes and 136 down-regulated genes were expressed in six kinds of expression patterns; 28 genes with significantly differentially expressed genes and transcription factor genes were selected during the cold acclimation process of ’71-3-150 ’, and the genes that regulate the perception and conduction of hypothermia Significantly up-regulated expression was observed between 0 and 2 h. The genes involved in osmotic adjustment, reactive oxygen metabolism, membrane and protein protection showed diverse expression patterns under low temperature treatment, indicating that the rootstock had rapid and diverse low temperature during cold acclimation Signaling, transduction and response of the molecular machine system.