低温胁迫下东南景天的生理响应及转录组和代谢组分析

Physiological responses of Sedum alfredii Hance to low temperature stress and analysis of transcriptome and metabolome

  • 摘要: 【目的】 解析低温胁迫对东南景天生理机制、转录及代谢调控网络的影响,为后续研究东南景天抗寒机制及其在镉污染农田修复的工程应用提供科学参考。【方法】 以东南景天标准苗为试验材料,采用盆栽方法将东南景天低温(10和-2℃)处理6 h,测定对照(22℃)及低温处理后叶片中丙二醛、可溶性糖和可溶性蛋白含量、相对电导率、抗氧化酶活性等生理指标,并进行转录组和代谢组测序分析。【结果】 低温胁迫导致东南景天丙二醛、可溶性糖和可溶性蛋白含量及相对电导率极显著增加(P<0.01,下同),抗氧化酶活性显著(P<0.05)或极显著提高。与对照相比,10℃低温处理的转录组测序获得7152个差异表达基因(DEGs),-2℃低温处理获得12972个DEGs,代谢组测序分别获得226和228个差异累积代谢物(DAMs)。转录组测序分析结果表明,在低温胁迫下,东南景天中类黄酮生物合成、MAPK信号通路、苯丙素生物合成,以及淀粉和蔗糖代谢均受明显影响。代谢组学分析结果显示,在低温胁迫下,黄酮和黄酮醇生物合成、甘油磷脂代谢、戊糖和葡萄糖醛酸相互转化,以及单萜生物合成等过程发挥关键的调控作用。转录组和代谢组富集通路存在重叠,关联分析表明戊糖和葡萄糖醛酸相互转化、类黄酮生物合成及单萜合成的代谢得到增强。实时荧光定量PCR结果验证了转录组数据的可靠性,戊糖和葡萄糖醛酸相互转化、类黄酮生物合成及单萜生物合成通路中的大多数DEGs表达极显著上调。【结论】 在低温胁迫下,东南景天生理指标发生变化,有多个转录和代谢通路共同响应低温胁迫,戊糖和葡萄糖醛酸相互转化、类黄酮生物合成增加、单萜合成代谢增强,均对东南景天抵抗低温胁迫发挥关键的调控作用。

     

    Abstract: 【Objective】 To analyze the impact of low temperature stress on the physiological mechanisms,transcription and metabolic regulatory network of Sedum alfredii Hance,which could provide scientific reference for subsequent research on its cold resistance mechanisms and engineering applications in cadmium-contaminated farmland remediation.【Method】Standard seedlings of S.alfredii were used as experimental materials by employing the potted method.These seedlings were then subjected to low temperature treatments(10 and-2℃)for 6 h,control(22℃)was also set,after which physiological indicators,including the contents of malondialdehyde,soluble sugar and soluble protein,the relative electrical conductivity and antioxidant enzyme activity in the leaves,were measured.Then,the transcriptome and metabolome sequencing analysis were performed. 【Result】 Low temperature stress led to extremely significant increase in the contents of malondialdehyde,soluble sugar and soluble protein,as well as relative conductivity(P<0.01,the same below),and significantly(P<0.05) or extremely significantly increased antioxidant enzyme activity in S.alfredii.Compared with the control,7152 differentially expressed genes(DEGs) were obtained by transcriptome sequencing under 10℃low temperature treatment,and 12972 DEGs were obtained under-2℃low temperature treatment.Metabolome sequencing obtained 226 and 228 differential accumulated metabolites(DAMs) respectively.Under low temperature stress,transcriptome sequencing analysis results showed that the pathway of flavonoid biosynthesis,MAPK signaling,phenylpropanoid biosynthesis,as well as starch and sucrose metabolism in S.alfredii were obviously impacted.Metabolomics analysis showed that the biosynthesis of flavonoids and flavonols,metabolism of glycerophospholipids,interconversion of pentose and glucuronic acid,and monoterpene biosynthesis played key regulatory roles under low temperature stress.There was overlap between transcriptome and metabolome enrichment pathways.Correlation analysis showed that the interconversion of pentose and glucuronic acid,biosynthesis of flavonoids,and metabolism of monoterpene synthesis were enhanced.The results of real-time fluorescence quantitative PCR validated the reliability of the transcriptome data,and most DEGs in the pathways of pentose and glucuronide interconversion,flavonoid biosynthesis and monoterpenoid biosynthesis were significantly up-regulated. 【Conclusion】 Under low temperature stress,the physiological indicators of S.alfredii undergo great changes,with multiple transcriptional and metabolic pathways collectively responding to low temperature stress,interconversion of pentose and glucuronic acid,increased biosynthesis of flavonoids,and enhanced monoterpenoid synthesis metabolism.These pathways play crucial regulatory roles in enabling S.alfredii to resist low temperature stress.

     

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