基于转录组测序筛选新疆野苹果组培苗应答冻害谷胱甘肽代谢相关的基因  

Screening of genes related to glutathione metabolism responding to freezing stress in Malus sieversii seedlings in vitro based on transcriptome sequencing

作　　者：苏永峰 刘俐君 马红喜 袁引燕 张德恩 鲁晓燕[1] SU Yongfeng;LIU Lijun;MA Hongxi;YUAN Yinyan;ZHANG De’en;LU Xiaoyan(College of Agriculure,Shihezi University/Xinjiang Production and Construction Corps Key Laboratory of Special Fruit Sand Vegetables Cultivation Physiology and Germplasm Resources Utilization,Shihezi 832000,Xinjiang,China)

机构地区：[1]石河子大学农学院·特色果蔬栽培生理与种质资源利用兵团重点实验室,新疆石河子832000

出　　处：《果树学报》2023年第5期829-840,共12页Journal of Fruit Science

基　　金：国家自然科学基金委NSFC-新疆联合基金(培育)项目(U1703116)。

摘　　要：【目的】探索新疆野苹果组培苗谷胱甘肽代谢对冻害胁迫的响应,筛选新疆野苹果谷胱代谢相关应答冻害基因,解析新疆野苹果组培苗抗冻的可能作用方式。【方法】以单株系的新疆野苹果组培苗为研究对象,在-3℃模拟冻害条件下,观察及检测CK、T6h、T12h、T36h、HF24h处理新疆野苹果组培苗的形态、荧光参数Fv/Fm值、相对电导率及丙二醛、过氧化氢、还原性谷胱甘肽含量,并对对照、T6h、T12h3个处理的幼苗叶片进行转录组测序分析。【结果】使用-3℃模拟冻害对新疆野苹果组培苗进行胁迫,处理12h时新疆野苹果组培苗叶尖卷缩,处理36h时整个植株完全萎蔫,在HF24h时萎蔫的植株完全恢复;与对照相比,随着冻害处理时间的延长,相对电导率及丙二醛、过氧化氢、还原性谷胱甘肽含量显著上升,荧光参数Fv/Fm值显著下降。在HF24h,相对电导率、丙二醛、过氧化氢的含量相比T36h呈现下降趋势,荧光参数恢复到与对照相当,还原性谷胱甘肽含量相比T36h没有显著变化。利用KEGG数据库对转录组数据进行分析,筛选到18个谷胱甘肽代谢途(map00480)差异基因,分别注释为GGCT、GSS、GGT1_5、CARP、speE和IDH1。它们参与GSH的降解与合成、GSH与GSSG的动态平衡和GSH消除氧化电位高的物质反应中。【结论】新疆野苹果组培苗在-3℃模拟冻害条件下,随着处理时间的延长,新疆野苹果组培苗受到的伤害不断加深,恢复处理24h后,组培苗有显著的恢复,在转录组水平,基因GST和GPX显著上调表达,表明新疆野苹果可以通过GSH代谢抵御低温胁迫。【Objective】To explore the response of glutathione metabolism and screen the glutathione metabolity-related genes responding to freezing stress,the possible pathway of freezing resistance was analyzed in Malus sieversii seedlings in vitro.【Methods】The tissue-cultured plantlets of M.sieversii were used as the material,the phenotype,maximum quantum yield of photosynthetic systemⅡ(Fv/Fm),electrolyte leakage,malondialdehyde(MDA),hydrogen peroxide(H2O2)and glutathione(GSH)con-tents were determined in M.sieversii seedlings in vitro,with the treatments of the control(CK),T6h,T12h,T36h and HF24h at-3℃.Transcriptome sequencing analysis was performed on leaves of M.sieversii seedlings treated with CK,T6h and T12h.【Results】Compared with CK,there were no signifi-cant changes in leaf shape and color after treatment at-3℃for 6 h,the edge of top leaf on M.sieversii seedlings showed reverse winding,the leaf color became dark,and the apical tip tender leaves were wilting after treatment at-3℃for 12 h,the leaves on M.sieversii seedlings treated were wilting and drooping,and the leaf color was further dark after treatment at-3℃for 36 h.After recovery at 25℃for 24 h,the leaf of wilting curl stretched,the leaf color also became bright,and the brown small spots appeared on the leaf.Compared with CK,Fv/Fm decreased by 20%,30%and 53%at T6h,T12h and T36h in leaves on M.sieversii seedlings under-3℃low temperature stress,and there was no signifi-cant difference between HF24h and CK.Compared with CK,the electrolyte leakage did not increase significantly at T6h,but increased significantly by 50%and 175%,respectively at T12h and T36h,and the electrolyte leakage at HF24h was 48%lower than T36h in leaves on M.sieversii seedlings un-der-3℃low temperature stress.Compared with CK,the content of MDAdid not change significantly atT6h,but increased by 57%and 86%atT12h andT36h,and the MDAcontent at HF24h was 23%low-er than T36h in leaves on M.sieversii seedlings under-3℃low temperature stress.Compared with CK,the hydrogen

关 键 词：新疆野苹果组培苗 冻害胁迫 谷胱甘肽代谢 转录组 

分 类 号：S661.1[农业科学—果树学]