浙江农业学报

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南方型紫花苜蓿叶片盐胁迫应答转录因子鉴定与分析

  

  1. (浙江农林大学 风景园林与建筑学院,浙江 临安 311300)
  • 出版日期:2016-04-25 发布日期:2016-04-27

Identification and analysis of salt stressresponsive transcription factor in leaf of southern type alfalfa

  1. (School of Landscape Architecture, Zhejiang A of F University, Linan 311300, China)
  • Online:2016-04-25 Published:2016-04-27

摘要: 以南方型紫花苜蓿‘Millennium’为材料,以正常培养(WT_CK2)和250 mmol·L-1 NaCl胁迫下72 h时(WT_N2)条件下的2个样品叶片进行转录组分析,鉴定紫花苜蓿叶片盐胁迫应答转录因子基因。同时随机挑选4个转录因子差异表达基因进行实时荧光定量qRTPCR验证RNASeq结果的可靠性。紫花苜蓿叶片在250 mmol·L-1 NaCl胁迫下72 h时,检测到表达量差异达到2倍以上的基因共7 497个,其中包括隶属于46个转录家族474个转录因子在盐胁迫下的差异表达,上调242个,下调232个。bHLH,NAC,WRKY和C2H2转录家族成员基因70%以上表达上调。实时荧光定量PCR分析表明4个随机选择的基因在胁迫前后的表达特点与表达谱测序结果一致。此外,还发现大量紫花苜蓿盐胁迫应答的转录因子候选基因如MsERF110,MsERF071,MsbHLH36,MsZFP,MsHSFB3,MsMYB,MsNAC和MsWRKY等。同时,也揭示了紫花苜蓿叶片对盐胁迫响应可能是多种转录因子家族共同参与的应答过程。

关键词: 南方型紫花苜蓿, 转录因子, 叶片转录组, 盐胁迫

Abstract: Illumina RNAsequencing was performed in two alfalfa samples of control (WT_CK2) and NaCltreated samples(WT_N2)in order to estimate a broad spectrum of transcription factor genes affected by salt stress,and the realtime fluorescent qRTPCR technique was used to verify the expression characteristics of random selected 4 genes. These results indicated that 7 497 of them showed two fold higher or lower difference under NaCl stress from the control, including 474 transcription factors (TFs) belonging to 46 transcription family, among which 242 genes were upregulated and 232 were downregulated. Transcription factor families including bHLH,NAC,WRKY and C2H2, among which more than 70% genes were upregulated. Expression patterns of random selected 4 genes confirmed the results of the digital gene expression profile. In addition, this study also identified a large number of candidate functional transcription factor genes such as MsERF110, MsERF071, Ms bHLH36, MsZFP, MsHSFB3, MsMYB, MsNAC, MsWRKY and so on, which appeared to be constitutively involved in the response to salt stress. This study revealed that many transcription factor family were involved in response to salt stress in leaf of southern type alfalfa.

Key words: southern type alfalfa, transcription factor, leaf transcriptome, salt stress