蒺藜苜蓿低温胁迫响应基因MtbHLH-1的克隆及功能分析

doi:10.3864/j.issn.0578-1752.2012.16.024

摘要/Abstract

摘要： 【目的】克隆蒺藜苜蓿（Medicago truncatula）低温胁迫响应基因MtbHLH-1，并对其进行序列特征分析，研究MtbHLH-1对牧草蒺藜苜蓿耐低温能力的影响。【方法】利用RT-PCR技术获得蒺藜苜蓿MtbHLH-1全长的cDNA序列。对该序列进行生物信息学分析，采用半定量RT-PCR研究MtbHLH-1的低温（4℃）表达模式。【结果】测序结果显示，cDNA片段全长778 bp，包含一个741 bp完整的开放阅读框，编码246个氨基酸残基，命名为MtbHLH-1，GenBank登录号为JN800447。氨基酸BLAST分析表明，MtbHLH-1氨基酸序列与已报道的其它植物（大豆、拟南芥、烟草、小麦和玉米等）bHLH氨基酸序列具有45%—73%相似性。半定量RT-PCR研究表明，MtbHLH-1在低温胁迫条件下上调表达。【结论】从蒺藜苜蓿中克隆了低温胁迫碱性/螺旋-环-螺旋转录因子MtbHLH-1，半定量分析表明其在蒺藜苜蓿抗冷过程中起作用。

关键词: 蒺藜苜蓿, MtbHLH-1, bHLH转录因子, 功能分析

Abstract: 【Objective】This study is aimed to characterize the low-temperature-responsive gene MtbHLH-1 in M. truncatula by clone and analyse its sequence, and study the function of MtbHLH-1 in low temperature response. 【Method】The full-length cDNA sequence of MtbHLH-1 was isolated by RT-PCR. The obtained cDNA sequence and the deduced amino acid sequence were analysed. Semi-quantitative RT-PCR was used to assess the expression of MtbHLH-1 in response to low temperature (4℃) stress. 【Result】The sequencing result showed that the cloned cDNA (designated as MtbHLH-1, GenBank Accession No. JN800447) was 778 bp, contains an open reading frame (ORF) of 741 bp, and encoding a protein with 246 amino acids. BLASTp search revealed 45% to 73% similarities between the MtbHLH-1 protein and those reported bHLHs from other species, such as Glycine max, Arabidopsis thaliana, Nicotiana tabacum, Triticum aestivum and Zea mays. The qRT-PCR result showed that MtbHLH-1 was expressed upregulatedly in response to low temperature stress.【Conclusion】The cold stress response transcription factor MtbHLH-1 was cloned from M. truncatula and the semi-quantitative analysis of the gene MtbHLH-1 under low temperature demonstrate that the MtbHLH-1 gene probably improved low temperature stress tolerant capacities.

Key words: Medicago truncatula, MtbHLH-1, bHLH transcription factors, functional analysis

李宇伟, 连瑞丽, 王新民. 蒺藜苜蓿低温胁迫响应基因MtbHLH-1的克隆及功能分析[J]. 中国农业科学, 2012, 45(16): 3430-3436.

LI Yu-Wei, LIAN Rui-Li, WANG Xin-Min. Isolation and Functional Analysis of the Low-Temperature-Responsive Gene MtbHLH-1 in Medicago truncatula[J]. Scientia Agricultura Sinica, 2012, 45(16): 3430-3436.

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