四个苹果<em>MdCBF</em>基因克隆与表达分析

doi:10.7668/hbnxb.2017.S1.004

摘要/Abstract

摘要： 为了筛选出苹果中的抗冻负调控基因，培育抗冻苹果新品种奠定基础，通过生物信息学的方法从苹果中克隆了4个具有完整开放阅读框的CBF基因，命名为MdCBFL1~MdCBFL4。通过与其他植物CBF/DREB蛋白氨基酸序列的多重比对发现，这4个蛋白具有CBF蛋白典型的结构特点，即1个AP2结构域，2个特征序列，即核定位信号区PKRPAGRTKFRETRHP和DSAWR保守序列。从系统进化树中可以看出苹果MdCBFL1~MdCBFL4蛋白与AtDREB1蛋白聚在A-1亚族，推测它与拟南芥A-1转录因子功能相似，MdCBFL1~MdCBFL4蛋白表达受干旱、盐碱、低温等逆境胁迫的诱导。采用半定量RT-PCR的方法检测分析苹果中MdCBFL1~MdCBFL4基因在高盐（200 mmol/L NaCl）、干旱和低温处理下基因的表达情况，结果表明，MdCBFL1基因对低温、干旱、盐胁迫3种逆境胁迫均有一定程度的响应，MdCBFL2~MdCBFL4基因受低温与高盐的诱导。为进一步阐明苹果MdCBF基因在逆境应答中的功能、机制提供了有益线索。

关键词: 苹果, CBF/DREB转录因子, 基因克隆, 非生物胁迫, 表达分析

Abstract: In order to select antifreeze negative gene in apple, lay a foundation for cultivating new varieties of antifreeze apples,4 open reading frame CBF genes were cloned from apple by the method of information biology,named as MdCBFL1-MdCBFL4. These genes(MdCBFL1-MdCBFL4) were shown to encode proteins that share a high similarity with plant cold stress-related CBF proteins,which contained a AP2 domain and two classic sequences that conserved PKRPAGRTKFRETRHP nuclear location signal area and DSAWR conservative sequence. Phylogenetic analysis revealed that the isolated apple genes could be classified into the Arabidopsis A-1 subgroup, it was speculated that it was similar to the function of Arabidopsis A-1 transcription factor, and the expression of MdCBFL1-MdCBFL4 proteins was induced by stress of drought, low temperature and salt stress.The expression of MdCBFL1-MdCBFL4 gene in apple was detected by semi-quantitative RT-PCR in high salt(200 mmol/L NaCl),drought and low temperature.The results showed that MdCBFL1 gene was response to the low temperature,drought and salt stress. MdCBFL2-MdCBFL4 gene was induced by low temperature and high salt. This provides useful clues to further elucidate the function and mechanism of apple MdCBF gene in stress response.

Key words: Apple, CBF/DREB protein, Gene cloning, Abiotic stress, Analysis of gene expression

中图分类号:

Q78
S661.03

靳芹, 代培红, 刘超, 胡芹华, 刘晓东, 李月. 四个苹果MdCBF基因克隆与表达分析[J]. 华北农学报, 2017, 32(S1): 17-22. doi: 10.7668/hbnxb.2017.S1.004.

JIN Qin, DAI Peihong, LIU Chao, HU Qinhua, LIU Xiaodong, LI Yue. Cloning and Expression Analysis of Four Apple MdCBF Genes[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2017, 32(S1): 17-22. doi: 10.7668/hbnxb.2017.S1.004.

http://www.hbnxb.net/CN/Y2017/V32/IS1/17

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四个苹果MdCBF基因克隆与表达分析

Cloning and Expression Analysis of Four Apple MdCBF Genes

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