沙棘PEPCK基因的克隆及表达研究
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自治区自然科学基金项目(2014211B037)


Clone and Expression of PEPCK Gene in Hippophae rhamnoides L.
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    摘要:

    以沙棘为研究对象,通过RACE、基因克隆和定量PCR技术,克隆并分析沙棘磷酸烯醇式丙酮酸羧激酶基因(PEPCK)的序列信息,以及进化树构建和氨基酸的二级结构分析,比较NaCl胁迫后PEPCK基因的表达变化,为揭示PEPCK基因在沙棘抗性方面的功能研究奠定基础。结果显示:(1)成功克隆了沙棘PEPCK基因,该基因开放阅读框(ORF)为2 001 bp,编码666个氨基酸。(2)序列分析表明,沙棘PEPCK与蓖麻、黄顶菊、核桃等序列一致性高达85%以上;进化上沙棘PEPCK基因与蓖麻和狭叶羽扇豆较近,与烟草、拟南芥进化关系较远。(3)在300 mmol·L-1 NaCl盐胁迫下,沙棘PEPCK基因表达在胁迫组中显著上升,胁迫后7 d时为未处理的2.5倍,15 d时达到未处理的3.2倍。研究表明,沙棘PEPCK基因在沙棘适应外源盐胁迫的过程中可能发挥了重要作用。

    Abstract:

    For exploring Hippophae rhamnoides L., using RACE and RTqPCR technique, we cloned phosphoenolpyruvate carboxykinase (PEPCK) gene. The sequence analysis was performed to determine the homology, phylogenetic tree and secondary structure for PEPCK protein in H. rhamnoides L., compare the expression differentiation of PEPCK under NaCl stress. This study laid the foundation in order to reveal the function of PEPCK gene resistant to salt stress in H. rhamnoides L. The results indicated that: (1) the open reading frame (ORF) of H. rhamnoides L. was 2 001 bp, and 666 amino acids were encoded. (2) Sequence analysis showed that the consistency of H. rhamnoides L. was as high as 85% with Ricinus communis, Flaveria pringlei and Juglans regia and the evolution relationship of PEPCK is relatively close to that of R. communis and Lupinus angustifolius, far from Nicotiana tabacum and Arabidopsis thaliana. (3) Under 300 mmol·L-1 NaCl stress, PEPCK gene expression of H. rhamnoides L. increased significantly in the stress group with 2.5 times higher than that of normal group at 7th day, and 3.2 times higher at 15th day, which showed a significant rise in PEPCK gene expression under salt stress. Our study showed that the complete coding sequence of PEPCK gene in H. rhamnoides L. was successfully cloned, and PEPCK gene may play an important role in the process of adaptation exogenous salt stress. The result has laid a theoretical basis for revealing the molecular mechanism of H. rhamnoides L. in resistance to salt stress.

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宋 彬,胡安鸿,田永芝,等.沙棘PEPCK基因的克隆及表达研究[J].西北植物学报,2017,37(10):1934-1940

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  • 在线发布日期: 2017-12-01
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