水稻干尖线虫海藻糖酶Ab-tre-1基因克隆与逆境条件下的表达分析

发布时间：2018-04-09 13:20

  本文选题：水稻干尖线虫　切入点：海藻糖酶　出处：《核农学报》2016年12期

【摘要】：海藻糖酶参与生物体内能量代谢,在抵御逆境胁迫过程中发挥关键作用。水稻干尖线虫是水稻上重要的寄生线虫,能够在多种逆境中存活,严重危害水稻生长。为探究水稻干尖线虫海藻糖酶基因的功能,利用RACE-PCR技术获得了该线虫海藻糖酶基因全长,将其命名为Ab-tre-1,并利用实时荧光定量PCR技术分析在线虫卵、不同龄期和若干逆境条件下该基因的表达情况。结果表明,该基因DNA序列长2 083bp,包含9个内含子,开放阅读框1 743 bp,编码580个氨基酸,含有2个典型的海藻糖酶标签基序。进化树结果表明,水稻干尖线虫海藻糖酶与自由生活线虫和动物寄生线虫位于不同进化分支。Ab-tre-1在线虫卵中的表达最高,二龄幼虫和成虫次之,三、四龄幼虫表达水平较低。此外,当水稻干尖线虫暴露于干燥、活性氧、高温和药剂胁迫环境中,Ab-tre-1表达水平均显著提高,表明该基因可能与水稻干尖线虫的抗逆过程有关。本研究结果为阐明水稻干尖线虫海藻糖酶基因抗逆表达提供了理论依据。
[Abstract]:Trehalase is involved in energy metabolism and plays a key role in resisting stress.Rice dry tip nematode is an important parasitic nematode in rice. It can survive many kinds of stresses and seriously harm the growth of rice.In order to study the function of trehalase gene of rice dry tip nematode, the full length of trehalase gene was obtained by RACE-PCR technique, named Ab-tre-1, and real-time fluorescence quantitative PCR was used to analyze the eggs.The expression of the gene in different ages and some stress conditions.The results showed that the DNA sequence of the gene was 208.3 BP, containing 9 introns, open reading frame 1,743 BP, encoding 580amino acids, and containing two typical trehalase tag motifs.The results of phylogenetic tree showed that the expression of trehalase, free living nematodes and parasitic nematodes were the highest in the eggs of different evolutionary branches. The second instar larvae and adults were the second, and the third and fourth instar larvae had lower expression level.In addition, the expression level of Ab-tre-1 was significantly increased in dry, reactive oxygen species, high temperature and pesticide stress environments, suggesting that the gene might be related to the stress resistance process of rice dry tip nematode.The results of this study provide a theoretical basis for clarifying the stress resistance of trehalase gene of rice dry tip nematode.
【作者单位】： 江苏省农业科学院植物保护研究所;江苏丘陵地区镇江农业科学研究所;
【基金】：国家青年科学基金项目(31401728) 镇江市农业重点研发计划项目(2015021)
【分类号】：S435.111.48
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本文编号：1726596