拟南芥lncRNAAt5NC056820 过表达载体构建及其转基因植株的抗旱性研究
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神华集团科技创新项目(01300/K403021408)


Construction of lncRNAAt5NC056820 Overexpression Vector in Arabidopsis thaliana and Study on Drought Resistance of Transgenic Plants
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    摘要:

    以拟南芥At5NC056820 为研究对象,构建其过表达载体,以蘸花法侵染拟南芥,经半定量PCR鉴定成功获得了12株过表达株系,并测定转基因植株在干旱胁迫下的生理指标,分析转基因植株的抗旱性,以明确At5NC056820 对干旱胁迫的响应,为进一步揭示lncRNA影响植物抗旱性的分子机制提供依据。结果显示:(1)成功构建出含有At5NC056820的载体,并得到了12株阳性植株。(2)阳性植株经RCR验证,证实At5NC056820 已在拟南芥中过表达;转基因拟南芥A3、A7和A8在干旱处理下均较野生型长势良好。(3)干旱处理10 d后,转基因拟南芥的游离脯氨酸含量为2 429.58 μg·g-1,比野生型高2.2~2.5倍;转基因拟南芥叶绿素含量为0.82 mg·g-1,平均下降了48.9%,而野生型拟南芥下降了63.0%;转基因拟南芥丙二醛含量为2.15 mmol·L-1·g-1,其上升量较野生型少1.19 mmol·L-1·g-1;转基因拟南芥的植物组织相对含水量为53.73%,其下降量较野生型少19.0%。研究表明,lncRNAAt5NC056820 能够在一定程度上提高拟南芥的耐旱性。

    Abstract:

    At5NC056820 of Arabidopsis thaliana overexpressing vector was constructed by dipping method and 12 strains of overexpression lines were obtained by semiquantitative PCR. The physiological indexes of transgenic plants under drought stress were analyzed to research the relationship between At5NC056820 and drought stress, which provided the basis for revealing the molecular mechanism of lncRNA affecting plant drought resistance. The results showed that: (1) the vector containing At5NC056820 was sucessfully constructed and 12 positive plants were obtained. (2) At5NC056820 was overexpressed in Arabidopsis thaliana which were confirmed by RCR; the transgenic plants A3, A7 and A8 grow better than wild plants under drought treatment. (3) After 10 days of drought treatment, the free proline content of transgenic Arabidopsis was 2 429.58 μg·g-1, which was 2.2-2.5 times higher than that of wild type. The chlorophyll content of transgenic Arabidopsis was 0.82 mg·g-1, which decreased by 48.9% on average, while that of wild type Arabidopsis decreased by 63.0%. The content of malondialdehyde (MDA) was 2.15 mmol·L-1·g-1, which was 1.19 mmol·L-1·g-1 ouer than that of wild type, and the relative water content of transgenic Arabidopsis was 53.73% and 19.0% lower than that of wild type. The result shows that lncRNAAt5NC056820 could increase the drought resistance of A. thaliana.

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毋若楠,王 红,杨成成,等.拟南芥lncRNAAt5NC056820 过表达载体构建及其转基因植株的抗旱性研究[J].西北植物学报,2017,37(10):1904-1909

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  • 在线发布日期: 2017-12-01
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