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

发布时间：2018-11-15 21:12

【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10d后,转基因拟南芥的游离脯氨酸含量为2 429.58μg·g~(-1),比野生型高2.2~2.5倍;转基因拟南芥叶绿素含量为0.82mg·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1.19mmol·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]:The overexpression vector of Arabidopsis thaliana At5NC056820 was constructed, and infected Arabidopsis thaliana with flower dipping method. Twelve over-expressed lines were successfully identified by semi-quantitative PCR, and the physiological indexes of transgenic plants under drought stress were determined. The drought resistance of transgenic plants was analyzed in order to clarify the response of At5NC056820 to drought stress and to provide the basis for further revealing the molecular mechanism of lncRNA affecting drought resistance of plants. The results showed that: (1) the vector containing At5NC056820 was successfully constructed and 12 positive plants were obtained. (2) the positive plants were verified by RCR that At5NC056820 had been overexpressed in Arabidopsis thaliana. The transgenic Arabidopsis thaliana A-3, A-7 and A-8 had better growth than wild type under drought treatment. (3) after drought treatment for 10 days, the free proline content of transgenic Arabidopsis thaliana was 2 429.58 渭 g / g ~ (-1). It was 2.5 times higher than the wild type. The chlorophyll content of transgenic Arabidopsis thaliana was 0.82mg g-1, which decreased by 48.9% on average and 63.0% by wild-type Arabidopsis thaliana. The malondialdehyde content of transgenic Arabidopsis thaliana was 2.15 mmol L ~ (-1) g ~ (-1), which was less 1.19mmol L ~ (-1) g ~ (-1) than that of wild type. The relative water content of transgenic Arabidopsis thaliana was 53.73, which was 19.0% less than that of wild type. The results show that lncRNA-At5NC056820 can improve drought tolerance of Arabidopsis thaliana to some extent.
【作者单位】：西北农林科技大学生命科学学院;
【基金】：神华集团科技创新项目(01300/K403021408)
【分类号】：Q943.2

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