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| 砂藓SMPD基因的克隆及对干旱和高温胁迫响应分析 |
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鲍薇1,2,王晶晶1,2,沙伟1,2,张梅娟1,2,彭疑芳1,2,马天意1,2*
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1.齐齐哈尔大学 生命科学与农林学院,黑龙江省 齐齐哈尔 161000；2. 抗性基因工程与寒地生物多样性保护黑龙江重点实验室,黑龙江省 齐齐哈尔 161000
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| 摘要: |
| 鞘磷脂磷酸二酯酶（sphingomyelin phosphodiesterase，SMPD）能够水解鞘磷脂，形成磷脂胆碱和神经酰胺，在鞘脂代谢中起关键作用。为研究强耐脱水性植物砂藓（Racomitrium canescens）中一个SMPD基因（RcSMPD）的耐旱和抗高温相关功能，该文利用实时荧光定量PCR检测RcSMPD在砂藓脱水胁迫及高温处理下的表达量，对其编码序列进行克隆，对其编码蛋白质进行生物信息学预测，构建其过表达转基因拟南芥（Arabidopsis thaliana）株系以检测RcSMPD的耐旱及抗高温能力。结果表明：（1）RcSMPD能够响应砂藓的脱水胁迫及高温处理。（2）RcSMPD总氨基酸残基数为326，为亲水性蛋白质，可能为酸性或中性SMPD并定位于叶绿体或液泡。（3）RcSMPD过表达转基因拟南芥与野生型植株相比形态发生改变，但在模拟干旱胁迫和高温胁迫处理下的抗逆能力无显著差异。综上，RcSMPD能够参与砂藓对脱水和高温胁迫的应答，过表达RcSMPD能够改变拟南芥形态，但未对其耐旱性和抗高温性产生显著影响。 |
| 关键词: 砂藓，鞘磷脂磷酸二酯酶基因，表达分析，干旱胁迫，高温胁迫 |
| DOI：10.11931/guihaia.gxzw202410003 |
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| Fund project:黑龙江省属高等学校基本科研业务费青年创新人才项目(145109212) |
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| Cloning and analysis of the responses to drought and high-temperature stress of SMPD in Racomitrium canescens |
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BAO Wei1,2 , WANG Jingjing1,2, SHA Wei1,2, ZHANG Meijuan1,2, PENG Yifang1,2, MA Tianyi1,2*
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1.College of Life Sciences, Agriculture and Forestry, Qiqihar University, Qiqihar 161000, Heilongjiang, China; 2. Heilongjiang Key Laboratory of Resistance Genetic Engineering and Coldland Biodiversity Conservation, Qiqihar 161000, Heilongjiang, China
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| Abstract: |
| Sphingomyelin phosphodiesterase (SMPD) plays a crucial role in the sphingolipid metabolism process by hydrolyzing sphingomyelin into phosphatidylcholines and ceramides. To investigate the functions related to drought tolerance and high-temperature resistance of an SMPD gene (RcSMPD) in the strongly desiccation-tolerant plant Racomitrium canescens, real-time fluorescence quantitative PCR was employed to detect the expression of RcSMPD under dehydration stress and high-temperature treatments. The coding sequence of RcSMPD was cloned, and bioinformatics analysis was carried out to predict the protein encoded by RcSMPD. Furthermore, transgenic Arabidopsis thaliana lines overexpressing RcSMPD were constructed to evaluate the drought tolerance and high-temperature resistance of RcSMPD. The results were as follows: (1) RcSMPD was found to respond to the dehydration stress and high-temperature treatments of R. canescens to different degrees. (2) RcSMPD had a total of 326 amino acid residues and was a hydrophilic protein. RcSMPD could be an acidic or neutral SMPD and was predicted located in chloroplasts or vacuoles. (3) Compared with wild-type plants, the morphology of the RcSMPD overexpression transgenic A. thaliana showed differences, yet there was no significant difference in stress resistance under simulated drought stress and high-temperature stress treatments among the A. thaliana lines. In conclusion, RcSMPD can participate in the response of R canescens to dehydration and high-temperature stress. Overexpression of RcSMPD can change the morphology of A. thaliana, but has no significant impact on its drought tolerance and high-temperature resistance. |
| Key words: Racomitrium canescens, sphingomyelin phosphodiesterase gene, expression analysis, drought stress, high-temperature stress |
