马铃薯StAVP1基因提高拟南芥抗旱性的功能鉴定
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1.<2.sup>3.宁夏农林科学院农业生物技术研究中心<4./sup>5.宁夏农业技术推广总站<

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宁夏自然科学基金(2022AAC03431);宁夏回族自治区农业育种专项(2019NYYZ01);国家自然科学基金(32260504);宁夏自然科学基金(2021AAC05015);宁夏自然科学基金(2022AAC03429)


Functional Characterization of Potato StAVP1 to Enhance Drought Resistance in Arabidopsis
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Natural Science Foundation Project of Ningxia(2022AAC03431),Agricultural Breeding Project of Ningxia(2019NYYZ01),National Natural Science Foundation of China(32260504),Natural Science Foundation Project of Ningxia(2021AAC05015),Natural Science Foundation Project of Ningxia(2022AAC03429)

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    摘要:

    马铃薯(Solanum tuberosum L.)是全球第四大粮食作物,挖掘鉴定抗旱基因生物学基础,是培育抗旱马铃薯新品种的重要途径。马铃薯StAVP1基因参与跨液泡膜物质运输,在植物应对物质运输和非生物胁迫中发挥重要作用。本研究从‘大西洋’马铃薯叶片中克隆得到编码区全长2301 bp的StAVP1基因,生物信息学分析表明,包含典型的H+焦磷酸酶结构域和13个跨膜结构域,与近缘物种的AVP1蛋白具有较高的序列相似性,预测定位于液泡膜上;组织表达模式分析显示,StAVP1基因在花中表达量最高,块茎中表达量最低;StAVP1稳定过表达拟南芥株系的生物量和根长显著高于受体,控水干旱胁迫后,过表达StAVP1拟南芥株系叶丙二醛(MDA)含量显著低于受体,超氧化物歧化酶(SOD)、过氧化物酶(POD)和脯氨酸(Pro)含量显著高于受体;DREB等逆境应答基因表达量显著上调;CoIP试验结果显示,StAVP1与StRAB发生直接互作,且干旱胁迫下StAVP1过表达拟南芥株系中StRAB基因表达量也显著上调。本研究通过生物信息分析、异源表达表型、生理、基因表达方面功能验证、CoIP互作蛋白分析,表明马铃薯StAVP1基因是马铃薯遗传改良的重要基因来源。

    Abstract:

    Potato (Solanum tuberosum L.) is the fourth largest food crop in the world. To explore and identify the biological basis of drought resistance genes is an important way to cultivate new varieties of drought-resistant potato. Potato StAVP1 gene is involved in material transport across the vacuolar membrane and plays an important role in plant response to material transport and abiotic stress. In this study, the StAVP1 gene with the full length of 2301 bp coding region was cloned from the leaves of "Atlantic" potato. Bioinformatics analysis showed that the StAVP1 gene contained typical H+ pyrophosphatase domain and 13 transmembrane domains, which had high sequence similarity with AVP1 protein of related species, and was predicted to be located on the vesicular membrane. Tissue expression pattern analysis showed that StAVP1 gene expression was the highest in flowers and the lowest in tubers. The biomass and root length of Arabidopsis Thaliana plants overexpressed in StAVP1 were significantly higher than those of receptors, and the contents of malondialdehyde (MDA) in leaves of Arabidopsis Thaliana plants overexpressed in StAVP1 were significantly lower than those of receptors under water-controlled drought stress, while the contents of superoxide dismutase (SOD), peroxidase (POD) and proline (Pro) were significantly higher than those of receptors. Stress response genes such as DREB were significantly up-regulated. The results of CoIP test showed that StAVP1 directly interacted with StRAB, and StRAB gene expression was significantly up-regulated in Arabidopsis Thaliana strains overexpressed in StAVP1 under drought stress. In this study, through biological information analysis, heterologous expression phenotype, physiological, functional verification of gene expression, CoIP interaction protein analysis, potato StAVP1 gene is an important gene source for potato genetic improvement.

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  • 收稿日期:2024-01-02
  • 最后修改日期:2024-05-31
  • 录用日期:2024-07-17
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