苦荞基因FtAPDSLX1的克隆及其抗立枯病功能分析
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1.湖南科技大学生命科学与健康学院;2.中国农业科学院作物科学研究所;3.海南省种业实验室

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国家自然科学基金青年项目(32301813)


Cloning of the FtAPDSLX1 Gene in Tartary Buckwheat and Analysis of Its Anti-Phytophthora Function
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National Natural Science Foundation Youth Program (32301813)

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    摘要:

    苦荞(Fagopyrum tataricum)是蓼科(Polygonaceae)荞麦属(Fagopyrum)的一年生草本植物,是一种药食同源的优质作物。荞麦立枯病的发生严重影响到了苦荞种植的推广与发展。本研究对茉莉酸或水杨酸处理后的苦荞进行转录组分析,发现天冬氨酸蛋白酶基因FtAPDSLX1的表达模式受到茉莉酸或水杨酸的诱导。对不同抗性水平苦荞中的FtAPDSLX1表达量分析结果显示立枯丝核菌抗性材料中FtAPDSLX1的表达量显著高于立枯丝核菌易感材料。以苦荞代表性品种川荞一号为材料,克隆得到FtAPDSLX1基因。生物信息学分析结果显示,FtAPDSLX1基因的CDS序列中包含1143 个碱基对,编码380个氨基酸构成的蛋白质,该基因启动子具有多个与抗病相关的顺式元件。对FtAPDSLX1的表达模式进行分析发现,苦荞幼苗中的FtAPDSLX1基因表达量受到立枯丝核菌侵染的诱导,且具有显著的组织特异性。对转FtAPDSLX1基因拟南芥株系进行抗病表型分析,结果表明FtAPDSLX1提高了转基因拟南芥的立枯丝核菌抗性,同时病菌胁迫下转基因拟南芥中FtAPDSLX1的表达量显著提高,并且FtAPDSLX1提高转基因拟南芥中SOD、POD的酶活性。以上结果显示,苦荞受到立枯丝核菌侵染时,FtAPDSLX1可能参与了对立枯丝核菌的防御机制。FtAPDSLX1的功能验证及相关研究为苦荞抗立枯丝核菌的分子机制解析以及抗病种质资源的筛选奠定了工作基础。

    Abstract:

    Tartary buckwheat (Fagopyrum tataricum) is an annual herbaceous plant of Fagopyrum in Polygonaceae, which is a high-quality crop that is homologous of medicine and food. The occurrence of Fagopyrum damping-off has seriously affected the promotion and development of Tartary buckwheat planting.This study conducted transcriptome analysis on Fagopyrum tataricum treated with jasmonic acid or salicylic acid, revealing that the expression pattern of the aspartic protease gene FtAPDSLX1 is induced by jasmonic acid or salicylic acid. Further analysis of FtAPDSLX1 expression levels in Fagopyrum tataricum with different resistance degrees revealed a significantly higher expression level of FtAPDSLX1 in Fagopyrum tataricum resistant to Rhizoctonia solanicompared than in susceptible materials. Using the representative variety of Fagopyrum tataricum, Chuanqiao No. 1, the FtAPDSLX1 gene was cloned. Bioinformatics analysis revealed that the coding sequence of the FtAPDSLX1 gene contains 1143 base pairs, encoding the protein consisting of 380 amino acids,while its promoter harbors multiple cis-elements associated with disease resistance. Examination of FtAPDSLX1 expression patterns unveiled its upregulation in Tartary buckwheat seedlings upon Rhizoctonia solaniand invasion, exhibiting pronounced tissue specificity. Phenotypic analysis of disease resistance in transgenic Arabidopsis lines harboring the FtAPDSLX1 gene underscored its role in enhancing resistance against Rhizoctonia solaniand .Furthermore, under pathogen stress, FtAPDSLX1 expression substantially increased in transgenic Arabidopsis, concomitant with heightened enzymatic activities of SOD and POD. These findings suggest that FtAPDSLX1 may participate in the defense mechanism against Rhizoctonia solaniin Fagopyrum tataricum. The functional validation and related research of FtAPDSLX1 provide a foundation for unraveling the molecular mechanisms underlying Fagopyrum tataricum"s resistance to Rhizoctonia solaniand for screening disease-resistant germplasm resources.

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  • 收稿日期:2024-04-10
  • 最后修改日期:2024-05-09
  • 录用日期:2024-10-18
  • 在线发布日期: 2024-11-11
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