野生大豆GsSnRK1.1蛋白激酶在淹水胁迫下功能分析

Functional analysis of Glycine soja GsSnRK1.1 protein kinase in plant resistance to waterlogging stress

为明确GsSnRK1.1蛋白激酶在植物淹水胁迫中的作用与功能,在野生型拟南芥(Col-0)中过量表达GsSnRK1.1和突变基因GsSnRK1.1(T176A)、GsSnRK1.1(T176E)、GsSnRK1.1(K49M)。当转基因拟南芥植物生长至四叶期时,对其淹水胁迫处理后,测定各种基因型植物叶绿素、SOD和Pro等生理指标,以及ADH1、PDC1等淹水应激反应基因表达模式,同时对proGsSnRK1.1::GUS转基因拟南芥淹水胁迫并染色。结果表明,野生大豆GsSnRK1.1基因对淹水胁迫有响应,且其启动子可感知淹水胁迫。GsSnRK1.1和GsSnRK1.1(T176E)转基因植株在淹水胁迫下,叶绿素、SOD和Pro含量升高,ADH1及PDC1基因表达上调;然而GsSnRK1.1(T176A)和GsSnRK1.1(K49M)转基因植株在淹水胁迫下,叶绿素、SOD和Pro含量降低,ADH1及PDC1基因表达未发生明显变化。研究为进一步探究大豆抗涝品种选育提供理论依据。

In order to clarify the role and function of GsSnRK1.1 protein kinase in plant waterlogging stress.GsSnRK1.1,GsSnRK1.1(T176A),GsSnRK1.1(T176E)and GsSnRK1.1(K49M)were overexpressed in Col-0 Arabidopsis thaliana.When Arabidopsis thaliana transgenic plants grew to the four-leaf stage,the plants were treated by water flood.Subsequently,chlorophyll,SOD and Pro contents were measured,and the gene expression patterns of ADH1 and PDC1 hypoxic stress responsive marker genes in Col 0 and transgenic Arabidopsis plants were determined.At the same time,the proGsSnRK1.1::GUS transgenic Arabidopsis thaliana plants were subjected to waterlogging stress to observe the staining.The results showed that the Glycine soja GsSnRK1.1 gene responded to waterlogging stress,and its promoter could sense waterlogging stress.GsSnRK1.1 and GsSnRK1.1(T176E)transgenic plants had increased chlorophyll,SOD and Pro contents under waterlogging stress,and the expressions of ADH1 and PDC1 genes were up-regulated.However,GsSnRK1.1(T176A)and GsSnRK1(K49M)transgenic plants under waterlogging stress had reduced chlorophyll,SOD and Pro,and the expressions of ADH1 and PDC1 genes did not change significantly.The research provided a theoretical basis for further exploring the breeding of waterlogging-resistant soy-bean varieties.