线虫胁迫下大豆钙调蛋白基因的表达分析

发布时间：2018-04-13 14:41

本文选题：大豆胞囊线虫 + 钙调蛋白　；参考：《中国油料作物学报》2017年01期

【摘要】：为明确受到大豆胞囊线虫侵染后大豆根部钙调蛋白基因的表达水平,以感病品种辽豆15和抗病品种灰皮支黑豆(ZDD2315)为材料,分别在接种线虫(SCN3)1d,3d,5d,9d,15d,20d取样,提取接种和未接种的大豆根部总RNA,利用实时荧光定量PCR检测大豆钙调蛋白家族:SCaM1、SCaM2、SCaM3、SCaM4和SCaM5基因的表达量。通过扩增SCaM4基因编码区序列,构建与报告基因GFP融合表达的细胞定位检测载体p COMBIA1303-SCaM4,采用农杆菌介导的方法将其瞬时表达在本氏烟(Nicotiana benthamiana)叶片上,激光共聚焦显微镜观察。结果表明无论是抗性品种灰皮支黑豆还是感病品种辽豆15,在线虫侵染后5个钙调蛋白基因的表达量都在接种后第5d达最大值,其中灰皮支黑豆SCaM4基因接种后表达量高达12.73,是未接种的1.85倍,为感病品种的1.96倍,初步认为SCaM4可能参与了灰皮支黑豆抗线虫的过程。亚细胞定位的结果表明SCaM4蛋白定位在细胞膜上。
[Abstract]:The total RNAs of soybean roots were extracted from inoculated and uninoculated soybean, and the expression of SCAM4 and SCaM5 genes were detected by real-time fluorescence quantitative PCR in the soybean calmodulin family: SCaM1, SCaM2, SCaM3, SCaM4 and SCAM4.By amplifying the coding region of SCaM4 gene, a cell localization detection vector pCOMBIA1303-SCaM4 was constructed, which was fused with the reporter gene GFP. It was transiently expressed on the leaves of Nicotiana benthamiana by Agrobacterium tumefaciens and observed by confocal laser microscope.The results showed that the expression of 5 calmodulin genes reached the maximum at the 5th day after inoculation, whether the resistant variety Liaopizhi black bean or the susceptible cultivar Liaodou 15, was infected with online insect, and the expression of calmodulin gene reached the maximum level on the 5th day after inoculation.After inoculation, the expression of SCaM4 gene in black bean was as high as 12.73, 1.85 times higher than that in non-inoculated black bean, and 1.96 times of susceptible variety. It was suggested that SCaM4 might be involved in the process of resistance to nematode.The results of subcellular localization showed that SCaM4 protein was located on the cell membrane.
【作者单位】：沈阳农业大学植物保护学院北方线虫研究所;
【基金】：国家自然科学基金重点项目(31330063)
【分类号】：S435.651

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