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线虫胁迫下大豆钙调蛋白基因的表达分析
Analysis of calmodulins relative gene expression from different soybeans under SCN infection
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，本研究以感病品种辽豆１５和抗病
品种灰皮支黑豆（ＺＤＤ２３１５）为试验材料，分别在接种线虫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，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Ｐｃｏｍｂｉａ１３０３－ＳＣａＭ４，采
用农杆菌介导的方法将其瞬时表达在本氏烟（Ｎｉｃｏｔｉａｎａｂｅｎｔｈａｍｉａｎａ）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基因接种后基因表达量高达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。
A susceptible cultivar Liaodou 15 and a resistant cultivar Huipizhi heidou (ZDD2315) were selected to research the gene expression of calmodulins from soybeans roots after SCN3 infection. RNA from infected and uninfected roots was extracted at 1d, 3d, 5d, 9d, 15d and 20d and real-time PCR was carried out respectively. The complete SCaM4 gene was amplified into binary expression vector to construct Pcombia1303- SCaM4. LSCM was used to observe the location. The results demonstrated that in both cultivars the five calmodulins genes expression levels reached the maximum at the fifth day after infection among six time points. The gene expression of SCaM4 after infected was 1.85 fold of uninfected. And in resistant cultivar the relative expression of SCaM4 was 1.96 times of that in susceptible cultivar. The result of subcellular localization showed that SCaM4 protein was located on the cytomembrane. In summary SCaM4 may participate in the interaction of soybean and SCN3.
大豆胞囊线虫 / 钙调蛋白 / 基因相对表达量 {{custom_keyword}} /
Hereodera glycines / calmodulin / relative gene expression {{custom_keyword}} /
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