大豆肌醇加氧酶基因响应线虫胁迫的表达分析

发布时间：2018-05-12 19:55

本文选题：大豆肌醇加氧酶 + 大豆胞囊线虫　；参考：《中国油料作物学报》2017年06期

【摘要】：肌醇加氧酶通常与植物细胞壁的合成以及诱导合胞体发育相关。为确定大豆胞囊线虫侵染大豆根部后,肌醇加氧酶基因表达水平的变化,以感病品种辽豆15和抗病品种灰皮支黑豆、哈尔滨小黑豆和小粒黑豆为试验材料,在人工接种大豆胞囊线虫3号生理小种后,利用实时荧光定量PCR检测肌醇加氧酶家族基因表达量。结果表明,抗病品种中GmMIOX2基因均在接种后5d表达量达到最大,而感病品种变化并不明显。GmMIOX4基因在抗病品种灰皮支黑豆、哈尔滨小黑豆和小粒黑豆中表达量分别在接种后5d,20d和10d显著高于感病品种的表达量。表明GmMIOX2和GmMIOX4基因与大豆胞囊线虫发育调控相关。通过扩增Gm MIOX基因的CDS序列,构建GFP融合表达载体p CAMBIA1303-MIOX2和p CAMBIA1303-MIOX4,农杆菌浸润法注射本氏烟叶片进行亚细胞定位观察,GmMIOX2蛋白主要分布在细胞膜,GmMIOX4蛋白主要分布在细胞质和细胞膜。生物信息学分析表明,GmMIOX2和GmMIOX4蛋白结构域主要由α螺旋构成,蛋白序列进化分析发现GmMIOX2和GmMIOX4分别与拟南芥At MIOX1、At MIOX4亲缘性较近。
[Abstract]:Inositol oxygenase is usually associated with the synthesis of plant cell walls and the induction of syncytial development. In order to determine the changes of inositol oxygenase gene expression in soybean roots infected by soybean cyst nematode, the susceptible cultivar Liaodou 15, the disease-resistant variety Liaopizhi black bean, Harbin small black bean and small black bean were used as experimental materials. After inoculation of soybean cyst nematode No. 3 physiological race, real-time fluorescence quantitative PCR was used to detect the gene expression of inositol oxygenase family. The results showed that the expression of GmMIOX2 gene reached the maximum at 5 days after inoculation, while the expression of GmMIOX4 gene was not obvious in the resistant variety. The expression levels in Harbin small black bean and small black bean were significantly higher than those in susceptible varieties at 20 days and 10 days after inoculation respectively. The results showed that GmMIOX2 and GmMIOX4 genes were related to the development and regulation of soybean cyst nematode. By amplifying the CDS sequence of GM MIOX gene, the GFP fusion expression vectors p CAMBIA1303-MIOX2 and p CAMBIA1303-MIOX4 were constructed. The subcellular localization of GmMIOX2 protein was observed by Agrobacterium tumefaciens infiltration method. The GmMIOX4 protein mainly distributed in the cytoplasm and cell membrane. Bioinformatics analysis showed that the protein domains of GmMIOX2 and GmMIOX4 were mainly composed of 伪 helix. The phylogenetic analysis of GmMIOX2 and GmMIOX4 showed that GmMIOX2 and GmMIOX4 were close to Arabidopsis thaliana at MIOX4.
【作者单位】：沈阳农业大学生物科学技术学院;黑龙江省农业科学院;沈阳农业大学植物保护学院;
【基金】：国家自然科学基金重点项目资助(31330063)
【分类号】：S435.651

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