野生大豆碳酸盐胁迫应答基因GsMIPS2的克隆及功能分析

摘要

To identify the key functional genes in response to bicarbonate stress inGlycine soja L. G07256, we constructed a gene expression profile under bicarbonate treatment using high throughput RNA-seq data, from which we selected a myo-inositol- 1-phosphate synthase gene whose expression was significantly induced by bicarbonate stress. By homology-based cloning, we acquired its full length cDNA and termed asGsMIPS2. The results of quantitative real-time PCR demonstrated the bicarbonate stress induced expression ofGsMIPS2 and its tissue expression specificity. We also verified the function ofGsMIPS2 in bicarbon-ate responses by using theGsMIPS2 transgenicArabidopsis combined with the T-DNA insertion lineatmips2. We revealed that the germination rate ofGsMIPS2 overexpression lines was significantly higher, while that ofatmips2 mutant line was much lower than that of wild type under bicarbonate stress. These results indicate the positive role ofGsMIPS2 in plant bicarbonate stress responses.%为挖掘野生大豆(Glycine sojaL. G07256)耐碳酸盐关键功能基因,利用前期高通量转录组测序数据,从构建的碳酸盐胁迫基因表达谱中,选取了一个碳酸盐胁迫下显著上调表达的肌醇-1-磷酸合酶类基因。采用同源克隆的方法,获得该基因的全长 cDNA,命名为 GsMIPS2。实时荧光定量 PCR 结果显示该基因受碳酸盐胁迫诱导表达,并且其表达量具有组织特异性。将GsMIPS2基因转化拟南芥,并结合拟南芥中T-DNA插入突变体atmips2来验证其耐碳酸盐功能。结果表明,碳酸盐胁迫条件下, GsMIPS2超量表达拟南芥种子萌发率显著高于野生型,而拟南芥突变体atmips2种子萌发率显著低于野生型。上述结果表明, GsMIPS2基因在植物应答碳酸盐胁迫过程中起重要作用。