REN Qiu-yan,GAI Jun-yi,LI Kai.Cloning and Expression Analysis of Soybean Stress-tolerant Gene GmMADS[J].Soybean Science,2020,39(01):30-38.[doi:10.11861/j.issn.1000-9841.2020.01.0030]
大豆GmMADS基因的克隆及表达分析
- Title:
- Cloning and Expression Analysis of Soybean Stress-tolerant Gene GmMADS
- Keywords:
- Soybean［Glycine max (L.) Merr.］; GmMADS; Cis-acting element; Subcellular localization; Soybean mosaic virus (SMV); Abiotic stress
- 文献标志码:
- A
- 摘要:
- 为了探究大豆MADS-box编码基因在生物和非生物胁迫中的调控作用，以SMV抗感大豆品种为试验材料，从大豆中克隆GmMADS基因Glyma.02g121500，进行生物信息学分析和表达特点分析，同时检测不同逆境胁迫下GmMADS基因表达量变化情况。结果表明：GmMADS基因完整ORF长度为735 bp，编码244个氨基酸，pI 6.68，相对分子质量为28.2 kD。抗感品种间基因CDS序列存在1个SNP差异，氨基酸序列无差异。启动子序列包含防卫和胁迫响应元件、植物激素应答元件、光应答元件等许多顺式作用元件。GmMADS与木豆、花生、豇豆、羽扇豆等物种中的同源基因亲缘关系较近。亚细胞定位结果显示GmMADS在细胞核上表达。组织特异性表达分析显示GmMADS在花中的表达量最高。接种大豆花叶病毒后，抗病品种科丰1号GmMADS在2 h表达最高且显著高于感病品种南农1138-2；低温胁迫时，GmMADS表达在2 h上调；盐胁迫时，GmMADS表达在1 h下调；干旱胁迫时，GmMADS表达在2 h下调。本研究对下一步分析该基因功能、阐明该基因在调控大豆抗病和耐逆的分子机制具有重要意义。
- Abstract:
- In order to explore the regulatory role of soybean GmMADS gene in biotic and abiotic stresses, we cloned gene glyma.02g121500 from SMV resistant and susceptible soybean varieties. We analyzed the gene structure with bioinformatics, analyzed the expression characteristics, at the same time, we detected the expression of the gene under different stress. The results revealed: Gene glyma.02g121500 consisted a complete ORF of 735 bp encoding 244 amino acids. The theoretical isoelectric point is 6.68 and the relative molecular mass is 28.2 kD. The sequence alignment showed that there was one SNP difference in the CDS sequence between the resistant and susceptible varieties, and there was no difference in the amino acid sequence. Its promoter sequence contains a number of cis-acting elements such as defense and stress response elements, plant hormone response elements, photo responsive elements, and so on. The homologous genes in the species such as pigeonpea, peanut, cowpea and lupin were closely related. Subcellular localization results showed that GmMADS was expressed on the nucleus. Tissue-specific expression analysis showed that GmMADS had the highest expression in flowers. After soybean mosaic virus inoculation, the expression of GmMADS in resistant Kefeng 1 was highest at 2 h and significantly higher than the susceptible Nannong 1138-2. The expression of GmMADS was up-regulated at 2 h at low temperature stress, the expression of GmMADS was down-regulated at 1 h under salt stress and down-regulated at 2 h under drought stress. This study is important for the next step in analyzing the function of this gene and elucidating its molecular mechanism for regulating soybean disease resistance and tolerance.
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备注/Memo
收稿日期：2019-08-02