LIU Wei,ZHANG Yan-wei,WANG Yu-bin,et al.Screening of Soybean Drought Responsive GRAS Genes and Bioinformatics and Adversity Stress Expression Analysis on GmGRAS27[J].Soybean Science,2022,41(01):36-042.[doi:10.11861/j.issn.1000-9841.2022.01.0036]
大豆干旱响应GRAS基因筛选及GmGRAS27的生物信息学和逆境表达分析
- Title:
- Screening of Soybean Drought Responsive GRAS Genes and Bioinformatics and Adversity Stress Expression Analysis on GmGRAS27
- Keywords:
- soybean; drought; GRAS; GmGRAS27; bioinformatics; adversity stress; expression analysis
- 文献标志码:
- A
- 摘要:
- GRAS转录因子在植物抵抗非生物胁迫过程中发挥了重要作用，为了研究大豆GRAS基因在干旱胁迫中的响应，以挖掘大豆GRAS转录因子在干旱等非生物胁迫响应中的功能和分子机制提供分子基础，本研究利用基因表达综合数据库（Gene Expression Omnibus， GEO）筛选响应干旱胁迫的大豆GRAS基因，利用PlantCARE分析其启动子中的顺式作用元件。进一步对在营养生长时期（V6）和盛花期（R2）均受到干旱胁迫强烈诱导的GRAS基因GmGRAS27进行克隆和生物信息学分析，并采用实时荧光定量PCR方法分析干旱、盐和ABA胁迫下GmGRAS27基因的表达情况。结果显示：从大豆117个GRAS基因中筛选出10个响应干旱胁迫的GRAS基因，其启动子序列中均至少含有1个激素或逆境胁迫应答相关元件，GmGRAS27（Glyma.06G265500）基因的启动子同时含有ABRE应答元件和MYB/MYC结合元件。GmGRAS27在V6和R2期均受到干旱胁迫的强烈诱导，其编码的蛋白质GmGRAS27含有典型的GRAS基因家族结构域，高级结构以α-螺旋和无规则卷曲为主。实时荧光定量PCR分析显示GmGRAS27不仅受到干旱胁迫的强烈诱导，还受到盐胁迫和ABA的正向调控。推测GmGRAS27不仅是大豆干旱胁迫的重要候选基因也可能响应其他非生物胁迫。
- Abstract:
- GRAS transcription factors play important roles in resisting abiotic stresses in plants. In order to study the response of GRAS genes to drought stress in soybean， and to provide a molecular basis for mining the function and molecular mechanism of soybean GRAS transcription factors in response to drought stress， we analyzed the microarray data of soybean in Gene Expression Omnibus (GEO) database to identify drought responsive GRAS genes. The cis-acting elements in the promoters of the drought responsive GRAS genes were analyzed by PlantCARE. Then we cloned and performed bioinformatics analysis of a GRAS gene， GmGRAS27， which was up-regulated by drought stress in both the vegetative stage(V6) and the full flowering stage(R2). The promoter of GMGRAS27(Glyma.06g265500) gene contains both ABRE responsive element and MYB/MYC binding element. Furthermore， the expression of GmGRAS27 under drought， salt and ABA stress was analyzed by using the qRT-PCR method. The results displayed that， in the 117 soybean GRAS genes， 10 genes responded to the drought stress. All of the ten GRAS genes were found to possess at least one stress or hormone responsive element. GmGRAS27 was up-regulated by drought stress in both the vegetative stage (V6) and the full flowering stage (R2)， and it encoded the product GmGRAS27 containing a conserved GRAS domain. The structure of GmGRAS27 was found to be mainly composed of α-helixes and random coils. GmGRAS27 was not only positively regulated by drought stress， but also induced by salt and ABA stress. These results suggested that GmGRAS27 might be an important candidate gene responding to abiotic stresses in soybean.
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备注/Memo
收稿日期：2021-07-30