LIU Su-shuang,QIU Ying-sheng,LIU Yan-min,et al.Analysis of Soybean GmPM31 Bioinformatics，Tissue Expression and Response to High Temperature and High Humidity[J].Soybean Science,2021,40(05):612-619.[doi:10.11861/j.issn.1000-9841.2021.05.0612]
大豆GmPM31基因生物信息学、组织表达及高温高湿响应分析
- Title:
- Analysis of Soybean GmPM31 Bioinformatics，Tissue Expression and Response to High Temperature and High Humidity
- Keywords:
- Spring soybean; GmPM31; Bioinformatics analysis; Cis-element; Expression; Seed; High temperature and high humidity stress
- 文献标志码:
- A
- 摘要:
- sHSPs家族基因GmPM31具有典型的ACD结构域，为了预测和研究GmPM31的功能，本研究以田间劣变抗性春大豆湘豆3号为材料，分离GmPM31及其启动子序列，对该基因的结构及启动子上的顺式作用元件进行生物信息学分析，采用qRT-PCR方法对GmPM31的组织表达模式及高温高湿处理下的表达量变化进行分析。结果表明：GmPM31基因的ORF全长为459 bp，编码153个氨基酸，包含1个ACD保守域结构。蛋白质系统进化树分析结果显示，GmPM31与CI(Class I)亚家族的sHSPs同源性较高，表明GmPM31属于Class I sHSPs家族成员。GmPM31基因启动子中有多个逆境胁迫响应元件，包括激素应答相关元件ABRE、ERE和AAGAA-motif等，干旱和冷胁迫相关的MYB和MYC转录因子作用元件，高盐胁迫相关元件Box III，厌氧胁迫相关元件ARE等。qRT-PCR结果显示，GmPM31在成熟种子中大量表达，其次在幼荚和叶片中表达，在根、茎和花中表达量极低。种子发育过程中，GmPM31表达量呈现先上升后下降的趋势，在开花后第45天表达量达到最大。高温高湿胁迫处理96和168 h时种子中GmPM31的表达量显著高于对照组，说明高温高湿胁迫诱导种子中GmPM31上调表达。研究结果表明GmPM31参与了春大豆种子的发育以及高温高湿胁迫响应过程。
- Abstract:
- GmPM31 contains a typical ACD domain of the sHSPs family.In order to predict and study the function of GmPM31，we isolated the sequence of GmPM31 and its promoter from spring field deterioration resistant soybean Xiangdou 3，and resistance the structure and cis-regulatory element of GmPM31 by bioinformatics.In addition，we analyzed the tissue expression pattern and the expression levels of GmPM31 under high temperature and high humidity by qRT-PCR.The results showed that the complete ORF of GmPM31 was 459 bp.GmPM31 encoded 153 amino acids and contained a highly conserved ACD domain.The results of the evolutionary tree showed that GmPM31 has high homology with the Class I sHSPs，so we speculated GmPM31 may belong to the Class I sHSPs family.The prediction result of promoter cis-elements showed that the GmPM31 promoter contained many stress responsive elements such as hormone responsive element ABRE，ERE，AAGAA-motif，element involoved in MYB and MYC transcription factors related to drought and cold stress，high salt stress responsive element Box III，and anaerobic stress responsive element ARE.qRT-PCR results showed that the highest expression level of GmPM31 was found in mature seeds，followed by in young pods and leaves,and a lowest expression level in root，stems，leaves and flowers.Over the seed developing，the expression of GmPM31 increased at first and then decreased，and reached higher levels at 45 d after flowering.Compared to the corresponding control，the expression of GmPM31 in seeds was significantly higher at 96 and 168 h under high temperature and high humidity stress，which indicated that GmPM31 in seeds was up-regulated by high temperature and high humidity stress.These results showed that GmPM31 was involved in the development of spring soybean seeds and the response to high temperature and high humidity stress.
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备注/Memo
收稿日期：2021-04-14