| 孙 璐，蔡浩宇，李 明，曹高燚，丁 博，包曙光，王俊斌，谢晓东，陈小强.大麦气孔发育相关基因HvFAMA的克隆及功能初步分析[J].麦类作物学报,2024,(11):1397 |
| 大麦气孔发育相关基因HvFAMA的克隆及功能初步分析 |
| Cloning and Preliminary Functional Analysis of HvFAMA Gene Related to Stomatal Development in Hordeum vulgare |
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| DOI： |
| 中文关键词: 大麦 气孔发育 HvFAMA基因 bHLH转录因子 基因克隆 |
| 英文关键词:Hordeum vulgare Stomatal development FAMA gene bHLH transcription factors Gene cloning |
| 基金项目:天津市科技计划项目（22ZXZYSN00020）；天津市科技计划项目（22ZYCGSN00020）；天津市大学生创新创业训练计划项目（202410061102）;天津市自然科学基金（22JCQNJC01470） |
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| 中文摘要: |
| 气孔是植物吸收CO2、散失水分的通道，叶片内气孔数目的多少、气孔的开闭程度都直接影响其蒸腾强度和光合作用的强弱。FAMA为碱性的螺旋-环-螺旋转录因子，是促进气孔保卫细胞分化和阻止其直接前体的增殖分裂所必需的，与气孔初始发育关系密切。为揭示其在禾本科作物气孔发育中的功能，本研究以大麦Morex为材料，克隆了大麦FAMA（HvFAMA）基因，并对该基因生物学特性及表达模式进行了分析。结果表明，HvFAMA基因编码区长为1 260 bp，由420个氨基酸构成，相对分子量为45 320.37 Da，理论等电点为5.14，属于亲水性不稳定蛋白；HvFAMA基因编码蛋白属于bHLH 家族成员，其三维结构含有螺旋-环-螺旋（HLH）结构；HvFAMA蛋白二级结构主要含有α-螺旋、延伸链、β-折叠和无规则卷曲，分别占36.04%、10.74%、2.86%和50.36%；HvFAMA蛋白的氨基酸序列含有44个磷酸化位点和65个糖基化位点；HvFAMA∶GFP融合蛋白亚细胞定位于细胞核。系统进化分析显示，大麦HvFAMA蛋白与小麦、二粒小麦、二穗短柄草和山羊草的FAMA亲缘关系相对较近。qRT-PCR结果显示，HvFAMA基因在大麦幼叶中的表达量达到峰值。综上所述，HvFAMA基因可能与大麦气孔初始发育有关。 |
| 英文摘要: |
| Stomata are the channels for plants to absorb CO2 and dissipate water. The number of stomata in leaves and the degree of opening and closing of stomata directly affect the intensity of transpiration and photosynthesis. FAMA is an alkaline helix-loop-helix transcription factor, which is necessary to promote the differentiation of stomatal guard cells and prevent the proliferation and division of their direct precursors. FAMA is closely related to initial development of stomata. In order to reveal its function in barley stomatal development, the barley variety Morex was used as material in this study to clone barley FAMA(HvFAMA) gene and its biological characteristics and expression pattern were analyzed. The results showed that the coding region of HvFAMA was 1 260 bp, which was composed of 420 amino acids. The relative molecular weight was 45 320.37 Da, and the theoretical isoelectric point was 5.14, which belonged to hydrophilic unstable protein. HvFAMA was a member of the bHLH family, and its three-dimensional structure contained a helix-loop-helix(HLH) structure; the secondary structure of HvFAMA protein mainly contained α-helix, extended strand, β-sheet and random coil, accounting for 36.04%, 10.74 %, 2.86% and 50.36%, respectively. The amino acid sequence of HvFAMA protein contained 44 phosphorylation sites and 65 glycosylation sites. HvFAMA:GFP fusion protein was localized in the nucleus. Phylogenetic analysis showed that the HvFAMA protein of barley was closely related to the FAMA of subspecies of Triticum aestivum, Triticum dicoccoides, Brachypodium distachyon, and Aegilops tauschii. The results of qRT-PCR showed that the expression level of HvFAMA gene in barley young leaves reached the peak. In summary, it is suggested that HvFAMA gene may be related to the initial development of stomata in Hordeum vulgare. |
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