论文部分内容阅读
生长素响应因子(auxin response factor,ARF)在植物生长和发育中起到了非常关键的作用。本研究利用实验室前期转录组测序技术得到的差异表达基因Gm ARF,并应用PLACE数据库、Interpro、expasy数据库、SOPMA、Phyre、Protscale、MEGA5等工具对该基因的启动子元件、编码氨基酸的功能、亲水性、等电点、二级结构、三级结构、同源进化树及KEGG通路进行分析。并研究了对其在外源激素处理下的表达分析。结果表明:其前体序列具有很多与再生相关的响应元件,理论等电点为6.28,稳定系数为60.45,属于不稳定蛋白,Gm ARF编码的蛋白属于生长素蛋白,有一个B3超家族的保守结构域,一个生长素响应因子,以及一个PB1结构域,二级结构中23.33%为α螺旋结构,49.67%为不规则卷曲,8.59%为α转角,18.42%为延伸链,三级结构共有194个氢键,9个螺旋结构,32个转角结构,亲水性平均系数为-0.497,Gm ARF基因与苜蓿亲缘较近,其KEGG通路属于植物激素信号调节通路中色氨酸代谢途径,q RT-PCR结果表明在外源激素处理下,Gm ARF的表达量显著提高,说明Gm ARF基因作为正调控因子参与大豆再生过程。本研究的初步结果对今后进一步研究该基因的功能,提高大豆再生率,建立稳定的大豆再生体系提供了理论基础。
The auxin response factor (ARF) plays a crucial role in plant growth and development. In this study, differential expression gene Gm-ARF was obtained by pre-PCR transcriptome sequencing. The promoter elements, the function of amino acid encoding, Hydrophilicity, isoelectric point, secondary structure, tertiary structure, homologous phylogenetic tree and KEGG pathway. And analyzed its expression analysis under the treatment of exogenous hormones. The results showed that the precursor sequence had many regeneration-related response elements with a theoretical isoelectric point of 6.28 and a stability coefficient of 60.45, belonging to the unstable protein. The Gm-ARF-encoded protein belongs to the auxin protein and has a conserved B3 superfamily Domain, an auxin-responsive factor and a PB1 domain. 23.33% of the secondary structure is a-helical structure, 49.67% is irregular curly, 8.59% is α-turn and 18.42% is an extended chain. A hydrogen bond, nine helical structures and 32 corner structures. The average hydrophilicity coefficient was -0.497. The Gm ARF gene was closely related to alfalfa, and the KEGG pathway belonged to the tryptophan metabolic pathway in phytohormones signaling pathway. Q RT The results of PCR showed that Gm ARF expression increased significantly under exogenous hormone treatment, indicating that Gm ARF gene is involved in the soybean regeneration process as a positive regulator. The preliminary results of this study provide a theoretical basis for further study on the function of this gene, increase the soybean regeneration rate and establish a stable soybean regeneration system.