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分蘖角度影响植物群体结构、光合效率以及植株的形态建成,最终影响其产量及品质。小麦中关于分蘖角度的研究鲜有报道。前人研究显示,水稻OsTAC1正向调控分蘖角度的大小,玉米Zm TAC1与叶片角度呈正相关。本研究的目的是解析Ta TAC1的表达模式并初步了解该基因的分子遗传机制及其与分蘖角度的遗传关系。本研究以CN16、SM969、Lan2399、SHW-1为材料,利用同源克隆分离获得Ta TAC1,利用生物信息学软件对Ta TAC1进行序列特征分析,应用实时荧光定量PCR对其表达模式进行分析,并进行Ta TAC1蛋白的亚细胞定位分析。结果显示,Ta TAC1长度约1.1~1.2 kb,包括780 bp的完整开放阅读框和320~370 bp的3’-UTR。Ta TAC1的c DNA序列分为2类,其中CN16-2、SM969-2的第10号碱基发生突变,引起终止子提前,导致Ta TAC1表达受阻;Lan2399-2和SHW-1-2在109~115碱基位置有“CGCGCG”片段插入,导致蛋白质β-折叠片减少。表达分析表明,Ta TAC1在分蘖期的叶鞘、茎高效表达,分蘖节其次,叶与根表达量最低。相关分析表明该基因在分蘖节各时期表达量与分蘖角度呈显著正相关,Pearson相关系数为0.677,其他组织表达量与分蘖角度无显著相关性。亚细胞定位分析显示Ta TAC1定位于细胞膜。从上述结果可以推测Ta TAC1在mRNA水平上正向调控分蘖角度,且可能参与生长素极性运输过程从而改变分蘖角度大小。
The tillering angle affects plant community structure, photosynthetic efficiency and plant morphogenesis, ultimately affecting its yield and quality. Little research has been reported on the tillering angle in wheat. Previous studies showed that rice OsTAC1 positively regulated the tillering angle and the correlation between Zm TAC1 and leaf angle. The purpose of this study is to analyze the expression pattern of Ta TAC1 and to understand the molecular genetic mechanism of the gene and its genetic relationship with tiller angle. In this study, Ta TAC1 was isolated by homologous cloning using CN16, SM969, Lan2399 and SHW-1 as materials, Ta TAC1 was analyzed by bioinformatics software, and its expression pattern was analyzed by real-time fluorescence quantitative PCR Subcellular localization analysis of Ta TAC1 protein was performed. The results showed that Ta TAC1 was about 1.1-1.2 kb in length, including 780 bp complete open reading frame and 320-370 bp 3’-UTR. Ta TAC1 c DNA sequences are divided into two categories, including CN16-2, SM969-2 No. 10 base mutation, causing the terminator advance, resulting in Ta TAC1 expression blocked; Lan2399-2 and SHW-1-2 at 109 The insertion of a “CGCGCG” fragment at a ~ 115 base position resulted in a decrease of the protein β-sheet. Expression analysis showed that Ta TAC1 was highly expressed in leaf sheaths and stems at tillering stage, followed by tillers, followed by leaves and roots. Correlation analysis showed that there was a significant positive correlation between the expression level of tiller node and the tillering angle, the Pearson correlation coefficient was 0.677 and the expression level of other genes had no significant correlation with the tillering angle. Subcellular localization analysis showed that Ta TAC1 localized to the cell membrane. From the above results, it can be inferred that Ta TAC1 positively regulates tiller angle at the mRNA level, and may be involved in the polarity transport process of auxin to change the tiller angle.