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以本实验室前期建立的落叶松(Larix kaempferi)转录组数据库为基础,利用RT-PCR从落叶松中克隆获得一个1 725 bp的生长素受体基因全长编码序列,命名为Lk TIR1。蛋白结构特性分析发现Lk TIR1编码574个氨基酸,蛋白分子质量为64 397Da,等电点为6.97,为亲水性蛋白。蛋白二级结构主要包括α-螺旋、β-折叠和环肽链。功能结构域分析显示该蛋白包含6个富含亮氨酸重复基序和1个F-box蛋白结构域,推测其属于F-box基因家族。多重序列比对和系统进化分析表明Lk TIR1所编码的氨基酸与裸子植物火炬松(Pinus taeda)亲缘关系最近。同时,通过构建植物表达载体,利用浸花法转化模式植物拟南芥的功能研究发现,过量表达落叶松Lk TIR1的转基因拟南芥叶片显著变大,叶片数目明显增多,这表明落叶松生长素受体基因Lk TIR1在植物的生长发育过程中起着重要的调节作用。
Based on the Larix kaempferi transcriptome database previously established in our laboratory, a full-length coding sequence of 1 725 bp auxin receptor gene was cloned from larch by RT-PCR and named Lk TIR1. Lk TIR1 encoded 574 amino acids. The molecular mass of the protein was 64 397 Da and the isoelectric point was 6.97. It was a hydrophilic protein. Protein secondary structure mainly includes α-helix, β-sheet and cyclopeptide chains. Functional domain analysis showed that the protein contains six leucine-rich repeat motifs and one F-box protein domain, suggesting that it belongs to the F-box gene family. Multiple sequence alignment and phylogenetic analysis indicated that the amino acids encoded by Lk TIR1 were the closest to Pinus taeda. At the same time, we found that transgenic Arabidopsis plants overexpressing Lk TIR1 significantly increased the number of leaves and significantly increased the number of leaves by constructing a plant expression vector. The results showed that larch auxin The receptor gene Lk TIR1 plays an important regulatory role in plant growth and development.