马尾松Aux/IAA家族相关基因克隆及初步分析

Cloning and Premilinary Analysis of Aux/IAA Family Genes from Pinus massoniana

生长素信号转导途径在植物生长发育过程中起到至关重要的作用,Aux/IAA(auxin/indoleacetic acids)家族基因可以直接和生长素相连而发生作用。研究马尾松Aux/IAA家族相关基因,可为马尾松体内生长素信号转导途径提供理论支持。本研究克隆出3个Aux/IAA家族基因,分别命名为Pm IAA1、Pm IAA2和Pm IAA3。Pm IAA1基因,c DNA全长1 330 bp,包含711 bp完整的ORF,能够编码含有236个氨基酸的蛋白质,此蛋白相对分子质量为26 015.6 u,等电点为7.01;Pm IAA2基因,c DNA全长1 628 bp,包含909 bp完整的ORF,能够编码含有302个氨基酸的蛋白质,此蛋白相对分子质量为41 176.1 u,等电点为5.25;Pm IAA3基因,c DNA全长1 316 bp,包含759 bp完整的ORF,能够编码含有252个氨基酸的蛋白质,此蛋白相对分子质量为28 060.1 u,等电点为6.01。Pm IAA1蛋白与火距松IAA1蛋白同源性高达95%,Pm IAA2蛋白与火距松IAA2蛋白同源性高达97%,Pm IAA3蛋白与火距松IAA3蛋白同源性高达98%。Pm IAA1、Pm IAA2、Pm IAA3蛋白都含有Aux/IAA结构域,此结构域可以与ARFs(auxin response factors)结合而发生作用,这符合Aux/IAA家族的特点。利用实时定量技术得知,Pm IAA1、Pm IAA2和Pm IAA3在马尾松嫩根、嫩芽、嫩叶中都有表达。Pm IAA1在嫩叶中表达量最高、嫩根中次之、嫩茎中最低;Pm IAA2和Pm IAA3在嫩根中表达最高、嫩叶和嫩茎中次之。本研究可为生长素调控马尾松生长发育相关机理提供依据。

Auxin signal transduction pathways plays a crucial role in plant growth and development. Aux/IAA family genes can be directly connected to auxin and works. Studying Aux/IAA(auxin/indoleacetic acids) family genes can provide theoretical support for auxin signal transduction pathways from P. massoniana. In this study,three genes from Aux/IAA gene family were cloned,named Pm IAA1,Pm IAA2 and Pm IAA3. Pm IAA1,having1 330 bp in full length,contains an opening frame of 711 bp that encodes a protein of 236 amino acids with predicted relative molecular mass of 26 015.6 u and PI of 7.01; Pm IAA2,having 1 628 bp in full length,contains an opening frame of 909 bp that encodes a protein of 302 amino acids with predicted relative molecular mass of41 176.1 u and PI of 5.25; Pm IAA3,having 1 316 bp in full length,contains an opening frame of 759 bp that encodes a protein of 252 amino acids with predicted relative molecular mass of 28 060.1 u and PI of 6.01. The homology of Pm IAA1 protein from P. massoniana and IAA1 protein from P. taeda was as high as 95%,the homology of Pm IAA2 protein from P. massoniana and IAA2 protein from P. taeda was as high as 97%,and the homology of Pm IAA3 protein from P. massoniana and IAA3 protein from P. taeda was as high as 98%. Pm IAA1,Pm IAA2 and Pm IAA3 contain Aux/IAA domain,the domain may combine with ARFs(auxin response factors),which is consistent with the characteristics of the family of Aux/IAA. Using real-time quantitative techniques,we identified the amount of Pm IAA1,Pm IAA2 and Pm IAA3 expression,the amount of Pm IAA1 expression were the highest in leaf and the lowest in stem from P. massoniana,the amount of Pm IAA2 and Pm IAA3 expression were the highest in root and the lower in leaf and stem from P. massoniana. The study can help to discover how the auxin regulating growth and development of P. massoniana.