毛竹PeGRF1基因克隆及初步功能分析

Cloning and Primary Functional Analysis of Pe GRF1 Gene in Phyllostachys edulis

为了探究生长调控因子(GRF)转录因子在竹笋-茎秆生长中的作用,本研究以毛竹(Phyllostachys edulis)为材料,通过RT-PCR方法进行GRF基因的克隆,应用生物信息学方法对其序列进行分析,利用RT-qPCR对其在毛竹竹笋-幼竹的生长过程中的表达模式进行分析,并通过在拟南芥中异位表达初步分析其功能。结果表明,从毛竹中克隆获得1个1 164 bp的GRF基因,命名PeGRF1,其编码387个氨基酸;蛋白序列分析表明PeGRF1具有完整的GRF特征结构域WRC和QLQ。进化分析显示,PeGRF1与水稻等单子叶植物的GRFs聚类在较近的分支,亲缘关系较近。在毛竹竹笋生长过程中,PeGRF1主要在幼嫩竹笋中表达,在快速生长阶段的表达量明显高于生长缓慢时期,在竹笋顶端及中部的表达量明显高于基部。过量表达PeGRF1能增加转基因拟南芥的株高。本研究结果可为阐明GRF基因在竹子的生物学功能提供了参考。

To explore the role of growth-regulating-factor(GRF) in regulating growth of bamboo shoots, one GRF gene was isolated from moso bamboo(Phyllostachys edulis) by RT-PCR. Bioinformatics methods were used for gene sequence analysis, the expression pattern of the GRF gene in bamboo shoots at different development stages was analyzed by RT-qPCR. Simultaneously, ectopic expression of the GRF gene in Arabidopsis was conducted to validate its function. The results showed that the isolated gene from moso bamboo was 1 164 bp, encoding 387 amino acids, which was named as PeGRF1. Protein sequence analysis showed that PeGRF1 had the complete typical domains(WRC and QLQ) of GRF family. The phylogenetic analysis demonstrated that PeGRF1 was clustered closer to the GRFs of monocotyledonous plants such as Oryza sativa, indicating they had close relationship. PeGRF1 expressed predominantly in young bamboo shoots. Meanwhile, the expression level of PeGRF1 was significantly higher during rapid growth stage than that during slow growth stage of bamboo shoots, which was significantly higher in the upper and middle of bamboo shoots than that in the base. Overexpression of PeGRF1 could increase the plant height of transgenic Arabidopsis. Taken together, The results of PeGRF1 could provide references for elucidating the biological functions of GRF genes in bamboo.