毛竹PeGRF1基因的克隆及初步功能分析

Yongfeng Lou, X. Song, Huayu Sun, Z. Gao
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摘要

为探讨生长调节因子(GRF)在竹笋生长中的作用,采用RT-PCR方法从毛竹中分离到一个GRF基因。采用生物信息学方法进行基因序列分析,并利用qRT-PCR分析GRF基因在竹笋不同发育阶段的表达规律。同时,在拟南芥中进行了异位表达以验证该基因的功能。结果表明,从毛竹中分离到的基因长度为1 164 bp,编码387个氨基酸,命名为PeGRF1。蛋白序列分析表明,PeGRF1具有GRF家族完整的典型结构域(WRC和QLQ)。系统发育分析表明,PeGRF1与单子叶植物(如Oryza sativa)的GRFs聚类更接近,表明二者亲缘关系较近。PeGRF1主要在幼芽中表达。同时,竹笋快速生长期PeGRF1的表达量显著高于慢生长期,竹笋上部和中部PeGRF1的表达量显著高于基部。过表达PeGRF1可以提高转基因拟南芥的株高。综上所述,我们的研究结果表明PeGRF1参与了竹笋的发育,这为阐明GRF基因在竹子中的生物学功能提供了参考。
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Cloning and Primary Functional Analysis of PeGRF1 in Phyllostachys edulis
To explore the role of Growth regulating-factor (GRF) in the 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, and the expression pattern of the GRF gene in bamboo shoots at different development stages was analyzed by qRT-PCR. Simultaneously, ectopic expression in  Arabidopsis  was conducted to validate the gene function. The result 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 were 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, and the expression level of  PeGRF1  in the upper and middle of bamboo shoots was significantly higher than that in the base. Overexpression of  PeGRF1  could increase the plant height of transgenic  Arabidopsis . Taken together, our results demonstrated that  PeGRF1  was involved in development of bamboo shoots, which provided references for elucidating the biological functions of GRF genes in bamboo.
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