二穗短柄草BdTOR基因的克隆及功能分析

在酵母、植物、动物和人类中,TOR是一个进化保守的重要调节因子,它通过整合营养和能量信号促进细胞增殖和生长。从二穗短柄草中克隆得到全长7 401 bp的Bd TOR基因,编码2 466个氨基酸残基,预测其具有HEAT重复域、FAT域、FRB域、激酶域、FATC域等保守结构域。实时定量PCR表明Bd TOR在二穗短柄草组织器官中相对表达量由高到低依次为幼穗幼苗根叶茎;Bd TOR及其激酶域的瞬时表达表明其定位于拟南芥原生质体的细胞核和细胞质,并在细胞质中区域化分布;Bd TOR具有促进根、叶生长,增加生物量,提高生长速率的功能;过表达Bd TOR可以回补雷帕霉素对Sc FKBP12过表达株系BP12的抑制;在雷帕霉素处理下Bd TOR能够与Sc FKBP12互作。

Cloning and Function Analysis of a BdTOR Gene from Brachypodium distachyon

TOR, a master regulator, is evolutionarily conserved among yeast, plant, animal and human. It can promote cell proliferation and growth through integrating nutrient and energy signaling. A 7 401 bp gene BdTOR was cloned from Brachypodium distachyon, which code a 2 466 aa protein, and had conservative domains like HEAT repeats, FAT domain, FRB domain, kinase domain, FATC domain. Real-Time PCR showed that the relative expression level of BdTOR in Brachypodium distachyon was young ear > young seedling > root > leave > stem. BdTOR and its kinase domain located in Arabidopsis protoplasts cell nuclear and cytoplasmic, and it was compartmentalization in cytoplasmic. BdTOR had the functions of promoting root and leaf growth, increasing plant biomass, and improving growth rate. Overexpression of BdTOR in BP12, a line over expressed ScFKBP12, could recover the inhibition by rapamycin. BdTOR can be interacted with ScFKBP12 under the treatment of rapamycin.