Functional identification and analysis of PtNRT2.7 gene from Populus trichocarpa.
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摘要: 本研究从毛果杨基因组中克隆一个NRT2家族的新基因PtNRT2.7。生物信息学分析显示,其编码的蛋白不存在信号肽及切割位点,属于非分泌蛋白;具有11个跨膜结构域,亲水区与输水区交替分布,主要分布于细胞膜,而且该基因的亚细胞定位分析显示该蛋白定位于细胞膜,综合以上结果可判断此蛋白为膜蛋白。对其在毛果杨的表达特征分析显示,PtNRT2.7基因主要在叶片中表达,其中成熟叶中的表达量最高;PtNRT2.7基因受硝酸盐诱导并在1 h时达到表达峰值;SA、JA、GA或IAA激素处理可诱导PtNRT2.7基因表达,而在Eth、ABA或NaCl处理下该基因的表达受抑制。利用农杆菌花序侵染法转化野生型拟南芥和突变体(nrt2.7),显示在KNO3浓度大于0.1 mmol/L时,PtNRT2.7过表达株系相比较拟南芥野生型和突变体株系促进了根的生长,而atnrt2.7突变体在KNO3浓度大于1 mmol/L的条件下抑制了根的生长,且PtNRT2.7过表达atnrt2.7突变体株系可恢复正常生长。研究结果表明,PtNRT2.7可以由硝酸盐和激素诱导并增强植物根系的生长,提高氮素的吸收利用。
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关键词:
- 毛果杨 /
- PtNRT2.7基因 /
- 序列分析 /
- 功能鉴定
Abstract: In present study, we successfully isolated a gene from NRT2 family named PtNRT2.7. Bioinformatic analyses showed that PtNRT2.7 had 11 transmembrane regions, and hydrophobic regions and hydrophilic regions are distributed alternately on cell membrane without signal peptide. Subcellular localization analysis showed that the PtNRT2.7-GFP fusion protein is distributed on cell membrane. According to above findings, it belongs to a membrane protein. The expression features of Populus trichocarpa showed that PtNRT2.7 expresses in leaves, with the highest expression in mature leaves; the highest expression appears at 1 h under the nitrate stress; under the stress of SA/JA/GA/IAA, the expression of PtNRT2.7 is up-regulated in both above- and underground parts; under the stress of Eth/ABA/NaCl, the expression of PtNRT2.7 is down-regulated. The expression vector was constructed and transformed into Arabidopsis thaliana wild type (Col-0) and atnrt2.7 mutant using floral dip method. The stress of different concentrations of KNO3 showed that, when the concentration of KNO3 is more than 0.1 mmol/L, the root enlongation of overexpression lines is longer than others; the mutant lines are inhibited when the concentration is more than 1 mmol/L but the complementary lines can remove the inhibition. The result showed that hormones and NO-3 can induce PtNRT2.7, enhance the root growth and promote the absorption of nitrogen.-
Keywords:
- Populus trichocarpa /
- PtNRT2.7 /
- sequence analysis /
- functional identification
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