《植物生理学报》 2018, 54(8): 1356-1364

小黑杨PnGRAS47基因克隆及氮素诱导下的表达模式分析

张翔, 刘野, 曲春浦, 宋长江, 崔帅康, 李桂英, 刘关君, 杨成君^*

东北林业大学林学院, 哈尔滨150040

通信作者：杨成君；E-mail: nxyycj@163.com

摘　要：

GRAS转录因子是一种植物特有且广泛分布的转录因子, 参与调控植物生长发育及在植物响应环境胁迫中发挥重要的功能。本研究通过克隆小黑杨(Populus simonii ×P. nigra) PnGRAS47基因序列, 分析其基因及蛋白序列的基本特征; 通过实时定量PCR技术检测在不同形态和不同浓度的氮素处理下小黑杨PnGRAS47基因的组织表达模式。研究结果显示: PnGRAS47基因全长1 528 bp, 编码452个氨基酸。PnGRAS47蛋白分子质量为50 812.19 Da, 理论等电点为5.41, 为不稳定蛋白, 不存在信号肽, 有2段跨膜区域。氨态氮(NH₄⁺-N)和硝态氮(NO₃ ⁻ -N)处理下, 小黑杨PnGRAS47基因呈现组织特异性表达。0.1和10 mmol·L^-1 NO₃⁻-N诱导了PnGRAS47基因在叶和根中的表达, 0.1和10 mmol·L^-1的NH₄⁺-N处理诱导了PnGRAS47基因在叶和茎中的表达, 但抑制了其在根中表达。因此, 小黑杨PnGRAS47基因受不同氮素的诱导, 在不同组织中存在差异, GRAS基因可能在小黑杨对氮素的吸收与利用中发挥作用。

关键词：小黑杨; PnGRAS47基因; 生物信息学分析; 氮素诱导; 表达模式

收稿：2018-03-26   修定：2018-08-03

资助：东北林业大学大学生创新训练计划项目(201610225016)和黑龙江省博士后科研启动金(LBH-Q15005)。

Cloning and expression analysis of PnGRAS47 gene induced by nitrogen in Populus simonii ×Populus nigra

ZHANG Xiang, LIU Ye, QU Chun-Pu, SONG Chang-Jiang, CUI Shuai-Kang, LI Gui-Ying, LIU Guan-Jun, YANG Cheng-Jun^*

College of Forestry, Northeast Forestry University, Harbin 150040, China

Corresponding author: YANG Cheng-Jun; E-mail: nxyycj@163.com

Abstract:

GRAS transcription factor is one of the plant-specific transcription factors, widely distributed in plants, involves in the regulation of plant growth and development, and plays an important role in the plant’s response to environmental stresses. In this study, we cloned the PnGRAS47 gene sequence from Populus simonii ×P. nigra and analyzed the basic characteristics of its gene and protein sequence. Expression pattern of PnGRAS47 gene was detected by quantitative real-time PCR under different morphological nitrogens and different nitrogen concentrations. The results show that the full length of PnGRAS47 gene is 1 528 bp and the gene encodes 452 amino acids. The molecular mass of PnGRAS47 is 50 812.19 Da and the pI value is 5.41. It is an unstable protein that has no signal peptide and two transmembrane domains. Under the treatments of ammonia nitrogen and nitrate nitrogen, PnGRAS47 gene showed tissue-specific expressions. 0.1 and 10 mmol·L^-1 NO₃ ⁻ -N induced the expression of PnGRAS47 gene in leaves and roots; 0.1 and 10 mmol·L^-1 NH₄⁺-N induced the expression of PnGRAS47 gene in leaves and stems, but inhibited that in roots. Therefore, the PnGRAS47 gene was induced by nitrogen and was differentially induced in various tissues. GRAS may play a role in the absorption and utilization of nitrogen in P. simonii ×P. nigra.

Key words: Populus simonii ×Populus nigra; PnGRAS47 gene; bioinformatic analysis; nitrogen induction; expressional pattern