谷子MYB类转录因子SiMYB42提高转基因拟南芥低氮胁迫耐性

摘要

Myeloblastosis (MYB)类转录因子是高等植物中最大的转录因子家族之一,在植物发育及防御反应过程中发挥重要作用,还参与植物对干旱等非生物胁迫的响应.谷子(Setaria italica L.)起源于中国,具有抗旱、耐瘠薄的特性,是研究单子叶作物非生物胁迫抗性的理想材料.本研究对耐低氮胁迫谷子品种郑204经低氮处理后进行转录组分析,鉴定出一个在低氮胁迫条件下明显上调的MYB类转录因子SiMYB42.系统发育树结果表明, SiMYB42属于R2R3-MYB亚族,具有2个MYB保守域;表达模式分析显示,SiMYB42在低氮、高盐、干旱和ABA 胁迫条件下表达量显著上调;亚细胞定位、quantitative real-time PCR 及转录激活活性分析结果表明, SiMYB42蛋白定位于植物的细胞核和细胞膜中,主要在谷子的叶部或根部表达,具有转录激活活性;基因功能分析结果表明,在正常条件下,转SiMYB42基因拟南芥与野生型Columbia-0拟南芥(WT)无明显差异,但在低氮条件下,转SiMYB42基因拟南芥的主根长、根系表面积及鲜重均显著高于 WT,结果证明SiMYB42基因可以提高转基因植物对低氮胁迫的耐性;下游基因表达分析结果显示,在转 SiMYB42基因拟南芥中,参与植物氮素转运的硝酸盐转运基因NRT2.1、NRT2.4和NRT2.5的表达水平均高于WT,启动子分析结果显示NRT2.1、NRT2.4和NRT2.5基因启动子序列中均具有MYB结合位点.以上结果证明,SiMYB42可以通过调控下游硝酸盐转运体基因的表达提高植物在低氮条件下的耐性.本研究揭示了 SiMYB42基因在低氮胁迫反应途径中的作用,为进一步了解谷子低氮胁迫响应的调控网络奠定了基础.%Myeloblastosis (MYB) transcription factors are one of the largest families of transcription factors in higher plants. They play an important role in plant development, defense response processes, and non-biological stresses, i.e., drought stress.Foxtail millet(Setaria italica L.),originated in China,is resistant to drought and low nutrition stresses and has been regarded as an ideal material for studying abiotic stress resistance in monocotyledon. In this study, we ran a tran-scription profile analysis of zheng 204 under low-nitrogen conditions and identified a MYB-like transcription factor SiMYB42,which was up-regulated under low-nitrogen stress.Phylogenetic tree analysis showed that SiMYB42 belongs to R2R3-MYB subfamily and has two MYB conserved domains.Expression pattern analysis showed that SiMYB42 was sig-nificantly up-regulated under various stress conditions, including low-nitrogen stress, high salt, drought and ABA conditions. The results of subcellular localization, quantitative real-time PCR and transcriptional activation analysis indicated that SiMYB42 protein localizes to the nucleus and cell membrane of plant cells, mainly expressed in the leaf or root of foxtail millet, and has transcription activation activity. Functional analysis showed that there was no significant difference between transgenic SiMYB42 Arabidopsis and wild-type (WT) Arabidopsis under normal conditions; however, under low-nit-rogen condition,the root length,surface area and seedling fresh weight in transgenic SiMYB42 Arabidopsis,were signifi-cantly higher than their counterparts in WT.These results suggest that SiMYB42 transgenic plants exhibit higher tolerance to low-nitrogen stress.Expression levels of nitrate transporters genes NRT2.1,NRT2.4 and NRT2.5,which are the transcrip-tional targets of SiMYB42,were higher in transgenic SiMYB42 Arabidopsis plants than those in WT;the promoter regions of NRT2.1,NRT2.4 and NRT2.5 all have MYB binding sites.These results indicate that SiMYB42 might enhance foxtail millet tolerance to low-nitrogen condition through regulating the expression of nitrate transporter genes. This study reveals the possible functions of SiMYB42 in a low-nitrogen stress response pathway, and provides a foundation for further under-standing the entire regulation network of foxtail millet in response to low-nitrogen stress.