Ma Xiaoli, Liang Wenji, Gu Peihan, Huang Zhanjing
College of Life Science, Hebei Normal University, Shijiazhuang 050024, People's Republic of China.
College of Life Science, Hebei Normal University, Shijiazhuang 050024, People's Republic of China; College of Clinical Medicine, North China University of Science and Technology, Tangshan 063000, People's Republic of China.
Plant Physiol Biochem. 2016 Sep;106:129-40. doi: 10.1016/j.plaphy.2016.04.033. Epub 2016 Apr 29.
The expression profile chip of the wheat salt-tolerant mutant RH8706-49 was investigated under salt stress in our laboratory. Results revealed a novel gene induced by salt stress with unknown functions. The gene was named as TaZNF (Triticum aestivum predicted Dof zinc finger protein) because it contains the zf-Dof superfamily and was deposited in GenBank (accession no. KF307327). Further analysis showed that TaZNF significantly improved the salt-tolerance of transgenic Arabidopsis. Various physiological indices of the transgenic plant were improved compared with those of the control after salt stress. Non-invasive micro-test (NMT) detection showed that the root tip of transgenic Arabidopsis significantly expressed Na(+) excretion. TaZNF is mainly localized in the nucleus and exhibited transcriptional activity. Hence, this protein was considered a transcription factor. The TaZNF upstream promoter was then cloned and was found to contain three salts, one jasmonic acid methyl ester (MeJA), and several ABA-responsive elements. The GUS staining and quantitative results of different tissues in the full-length promoter in the transgenic plants showed that the promoter was not tissue specific. The promoter activity in the root, leaf, and flower was enhanced after induction by salt stress. Moreover, GUS staining and quantitative measurement of GUS activity showed that the promoter sequence contained the positive regulatory element of salt and MeJA after their respective elements were mutated in the full-length promoter. RNA-Seq result showed that 2727 genes were differentially expressed; most of these genes were involved in the metabolic pathway and biosynthesis of secondary metabolite pathway.
本实验室对小麦耐盐突变体RH8706 - 49在盐胁迫下的表达谱芯片进行了研究。结果揭示了一个受盐胁迫诱导的功能未知的新基因。该基因被命名为TaZNF（普通小麦预测的Dof锌指蛋白），因为它含有zf - Dof超家族，并已存入GenBank（登录号：KF307327）。进一步分析表明，TaZNF显著提高了转基因拟南芥的耐盐性。盐胁迫后，转基因植株的各项生理指标均优于对照。非损伤微测（NMT）检测表明，转基因拟南芥根尖显著表达Na(+)外排。TaZNF主要定位于细胞核并表现出转录活性。因此，该蛋白被认为是一种转录因子。随后克隆了TaZNF上游启动子，发现其含有三个盐响应元件、一个茉莉酸甲酯（MeJA）响应元件和几个脱落酸（ABA）响应元件。转基因植株中全长启动子不同组织的GUS染色和定量结果表明，该启动子无组织特异性。盐胁迫诱导后，根、叶和花中的启动子活性增强。此外，GUS染色和GUS活性定量测定表明，在全长启动子中各自元件突变后，启动子序列含有盐和MeJA的正调控元件。RNA - Seq结果显示有2727个基因差异表达；这些基因大多参与代谢途径和次生代谢物生物合成途径。
