JIPB：山东农大、枣庄学院丨TaPUB1提升小麦耐盐性机制（附NMT实验体系）

期刊：J Integr Plant Biol

主题：TaPUB1提升小麦耐盐性机制

标题：The involvement of wheat (Triticum aestivum L.) U-box E3 ubiquitin ligase TaPUB1 in salt stress tolerance

影响因子：3.129

检测指标：Na+、K+、H+流速

检测部位：根部分生区

Na+、K+、H+流实验处理方法：

7日龄小麦，200mM NaCl瞬时胁迫处理/200uM NaC处理7天

Na+、K+、H+流实验测试液成份：

文献无

推荐测试液成份：

• Na+/H+：0.5mM NaCl ，0.3mM MES，pH6.0

• K+/H+：0.1mM KCl ，0.3mM MES，pH6.0

通讯作者：山东农业大学王玮、枣庄学院王文强

英文摘要

U‐box E3 ubiquitin ligases play important roles in the ubiquitin/26S proteasome machinery and in abiotic stress responses. TaPUB1‐overexpressing wheat (Triticum aestivum L.) were generated to evaluate its function in salt tolerance. These plants were more salt stress tolerance during seedling and flowering stages, whereas the TaPUB1‐RNAi‐mediated knock‐down transgenic wheat showed more salt stress sensitivity than the wild type (WT).

TaPUB1 overexpression up‐regulated the expression of genes related to ion channels and increased the net root Na+ efflux, but decreased the net K+ efflux and H+ influx, thereby maintaining a low cytosolic Na+/K+ ratio, compared with the WT. However, RNAi‐mediated knock‐down plants showed the opposite response to salt stress. TaPUB1 could induce the expression of some genes that improved the antioxidant capacity of plants under salt stress. TaPUB1 also interacted with TaMP (Triticum aestivum.α‐mannosidase protein), a regulator playing an important role in salt response in yeast and in plants.

Thus, low cytosolic Na+/K+ ratios and better antioxidant enzyme activities could be maintained in wheat with overexpression of TaPUB1 under salt stress. Therefore, we conclude that the U‐box E3 ubiquitin ligase TaPUB1 positively regulates salt stress tolerance in wheat.

 中文摘要（谷歌机翻）

U-box E3泛素连接酶在泛素/ 26S蛋白酶体机制和非生物应激反应中起重要作用。产生TaPUB1-过表达的小麦（Triticum aestivum L.）以评估其在耐盐性中的功能。这些植物在幼苗和开花期间具有更多的盐胁迫耐受性，而TaPUB1-RNAi介导的敲低转基因小麦显示出比野生型（WT）更多的盐胁迫敏感性。

与WT相比，TaPUB1过表达上调与离子通道相关的基因的表达并增加净根Na +流出，但降低净K +流出和H +流入，从而维持低细胞溶质Na + / K +比率。然而，RNAi介导的敲低植物显示出对盐胁迫的相反反应。TaPUB1可诱导某些基因的表达，从而提高盐胁迫下植物的抗氧化能力。TaPUB1还与TaMP（Triticumaestivum.α-甘露糖苷酶蛋白）相互作用，TaMPUB是一种在酵母和植物中的盐响应中起重要作用的调节剂。

因此，在盐胁迫下过表达TaPUB1的小麦中可以维持较低的细胞溶质Na + / K +比率和较好的抗氧化酶活性。因此，我们得出结论，U-box E3泛素连接酶TaPUB1正调节小麦的盐胁迫耐受性。

Figure 6. The fluxes of Na+, K+ and H+ in response to 200 mM NaCl shock

(A, C and E) The transient kinetics of Na+, K+ and H+ fluxes in roots of transgenic and WT plants before and after salt shock (Before salt shock, steady fluxes for 2 minutes). (B, D, and F) The mean net Na+, K+, and H+ fluxes in roots of transgenic and WT plants during a short period of salt shock. The data were presented as the mean ±SD of three independent experiments. Asterisks above each column indicate statistical differences to the WT plants (*P<0.05; **P<0.01).