JIPB：山東農大、棗莊學院丨TaPUB1提升小麥耐鹽性機制（附NMT實驗體系）

期刊：J Integr Plant Biol

主題：TaPUB1提升小麥耐鹽性機制

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

影響因子：3.129

檢測指標：Na+、K+、H+流速

檢測部位：根部分生區

Na+、K+、H+流實驗處理方法：

7日齡小麥，200mM NaCl瞬時脅迫處理/200uM NaC處理7天

Na+、K+、H+流實驗測試液成份：

文獻無

推薦測試液成份：

• Na+/H+：0.5mM NaCl ，0.3mM MES，pH6.0

• K+/H+：0.1mM KCl ，0.3mM MES，pH6.0

通訊作者：山東農業大學王瑋、棗莊學院王文強

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（谷歌機翻）

U-box E3泛素連接酶在泛素/ 26S蛋白酶體機制和非生物應激反應中起重要作用。產生TaPUB1-過表達的小麥（Triticum aestivum L.）以評估其在耐鹽性中的功能。這些植物在幼苗和開花期間具有更多的鹽脅迫耐受性，而TaPUB1-RNAi介導的敲低轉基因小麥顯示出比野生型（WT）更多的鹽脅迫敏感性。

與WT相比，TaPUB1過表達上調與離子通道相關的基因的表達并增加凈根Na +流出，但降低凈K +流出和H +流入，從而維持低細胞溶質Na + / K +比率。然而，RNAi介導的敲低植物顯示出對鹽脅迫的相反反應。TaPUB1可誘導某些基因的表達，從而提高鹽脅迫下植物的抗氧化能力。TaPUB1還與TaMP（Triticumaestivum.α-甘露糖苷酶蛋白）相互作用，TaMPUB是一種在酵母和植物中的鹽響應中起重要作用的調節劑。

因此，在鹽脅迫下過表達TaPUB1的小麥中可以維持較低的細胞溶質Na + / K +比率和較好的抗氧化酶活性。因此，我們得出結論，U-box E3泛素連接酶TaPUB1正調節小麥的鹽脅迫耐受性。

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).

文章鏈接：

https://onlinelibrary.wiley.com/doi/abs/10.1111/jipb.12842