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).

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

期刊：BMC Plant Biology
主题：BjHMA4R通过结合胞质Cd²⁺促植物耐镉
标题：A repeat region from the Brassica juncea HMA4 gene BjHMA4R is specifically involved in Cd²⁺ binding in the cytosol under low heavy metal concentrations
影响因子：3.930
检测指标： Cd²⁺流速
检测部位：大肠杆菌细胞，酵母细胞
Cd²⁺流实验处理方法：
大肠杆菌，酵母细胞，30μMCdCl2胁迫

Cd²⁺流实验测试液成份：
文献无
推荐测试液成份：
0.03 mM CdCl2, 0.3 mM MES, 10 mM glucose, pH 6.0
通讯作者：中国科学院大学生命科学学院柴团耀、梁爽

英文摘要

HMA4 transporters are involved in the transport and binding of divalent heavy metals (Cd, Zn, Pb [lead] and Co [cobalt]). In general, as efflux pumps, HMA4 transporters can increase the heavy metal tolerance of yeast and Escherichia coli. Additional research has shown that the C-terminus of HMA4 contains a heavy metal-binding domain and that heterologous expression of a portion of peptides from this C-terminal domain in yeast provides a high level of Cd tolerance and Cd hyperaccumulation.

We cloned BjHMA4 from Brassica juncea, and quantitative real-time PCR analysis revealed that BjHMA4 was upregulated by Zn and Cd in the roots, stems and leaves. Overexpression of BjHMA4 dramatically affects Zn/Cd distribution in rice and wheat seedlings. Interestingly, BjHMA4 contains a repeat region named BjHMA4R within the C-terminal region; this repeat region is not far from the last transmembrane domain. We further characterized the detailed function of BjHMA4R via yeast and E. coli experiments. Notably, BjHMA4R greatly and specifically improved Cd tolerance, and BjHMA4R transformants both grew on solid media that contained 500 μM CdCl2 and presented improved Cd accumulation (approximately twice that of wild-type [WT] strains). Additionally, visualization via fluorescence microscopy indicated that BjHMA4R clearly localizes in the cytosol of yeast. Overall, these findings suggest that BjHMA4R specifically improves Cd tolerance and Cd accumulation in yeast by specifically binding Cd2+ in the cytosol under low heavy metal concentrations. Moreover, similar results in E. coli experiments corroborate this postulation.

BjHMA4R can specifically bind Cd2+ in the cytosol, thereby substantially and specifically improving Cd tolerance and accumulation under low heavy metal concentrations.

中文摘要（谷歌机翻）

HMA4转运蛋白参与二价重金属（Cd，Zn，Pb [铅]和Co [钴]）的转运和结合。通常，作为外排泵，HMA4转运蛋白可以增加酵母和大肠杆菌的重金属耐受性。另外的研究表明，HMA4的C末端含有重金属结合结构域，并且酵母中来自该C末端结构域的一部分肽的异源表达提供了高水平的Cd耐受性和Cd超积累。

我们从芥菜中克隆了BjHMA4，定量实时PCR分析表明BjHMA4在根，茎和叶中被Zn和Cd上调。BjHMA4的过表达显着影响水稻和小麦幼苗的Zn / Cd分布。有趣的是，BjHMA4在C末端区域内含有一个名为BjHMA4R的重复区域;这个重复区域距离最后一个跨膜结构域不远。我们通过酵母和大肠杆菌实验进一步表征了BjHMA4R的详细功能。值得注意的是，BjHMA4R极大地且特异性地改善了Cd耐受性，并且BjHMA4R转化体均在含有500μMCdCl2的固体培养基上生长并且呈现出改善的Cd积累（大约是野生型[WT]菌株的两倍）。另外，通过荧光显微镜观察表明BjHMA4R清楚地定位于酵母的胞质溶胶中。总体而言，这些发现表明BjHMA4R通过在低重金属浓度下特异性结合胞质溶胶中的Cd2+，特异性地改善了酵母中的Cd耐受性和Cd积累。此外，大肠杆菌实验中的类似结果证实了这种假设。

BjHMA4R可特异性结合胞质溶胶中的Cd2+，从而在低重金属浓度下基本上和特异性地改善Cd耐受性和积累。

Fig. 12. Detection of BjHMA4R activity by Cd²⁺ flux measurements. Transgenic BjHMA4R and control cells were exposed to 30 μM Cd²⁺ and measurements were taken for 600 s, using a vibrating probe, after the flux became ready. a yeast cells, b E. coli cells. The data are expressed as the mean ± SE of three replicates; * and ** indicate significant levels at 5 and 1% (evaluated by Student’s t test), respectively

文章链接：https://bmcplantbiol.biomedcentral.com/articles/10.1186/s12870-019-1674-5

期刊：Environmental and Experimental Botany（EEB）
主题：MsPIP2; 2提升转基因拟南芥耐盐性
标题：MsPIP2; 2, a novel aquaporin gene from Medicago sativa, confers salt tolerance in transgenic Arabidopsis
影响因子：3.712
检测指标： Na+、K+流速
检测部位：伸长区，距离根尖550-600μm
Na+、K+流实验处理方法：
7日龄拟南芥，150mM NaCl处理24小时

Na+、K+流实验测试液成份：
文献无
推荐测试液成份：
Na+：0.5mMNaCl ，0.3mM MES，pH6.0
K+：0.1mMKCl ，0.3mM MES，pH6.0
通讯作者：西北农林科技大学杨培志

英文摘要

Aquaporins (AQPs) are channel proteins that facilitate water transport across cell membranes and play important roles in many biological processes. However, most AQP functions are still poorly understood in the plant kingdom. Here, MsPIP2;2 was isolated and identified from alfalfa (Medicago sativa). MsPIP2;2 was localized to the plasma membrane, and its expression was induced by salt and abscisic acid (ABA) treatment. Overexpression of MsPIP2;2 in Arabidopsis increased the seed germination rate, seedling root length, survival rate, proline content and antioxidant defence activity and decreased cell membrane damage and reactive oxygen species (ROS) accumulation compared to those in WT under salt stress.

The salt tolerance of MsPIP2;2 was affected by Ca2+ and pH in transgenic Arabidopsis plants. MsPIP2;2-overexpressing plants maintained a better K+/Na+ ratio and higher Ca2+ content under salt stress. The higher K+/Na+ maintenance in transgenic plants was mainly achieved by increasing Na+ efflux and K+ retention in roots via regulating the expression of the related ion channel genes. Stress-responsive genes, including P5CS1, RD29A, DREB2 and KIN2, were upregulated in transgenic plants under salt stress.

These results suggest that MsPIP2;2 confers salt tolerance by regulating antioxidant defence system-mediated ROS scavenging, K/Na ion homeostasis and stress-responsive gene expression in plants.

中文摘要（谷歌机翻）

水通道蛋白（AQPs）是促进水跨细胞膜转运的通道蛋白，在许多生物过程中起重要作用。然而，大多数AQP功能在植物界仍然知之甚少。在此，从紫花苜蓿（Medicago sativa）中分离并鉴定了MsPIP2; 2。MsPIP2; 2定位于质膜，其表达由盐和脱落酸（ABA）处理诱导。与盐胁迫下WT相比，拟南芥中MsPIP2; 2的过量表达提高了种子萌发率，幼苗根长，存活率，脯氨酸含量和抗氧化防御活性，降低了细胞膜损伤和活性氧（ROS）积累。

MsPIP2; 2的耐盐性受转基因拟南芥植物中Ca2+和pH的影响。MsPIP2; 2过表达植物在盐胁迫下保持更好的K+ / Na+比率和更高的Ca2+含量。转基因植物中较高的K+ / Na+维持率主要通过调节相关离子通道基因的表达来增加根中的Na+流出和K+保留来实现。应激反应基因，包括P5CS1，RD29A，DREB2和KIN2，在盐胁迫下在转基因植物中上调。

这些结果表明，MsPIP2; 2通过调节抗氧化防御系统介导的ROS清除，K / Na离子稳态和植物中的应激反应基因表达来赋予耐盐性。

Fig. 9. The net flux of Na⁺ and K⁺ at the root tip elongation zones of 7-day-old Arabidopsis seedlings. (A) The net Na⁺ flux under normal conditions (0 mM NaCl). (B) The net Na⁺ flux after 150 mM NaCl stress. (C) The mean Na⁺ flux. (D) The net K⁺ flux under normal conditions (0 mM NaCl). (E) The net K⁺ flux after 150 mM NaCl stress. (F) The mean K⁺ flux. Bars represent the mean ± SE (n = 3). Asterisks and double asterisks above the bars indicate significant differences between the transgenic lines and the WT under the same growth conditions: *, P < 0.05; **, P < 0.01.

文章链接：https://www.sciencedirect.com/science/article/abs/pii/S0098847219300036

期刊：plant science

主题：氮素利用效率与镉耐受性的平衡

标题：Balance between nitrogen use efficiency and cadmium tolerance in Brassica napus and Arabidopsis thaliana

影响因子：3.785

检测指标： H⁺、Cd²⁺、NO₃^-流速

检测部位：根部液泡

H⁺、Cd²⁺、NO₃^-流实验处理方法：

4周龄拟南芥，20 μM CdCl2处理3天
21日龄油菜，100 μM CdCl2处理7天

H⁺、Cd²⁺、NO₃^-流实验测试液成份：

文献无

推荐测试液成份：

拟南芥根部液泡：0.002mM CdCl2，0.1 mM KNO3 , 600 mM mannitol, 0.05mM MES, pH 7.2

油菜根部液泡：0.01mM CdCl2，0.1 mM KNO3 , 600 mM mannitol, 0.05mM MES, pH 7.2

作者：湖南农业大学张振华、廖琼

英文摘要

The transmembrane transport of NO3− andCd2+ into plant cell vacuoles relies on the energy from their tonoplast protonpumps, V-ATPase and V-PPase. If the activity of these pumps is reduced, itresults in less NO3− and Cd2+ being transported into the vacuoles, whichcontributes to better nitrogen use efficiency (NUE) and lower Cd2+ tolerance inplants.

The physiological mechanisms that regulatethe balance between NUE and Cd2+ tolerance remain unknown. In our study, twoBrassica napus genotypes with differential NUEs, xiangyou 15 and 814, andAtclca-2 mutant and AtCAX4 over-expression line (AtCAX4-OE) of Arabidopsisthaliana, were used to investigate Cd2+ stress responses.

We found that the Brassica napus genotype,with higher NUE, was more sensitive to Cd2+ stress. The AtCAX4-OE mutant, withhigher Cd2+ vacuolar sequestration capacity (VSC), limited NO3− sequestrationinto root vacuoles and promoted NUE. Atclca-2 mutants, with decreased NO3− VSC,enhanced Cd2+ sequestration into root vacuoles and conferred greater Cd2+tolerance than the WT. This may be due to the competition between Cd2+ andNO3−in the vacuoles for the energy provided by V-ATPase and V-PPase. Regulating thebalance between Cd2+ and NO3− vacuolar accumulation by inhibiting the activityof CLCa transporter and increasing the activity of CAX4 transporter willsimultaneously enhance both the NUE and Cd2+ tolerance of Brassica napus,essential for improving its Cd2+ phytoremediation potential.

中文摘要（谷歌机翻）

NO3-和Cd2+跨膜转运到植物细胞液泡中依赖于其液泡膜质子泵，V-ATP酶和V-PPase的能量。如果这些泵的活性降低，则导致较少的NO3-和Cd2+被输送到液泡中，这有助于提高植物的氮利用效率（NUE）和降低Cd2+耐受性。

调节NUE和Cd2+耐受性之间平衡的生理机制尚不清楚。在我们的研究中，使用具有差异NUE，香优15和814的两种甘蓝型油菜基因型和拟南芥的Atclca-2突变体和AtCAX4过表达系（AtCAX4-OE）来研究Cd2+应激反应。

我们发现甘蓝型油菜基因型具有较高的NUE，对Cd2+胁迫更敏感。具有较高Cd2+液泡隔离能力（VSC）的AtCAX4-OE突变体限制NO3-隔离到根空泡中并促进NUE。具有降低的NO3-VSC的Atclca-2突变体增强了Cd2+隔离到根空泡中并且赋予比WT更大的Cd2+耐受性。这可能是由于V-ATP酶和V-PPase提供的能量在液泡中Cd2+和NO3-之间的竞争。通过抑制CLCa转运蛋白的活性和增加CAX4转运蛋白的活性来调节Cd2+和NO3-液泡积累之间的平衡将同时增强甘蓝型油菜的NUE和Cd2+耐受性，这对于改善其Cd2+植物修复潜力是必需的。

Fig. 2. The Arabidopsis thaliana vha-a2, vha-a3, and avp1 mutants were more sensitive to Cd2+ stress than the wild-type Col-0. (A) The phenotype and (B) the chlorophyll loss, in relation to the controls, of Col-0 and the vha-a2, vha-a3, and avp1 mutants grown for 4 weeks and then exposed to 20 μM CdCl2 for 3 days. Mean rates of (C) H+ fluxes and (D) Cd2+ fluxes during the first 160 s of measurements within the root vacuoles.

文章链接：https://www.sciencedirect.com/science/article/pii/S0168945218314754?via%3Dihub#!