转自中关村旭月非损伤微测技术产业联盟

• 期刊：Ecotoxicology and Environmental Safety
• 主题：镉胁迫下Na+/H+逆向转运蛋白（NHX1）转基因浮萍中镉的积累减少
• 标题：Declined cadmium accumulation in Na⁺/H⁺ antiporter (NHX1) transgenic duckweed under cadmium stress
• 影响因子：4.527
• 检测指标：Cd²⁺、H⁺流速
• 检测部位：浮萍距离根尖100μm
• Cd²⁺、H⁺流速流实验处理方法：浮萍在50μM CdCl2和50μM CdC+0.5mM阿米洛利分别处理2分钟，20分钟和2小时
• Cd²⁺、H⁺流速流实验测试液成份：0.05mM CdCl2, 0.1mM CaCl2, 0.1mM KCl, 0.3mM MES, pH 5.5
• 通讯作者：天津师范大学生命科学学院孙金生、杨琳

英文摘要

Cadmium (Cd) is a serious threat to plants health. Though some genes have been reported to get involved in the regulation of tolerance to Cd, the mechanisms underlying this process are not fully understood. Na⁺/H⁺ antiporter (NHX1) plays an important role in Na⁺/H⁺ trafficking. The salt and cadmium stress tolerance were found to be enhanced by NHX1 in duckweed according to our previous study, however, its function in Cd²⁺ flux under Cd stress has not been studied.

Here we explored the Cd²⁺ flux in wild type (WT) and NHX1 transgenic duckweed (NHX1) under Cd stress. We found that the Cd²⁺ influx in NHX1 duckweed was significantly declined, followed by an increased Cd²⁺ efflux after 20 min treatment of Cd, which resulted a less accumulation of Cd in NHX1.

Reversely, inhibition of NHX1 by amiloride treatment, enhanced Cd²⁺ influx in NHX1 duckweed, subsequently delayed Cd²⁺ efflux in both genotypes of duckweed under Cd²⁺ shock. H⁺ efflux in NHX1 duckweed was lower compare with that in WT with 20 min Cd²⁺ shock. NHX1 also increased the pH value with Cd²⁺ stress in the transgenic rhizoid. These finding suggested a new function of NHX1 in regulation of Cd²⁺ and H⁺ flow during short-term Cd²⁺ shock.

中文摘要（谷歌机翻）

镉（Cd）是对植物健康的严重威胁。虽然据报道一些基因参与调节对Cd的耐受性，但这一过程背后的机制尚不完全清楚。 Na⁺/H⁺逆向转运蛋白（NHX1）在Na⁺/H⁺运输中起重要作用。根据我们之前的研究，浮萍中的NHX1对盐和镉胁迫耐受性有增强作用，但其在Cd胁迫下对Cd²⁺ 通量的作用尚未得到研究。

在这里我们探讨了Cd胁迫下野生型（WT）和NHX1转基因浮萍（NHX1）中的Cd²⁺ 通量。我们发现NHX1浮萍中的Cd²⁺ 流入量显着下降，随后Cd处理20min后Cd²⁺ 流出量增加，导致NHX1中Cd的积累减少。

相反，阿米洛利处理对NHX1的抑制作用增强了NHX1浮萍中Cd2+ 的流入，延缓了Cd²⁺ 冲击下浮萍基因型中Cd²⁺ 的流出。 NHX1浮萍的H⁺流出量与WT相比，20min Cd²⁺ 冲击的H⁺流出量较低。 NHX1也增加了转基因根状茎中Cd²⁺ 胁迫的pH值。这些发现表明NHX1在短期Cd²⁺ 休克期间调节Cd²⁺ 和H⁺流动的新功能。

结果表明：在5分钟的镉处理后，如图a所示，在NHX1浮萍和WT浮萍中均检测到Cd²⁺吸收。有趣的是，NHX1浮萍中的Cd2+吸收量明显低于WT中的Cd²⁺吸收量。在5min处理镉时，NHX1浮萍中积累的镉含量较少。 阿米洛利是NHX1的抑制剂，用于研究瞬态Cd2+通量，在CdCl2（50μM）处理下NHX1活性下降。如图b所示，NHX1浮萍中的Cd²⁺吸收显着增加，与用镉和没有阿米洛利处理的WT中的Cd²⁺流入大致相同。该结果显示NHX1在短时间镉中在Cd²⁺流入中的作用。

文章链接：https://www.sciencedirect.com/science/article/pii/S0147651319307183?via%3Dihub

转自中关村旭月非损伤微测技术产业联盟

• 期刊：international journal of molecular sciences
• 主题：杨树ABC转运蛋白的异位表达赋予拟南芥Cd耐受性
• 标题：Ectopic Expression of Poplar ABC Transporter PtoABCG36 Confers Cd Tolerance in Arabidopsis thaliana
• 影响因子：4.183
• 检测指标： Cd²⁺流速
• 检测部位：拟南芥距离根尖800μm
• Cd²⁺流实验处理方法：15日龄幼苗，50μM CdCl2处理24小时
• Cd²⁺流实验测试液成份：0.1 mM KCl, 0.1 mM CaCl2, 0.05 mM CdCl2, 0.3 mM 2-(N-morpholino)ethane sulfonic acid, pH 5.8
• 通讯作者：西南大学王邦俊

英文摘要

Cadmium (Cd) is one of the most toxic heavy metals for plant growth in soil. ATP-binding cassette (ABC) transporters play important roles in biotic and abiotic stresses. However, few ABC transporters have been characterized in poplar.

In this study, we isolated an ABC transporter gene PtoABCG36 from Populus tomentosa. The PtoABCG36 transcript can be detected in leaves, stems and roots, and the expression in the root was 3.8 and 2 times that in stems and leaves, respectively. The PtoABCG36 expression was induced and peaked at 12 h after exposure to Cd stress. Transient expression of PtoABCG36 in tobacco showed that PtoABCG36 is localized at the plasma membrane. When overexpressed in yeast and Arabidopsis, PtoABCG36 could decrease Cd accumulation and confer higher Cd tolerance in transgenic lines than in wild-type (WT) lines. Net Cd²⁺ efflux measurements showed a decreasing Cd uptake in transgenic Arabidopsis roots than WT.

These results demonstrated that PtoABCG36 functions as a cadmium extrusion pump participating in enhancing tolerance to Cd through decreasing Cd content in plants, which provides a promising way for making heavy metal tolerant poplar by manipulating ABC transporters in cadmium polluted areas.

中文摘要（谷歌机翻）

镉（Cd）是土壤中植物生长最毒的重金属之一。 ATP结合盒（ABC）转运蛋白在生物和非生物胁迫中起重要作用。然而，很少有ABC转运蛋白在杨树中具有特征。

在这项研究中，我们从毛白杨（Populus tomentosa）中分离出一个ABC转运蛋白基因PtoABCG36。可以在叶，茎和根中检测到PtoABCG36转录物，并且根中的表达分别是茎和叶中的3.8和2倍。诱导PtoABCG36表达并在暴露于Cd胁迫后12小时达到峰值。 PtoABCG36在烟草中的瞬时表达显示PtoABCG36位于质膜上。当在酵母和拟南芥中过表达时，PtoABCG36可以减少Cd积累并赋予转基因品系中比野生型（WT）品系更高的Cd耐受性。净Cd²⁺外排测量显示转基因拟南芥根中的Cd摄取减少比WT。

这些结果表明，PtoABCG36作为镉挤压泵发挥作用，通过降低植物中的Cd含量来增强对Cd的耐受性，这为通过操纵镉污染区域的ABC转运蛋白制备重金属耐受性杨树提供了一种有前景的方法。

结果表明：在50 M CdCl2存在下，OX-1，OX-2和OX-3系的Cd²⁺净吸收量低于WT植物（分别为62.39％，54.50％和53.30％）。 相比之下，突变体abcg36具有比WT植物更高的Cd²⁺净吸收量。

文章链接：https://www.mdpi.com/1422-0067/20/13/3293

转自中关村旭月非损伤微测技术产业联盟

• 期刊：Ecotoxicology and Environmental Safety
• 主题：Ca²⁺流可作为防污涂料性能评价指标
• 标题：Synthesis and antifouling evaluation of indole derivatives
• 影响因子：4.527
• 检测指标： Ca²⁺流速
• 检测部位：藻细胞
• Ca²⁺流实验处理方法：亚心形扁藻，吲哚衍生物瞬时处理
• Ca²⁺流实验测试液成份：0.1mM KCl, 0.1mM CaCl2, 0.1mM MgCl2, 0.5mM NaCl, 0.3mM MES,0.2mM Na2SO4 and 0.1% sucrose, pH 7.8
• 通讯作者：中国海洋大学李霞、冯康

英文摘要

Indole derivatives derived from the secondary metabolites of marine organisms possess the excellent antifouling property to inhibit the biofouling. These compounds and their analogues are simple in structure and have been proven to have low toxicity and bioaccumulation. Therefore, the active indole antifoulants are expected to replace the potentially toxic antifoulants which are widely used in current antifouling coatings.

Seven indole derivatives were synthesized via the Friedel-Crafts alkylation reaction and were characterized by IR spectra, 1H NMR, 13C NMR and elemental analysis. Inhibition experiments against marine algae and bacteria were conducted, and the partial inhibition rates of algae and bacteria were more than 90%. This outcome indicates that indole derivatives possess excellent properties suitable for use as targeting anti-fouling compound for algae and bacteria. Non-invasive Micro-test Technology (NMT) reveals that the Ca²⁺ efflux of Platymonas subcordiformis dramatically increased in the presence of indole derivatives, which is inferred to be the molecular mechanism for inhibiting the growth of marine algae.

The antifouling coatings containing indole derivatives were prepared and subjected to an antifouling test in a marine environment, and the results show that N-(1-H-5-bromo-indole-3-ylmethyl) benzamide and N-(1-H-2-phenyl-indole-3-ylmethyl) benzamide possess better antifouling performance compared to copper pyrithione (CuPT). According to these results, indole derivatives in this study might become novel and promising antifoulants.

中文摘要（谷歌机翻）

来自海洋生物次生代谢产物的吲哚衍生物具有优异的防污性能，可抑制生物污损。这些化合物及其类似物结构简单，已被证明具有低毒性和生物累积性。因此，有效的吲哚防污剂有望取代目前防污涂料中广泛使用的潜在有毒的防污剂。

通过Friedel-Crafts烷基化反应合成了7种吲哚衍生物，并通过IR光谱，1H NMR，13C NMR和元素分析进行了表征。对海藻和细菌进行了抑制实验，藻类和细菌的部分抑制率超过90％。该结果表明吲哚衍生物具有适合用作藻类和细菌的靶向防污化合物的优异性质。非侵入性微测试技术（NMT）揭示了在吲哚衍生物存在下，亚心形扁藻的Ca²⁺流出显着增加，这被推断为抑制海藻生长的分子机制。

制备含有吲哚衍生物的防污涂料并在海洋环境中进行防污试验，结果表明N-（1-H-5-溴 - 吲哚-3-基甲基）苯甲酰胺和N-（1-H-）与吡啶硫酮铜（CuPT）相比，2-苯基 - 吲哚-3-基甲基）苯甲酰胺具有更好的防污性能。根据这些结果，本研究中的吲哚衍生物可能成为新颖且有前景的防污剂。

结果表明：在吲哚衍生物存在下，Ca²⁺外排明显增加。在吲哚衍生物的影响下，藻细胞中Ca²⁺外排的峰值几乎是对照峰值的三倍。从分子机制推断，吲哚衍生物可以作为跨膜转运的抑制剂，并触发藻类细胞Ca²⁺外排，从而减少细胞内Ca²⁺的浓度。

文章链接：https://www.sciencedirect.com/science/article/pii/S0147651319307535?via%3Dihub

转自中关村旭月非损伤微测技术产业联盟

• 期刊：Aquatic toxicology
• 主题：盐污染淡水对昆虫渗透调节和气管鳃功能的影响（Na⁺流）
• 标题：Impact of salt-contaminated freshwater on osmoregulation and tracheal gill function in nymphs of the mayfly Hexagenia rigida
• 影响因子：3.884
• 检测指标： Na⁺流速
• 检测部位：气管鳃丝中心轴
• Na⁺流实验处理方法： 长约20mm的蜉蝣在FW（Freshwater）对照条件或SCW（Salt-contaminated water）处理7天
• Na⁺流实验测试液成份： 0.5mM NaCl，206.5mM N-甲基-D-葡糖胺（NMDG）
• 通讯作者：约克大学Andrew Donini、Fargol Nowghani

英文摘要

The impact of freshwater (FW) salinization on osmoregulation as well as tracheal gill morphology and function was examined in nymphs of the mayfly Hexagenia rigida following exposure to salt contaminated water (SCW, 7.25 g/l NaCl) for a 7-day period.

Ionoregulatory homeostasis was perturbed in SCW exposed H. rigida nymphs as indicated by increased hemolymph Na⁺, K⁺ and Cl⁻ levels as well as hemolymph pH and water content. Despite this, SCW did not alter gill Na⁺-K⁺-ATPase (NKA) or V-type H⁺-ATPase (VA) activity. In addition, NKA and VA immunolocalization in gill ionocytes did not show alterations in enzyme location or changes in ionocyte abundance.

The latter observation was confirmed using scanning electron microscopy (SEM) to examine exposed tracheal gill ionocyte numbers. Ionocyte surface morphometrics also revealed that SCW did not change individual ionocyte surface area or ionocyte fractional surface area. Nevertheless, analysis of Na⁺ movement across the tracheal gill of mayfly nymphs using scanning ion-selective electrode technique indicated that FW nymphs acquired Na⁺ from surrounding water, while tracheal gills of SCW nymphs had the capacity to secrete Na⁺.

Because Na⁺ secretion across the gill of SCW-exposed animals occurred in the absence of any change in (1) NKA and VA activity or (2) ionocyte numbers/surface exposure, it was reasoned that Na⁺ movement across the gill of SCW animals may be occurring, at least in part, through the paracellular pathway. The ultrastructure of tracheal gill septate junctions (SJs) supported this idea as they exhibited morphological alterations indicative of a leakier pathway. Data provide a first look at alterations in osmoregulatory mechanisms that allow H. rigida nymphs to tolerate sub-lethal salinization of their surroundings.

中文摘要（谷歌机翻）

在暴露于盐污染的水（SCW，7.25g / l NaCl）7天后，在may fly（Hexflynia rigida）的若虫中检查淡水（FW）盐化对渗透调节以及气管鳃形态和功能的影响。

在SCW暴露的H. rigida若虫中扰乱了离子调节的体内平衡，如血淋巴Na⁺，K⁺和Cl^-水平增加以及血淋巴pH和水含量所示。尽管如此，SCW没有改变鳃Na⁺ -K⁺ -ATP酶（NKA）或V型H⁺ -ATP酶（VA）活性。此外，鳃离子细胞中的NKA和VA免疫定位没有显示酶位置的改变或离子细胞丰度的变化。

使用扫描电子显微镜（SEM）确认后一观察结果以检查暴露的气管鳃离子细胞数。离子细胞表面形态测量也表明，SCW没有改变个体离子细胞表面积或离子细胞分数表面积。然而，使用扫描离子选择性电极技术分析穿过may may若虫的气管鳃的Na+运动表明FW若虫从周围水中获得Na⁺，而SCW若虫的气管鳃具有分泌Na⁺的能力。

因为在SCW暴露的动物的鳃上Na+分泌发生在（1）NKA和VA活性或（2）离子细胞数量/表面暴露没有任何变化的情况下，因此推测在SCW动物的鳃上Na⁺运动可能是至少部分地通过细胞旁路径发生。气管鳃隔膜交界处（SJs）的超微结构支持这一观点，因为它们表现出指示渗漏途径的形态学改变。数据首次提供了渗透调节机制的改变，这些改变使得H. rigida若虫能够耐受其周围环境的亚致死盐化。

结果表明，在RI至RVI六个不同区域，沿着蜉蝣的鳃在空间上检查Na⁺转运（图A）。在所测量的所有6个区域中，蜉蝣的鳃从周围的水中吸收Na⁺。在前4个鳃区域（RI-RIV）Na⁺吸收速率大于RV、RVI。相反，发现SCW蜉蝣的鳃在所检查的所有6个区域中分泌Na⁺，并且分泌速率的空间差异是显着的（图C），来自近端的前两个区域RI和RII比后4个区域的分泌速率要大很多。（图C）。

文章链接：https://www.sciencedirect.com/science/article/pii/S0166445X18310634?via%3Dihub