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SCI REP-UK ：山东农大丨磁处理对盐胁迫下杨树氮吸收与分布的影响（附NMT实验体系）

转自中关村旭月非损伤微测技术产业联盟

 

 

  • 期刊：Scientific Reports
  • 主题：磁处理对盐胁迫下杨树氮吸收与分布的影响
  • 标题：The effects of magnetic treatment on nitrogen absorption and distribution in seedlings of Populus × euramericana ‘Neva’ under NaCl stress
  • 影响因子：4.011
  • 检测指标：NH4+、NO3-流速
  • 检测部位：杨树根部（伸长区距离根尖15mm）叶肉细胞
  • NH4+、NO3-流速流实验处理方法：杨树幼苗，在0g/L NaCl+磁化（M0），0g/L NaCl+非磁化（NM0），4g/L NaCl+磁化（M4），4g/L NaCl+非磁化（NM4）中处理
  • NH4+、NO3-流速流实验测试液成份：NH4+: 0.1 mM NH4NO3, 0.1 mM CaCl2, 0.3 mM MES, pH 5.5 NO3-: 0.1 mM NH4NO3, 1.0 mM KCl, 0.1 mM CaCl2, 0.3 mM MES, pH 5.5
  • 作者：山东农业大学林学院马风云、刘秀梅

英文摘要

A potted experiment with Populus × euramericana ‘Neva’ was carried out to assess whether there are positive effects of magnetic treatment of saline water (MTSW) on nitrogen metabolism under controlled conditions in a greenhouse.

Growth properties, nitrogen contents, enzyme activities and metabolite concentrations were determined based on field experiments and laboratory analysis after a 30-day treatment.

The results were as follows: (1) Biomass accumulation, root morphological properties and total nitrogen content were improved by MTSW. (2) Magnetization led to a greater increase in nitrate-nitrogen (NO3−-N) content in roots than in leaves, accompanied by greater NO3− efflux and activated nitrate reductase. (3) MTSW led to a higher ammonium-nitrogen (NH4+-N) content and greater uptake of net NH4+ in the leaves than that in the roots. (4) Magnetization stimulated glutamine synthase, glutamate dehydrogenase and glutamate synthase activities, whereas the concentrations of glutathione and oxidized glutathione were increased in leaves but decreased in roots, and the total glutathione content was increased.

Overall, these results indicated some beneficial impacts of MTSW on nitrogen translocation under field conditions, especially for equilibrating the distribution of NO3−-N and NH4+-N. Moreover, these findings confirmed the potential of using low-quality water for agriculture.

中文摘要（谷歌机翻）

进行了欧洲杨（Populus×euramericana'Neva'）的盆栽试验，以评估在温室中受控条件下盐水（MTSW）磁处理对氮代谢的积极影响。

在30天处理后，基于田间实验和实验室分析确定生长特性，氮含量，酶活性和代谢物浓度。

结果如下：（1）MTSW提高了生物量积累，根系形态特征和总氮含量。（2）磁化导致根中硝态氮（NO3-N）含量比叶片更大，伴随着更多的NO3-外排和活化的硝酸还原酶。（3）MTSW导致叶片中铵态氮（NH4+-N）含量较高，净NH4+吸收量高于根系。 （4）磁化刺激谷氨酰胺合成酶，谷氨酸脱氢酶和谷氨酸合酶活性，而谷胱甘肽和氧化型谷胱甘肽的浓度在叶片中增加但在根中减少，总谷胱甘肽含量增加。

总体而言，这些结果表明MTSW对田间条件下氮转运的一些有益影响，特别是对于平衡NO3- N和NH4+-N的分布。此外，这些发现证实了使用劣质水进行农业的潜力。

结果表明：使用NMT技术监测叶肉细胞中和距离根尖15mm的伸长区NO3- 净流速，两者均显示外排。在测量的溶液中，叶肉细胞的NO3- 外排从235.09增加到290.23 pmol cm-2 s-1（图A），暴露于NaCl溶液中，细根中从123.53到157.51 pmol cm-2 s-1显着增加（p <0.05;图B）。在暴露于盐胁迫的植物中这些值比在对照中更高。当暴露于盐水溶液时，叶片和根部的NO3- 流速增加，并且叶肉细胞中的外排量大于根伸长区域中的外排量。与非磁处理组中的流速相比，NO3- 流速在暴露于磁场条件下的叶肉细胞中显示出显着更高的速率。吸收最大的为M4（290.23 pmol cm-2 s-1），最低吸收量为NM0（181.42 pmol cm-2 s-1）。然而，NO3- 流速在根中表现出相反的模式，在伸长区域中的外排显着降低;NO3- 最高外排为NM4（157.51 pmol cm-2 s-1），最低外排量为M0（38.05 pmol cm-2 s-1）

结果表明：NH4+的净流速不同于NO3-;叶肉细胞为吸收趋势（图A），根部为外排趋势（图B）。叶中NH4+的吸收显示出较低的绝对值。暴露于NaCl溶液（M4，NM4）的样品和暴露于对照处理（M0，NM0）的样品相比，M0显示出最大的流入量（-2518.85pmol cm-2 s-1），M4显示出第二大流出量（1164.15pmol cm-2 s-1），他们有着显着不同（p<0.05）。 此外，NM4中NH4+通量的绝对值与NM0非常接近，并且它们彼此没有差异。相反，盐胁迫下新生长的细根中NH4+的外排高于对照中观察到的，并且M4显示出最高的外排量（186.83 pmolcm-2 s-1），与M0相比显着不同（p <0.05）。如在叶中观察到的，NM4中的NH4+外排与根中的NM0相似，并且数值没有差异。与盐水处理的效果不同，磁处理诱导NH4+的外排比非磁处理更大，并且M4和M0都显示出比NM4和NM0更高的值。这两个因素之间也存在显着的相互作用（p <0.05）

文章链接：https://www.nature.com/articles/s41598-019-45719-6

 

 

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AQUAT TOXICOL ：台湾师大联盟专家 | 银铜纳米颗粒对斑马鱼胚胎侧线毛细胞的毒性作用（附NMT实验体系）

转自中关村旭月非损伤微测技术产业联盟

 

 

  • 期刊：Aquatic Toxicology
  • 主题：银铜纳米颗粒对斑马鱼胚胎侧线毛细胞的毒性作用
  • 标题：Toxic effects of silver and copper nanoparticles on lateral-line hair cells of zebrafish embryos
  • 影响因子：3.794
  • 检测指标：Ca2+流速
  • 检测部位：斑马鱼胚胎侧线毛细胞
  • Ca2+流速流实验处理方法：斑马鱼胚胎在受精后0-96 h（hpf），0, 0.9, 9.3, 27.8 μM的AgNPs，和0,0.16,1.6，15.8μM的CuNP分别进行处理
  • Ca2+流速流实验测试液成份：0.5mM NaCl, 0.2mM CaSO4, 0.2mM MgSO4, 0.16mM KH2PO4, and 0.16mM K2HPO4 (pH 7.0)，300μM MOPS，0.1 mg/l tricaine
  • 作者：台湾师范大学林豊益、Hsiu-Ju Yen

视频：NMT非损伤微测技术检测斑马鱼毛細胞离子流

视频：NMT非损伤微测技术检测斑马鱼皮肤离子细胞

英文摘要

The potential toxicity of nanoparticles (NPs) to the early stages of fish is still unclear. In this study, we investigated the toxic effects of silver (AgNPs) and copper nanoparticles (CuNPs) on lateral-line hair cells of zebrafish embryos.

Zebrafish embryos were incubated in different concentrations of AgNPs and CuNPs at 0˜96 h post-fertilization (hpf). Both AgNPs and CuNPs were found to cause toxic effects in zebrafish embryos in a dose-dependent manner. Values of the 96-h 50% lethal concentration (LC50) of AgNPs and CuNPs were 6.1 ppm (56.5 μM) and 2.61 ppm (41.1 μM), respectively.

The number of FM1-43-labeled hair cells and the microstructure of hair bundles were significantly impaired by AgNPs [≥1 ppm (9.3 μM)] and CuNPs [≥0.01 ppm (0.16 μM)]. Ca2+ influxes at hair bundles of hair cells were measured with a scanning ion-selective microelectrode technique to evaluate the function of hair cells.

AgNPs [≥0.1 ppm (0.9 μM)] and CuNPs [≥0.01 ppm (0.16 μM)] were both found to significantly reduce Ca2+ influxes. Similar toxic effects were also found in hatched embryos subjected to 4 h of exposure (96˜100 hpf) to AgNPs and CuNPs. This study revealed that lateral-line hair cells of zebrafish are susceptible to AgNPs and CuNPs, and these contaminants in aquatic environments could pose a threat to fish survival.

中文摘要（谷歌机翻）

纳米粒子（NPs）对鱼类早期的潜在毒性尚不清楚。在这项研究中，我们研究了银（AgNPs）和铜纳米颗粒（CuNPs）对斑马鱼胚胎侧线毛细胞的毒性作用。

在受精后0~96h（hpf），将斑马鱼胚胎在不同浓度的AgNPs和CuNPs中孵育。发现AgNPs和CuNPs都以剂量依赖的方式对斑马鱼胚胎产生毒性作用。 AgNPs和CuNPs的96小时50％致死浓度（LC50）的值分别为6.1ppm（56.5μM）和2.61ppm（41.1μM）。

AgNPs [≥1ppm（9.3μM）]和CuNPs [≥0.01ppm（0.16μM）]显着损害了FM1-43标记的毛细胞数量和发束的微观结构。用扫描离子选择性微电极技术测量毛细胞发束处的Ca2+流入，以评估毛细胞的功能。

发现AgNPs [≥0.1ppm（0.9μM）]和CuNPs [≥0.01ppm（0.16μM）]均可显着降低Ca2+流入量。在孵化4h（96〜100hpf）的AgNPs和CuNPs孵化的胚胎中也发现了类似的毒性效应。这项研究表明，斑马鱼的侧线毛细胞对AgNPs和CuNPs敏感，这些污染物在水生环境中可能对鱼类的存活构成威胁。

结果表明：在经历96小时AgNP或CuNP处理的胚胎中也发现了毛细胞的功能损伤。使用NM测量L1神经瘤的毛细胞的Ca2+流入量（图A），在0.1,1和3ppm（0.9,9.3和27.8μM）的AgNP组中，Ca2+流入量显着减少26％，46％和91％（图B）。在0.01 ppm（0.16μM）CuNP组中，Ca2+流入量减少了38％，而在0.1和1 ppm（1.6和15.8μM）CuNP组中，Ca2+流入几乎检测不到（图C）。

文章链接：https://www.sciencedirect.com/science/article/pii/S0166445X19304114?via%3Dihub

 

联盟专家暨本文作者介绍

 

林豊益，博士，教授。台湾师范大学生命科学学院生理组，主要从事鱼类环境生理、水生环境毒理、机械性感觉生理、斑马鱼疾病模式与药物筛选研究。

 

近期代表性成果

  • Lin, L. Y., Y. H. Yeh, G.Y. Hung, C. H. Lin, P. P. Hwang, J. L. Horng. 2018. Role of calcium-sensingreceptor in mechanotransducer-channel-mediated Ca2+ influx in hair cells ofzebrafish larvae. Front. Physiol. 649:9.
  • L. Horng, L. L. Yu, S. T. Liu, P. Y. Chen, L. Y. Lin. 2017. Potassiumregulation in medaka (Oryzias latipes) larvae acclimated to fresh water:passive uptake and active secretion by the skin cells. Sci. Rep. 7, 16215.
  • Liu, S. T., J. L. Horng, P. Y. Chen, P. P. Hwang, L.Y. Lin. 2016. Saltsecretion is linked to acid-base regulation of ionocytes in seawater-acclimatedmedaka: new insights into the salt-secreting mechanism. Sci. Rep. 6, 31433.
  • Horng, J. L., P. L. Chao, P. Y. Chen, T. H. Shih, L.Y. Lin. 2015.Aquaporin 1 is involved in acid secretion by ionocytes of zebrafish embryosthrough facilitating CO2 transport. Plos One 10(8):e0136440.
  • Lin, Y. H., G. Y. Hung, L. C. Wu, S. W. Chen, L. Y. Lin. J. L. Horng.2015. Anion exchanger 1b in sereocilia is required for the functioning ofmechanotransducer channels in lateral-line hair cells of zebrafish. Plos One10(2): e0117041.

 

洪君琳，博士，副教授。台北医学大学医学院医学科学研究所，斑马鱼模式核心服务实验室主任，主要从事以斑马鱼为模式研究毛细胞功能及药物影响的研究。

 

近期代表性成果

  • Lin LY, et al,. Role of Calcium-Sensing Receptor in Mechanotransducer Channel Mediated Ca2+ Influx in Hair Cells of Zebrafish Larvae . frontiers in physiology .2018 ;(9):649

  • Hung Giun Yi, et al,. Incidence of immune thrombocytopenia in Taiwan: a nationwide population-based study . Transfusion .2018 ;(58):2712-2719

  • Horng JL, et al,. Potassium Regulation in Medaka (Oryzias latipes) Larvae Acclimated to Fresh Water: Passive Uptake and Active Secretion by the Skin Cells. . Sci Rep. .2017 ;(7):16215

  • Hung GY, et al,. Cancer in adolescents: Incidences and trends during 1995-2009 in Taiwan. . Cancer Lett .2016 ;(372):110-117

  • Hung GY, et al,. Changing incidence patterns of hepatocellular carcinoma among age groups in Taiwan . J Hepatol .2015 ;(63):1390-1396

 

 

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PROTOPLASMA ：河南农大丨高温干旱胁迫致ABA和ROS积累影响水稻萌发（附NMT实验体系）

转自中关村旭月非损伤微测技术产业联盟

 

 

  • 期刊：PROTOPLASMA
  • 主题：高温和干旱胁迫导致水稻脱落酸和活性氧的积累，抑制种子萌发生长
  • 标题：High temperature and drought stress cause abscisic acid and reactive oxygen species accumulation and suppress seed germination growth in rice.
  • 影响因子：2.633
  • 检测指标：Ca2+、H2O2流速
  • 检测部位：水稻胚芽鞘出苗位点
  • Ca2+、H2O2流速流实验处理方法：种子分别在高温38摄氏度，20％PEG-6000处理5天
  • Ca2+、H2O2流速流实验测试液成份：0.1 mM CaCl2, 0.1 mM KCl,0.3mM MES, pH 6.0
  • 作者：河南农业大学赵全志、刘娟

英文摘要

Seed germination is one of the most important biological processes in the life cycle of plants, and temperature and water are the two most critical environmental factors that influence seed germination.

In the present study, we investigated the roles of the plant hormone abscisic acid (ABA) and reactive oxygen species (ROS) in high temperature (HT) and drought-induced inhibition of rice seed germination. HT and drought stress caused ABA accumulation in seeds and inhibited seed germination and seedling establishment. Quantitative real-time polymerase chain reaction analysis revealed that HT and drought stress induced the expression of OsNCED3, a key gene in ABA synthesis in rice seeds.

In addition, ROS (O2•- and H2O2) and malondialdehyde contents were increased in germinating seeds under HT and drought stress. Moreover, we adopted the non-invasive micro-test technique to detect H2O2 and Ca2+ fluxes at the site of coleoptile emergence. HT and drought stress resulted in a H2O2 efflux, but only drought stress significantly induced Ca2+ influx. Antioxidant enzyme assays revealed that superoxide dismutase (SOD), peroxidase, catalase (CAT), and ascorbate peroxidase (APX) activity were reduced by HT and drought stress, consistent with the expression of OsCu/ZnSOD, OsCATc, and OsAPX2 during seed germination.

Altogether, these results suggest that ABA and ROS accumulation under HT and drought conditions can inhibit rice seed germination and growth.

中文摘要（谷歌机翻）

种子萌发是植物生命周期中最重要的生物过程之一，温度和水是影响种子萌发的两个最关键的环境因素。

在本研究中，我们研究了植物激素脱落酸（ABA）和活性氧（ROS）在高温（HT）和干旱诱导的水稻种子萌发抑制中的作用。 HT和干旱胁迫导致种子中ABA积累，抑制种子萌发和幼苗建立。定量实时聚合酶链反应分析表明，HT和干旱胁迫诱导水稻种子ABA合成中关键基因OsNCED3的表达。

此外，在HT和干旱胁迫下，萌发种子中ROS（O2• - 和H2O2）和丙二醛含量增加。此外，我们采用非侵入性微测试技术检测胚芽鞘出现部位的H2O2和Ca2+通量。 HT和干旱胁迫导致H2O2流出，但只有干旱胁迫才能显着诱导Ca2+流入。抗氧化酶测定表明，HT和干旱胁迫使超氧化物歧化酶（SOD），过氧化物酶，过氧化氢酶（CAT）和抗坏血酸过氧化物酶（APX）活性降低，与种子萌发过程中OsCu / ZnSOD，OsCATc和OsAPX2的表达一致。

总之，这些结果表明，在HT和干旱条件下ABA和ROS的积累可以抑制水稻种子的萌发和生长。

结果表明：干旱胁迫显着加剧了发芽种子中胚芽鞘出苗位点的Ca2+净吸收（图b），Ca2+净吸收量比对照种子高61.2％。同时，HT对Ca2+净流入没有显着影响，尽管与对照相比略有下降。 H2O2流速在对照组和治疗组之间的方向和大小上显示出显着差异（图c）。对照种子表现出H2O2的吸收。然而，在HT和干旱胁迫下，H2O2从种子外排到测试溶液中，分别达到1.20和0.44pmol•cm-2•s-1。这些结果可能表明H2O2外排进一步意味着在HT和干旱胁迫下发芽种子中积累更多的H2O含量。

文章链接：https://link.springer.com/article/10.1007%2Fs00709-019-01354-6

 

 

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PP ：林金星IAA流成果丨NRT1.1磷酸化调节侧根发育的机制（附NMT实验体系）

转自中关村旭月非损伤微测技术产业联盟

 

 

  • 期刊：Plant Physiology
  • 主题：NRT1.1磷酸化调节侧根发育的机制
  • 标题：Phosphorylation-mediated dynamics of nitrate transceptor NRT1.1 regulate auxin flux and nitrate signaling in lateral root growth
  • 影响因子：6.305
  • 检测指标：IAA流速
  • 检测部位：酵母细胞
  • IAA流速流实验处理方法：水稻幼苗，10uM chitin或10uM flg22肽瞬时胁迫
  • IAA流速流实验测试液成份：0.5 μM IAA, 2% (m/v) galactose, 0.3 mM MES, pH 5.8
  • 作者：北京林业大学单晓昳、林金星、张曦

英文摘要

The dual-affinity nitrate transceptor NITRATE TRANSPORTER 1.1 (NRT1.1) has two modes of transport and signaling, governed by threonine101 (T101) phosphorylation. NRT1.1 regulates lateral root (LR) development by modulating nitrate-dependent basipetal auxin export and nitrate-mediated signal transduction.

Here, using the Arabidopsis thaliana NRT1.1T101D phosphomimetic and NRT1.1T101A non-phosphorylatable mutants, we found that the phosphorylation state of NRT1.1 plays a key role in NRT1.1 function during LR development. Single-particle tracking revealed that phosphorylation affected NRT1.1 spatiotemporal dynamics. The phosphomimetic NRT1.1T101D form showed fast lateral mobility and membrane partitioning that facilitated auxin flux under low-nitrate conditions.

By contrast, non-phosphorylatable NRT1.1T101A showed low lateral mobility and oligomerized at the plasma membrane (PM), where it induced endocytosis via the clathrin-mediated endocytosis and microdomain-mediated endocytosis pathways under high-nitrate conditions.

These behaviors promoted LR development by suppressing NRT1.1-controlled auxin transport on the PM and stimulating Ca2+-ARABIDOPSIS NITRATE REGULATED 1 (ANR1) signaling from the endosome.

中文摘要（谷歌机翻）

双亲和硝酸盐转运蛋白NITRATE TRANSPORTER 1.1（NRT1.1）具有两种运输和信号传导模式，由苏氨酸101（T101）磷酸化控制。 NRT1.1通过调节硝酸盐依赖性碱基生长素输出和硝酸盐介导的信号转导来调节侧根（LR）发育。

在这里，使用拟南芥NRT1.1T101D磷酸化和NRT1.1T101A非磷酸化突变体，我们发现NRT1.1的磷酸化状态在LR发育期间在NRT1.1功能中起关键作用。单粒子追踪显示磷酸化影响NRT1.1时空动态。磷酸化模拟NRT1.1T101D形式显示出快速的侧向移动性和膜分配，其促进了在低硝酸盐条件下的生长素通量。

相比之下，不可磷酸化的NRT1.1T101A显示出低的侧向迁移率并且在质膜（PM）处寡聚化，其在高硝酸盐条件下通过网格蛋白介导的内吞作用和微区介导的胞吞作用途径诱导内吞作用。

这些行为通过抑制PM上的NRT1.1控制的生长素转运并刺激来自内体的Ca2 + -ARABIDOPSIS硝酸盐调节1（ANR1）信号传导来促进LR发展。

酵母细胞IAA流检测

对照、NRT1.1、T101A、T101D转化酵母菌的IAA流（外排）结果。

文章链接：http://www.plantphysiol.org/content/early/2019/08/20/pp.19.00346

 

 

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CELL RES ：万建民院士丨钙离子启动免疫系统的分子机制（附NMT实验体系）

转自中关村旭月非损伤微测技术产业联盟

 

重大进展 | 万建民连发Science及Cell Res，揭示钙离子启动免疫系统的分子机制

 

  • 期刊：cell research
  • 主题：钙离子启动免疫系统的分子机制
  • 标题：A cyclic nucleotide-gated channel mediates cytoplasmic calcium elevation and disease resistance in rice
  • 影响因子：17.848
  • 检测指标：Ca2+流速
  • 检测部位：水稻叶肉细胞
  • Ca2+流速流实验处理方法：水稻幼苗，10uM chitin或10uM flg22肽瞬时胁迫
  • Ca2+流速流实验测试液成份：0.2mM CaCl2, 0.1mM NaCl, 0.1mM MgCl2 and 0.1mM KCl,pH 5.2
  • 作者：中国农科院万建民、王家昌

英文摘要

The transient elevation of cytoplasmic calcium is essential for pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI). However, the calcium channels responsible for this process have remained unknown.

Here, we show that rice CDS1 (CELL DEATH and SUSCEPTIBLE to BLAST 1) encoding OsCNGC9, a cyclic nucleotide-gated channel protein, positively regulates the resistance to rice blast disease. We show that OsCNGC9 mediates PAMP-induced Ca2+ influx and that this event is critical for PAMPs-triggered ROS burst and induction of PTI-related defense gene expression. We further show that a PTI-related receptor-like cytoplasmic kinase OsRLCK185 physically interacts with and phosphorylates OsCNGC9 to activate its channel activity.

Our results suggest a signaling cascade linking pattern recognition to calcium channel activation, which is required for initiation of PTI and disease resistance in rice.

 

中文摘要（谷歌机翻）

细胞质钙的瞬时升高对病原体相关分子模式（PAMP）- 触发免疫（PTI）至关重要。然而，负责该过程的钙通道仍然未知。

在这里，我们显示编码OsCNGC9（环核苷酸门控通道蛋白）的水稻CDS1（CELL DEATH和SUSCEPTIBLE to BLAST 1）正向调节对稻瘟病的抗性。我们显示OsCNGC9介导PAMP诱导的Ca2+内流，并且该事件对于PAMPs触发的ROS爆发和诱导PTI相关的防御基因表达是至关重要的。我们进一步显示PTI相关受体样细胞质激酶OsRLCK185与OsCNGC9物理相互作用并使其磷酸化以激活其通道活性。

我们的研究结果表明信号级联将模式识别与钙通道激活联系起来，这是启动水稻PTI和抗病性所必需的。

结果表明：响应于几丁质或flg22刺激，WT叶肉细胞比cds1叶肉细胞表现出强烈且快速的Ca2+流入（图3a，b和补充信息，图S6）。这些结果表明OsCNGC9可以介导水稻PTI中的Ca2+流入，并且这种能力在cds1突变体中受损。

结果表明：响应几丁质刺激，Nipponbare叶肉细胞比Osrlck185/ 55双突变体叶肉细胞，表现出快速的Ca2+内流（图e）。此外，在几丁质处理Oscerk1敲除突变体后未观察到显着的Ca2+流入（图f）。这些结果共同表明OsRLCK185及其紧密同源物OsRLCK55参与水稻抗稻瘟病和PAMP诱导的Ca2+内流的调节。

结果表明：在PAMPs刺激后，与Kitaake植物相比，OsCNGC9-OE转基因植物的叶肉细胞显示出更强的Ca2+流入。结合其他实验表明，OsCNGC9是水稻PTI的限速正调控因子。

文章链接：https://www.nature.com/articles/s41422-019-0219-7

 

 

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