NMT是基因功能的活体检测技术，已被103位诺贝尔奖得主所在单位，及北大、清华、中科院使用。期刊：Aquatic Toxicology主题：AgNPs和CuNPs抑制斑马鱼胚胎侧线毛细胞钙吸收标题：Toxic e?ects of silver and copper nanoparticles on lateral-line hair cells of zebra?sh embryos影响因子：3.794检测指标：Ca2+流速检测样品：斑马鱼胚胎侧线毛细胞发束Ca2+流实验处理方法：斑马鱼胚胎经0-96hpf受精后，分别用0，0.1，1，3ppm的AgNPs和0，0.01，0.1，1ppm的CuNPs进行受精Ca2+流实验测试液成份：0.5mM NaCl，0.2mM CaSO4，0.2mM MgSO4，0.16mM KH2PO4，and0.16mM K2HPO4（pH 7.0），300μM MOPS，0.1mg/l tricaine作者：台湾师范大学林豊益斑马鱼毛细胞离子流检测英文摘要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 dosedependent 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 lateralline 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仍不清楚。在这项研究中，我们调查了银（AgNPs）和铜纳米颗粒（CuNPs）对斑马鱼胚胎侧线毛细胞的毒性作用。斑马鱼胚胎在受精后0?96   h（hpf）的不同浓度的AgNPs和CuNPs中孵育。发现AgNPs和CuNPs均以剂量依赖性方式引起斑马鱼胚胎的毒性作用。AgNP和CuNP的96小时50％致死浓度（LC50）值分别为6.1  ppm（56.5μM）和2.61 ppm（41.1μM）。AgNPs [≥1ppm（9.3μM）]和CuNPs   [≥0.01ppm（0.16μM）]严重损害了FM1-43标记的毛细胞数量和发束的微观结构。用扫描离子选择微电极技术测量在毛细胞的发束处的Ca  2+流入，以评估毛细胞的功能。发现AgNPs [≥0.1ppm（0.9μM）]和CuNPs  [≥0.01ppm（0.16μM）]均能显着减少Ca2 +的流入。在暴露于AgNPs和CuNPs 4小时（96?100   hpf）的孵化胚胎中也发现了类似的毒性作用。 这项研究表明，斑马鱼的侧线毛细胞对AgNPs和CuNPs敏感，而这些污染物在水生环境中可能对鱼类的生存构成威胁。结果表明：在AgNP处理组中，相比于0 ppm，Ca2+吸收速率在0.1,1和3   ppm时分别减少了26%,46%和91%;在CuNP处理组中，相比于0 ppm，Ca2+吸收速率在0.01 ppm时减少了38%，而在0.1和1  ppm时几乎检测不到Ca2+的吸收速率。由此可见，AgNPs （≥0.1  ppm）和 CuNPs（≥0.01ppm）都会显著抑制Ca2+的吸收。海洋环境中的污染物会威胁鱼类的生存。

NMT是基因功能的活体检测技术，已被103位诺贝尔奖得主所在单位，及北大、清华、中科院使用。NMT历史上的今天2015年12月27日，中科院遗传发育所童依平、何雪用NMT在Plant Physiology上发表了标题为The Nitrate-Inducible NAC Transcription Factor TaNAC2-5A Controls Nitrate Response and Increases Wheat Yield的研究成果。期刊：Plant Physiology主题：TaNAC2-5A控制**盐反应提高小麦产量标题：The Nitrate-Inducible NAC Transcription Factor TaNAC2-5A Controls Nitrate Response and Increases Wheat Yield影响因子：7.394检测指标：NO3-流速作者：中科院遗传发育所童依平、何雪英文摘要Nitrate is a major nitrogen resource for cereal   crops; thus, understanding nitrate signaling in cereal crops is valuable  for engineering crops with improved nitrogen use efficiency. Although several regulators have been identified in nitrate sensing and signaling  in Arabidopsis (Arabidopsis thaliana), the equivalent information in  cereals is missing.Here, we isolated a nitrate-inducible and  cereal-specific NAM, ATAF, and CUC (NAC) transcription factor,  TaNAC2-5A, from wheat (Triticum aestivum). A chromatin  immunoprecipitation assay showed that TaNAC2-5A could directly bind to  the promoter regions of the genes encoding nitrate transporter and  glutamine synthetase. Overexpression of TaNAC2-5A in wheat enhanced root  growth and nitrate influx rate and, hence, increased the root’s ability  to acquire nitrogen. Furthermore, we found that  TaNAC2-5A-overexpressing transgenic wheat lines had higher grain yield  and higher nitrogen accumulation in aerial parts and allocated more  nitrogen in grains in a field experiment.These results suggest  that TaNAC2-5A is involved in nitrate signaling and show that it is an  exciting gene resource for breeding crops with more efficient use of  fertilizer. 中文摘要（谷歌机翻）**盐是谷物作物的主要氮资源。因此，了解谷类作物中的**盐信号对于提高氮素利用效率的工程作物非常有价值。尽管已经在拟南芥（Arabidopsis thaliana）的**盐感测和信号传导中确定了几种调节剂，但谷物中的等效信息却缺失了。在这里，我们从小麦（Triticum   aestivum）中分离出了**盐诱导型和谷类特异性NAM，ATAF和CUC（NAC）转录因子TaNAC2-5A。染色质免疫沉淀分析表明，TaNAC2-5A可以直接结合到编码**盐转运蛋白和谷氨酰胺合成酶的基因的启动子区域。TaNAC2-5A在小麦中的过表达增强了根系的生长和**盐的流入速率，因此增加了根系获得氮的能力。此外，我们发现在田间试验中，过表达TaNAC2-5A的转基因小麦品系具有较高的籽粒产量和较高的氮素在地上部分的积累，并在籽粒中分配了更多的氮。这些结果表明，TaNAC2-5A参与**盐信号传导，并表明它是利用更有效地使用肥料来育种作物的令人兴奋的基因资源。D and E, Nitrate in?ux rates at the root tip surface (D) and expression levels of nitrate transporters in whole seedlings   (E). Seedlings were grown for 4 d on solid medium that contained 6 mM   nitrate. The roots were used to measure nitrate ?ux rate in the   measuring solution that contained 1 mM nitrate. Data are means 6 SE for six plants. Whole seedlings were used to analyze the expression of   nitrate trans-porters. The relative expression levels were normalized to  the expression of AtACTIN2 (AtACT2). Data are means 6 SE of three   replicates. Asterisks indicate that the difference between the means of the wild type and the transgenic lines was signi?cant at the P , 0.05   (*) and P , 0.01 (**) levels.

2019年10月厦门大学成功采购扬格/旭月品牌的第七代NMT系统，今天旭月公司顺利的完成了厦门大学NMT设备的操作培训。培训全程依照中关村NMT联盟颁发的《NMT系统操作培训标准》。一直以来，厦门大学环境与生态学院利用NMT主要开展红树抗盐机制研究，前期已经发表了一些NMT成果。（Sci Rep-UK.2015, 5: 125-116 ；PLOS ONE, 2013,8(8): e71543；Tree Physiol, 2010, 30(12): 1570 - 1585. ）。后续将继续利用NMT深入开展红树抗逆、抗氧化机制的研究。此次旭月派出的是美国扬格（旭月北京）售后部负责人，巨肖宇  中关村NMT联盟认证高级售后工程师巨肖宇。拥有5年以上非损伤微测系统操作应用、售后经验，累计售后服务近五十家单位，售后服务经验3000+小时。Cu2+、Pb2+流速传感器商业化研发团队骨干成员。                

受厦门大学环境与生态学院邀请，中关村NMT联盟将于2020年1月3日，在厦大举行非损伤微测技术（Non-invasive Micro-test Technology, NMT）植物抗逆基因功能研究培训。环境与生态学院在利用NMT开展红树抗盐机制研究上，已经取得了一定的前期成果。后续将继续利用NMT深入开展红树抗逆、抗氧化机制的研究。Chen J, et al. Scientific Reports, 2015, 5: 125-116.Chen J, et al.  PLOS ONE, 2013,8(8): e71543.Chen J, et al. Tree Physiology, 2010, 30(12): 1570 - 1585.报告人介绍孙 健  非损伤微测技术资深应用专家，江苏师范大学副教授，兼任中关村NMT联盟副理事长、NMT国际标准化委员会（NISC）委员。孙健副教授是国内首篇高水平NMT盐胁迫文章的第一作者（Sun  J, et al. Plant Physiol, 2009, 149:   1141-1153.），单篇引用次数已高达265次，发表的NMT文献总引用超过1500次（注意该内容下也要有下划线）。其博士期间（导师：陈少良教授）从事胡杨抗盐机制研究，目前专注于甘薯抗盐/抗病机制研究，现为国家甘薯产业技术体系“细胞遗传与倍性育种技术”岗位团队骨干成员。其所在实验室是国内首批“中关村NMT联盟创新与服务中心”。截止2019年，已利用NMT发表SCI文献20余篇（第一/通讯）。培训信息 培训内容：NMT理论应用培训、专家分享、上机演示时间：2020年1月3日  星期五  9：00～12：00地点：厦门大学（翔安校区）环境与生态学院金泉楼B105室联系人：程老师  18515811370本次报告提供网络录播，录播时间为：2020年1月6日 19：00，可微信扫下方二维码报名观看（无需注册）。NMT简介非损伤微测技术（Non-invasive Micro-test Technology, NMT）是研究功能基因的一种技术，通过检测活体植物根茎叶与内/外环境间Ca2+/Cd2+/Na+/K+/N/O2...交换量的实时变化，揭示：Na+/K+运输与植物耐盐性Ca2+运输与植物环境胁迫感知与信号转导NH4+运输参与甘薯钾钠平衡调控Cd2+运输与植物重金属耐受性NMT在快速验证作物抗性遗传改良的应用......其他培训人员刘蕴琦  NMT高级应用顾问中关村非损伤微测技术产业联盟秘书长，《NMT   101问》、《NMT论文集》副主编。协助中国农大武维华院士、中科院植物所种康院士等课题组，设计了Cell、Plant   Cell等研究成果中的非损伤离子流实验部分。近5年，直接协助国内学者发表的SCI文章共67篇，累计影响因子308.64。巨肖宇  中关村NMT联盟认证高级售后工程师拥有5年以上非损伤微测系统操作应用、售后经验，累计售后服务近五十家单位，售后服务经验3000+小时。Cu、Pb流速传感器商业化研发团队骨干成员。 非损伤微测技术目前可检测的指标有：H+、Ca2+、Na+、K+、Cl-、Mg2+、Cd2+、Cu2+、Pb2+、NH４+、NO3-、O2、H2O2、IAA。

NMT是基因功能的活体检测技术，已被103位诺贝尔奖得主所在单位，及北大、清华、中科院使用。期刊：International journal of molecular science主题：杨树响应盐胁迫的生理和分子机制标题：Progress in Understanding the Physiological and Molecular Responses of Populus to Salt Stress影响因子：4.183作者：广西农科院、中科院昆明植物所英文摘要Salt stress (SS) has become an important factor   limiting afforestation programs. Because of their salt tolerance and   fully sequenced genomes, poplars (Populus spp.) are used as model   species to study SS mechanisms in trees. Here, we review recent insights  into the physiological and molecular responses of Populus to SS,   including ion homeostasis and signaling pathways, such as the salt   overly sensitive (SOS) and reactive oxygen species (ROS) pathways. We   summarize the genes that can be targeted for the genetic improvement of salt tolerance and propose future research areas.中文摘要（谷歌机翻）盐胁迫（SS）已成为限制造林计划的重要因素。 由于其耐盐性和完整的基因组序列，白杨（Populus   spp。）被用作模型物种来研究树木中的SS机理。 在这里，我们回顾了对胡杨对SS的生理和分子反应的最新见解，包括离子稳态和信号传导途径，例如盐过度敏感（SOS）和活性氧（ROS）途径。 我们总结了可用于耐盐遗传改良的基因，并提出了未来的研究领域。                

旭月简介非损伤微测技术（NMT）是基因功能的活体检测技术，已被103位诺贝尔奖得主所在单位，及北大、清华、中科院使用。旭月公司是中国NMT全方位专业服务团队，旭月NMT设备市场占有率达九成以上，且中国学者发表的400多篇NMT文献有九成以上出自我们的设备。旭月公司拥有目前世界上唯一一款商业化的，可检测活体植物大分子IAA生长素的传感器。旭月公司的售后团队由多名经过美国实地培训且通过中关村NMT联盟认证的工程师组成。2019年旭月推出“流速云”，NMT学习/设计/操作/采购一站式整合 。继2018年扬格/旭月IAA活体生理检测仪入选 Science 推荐产品版块（Science推荐产品：IAA活体生理检测），2019年“旭月科技”入围科学仪器行业成长潜力企业。一路走来，旭月发展到今天离不开每位老师的支持与鼓励。感谢大家见证旭月的成长。“2019年度科学仪器行业成长潜力企业”评选活动还在继续，还希望各位老师继续支持旭月，为旭月投上您宝贵的一票。

NMT是基因功能的活体检测技术，已被103位诺贝尔奖得主所在单位，及北大、清华、中科院使用。期刊：International Journal of Molecular Sciences主题：OsCIPK2过表达促进低氮条件下水稻NO3-吸收标题：Exploring the Potential of Overexpressed OsCIPK2   Rice as a Nitrogen Utilization E cient Crop and Analysis of Its   Associated Rhizo-Compartmental Microbial Communities影响因子：4.183检测指标：Ca2+、NH4+、NO3-流速检测样品：水稻根尖Ca2+、NH4+、NO3-流实验处理方法：幼苗长至三叶期，分别在正常氮（1.44 mM NH4NO3）和低氮（0.24 mM NH4NO3）处理Ca2+、NH4+、NO3-流实验测试液成份：Ca2+: 0.1 mmol/L KCl, 0.1 mmol/L CaCl2, 0.1 mmol/L MgCl2, 0.5 mol/L NaCl, 0.3 mmol/L MES, 0.2 mmol/L Na2SO4, pH 6.0;NO3-:  0.1 mmol/L NH4NO3, 0.1 mmol/L NH4NO3, 0.1 mmol/L KCl, 0.1 mmol/L KCl,  0.1 mmol/L KCl2, 0.3 mmol/CaCl2, 0.3 mmol/L MgCl2, 0.3 mmol；NH4+: 0.1 mmol/L NH4NO3, 0.1 mmol/L CaCl2, 0.3 mmol/L MES, pH 6.0作者：福建农林大学林文雄英文摘要Nitrogen (N) is one of the indispensable factors in   rice growth and development. China holds a premier position in the   production of rice and at the same time also faces higher N fertilizer   costs along with serious damage to the environment. A better solution is  much needed to address these issues, without disrupting the production  of rice as an important cereal, while minimizing all the deleterious   effects on the environment.Two isogenic lines Kitaake (WT) and  its genetically modified line CIPK2 (RC), overexpressing the gene for  Calcineurin B-like interacting protein kinase 2 (OsCIPK2) with better  nitrogen use efficiency (NUE), were compared for their growth and  development under low versus normal levels of N. NUE is a complex trait  mainly related to a plant’s efficiency in extraction, assimilation, and  recycling of N from soil. The microbial population was analyzed using  high-throughput Illumina Miseq 16S rRNA sequencing and found that RC  with CIPK2, specifically expressed in rice root, not only performed  better without nitrogen fertilizer (LN) but also increased the diversity  of bacterial communities in rice rhizosphere compartments (rhizosphere,  rhizoplane, and endosphere).The relative abundance of  beneficial bacteria phyla increased, which are known to promote the  circulation and transformation of N in rhizosphere soil. To further  explore the potential of RC regarding better performance under LN, the  ion fluxes in root apical were detected by non-invasive micro-test  technique (NMT). We found that RC can absorb more Ca2+ and NO3? under LN  as compared to WT. Finally, compared to WT, RC plants exhibited better  growth of root and shoot, and increased yield and N uptake under LN,  whereas there was no significant difference in the growth of two rice  lines under normal nitrogen (NN) treatment.We are able to get  preliminary results, dealing with the OsCIPK2 overexpressed rice line,  by studying the rice molecular, physiological, and chemical parameters  related to NUE. The results laid the foundation for further research on N  absorption and utilization in rice from the soil and the interaction  with microbial communities. 中文摘要（谷歌机翻）氮（N）是水稻生长发育不可或缺的因素之一。中国在大米生产中处于领先地位，同时也面临着更高的氮肥成本以及对环境的严重破坏。迫切需要一种更好的解决方案来解决这些问题，同时又不中断作为重要谷物的大米的生产，同时将对环境的所有有害影响降至最低。比较了两个同基因系Kitaake（WT）及其基因改造品系CIPK2（RC），它们以较低的氮素利用效率（NUE）过量表达了钙调磷酸酶B样相互作用蛋白激酶2（OsCIPK2）的基因，并在低温下对其生长发育进行了比较。与正常水平的氮相比。NUE是一个复杂的性状，主要与植物从土壤中提取，吸收和循环利用N的效率有关。使用高通量Illumina  Miseq 16S   rRNA测序分析了微生物种群，发现带有CIPK2的RC在水稻根部特异表达，不仅在不使用氮肥（LN）的情况下表现更好，而且增加了水稻根际区室细菌群落的多样性（根际，根际和内球）。有益菌门的相对丰度增加，已知它可以促进根际土壤中氮的循环和转化。为了进一步探索RC在LN下获得更好性能的潜力，通过无创微测试技术（NMT）检测了根尖中的离子通量。我们发现，与野生型相比，RC在LN下可以吸收更多的Ca2+和NO3-。最后，与WT相比，RC植物在LN下表现出更好的根和茎生长，并增加了产量和氮素吸收，而在正常氮（NN）处理下，两个水稻品系的生长没有显着差异。通过研究与NUE相关的水稻分子，生理和化学参数，我们能够获得初步结果，处理过表达OsCIPK2的水稻品系。研究结果为进一步研究水稻从土壤中吸收和利用氮以及与微生物群落的相互作用奠定了基础。结果表明：LN（low   nitrogen）处理时，RC（过表达型）根系的Ca2+吸收显著增加，而NN（normal   nitrogen）处理时无显著差异；LN处理时，RC根系的NO3-吸收增加，而NN条件下RC和WT（野生型）根系的NO3-表现为外排，且二者并无显著差异；至于NH4+，RC和WT均表现为外排，且无显著差异。这说明，LN条件下，根系CIPK2基因过表达促进了根系吸收NO3-，与RC根际的硝化作用增加是符合的。

NMT是基因功能的活体检测技术，已被103位诺贝尔奖得主所在单位，及北大、清华、中科院使用。期刊：内蒙古农业大学学报(自然科学版)标题：拟南芥VHA＿c3基因沉默株系对ABA的响应检测指标：H+流速检测样品：拟南芥分生区（距根尖200-400μm）H+流速流实验处理方法：6日龄幼苗，100μM ABA 处理5minH+流速流实验测试液成份：无作者：中国农业科学院草原研究所王瑞刚、韩晓东摘 要为了研究VHA-c3基因在拟南芥响应非生物胁迫中的作用。通过农杆菌介导,将设计的dsDN**段整合在拟南芥基因组上,筛选获得了能够不同程度特异沉默VHA-c3基因的9个转基因株系c3-1～c3-9,并对VHA-c3基因沉默株系进行ABA和葡萄糖胁迫处理。在ABA和6%葡糖糖处理后,c3-1株系主根的相对伸长量和种子的萌发率都有一定程度的抑制,但是,其被抑制效果远远低于野生型株系。并且,随着VHA-c3基因沉默效率的增加,沉默株系受ABA或葡糖糖的抑制效果越不明显;非损伤微测技术测定结果表明,ABA促进了c3-1株系的H+内流能力。综上, VHA-c3基因沉默株系表现出对ABA抑制作用不敏感的现象

NMT是基因功能的活体检测技术，已被103位诺贝尔奖得主所在单位，及北大、清华、中科院使用。NMT历史上的今天2014年12月24日，湖南大学曾光明、陈桂秋、谭琼用NMT在Chemosphere上发表了标题为Physiological  fluxes and antioxidative enzymes activities of immobilized   Phanerochaete chrysosporium loaded with TiO2 nanoparticles after   exposure to toxic pollutants in solution的研究成果。期刊：Chemosphere主题：TiO2提升黄孢原毛平革菌镉耐受力的生理证据标题：Physiological fluxes and antioxidative enzymes   activities of immobilized Phanerochaete chrysosporium loaded with  TiO2 nanoparticles after exposure to toxic pollutants in solution影响因子：4.208检测指标：H+、O2、Cd2+流速作者：湖南大学曾光明、陈桂秋、谭琼英文摘要Immobilized Phanerochaete chrysosporium loaded with   TiO2 nanoparticles (PTNs) are novel high-value bioremediation materials for adsorbing cadmium and for degrading 2,4-dichlorophenol (2,4-DCP).   The real-time changes in H+ and O2 fluxes were measured using the   noninvasive microtest technique (NMT).The H+ influx increased  after the addition of 2,4-DCP, and shifted to efflux following the  addition of Cd2+. The O2 flux decreased after the addition of both  2,4-DCP and Cd2+. A larger Cd2+ flux was immediately observed after  exposure to 0.5 mM Cd2+ (?351.25 pmol cm?2 s?1) than to 0.1 mM Cd2+  (?107.47 pmol cm?2 s?1). The removal of Cd2+ by the PTNs increased more  after treatment with the 0.5 mM exposure solution (27.6 mg g?1) than  with the 0.1 mM exposure solution (3.49 mg g?1).The enzyme  activities were analyzed to review the antioxidative defense system of  PTNs in a solution containing various concentrations of Cd2+. The  activities of the coenzyme nicotinamide adenine dinucleotide (NADH)  oxidase as well as the enzyme catalase (CAT) plateaued at 6.5 U g?1 FW  and 9.7 U g?1 FW, respectively, after exposure to 0.25 mM Cd2+. The  activity of superoxide dismutase (SOD) increased gradually in solutions  containing 0.1–0.6 mM Cd2+, and eventually reached a maximum (68.86 U  g?1 FW).These results illustrate how the antioxidative defense  system and the physiological fluxes of PTNs respond to the stress caused  by toxic pollutants.中文摘要（谷歌机翻）负载有TiO2纳米粒子（PTNs）的固定化Phanerochaete chsssporium是用于吸附镉和降解2,4-二氯苯酚（2,4-DCP）的新型高价值生物修复材料。使用非损伤微测技术（NMT）测量H+和O2通量的实时变化。加入2,4-DCP后，H+流入量增加，而加入Cd2+后，H+流入量移出。同时添加2,4-DCP和Cd2+后，O2通量下降。暴露于0.5  mM Cd2+（-351.25 pmol cm-2 s-1）后立即观察到更大的Cd2+通量，而不是暴露于0.1 mM Cd2+（-107.47   pmol cm-2 2 s-1）。用0.5 mM暴露溶液（27.6 mg g-1）处理后，PTN去除Cd2+的增加比使用0.1   mM暴露溶液（3.49 mg g-1）处理的增加更多。分析了酶的活性，以审查包含各种浓度的Cd2+的溶液中PTN的抗氧化防御系统。暴露于0.25  mM Cd2+后，辅酶烟酰胺腺嘌呤二核苷酸（NADH）氧化酶和过氧化氢酶（CAT）的活性分别稳定在6.5 U g-1 FW和9.7 U g-1   FW上。超氧化物歧化酶（SOD）的活性在含0.1–0.6 mM Cd2+的溶液中逐渐增加，并最终达到最大值（68.86 U g-1 FW）。这些结果说明了PTN的抗氧化防御系统和生理通量如何响应有毒污染物引起的压力。Fig. 2. (A) Real-time Cd2+ ?ux of PTNs exposed to 0.1 mM Cd(NO3)2 and 10 mg L??1 2,4-DCP.

NMT是基因功能的活体检测技术，已被103位诺贝尔奖得主所在单位，及北大、清华、中科院使用。期刊：Scientific Report主题：NaCl胁迫下磁处理可调控杨树N代谢标题：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-流速检测样品：杨树根伸长区（距根尖15mm）和叶肉细胞NH4+、NO3-流实验处理方法：4周龄杨树幼苗，分别经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）处理NH4+、NO3-流实验测试液成份：NH4+: 0.1 mM NH4NO3, 0.1 mM CaCl2, 0.3 mM MES, pH 5.5NO3-: 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）进行了盆栽的“ Neva”杨树的盆栽实验，以评估在受控条件下温室中磁化盐水（MTSW）对氮素代谢是否有积极作用。经过30天的处理后，根据田间实验和实验室分析确定了生长特性，氮含量，酶活性和代谢物浓度。结果表明：（1）MTSW提高了生物量积累，根系形态特征和总氮含量。（2）磁化作用导致根部的**盐氮（NO3-N）含量比叶片中的增加更大，同时伴随着更大的NO3-流出量和活化的**还原酶。（3）MTSW导致叶片中的铵态氮（NH4+  -N）含量更高，并且比根部更高地吸收了净NH4   +。（4）磁化刺激谷氨酰胺合酶，谷氨酸脱氢酶和谷氨酸合酶活性，而叶片中谷胱甘肽和氧化型谷胱甘肽的浓度增加，而根部减少，谷胱甘肽总含量增加。总体而言，这些结果表明，MTSW对田间条件下的氮转运有一些有益的影响，尤其是对于平衡NO3- -N和NH4+ -N的分布。此外，这些发现证实了将低质水用于农业的潜力。结果表明：对于NO3-，叶肉细胞和根伸长区均表现为外排。存在NaCl胁迫时，磁疗前后，叶肉细胞的NO3-外排速率从235.09  pmol·cm-2·s-1（NM4）增加到290.23   pmol·cm-2·s-1（M4）；经NaCl胁迫后，根伸长区的NO3-外排速率从123.53   pmol·cm-2·s-1（M4）增加到157.51 pmol·cm-2·s-1（NM4）。至于NH4+，叶肉细胞表现为吸收而根伸长区表现为外排。经NaCl胁迫后，叶肉细胞的NH4+吸收速率降低，NM0和NM4并无显著差异。但在根伸长区，NaCl胁迫后NH4+外排速率更高，M4的外排速率达到了186.83 pmol·cm-2·s-1。

旭月公司2020年元旦放假安排如下：1月1日（周三）放假1天，不调休。请广大客户提前安排好实验，放假间如有问题咨询，可以拨打我公司电话进行留言，或点击公司主页在线客服进行留言，我们会在上班后第一时间回复您。

NMT是基因功能的活体检测技术，已被103位诺贝尔奖得主所在单位，及北大、清华、中科院使用。期刊：Bone主题：DAla2GIP抑制钙吸收拮抗软骨细胞凋亡标题：DAla2GIP antagonizes H2O2-induced chondrocyte apoptosis and inflammatory factor secretion影响因子：4.360检测指标：Ca2+流速检测样品：7日龄小鼠，1.0ml乙醚麻醉后处死，第三代胸部软骨组织细胞Ca2+流实验处理方法：Control，300μM H2O2 ，100pM DAla2GIP 和 300μM H2O2+ 100pM DAla2GIPCa2+流实验测试液成份：无作者：山西医科大学第二医院卫小春、王宇泽英文摘要To investigate the protective effects of DAla2GIP   against the apoptosis and inflammatory factor secretion in H2O2-induced chondrocyte, and explore the possible mechanisms of DAla2GIP underlying  its protection.The chondrocytes were divided into the following  four groups: Control, 300?μM H2O2, 100?pM DAla2GIP and 300?μM  H2O2?+?100?pM DAla2GIP. The apoptosis of chondrocyte was measured by  using mitochondrial membrane potential assay kit (JC-1) and TUNEL assay,  the inflammatory factor secretion were assessed by ELISA assay, and the  cellular and molecular mechanisms of DAla2GIP protection were  investigated by using Real time-PCR, flow cytometry, Non- invasive  calcium detection and western blotting techniques.(1) DAPla2GIP  prevents apoptosis of chondrocyte induced by H2O2. (2) DAla2GIP  alleviated the inflammation of chondrocyte induced by H2O2. (3) DAla2GIP  prevents chondrocyte apoptosis by inhibiting calcium influx of  chondrocyte and regulating expression of Bcl-2 and Caspase-3induced by  H2O2. (4) DAla2GIP inhibited the H2O2 mediated inflammation by up-  regulating the expressions of Sox9 and Col2a1 and inhibiting  PI3K/Akt/NF-κB pathway.Our experimental results revealed that  DAla2GIP prevents chondrocyte apoptosis by inhibiting calcium influx of  chondrocyte and induced regulating expression of Bcl-2 and Casp ase-3by  H2O2. Further, molecular biology experiments confirmed that DAla2GIP  inhibited the H2O2 mediated inflammation vis up-regulating the  expressions of Sox9 and Col2a1 and inhibiting PI3K/Akt/NF-κB pathway.  The results demonstrate that DAla2GIP has protective properties in  H2O2-induced chondrocyte injury, this finding shows that novel GIP  analogues have the potential as a novel therapeutic for osteoarthritis  patients.中文摘要（谷歌机翻）研究DAla2GIP对H2O2诱导的软骨细胞凋亡和炎性因子分泌的保护作用，并探讨DAla2GIP保护其的可能机制。软骨细胞分为以下四组：对照组，300μmH2O2、100μmM  DAla2GIP和300μmH2O2n   +100μpMDAla2GIP。用线粒体膜电位分析试剂盒（JC-1）和TUNEL法检测软骨细胞的凋亡，用ELISA法评估炎性因子的分泌，并通过实时PCR研究DAla2GIP保护的细胞和分子机制，流式细胞仪，无创钙离子检测和蛋白质印迹技术。（1）DAPla2GIP可防止H2O2诱导的软骨细胞凋亡。（2）DAla2GIP减轻了H2O2诱导的软骨细胞炎症。 （3）DAla2GIP通过抑制软骨细胞的钙内流并调节过氧化氢诱导的Bcl-2和Caspase-3的表达来防止软骨细胞凋亡。（4）DAla2GIP通过上调Sox9和Col2a1的表达并抑制PI3K  / Akt /NF-κB通路来抑制H2O2介导的炎症。我们的实验结果表明，DAla2GIP通过抑制软骨细胞的钙内流并诱导H2O2调节Bcl-2和Casp   ase-3的表达来防止软骨细胞凋亡。此外，分子生物学实验证实，DAla2GIP通过上调Sox9和Col2a1的表达并抑制PI3K / Akt   /NF-κB通路来抑制H2O2介导的炎症。结果表明，DAla2GIP在H2O2诱导的软骨细胞损伤中具有保护性，这一发现表明，新型GIP类似物具有作为骨关节炎患者的新型治疗剂的潜力。结果表明：100 pM   DAla2-GIP组和Control组的Ca2+吸收速率并无显著差异，平均值分别是1.94 pmol·cm-2·s-1和1.42   pmol·cm-2·s-1；经300 μM H2O2处理后，Ca2+吸收速率明显增加，平均值为139.61 pmol·cm-2·s-1；经300  μM H2O2和100 pM DAla2-GIP共同处理后，Ca2+吸收速率明显低于300 μM H2O2处理组，平均值为26.68   pmol·cm-2·s-1，说明DAla2-GIP抑制了H2O2诱导的Ca2+吸收。而Ca2+吸收是软骨细胞凋亡早期的标志。

受联盟委托，旭月公司的联盟认证工程师于2019年12月17日，前往北京大学生命科学学院进行了上机演示。非损伤微测技术资深应用专家、江苏师范大学孙健副教授受邀做现场报告。        

NMT是基因功能的活体检测技术，已被103位诺贝尔奖得主所在单位，及北大、清华、中科院使用。NMT历史上的今天2014年12月16日，山东省农科院孔令安用NMT在Frontiers in Plant Science上发表了标题为Effects of high NH4+ on K+ uptake, culm mechanical strength and grain filling in wheat的研究成果。期刊：Frontiers in Plant Science主题：高铵抑制小麦根部钾吸收标题：Effects of high NH4+on K+ uptake, culm mechanical strength and grain filling in wheat影响因子：4.495检测指标：K+流速作者：山东省农科院孔令安英文摘要It is well established that a high external  NH4+ concentration depresses many processes in plant development, but  the underlying mechanisms are still not well understood. To determine   whether the negative effects of high levels of NH4+ are related to   competitive cation uptake, wheat was grown in a field with moderate (18 g  N m?2) and high (30 g N m?2) supplies ofNH4+ in the presence or absence  of additional K+ (6 g K2O m?2) to examine culm mechanical strength, the  main components of the vascular bundle, nitrogen (N) remobilization and  the grain-filling rate.The results indicated that an excessive  supply of NH4+ significantly decreased culm mechanical strength, the  cellulose and lignin contents of vascular bundles, the N remobilization  efficiency (NRE) and the grain-filling rate compared with a moderate  level of NH4+. The additional provision of K+ considerably alleviated  these negative effects of high NH4+, resulting in a 19.41–26.95%  increase in culm mechanical strength during grain filling and a 34.59%  increase in the NRE. An assay using the scanning ion-selective electrode  technique (SIET) showed that the net rate of transmembrane K+ influx  decreased by 84.62%, and measurements using flame photometry  demonstrated that the K+ content decreased by 36.13% in wheat plants  subjected to high NH4+.This study indicates that the effects of  high NH4+ on culm mechanical strength, cellulose and lignin contents,  the NRE and the grain-filling rate are probably associated with  inhibition of K+ uptake in wheat. 中文摘要（谷歌机翻）众所周知，外部NH4+   的高浓度会抑制植物发育中的许多过程，但其潜在机理仍未得到很好的理解。为了确定高水平的NH4+的负面影响是否与竞争性阳离子摄入相关，将小麦种植在提供适量（18  g N m-2）和大量（30 g N m-2）NH4+的田地上在存在或不存在额外的K+（6 g K2O   m-2）的情况下进行图4的测试，以检查茎秆机械强度，维管束的主要成分，氮（N）迁移和谷粒填充率。结果表明，与中等水平的NH4+相比，过量供应NH4+会显着降低茎秆机械强度，维管束中纤维素和木质素的含量，氮的固定效率（NRE）和籽粒填充率。额外提供的K+大大减轻了高NH4+的这些负面影响，导致籽粒灌浆期间茎秆机械强度提高了19.41–26.95％，NRE增加了34.59％。使用扫描离子选择电极技术（SIET）进行的分析表明，跨膜K  +流入的净速率降低了84.62％，而使用火焰光度法的测量结果表明，在高NH4+的小麦植株中，K+含量降低了36.13％。。这项研究表明，高NH4+对小麦茎秆机械强度，纤维素和木质素含量，NRE和籽粒充实率的影响可能与抑制小麦吸收K+有关。(B) The effects of 10 mM NH4+ on the kinetics of the net K+ ?uxes in  wheat root epidermal cells; each point is the mean from six individual  plants. (C) Each column presentsthe mean values of the K+ ?uxes  measured in the root epidermal cells over a recording period of 30 min.  Nm: moderate NH4+,Nh:10mM NH4+ treatment. The bars represent the  standard deviation for six individual plants. The different letters  indicated above each column refer to signi?cant differences at the P  

NMT是基因功能的活体检测技术，已被103位诺贝尔奖得主所在单位，及北大、清华、中科院使用。期刊：Genes主题：盐胁迫下钾转运体LrKUP8抑制K+外排标题：Potassium Transporter LrKUP8 Is Essential for K+Preservation in Lycium ruthenicum, A Salt-Resistant Desert Shrub影响因子：3.331检测指标：K+流速检测样品：黒枸杞愈伤组织K+流实验处理方法：有/无300 mM NaCl培养12hK+流实验测试液成份：0.5 mM KCl, 0.5 mM NaCl, 0.1mM MgCl2, 0.2 mM CaCl2, 2.5% sucrose，pH 5.7作者：北京林业大学陈金焕、戴逢斌英文摘要Salt stress is a major constraint for many crops and trees. A wild species of Goji named Lycium ruthenicum is an important   economic halophyte in China and has an extremely high tolerance to   salinity. L. ruthenicum grows in saline soil and is known as a   potash-rich species.However, its salt adaptation strategies and  ion balance mechanism remains poorly understood. Potassium (K+) is one  of the essential macronutrients for plant growth and development. In  this study, a putative salt stress-responsive gene encoding a HAK  (high-affinity K+)/KUP (K+ uptake)/KT (K+ transporter) transporter was  cloned and designated as LrKUP8.This gene belongs to the cluster  II group of the KT/HAK/KUP family. The expression of LrKUP8 was  strongly induced under high NaCl concentrations. The OE-LrKUP8 calli  grew significantly better than the vector control calli under salt  stress conditions. Further estimation by ion content and micro-electrode  ion flux indicated a relative weaker K+ efflux in the OE-LrKUP8 calli  than in the control.Thus, a key gene involved in K+ uptake under  salt condition was functionally characterized using a newly established  L. ruthenicum callus transformation system. The importance of K+  regulation in L. ruthenicum under salt tolerance was highlighted.中文摘要（谷歌机翻）盐胁迫是许多农作物和树木的主要限制因素。枸杞的一种野生种，称为枸杞，是中国重要的经济盐生植物，对盐分的耐受性极高。ruthenicum生长在盐渍土壤中，被称为富含钾肥的物种。然而，其盐适应策略和离子平衡机制仍然知之甚少。钾（K+）是植物生长和发育必不可少的大量营养素之一。在这项研究中，克隆了编码HAK（高亲和力K+）/ KUP（K+摄取）/ KT（K+转运蛋白）转运蛋白的假定盐胁迫响应基因，并将其命名为LrKUP8。该基因属于KT  / HAK /   KUP家族的II类。在高NaCl浓度下强烈诱导LrKUP8的表达。在盐胁迫条件下，OE-LrKUP8愈伤组织的生长明显好于载体对照的愈伤组织。通过离子含量和微电极离子通量的进一步估计表明，与对照相比，OE-LrKUP8愈伤组织中的K+外排相对较弱。因此，使用新建立的黑麦草愈伤组织转化系统对在盐条件下参与钾离子吸收的关键基因进行了功能鉴定。强调了在耐盐性条件下钌中的K+调节的重要性。结果表明：LrKUP8抑制了愈伤组织中K+外排。不存在NaCl胁迫时，两种愈伤组织的K+外排速率较小，且无显著差异。经盐胁迫后，K+外排速率远高于无胁迫组；对照组（pBI121）的K+外排速率明显高于过表达组（OE-LrKUP8）。过表达组由于其更强的保K+能力，因而维持高的K+/Na+比。保持最优的K+/Na+比是植物耐盐性的关键特征之一，证明LrKUP8的保K+能力促进了黒枸杞在盐胁迫下的生长发育。

NMT是基因功能的活体检测技术，已被103位诺贝尔奖得主所在单位，及北大、清华、中科院使用。2019年12月17日，由中关村NMT联盟组织的非损伤微测技术（Non-invasive Micro-test Technology, NMT）植物逆境信号转导研究培训，将在北京大学举行。主讲内容非损伤微测技术（Non-invasive Micro-test Technology, NMT）是研究功能基因的一种技术，通过检测活体植物根茎叶与内/外环境间Ca2+/Cd2+/Na+/K+/N/O2...交换量的实时变化，揭示：稻瘟病菌利用K通道抑制先天免疫的机制COLD1等位基因赋予水稻的耐寒新机制丙酮酸载体调控植物耐镉的机理干旱胁迫如何促进植物的氮吸收......培训内容：NMT理论应用培训、专家分享、上机演示时间：2019年12月17日  星期三  9：00～12：00地点：北京大学生命科学学院金光楼311孙 健  非损伤微测技术资深应用专家，江苏师范大学副教授，兼任中关村NMT联盟副理事长、NMT国际标准化委员会（NISC）委员。孙健副教授是国内首篇高水平NMT盐胁迫文章的第一作者（Sun  J, et al. Plant Physiol, 2009, 149:   1141-1153.），单篇引用次数已高达265次。其博士期间（导师：陈少良教授）从事胡杨抗盐机制研究，目前专注于甘薯抗盐/抗病机制研究，现为国家甘薯产业技术体系“细胞遗传与倍性育种技术”岗位团队骨干成员。其所在实验室是国内首批“中关村NMT联盟创新与服务中心”。截止2019年，已利用NMT发表SCI文献20余篇（第一/通讯）。刘蕴琦  NMT高级应用顾问中关村非损伤微测技术产业联盟秘书长，《NMT  101问》、《NMT论文集》副主编。协助中国农大武维华院士、中科院植物所种康院士等课题组，设计了Cell、Plant   Cell等研究成果中的非损伤离子流实验部分。近5年，直接协助国内学者发表的SCI文章共67篇，累计影响因子308.64。巨肖宇  中关村NMT联盟认证高级售后工程师拥有5年以上非损伤微测系统操作应用、售后经验，累计售后服务近五十家单位，售后服务经验3000+小时。Cu、Pb流速传感器商业化研发团队骨干成员。非损伤微测技术目前可检测的指标有：H+、Ca2+、Na+、K+、Cl-、Mg2+、Cd2+、Cu2+、Pb2+、NH４+、NO3-、O2、H2O2、IAA。        

NMT是基因功能的活体检测技术，已被103位诺贝尔奖得主所在单位，及北大、清华、中科院使用。期刊：Journal of Plant Nutrition And Soil Science主题：低氮提高云杉氮利用率新机制标题：Morphological and physiological responses of Picea asperata to different nitrogen availability and pH影响因子：2.057检测指标：NH4+、NO3-、H+流速检测样品：云杉幼苗根系NH4+、NO3-、H+流实验处理方法：50 or 1000 mM NH4NO3 with pH5 or pH7，发芽后生长8周用于测定NH4+、NO3-、H+流实验测试液成份：0.1mM KCl, 0.1mM CaCl2, pH5 or pH7作者：中科院成都生物所尹春英、唐波英文摘要Soil nitrogen (N) availability and pH are two   determinants affecting plant growth, both of which are influenced by   long‐term N deposition. However, the physiological mechanism of plants   response to the changes in soil N availability and pH are not fully   understood.To investigate the response of Picea asperata to both  factors, seedlings of P. asperata were exposed to 50 or 1000 μM NH4NO3  with pH 5 or pH 7. In the current study, P. asperata, regardless of N   availability and pH in growth medium, exhibited invariably a   urn:x-wiley:14368730:media:jpln201900103-math-0001 preference. Lower   root biomass, root : shoot mass ratio, total root length and area, and   root vitality were detected in high N condition compared to those in low  N supply, corresponding well to lower net influxes of   urn:x-wiley:14368730:media:jpln201900103-math-0002 and   urn:x-wiley:14368730:media:jpln201900103-math-0003 at the root surface   in both pH treatments. These results indicate that P. asperata may   employ an active‐forge strategy to exploit nutrient resources for growth  under low N availability, probably by increased below‐ground carbon   allocation and net influxes of   urn:x-wiley:14368730:media:jpln201900103-math-0004 and   urn:x-wiley:14368730:media:jpln201900103-math-0005.Although low  pH, to some extent may generate more malondialdehyde, P. asperata would  enhance pH tolerance by increased detoxification, i.e., antioxidant  enzymes (peroxidase), free proline and soluble protein as well as  improved carbohydrate status (i.e., soluble sugar and starch). 中文摘要（谷歌机翻）土壤氮（N）的有效性和pH是影响植物生长的两个决定因素，这两个因素均受长期氮沉降的影响。但是，尚未完全了解植物对土壤氮素利用率和pH值变化的生理机制。为了研究云杉对两种因素的响应，将云杉幼苗暴露于50或1000  μM NH4NO3，pH 5或pH   7。展示了urn：x-wiley：14368730：media：jpln201900103-math-0001偏好设置。与低氮供应相比，高氮条件下根系生物量，根：茎总质量比，总根长和面积以及根系活力较低，这与：x-wiley：14368730：培养基的净入渗量较低相对应。在两种pH处理中，根表面的jpln201900103-math-0002和urn：x-wiley：14368730：media：jpln201900103-math-0003这些结果表明，P。asperata可能采用主动锻造策略来开发养分资源，以在低氮可利用性下进行生长，这可能是由于地下碳分配增加以及骨净：x-wiley：14368730：media：jpln201900103-math-0004和urn：x-wiley：14368730：media：jpln201900103-math-0005。尽管低pH值在一定程度上可能产生更多的丙二醛，但曲霉假单胞菌会通过增加解毒作用（即抗氧化酶（过氧化物酶），游离脯氨酸和可溶性蛋白以及改善的碳水化合物状态（即可溶性糖和淀粉））提高pH耐受性。结果表明：无论pH是5或7，低N的NH4+吸收速率高于高N；不同pH的NH4+吸收速率并无显著差异（图b）。低N的NO3-吸收速率同样高于高N，pH   5时两种N的NO3-吸收速率均低于pH7（图c）。相同pH条件下，低N的H+外排速率高于高N；同样N水平下，不同pH的H+外排无显著差异（图d）。由此可见，云杉通过扩大吸收区域和提高吸收速率等形态和生理的一系列调控从而适应土壤中的限制性N环境。

        NMT是基因功能的活体检测技术，已被103位诺贝尔奖得主所在单位，及北大、清华、中科院使用。NMT历史上的今天2013年12月09日，华中农业大学张献龙、涂礼莉、唐文鑫用NMT在New Phytologist上发表了标题为The calcium sensor GhCaM7 promotes cotton fiber elongation by modulating reactive oxygen species (ROS) production的研究成果。期刊：New Phytologist主题：GhCaM7调节活性氧（ROS）促进棉纤维伸长标题：The calcium sensor GhCaM7 promotes cotton fiber elongation by modulating reactive oxygen species (ROS) production影响因子：6.736检测指标：Ca2+流速作者：华中农业大学张献龙、涂礼莉、唐文鑫英文摘要Fiber  elongation is the key determinant of ?ber quality and output in cotton  (Gossypium hirsutum). Although expression pro?ling and functional  genomics provide some data, the mechanism of ?ber development is still  not well understood.Here, a gene encoding a calcium sensor,  GhCaM7, was isolated based on its high expres-sion level relative to  other GhCaMs in ?ber cells at the fast elongation stage. The level of  expression of GhCaM7 in the wild-type and the fuzzless/lintless mutant  correspond to the presence and absence, respectively, of ?ber initials.Overexpressing  GhCaM7 promotes early ?ber elongation, whereas GhCaM7 suppression by  RNAi delays ?ber initiation and inhibits ?ber elongation. Reactive  oxygen species (ROS) play important roles in early ?ber development. ROS  induced by exogenous hydrogen perox-ide (H2O2) and Ca2+ starvation  promotes early ?ber elongation. GhCaM7 overexpression ?ber cells show  increased ROS concentrations compared with the wild-type, while GhCaM7  RNAi ?ber cells have reduced concentrations. Furthermore, we show that  H2O2 enhances Ca2+ in?ux into the ?ber and feedback-regulates the  expression of GhCaM7.We conclude that GhCaM7, Ca2+ and ROS are  three important regulators involved in early ?ber elongation. GhCaM7  might modulate ROS production and act as a molecular link between Ca2+  and ROS signal pathways in early ?ber development.中文摘要（谷歌机翻）纤维伸长率是决定棉花（陆地棉）纤维质量和产量的关键因素。尽管表达谱和功能基因组学提供了一些数据，但是纤维发育的机制仍未得到很好的理解。在此，基于钙离子传感器GhCaM7的表达水平相对于其在纤维细胞中处于快速延伸阶段的其他GhCaMs高表达水平而被分离出来。GhCaM7在野生型和无绒毛/无绒毛突变体中的表达水平分别对应于纤维首字母的存在与否。过表达的GhCaM7促进了早期纤维的伸长，而RNAi对GhCaM7的抑制则延迟了纤维的启动并抑制了纤维的伸长。活性氧（ROS）在纤维早期发育中起重要作用。外源过氧化氢（H2O2）和Ca2+饥饿诱导的ROS促进了早期纤维的伸长。与野生型相比，GhCaM7过表达纤维细胞的ROS浓度升高，而GhCaM7  RNAi纤维细胞的ROS浓度降低。此外，我们表明，H2O2增强了Ca2+的入流，并反馈调节GhCaM7的表达。我们得出结论，GhCaM7，Ca2+和ROS是参与早期纤维伸长的三个重要调控因子。GhCaM7可能调节ROS的产生，并在早期纤维发育中充当Ca2+和ROS信号通路之间的分子链接。(b) Ca2+ ?ux during ?ber development was measured by a  noninvasive scanning ion-selective electrode technique. During ?ber   elongation, there was an increasing Ca2+in?ux from the extracellular to the intracellular environment and the ?ux rate peaked during   plasmodesmatal closure from 10 to 15 d postanthesis (DPA). The rate  of Ca2+in?ux subsequently decreased during the secondary cell wall   synthesis. The inset (lower left) is a schematic diagram for the   measurement of Ca2+ ?ux in testing buffer by noninvasive scanning   ion-selective electrode. Error bars,  SD of nine samples.

2019年11月中关村NMT联盟全国测试服务网信息中关村NMN联盟创新与服务中心（测试中心）一览

NMT是基因功能的活体检测技术，已被103位诺贝尔奖得主所在单位，及北大、清华、中科院使用。2019年12月17日，由中关村NMT联盟组织的非损伤微测技术（Non-invasive Micro-test Technology, NMT）植物逆境信号转导研究培训，将在北京大学举行。主讲内容非损伤微测技术（Non-invasive Micro-test Technology, NMT）是研究功能基因的一种技术，通过检测活体植物根茎叶与内/外环境间Ca2+/Cd2+/Na+/K+/N/O2...交换量的实时变化，揭示：稻瘟病菌利用K通道抑制先天免疫的机制COLD1等位基因赋予水稻的耐寒新机制丙酮酸载体调控植物耐镉的机理干旱胁迫如何促进植物的氮吸收......培训内容：NMT理论应用培训、专家分享、上机演示时间：2019年12月17日  星期三  9：00～12：00地点：北京大学生命科学学院金光楼311联系人：程老师  18515811370孙 健  非损伤微测技术资深应用专家，江苏师范大学副教授，兼任中关村NMT联盟副理事长、NMT国际标准化委员会（NISC）委员。孙健副教授是国内首篇高水平NMT盐胁迫文章的第一作者（Sun J, et al. Plant Physiol, 2009, 149: 1141-1153.），单篇引用次数已高达265次。其博士期间（导师：陈少良教授）从事胡杨抗盐机制研究，目前专注于甘薯抗盐/抗病机制研究，现为国家甘薯产业技术体系“细胞遗传与倍性育种技术”岗位团队骨干成员。其所在实验室是国内首批“中关村NMT联盟创新与服务中心”。截止2019年，已利用NMT发表SCI文献20余篇（第一/通讯）。刘蕴琦  NMT高级应用顾问中关村非损伤微测技术产业联盟秘书长，《NMT 101问》、《NMT论文集》副主编。协助中国农大武维华院士、中科院植物所种康院士等课题组，设计了Cell、Plant Cell等研究成果中的非损伤离子流实验部分。近5年，直接协助国内学者发表的SCI文章共67篇，累计影响因子308.64。巨肖宇  中关村NMT联盟认证高级售后工程师拥有5年以上非损伤微测系统操作应用、售后经验，累计售后服务近五十家单位，售后服务经验3000+小时。Cu、Pb流速传感器商业化研发团队骨干成员。非损伤微测技术目前可检测的指标有：H+、Ca2+、Na+、K+、Cl-、Mg2+、Cd2+、Cu2+、Pb2+、NH４+、NO3-、O2、H2O2、IAA。        

NMT是基因功能的活体检测技术，已被103位诺贝尔奖得主所在单位，及北大、清华、中科院使用。NMT历史上的今天2011年12月02日，河北师范大学尚忠林、郝立华、王伟霞、陈忱利用NMT在Molecular Plant 上发表了标题为Extracellular ATP promotes stomatal opening of Arabidopsis thaliana through heterotrimeric G protein a subunit and reactive oxygen species的研究成果。期刊：Molecular Plant主题：胞外ATP通过异源G蛋白和ROS促进气孔开放标题：Extracellular ATP promotes stomatal opening of Arabidopsis thaliana through heterotrimeric G protein a subunit and reactive oxygen species影响因子：4.296检测指标：Ca2+、H+流速作者：河北师范大学尚忠林、郝立华、王伟霞、陈忱英文摘要In recent years, adenosine tri-phosphate (ATP) has been reported to exist in apoplasts of plant cells as a signal molecule. Extracellular ATP (eATP) plays important roles in plant growth, development, and stress tolerance.Here, extracellular ATP was found to promote stomatal opening of Arabidopsis thaliana in light and darkness. ADP, GTP, and weakly hydrolyzable ATP analogs (ATPγS, Bz-ATP, and 2meATP) showed similar effects, whereas AMP and adenosine did not affect stomatal movement. Apyrase inhibited stomatal opening. ATP-promoted stomatal opening was blocked by an NADPH oxidase inhibitor (diphenylene iodonium) or deoxidizer (dithiothreitol), and was impaired in null mutant of NADPH oxidase (atrbohD/F).Added ATP triggered ROS generation in guard cells via NADPH oxidase. ATP also induced Ca2+ influx and H+ efflux in guard cells. In atrbohD/F, ATP-induced ion flux was strongly suppressed. In null mutants of the heterotrimeric G protein α subunit, ATP-promoted stomatal opening, cytoplasmic ROS generation, Ca2+ influx, and H+ efflux were all suppressed.These results indicated that eATP-promoted stomatal opening possibly involves the heterotrimeric G protein, ROS, cytosolic Ca2+, and plasma membrane H+-ATPase.中文摘要（谷歌机翻）近年来，已经报道三磷酸腺苷（ATP）作为信号分子存在于植物细胞的质外体中。细胞外ATP（eATP）在植物生长，发育和胁迫耐受性中起重要作用。在这里，发现细胞外ATP在光与暗中促进拟南芥气孔的开放。ADP，GTP和弱水解性ATP类似物（ATPγS，Bz-ATP和2meATP）显示出相似的作用，而AMP和腺苷并不影响气孔运动。磷酸酶抑制气孔开放。ATP促进的气孔开放被NADPH氧化酶抑制剂（二亚苯基碘鎓）或脱氧剂（二硫苏糖醇）阻止，并且在NADPH氧化酶的无效突变体（atrbohD / F）中受损。添加的ATP通过NADPH氧化酶触发了保卫细胞中ROS的生成。ATP还诱导了保卫细胞中Ca2 +的流入和H +的流出。在atrbohD / F中，ATP诱导的离子通量被强烈抑制。在异源三聚体G蛋白α亚基的无效突变体中，ATP促进的气孔开放，细胞质ROS生成，Ca2 +涌入和H +外排均被抑制。这些结果表明，eATP促进的气孔开放可能涉及异三聚体G蛋白，ROS，胞质Ca2 +和质膜H + -ATPase。Figure 6ATP Stimulates Ca2+ Influx and H+Efflux in Guard Cells of Arabidopsis thaliana (Ecotype col-0).(A) Gadolinium chloride (50 μM) blocked ATP-promoted stomatal opening.(B) and (C) show time course of Ca2+ flux in MES buffer without (B) or with (C) epidermis before and after 0.6 mM ATP treatment, respectively. The arrow marks time points of ATP treatment.(D) The dose-dependence of ATP-promoted Ca2+ influx; data are the means ± SE (n = 30) of the peak Ca2+ influx value after ATP stimulation. The positive and negative values of ion flux represent ion influx and efflux, respectively.(E) Sodium vanadate (100 μM) blocked ATP-promoted stomatal opening.(F) and (G) show time courses of H+ flux in MES buffer without (F) or with (G) epidermis before and after 0.6 mM ATP treatment, respectively.The arrow marks time points of ATP treatment. In (A) and (E), data are means ± SE (n = 6) for stomatal aperture. In all figures, ‘control’ means treatment with MES buffer only. (H) The dose-dependence of ATP-promoted H+ influx; data are the means ± SE (n = 30) of the peak H+ efflux value after ATP stimulation.

NMT是基因功能的活体检测技术，已被103位诺贝尔奖得主所在单位，及北大、清华、中科院使用。期刊：Plant Cell & Environment主题：沙冬青虫害/盐害交互抗性机制研究标题：Herbivore exposure alters ion fluxes and improves salt tolerance in a desert shrub影响因子：6.125检测指标：Ca2+、K+、Na+、H+流速检测样品：矮沙冬青作者：北京林业大学沈应柏、陈迎迎文章简介沙冬青是西北荒漠唯一的常绿阔叶灌木，是第三纪孑遗植物，国家二级濒危物种。沙冬青生境极其恶劣，常伴随着极端温度、干旱、盐碱、虫害，但目前关于沙冬青生物与非生物交互抗性的研究甚少。因此，沈应柏教授研究团队利用非损伤微测、激光共聚焦等技术，系统研究了经昆虫取食的沙冬青在遭受盐胁迫时的根部离子跨膜转运模式。该研究发现，昆虫取食预处理可能通过激活OPR3活性诱导茉莉酸的积累，茉莉酸信号途径的激活诱导了胞内Ca2+的迅速积累，增强了质膜H+-ATPase活性，促进了胞内过多的Na+经由Na+/H+逆向转运体的外排，同时抑制K+的流失，有助于维持细胞内的K+/Na+平衡，最终增强沙冬青的耐盐性。博士研究生陈迎迎为该论文第一作者，沈应柏教授为通讯作者。该研究相关工作得到国家自然科学基金项目（31270655）的资助。英文摘要Plants have evolved complex mechanisms that allow them to withstand multiple environmental stresses, including biotic and abiotic stresses.Here, we investigated the interaction between herbivore exposure and salt stress of Ammopiptanthus nanus, a desert shrub. We found that jasmonic acid (JA) was involved in plant responses to both herbivore attack and salt stress, leading to an increased NaCl stress tolerance for herbivore-pretreated plants, and increase in K+/Na+ ratio in roots. Further evidence revealed the mechanism by which herbivore improved plant NaCl tolerance. Herbivore pretreatment reduced K+ efflux and increased Na+ efflux in plants subjected to long-term, short-term, or transient NaCl stress.Moreover, herbivore pretreatment promoted H+ efflux by increasing plasma membrane H+-ATPase activity. This H+ efflux creates a transmembrane proton motive force that drives the Na+/H+ antiporter to expel excess Na+ into the external medium. In addition, high cytosolic Ca2+ was observed in the roots of herbivore-treated plants exposed to NaCl, and this effect may be regulated by H+-ATPase.Taken together, herbivore exposure enhances A. nanus tolerance to salt stress by activating the JA signalling pathway, increasing plasma membrane H+-ATPase activity, promoting cytosolic Ca2+ accumulation, and then restricting K+ leakage and reducing Na+ accumulation in the cytosol.中文摘要（谷歌机翻）植物已经进化出复杂的机制，可以承受多种环境胁迫，包括生物和非生物胁迫。在这里，我们调查了草食动物暴露与沙漠灌木沙冬青盐胁迫之间的相互作用。我们发现茉莉酸（JA）参与了植物对食草动物侵袭和盐胁迫的反应，导致食草动物预处理过的植物对NaCl胁迫的耐受性增加，并且根中K+ / Na+比率增加。进一步的证据揭示了草食动物改善植物NaCl耐受性的机制。草食动物预处理可降低长期，短期或短暂NaCl胁迫下植物的K+流出量并增加Na +流出量。此外，草食动物预处理通过增加质膜H+ -ATPase活性来促进H +流出。这种H+外流产生跨膜质子原动力，该原动力驱动Na+ / H+反向转运蛋白将过量的Na+排出到外部介质中。此外，在暴露于NaCl的食草动物处理过的植物的根部中观察到高的胞质Ca2+，这种作用可能受H+ -ATPase调节。总之，通过激活JA信号传导途径，增加质膜H+ -ATPase活性，促进胞质Ca2 +积累，然后限制K+泄漏并减少细胞质中Na +的积累，草食动物暴露增强了南芥对盐胁迫的耐受性。Figure 2. Effects of NaCl on the stable and transient flux of K+ along the A. nanus root axis (from 0 to 2000 μm from the root apex) with or without herbivore pretreatment. (A) The stable K+ flux was recorded along the axis of the root apex (0~2000 μm from the root tip) at 200-μm intervals, after long-term (LT) combined stresses (24-h HE + 7-d NaCl), salt stress (7-d NaCl), or no stress (control). (B) The bar chart represents the mean K+ flux value of all points along the roots following the four treatments. (C) The transient K+ flux kinetics were measured at the surface of the root, 600 μm from the tip, before and after the application of 100 mM NaCl. Three minutes of baseline data were recorded before NaCl application. The arrow indicates the time point of NaCl addition. Ten minutes of data were recorded after NaCl application. Samples were pretreated with tetraethylammonium (TEA), a K+ channel blocker, for 30 min before test. (D) The mean K+ flux was calculated before (pre-exposure), immediately after (peak-response), and 10 min after (post-exposure) the NaCl treatment. Different letters indicate significant differences at P ≤ 0.05 (Student’s t-test). Data were obtained from 5–7 A. nanus individuals. Error bars represent SE.

北京大学NMT设备操作培训顺利完成

2019年9月北京大学通过单一来源的方式，采购了扬格/旭月的国产第七代自动化非损伤微测系统。本周一旭月公司顺利的完成了北京大学NMT设备的安装。安装过程按照中关村NMT联盟认证标准进行。2019年11月27日-2019年11月29日期间旭月将在北京大学进行NMT设备操作培训。培训全程依照中关村NMT联盟颁发的《NMT系统操作培训标准》 。此次旭月派出的是美国扬格（旭月北京）售后部负责人，中关村NMT联盟认证高级售后工程师巨肖宇。拥有5年以上非损伤微测系统操作应用、售后经验，累计售后服务近五十家单位，售后服务经验3000+小时。Cu、Pb流速传感器商业化研发团队骨干成员。        

NMT是基因功能的活体检测技术，已被103位诺贝尔奖得主所在单位，及北大、清华、中科院使用。期刊：Plant journal主题：水稻OsCBL10启动子天然变异影响种子萌发期的耐淹性标题：Natural variation in the promoter of rice   calcineurin B‐like protein10 (Os CBL 10) affects flooding tolerance   during seed germination among rice subspecies影响因子：5.775检测指标：Ca2+流速检测样品：水稻距胚芽鞘尖端不同距离（0、50、100、150、200、250、300、350、400、450、500和550μm）Ca2+流速流实验处理方法：水稻种子水淹胁迫Ca2+流速流实验测试液成份：0.1 mM CaCl2、0.1 mM KCl、0.3 mM MES，pH 6.0作者：香港中文大学张建华、山东农业大学刘鹰高、施璐文章简介2018年5月，香港中文大学张建华、山东农业大学刘鹰高教授课题组在Plant   Journal杂志上发表题为“Natural variation in the promoter of rice Calcineurin   B-like Protein10 (OsCBL10) affects flooding tolerance during seed   germination among rice subspecies”的研究论文。现代栽培水稻来源于旱稻的逐步演化，而这一演化过程的了解是揭示旱稻如何获得水淹抗性的关键，对于其它植物的耐水淹育种研究具有借鉴意义。该研究通过分析水稻和旱稻的钙调磷酸酶B类蛋白（OsCBL10）的启动子区域发现两者存在差异。耐水淹的水稻品种都存在同样的T-Type类型启动子序列，此类型启动子会通过影响OsCBL10表达进而调节水淹胁迫下的Ca2+信号。该项研究中，利用NMT技术检测了两个水稻栽培品种Low88和Up221胚芽鞘以及距离胚芽鞘尖端不同距离部位的Ca2+流速，结果显示两个品种直接的Ca2+流速存在差异。水淹胁迫下水稻胚芽鞘的Ca2+流速变化英文摘要Rice (Oryza sativa L.) has two ecotypes, upland and   lowland rice, that have been observed to show different tolerance levels  under flooding stress.In this study, two rice cultivars, upland  (Up221, flooding‐intolerant) and lowland (Low88, flooding‐tolerant),  were initially used to study their molecular mechanisms in response to  flooding germination. We observed that variations in the OsCBL10  promoter sequences in these two cultivars might contribute to this  divergence in flooding tolerance. Further analysis using another eight  rice cultivars revealed that the OsCBL10 promoter could be classified as  either a flooding‐tolerant type (T‐type) or a flooding‐intolerant type  (I‐type).The OsCBL10 T‐type promoter only existed in japonica  lowland cultivars, whereas the OsCBL10 I‐type promoter existed in  japonica upland, indica upland and indica lowland cultivars.  Flooding‐tolerant rice cultivars containing the OsCBL10 T‐type promoter  have shown lower Ca2+ flow and higher α‐amylase activities in comparison  to those in flooding‐intolerant cultivars.Furthermore, the  OsCBL10 overexpression lines were sensitive to both flooding and hypoxic  treatments during rice germination with enhanced Ca2+ flow in  comparison to wild‐type. Subsequent findings also indicate that OsCBL10  may affect OsCIPK15 protein abundance and its downstream pathways.In  summary, our results suggest that the adaptation to flooding stress during rice germination is associated with two different OsCBL10   promoters, which in turn affect OsCBL10 expression in different   cultivars and negatively affect OsCIPK15 protein accumulation and its   downstream cascade.中文摘要（谷歌机翻）水稻（Oryza sativa L.）有两种生态型，旱稻和低陆稻，在洪水胁迫下它们表现出不同的耐受水平。在这项研究中，最初使用两个水稻品种，高地（Up221，耐水）和低地（Low88，耐水）来研究其对洪水萌发的分子机制。我们观察到，在这两个品种中，OsCBL10启动子序列的变异可能导致这种泛洪耐受性的差异。使用另外八个水稻品种的进一步分析表明，OsCBL10启动子可以分为耐水型（T型）或耐水型（I型）。OsCBL10 T型启动子仅存在于粳稻低地品种中，而OsCBL10 I型启动子则存在于粳稻高地，in稻高地和low稻低地品种中。与耐水不耐旱品种相比，含有OsCBL10 T型启动子的耐水耐旱水稻品种显示出较低的Ca2+流量和较高的α-淀粉酶活性。此外，与野生型相比，OsCBL10过表达品系对水稻发芽期间的水淹和低氧处理均敏感，且Ca2+流量增加。随后的发现还表明，OsCBL10可能会影响OsCIPK15蛋白的丰度及其下游途径。总而言之，我们的结果表明，水稻发芽过程中对洪水胁迫的适应与两个不同的OsCBL10启动子有关，这反过来影响了OsCBL10在不同品种中的表达，并对OsCIPK15蛋白的积累及其下游级联反应产生了负面影响。

NMT是基因功能的活体检测技术，已被103位诺贝尔奖得主所在单位，及北大、清华、中科院使用。  NMT历史上的今天2011年11月25日，山东省农科院董合忠、孔祥强利用NMT在Journal of Experimental Botany 上发表了标题为Effects of non-uniform root zone salinity on water use Na+ recirculation, and Na+ and H+ flux in cotton的研究成果。期刊：Journal of Experimental Botany主题：根系不均一盐胁迫提升植物水分利用率的机制标题：Effects of non-uniform root zone salinity on water use Na+ recirculation, and Na+ and H+ flux in cotton影响因子：4.818检测指标：Na+、H+流速作者：山东省农科院董合忠、孔祥强英文摘要A new split-root system was established through   grafting to study cotton response to non-uniform salinity. Each root   half was treated with either uniform (100/100?mM) or non-uniform NaCl   concentrations (0/200 and 50/150?mM).In contrast to uniform  control, non-uniform salinity treatment improved plant growth and water  use, with more water absorbed from the non- and low salinity side.  Non-uniform treatments decreased Na+ concentrations in leaves. The [Na+]  in the ‘0’ side roots of the 0/200 treatment was significantly higher  than that in either side of the 0/0 control, but greatly decreased when  the ‘0’ side phloem was girdled, suggesting that the increased [Na+] in  the ‘0’ side roots was possibly due to transportation of foliar Na+ to  roots through phloem.Plants under non-uniform salinity extruded  more Na+ from the root than those under uniform salinity. Root Na+  efflux in the low salinity side was greatly enhanced by the higher  salinity side. NaCl-induced Na+ efflux and H+ influx were inhibited by  amiloride and sodium orthovanadate, suggesting that root Na+ extrusion  was probably due to active Na+/H+ antiport across the plasma membrane.Improved  plant growth under non-uniform salinity was thus attributed to  increased water use, reduced leaf Na+ concentration, transport of   excessive foliar Na+ to the low salinity side, and enhanced Na+ efflux   from the low salinity root.中文摘要（谷歌机翻）通过嫁接建立了一个新的分根系统，以研究棉花对盐度不均匀的响应。每个根部均用均一的（100/100 nonmM）或不均一的氯化钠浓度（0/200和50/150 mM）处理。与统一控制相反，非均匀盐度处理改善了植物生长和水分利用，从非盐度和低盐度侧吸收了更多的水。不均匀处理降低了叶片中的Na+浓度。0/200处理的'0'侧根中的[Na+]显着高于0/0对照中任一侧的[Na+]，但当'0'侧韧皮部被束腰时，其[Na+]大大降低，表明[  [0]侧根中的Na+]可能是由于叶面Na+通过韧皮部转运至根。盐度不均匀的植物比盐度均匀的植物从根部挤出的Na+更多。低盐度一侧的根Na+流出量被高盐度一侧大大增强。NaCl诱导的Na+流出和H+流入被阿米洛利和原钒酸钠抑制，这表明根Na+的挤出可能是由于跨质膜的活性Na+ / H+逆向转运所致。因此，在非均匀盐度下植物生长的改善归因于增加的水分利用，降低的叶片Na+浓度，过量的叶面Na+向低盐度侧的运输以及从低盐度根部提高的Na+外排。Effects of non-uniform (0/200 mM and 50/150 mM NaCl) and uniform (0/0, 50/50, 100/100, and 150/150 mM NaCl) root zone   salinity on net Na+ ?uxes in roots of cotton at 1 d (A) and 7 d (B)   after treatment. The data are main ?uxes of Na+ within the measuring   periods (15 min). Values are means 6SE (n?6). Bars with different   letters (a, b, and c) differ signi?cantly at P

NMT是基因功能的活体检测技术，已被103位诺贝尔奖得主所在单位，及北大、清华、中科院使用。近期，北京大学现代农学院向联盟提出理论应用培训申请，现将已确定信息对外发布，供大家参考。会员相关新闻                

今天上午，联盟认证的旭月售后工程师受联盟邀请，在西农旱区作物逆境生物学国家重点实验室进行了上机演示。现场不仅有旱区作物逆境生物学国家重点实验室的老师同学，还有来自植保学院、园艺学院、生科院等其他学院的老师同学参加了培训。此外，联盟还邀请了林学院鲁彦君老师，做题为《NMT在植物耐盐机理研究上的应用报告》的报告。                

NMT是基因功能的活体检测技术，已被31位诺贝尔奖得主所在单位，及北大、清华、中科院使用。期刊：Development主题：固醇调控FLS2蛋白胞吞的新机制（钙信号）标题：Sterols regulate endocytic pathways during flg22-induced defense responses in Arabidopsis影响因子：5.413检测指标：Ca2+流速检测样品：拟南芥叶片Ca2+流速流实验处理方法：7日龄拟南芥，0.1 μM flg22处理Ca2+流速流实验测试液成份：无推荐测试液：0.1mM CaCl2，pH 6.0作者：北京林业大学林金星、李晓娟英文摘要The plant transmembrane receptor kinase FLAGELLIN   SENSING 2 (FLS2) is crucial for innate immunity. Although previous   studies have reported FLS2-mediated signal transduction and endocytosis via the clathrin-mediated pathway, whether additional endocytic pathways  affect FLS2-mediated defense responses remains unclear.Here, we  show that the Arabidopsis thaliana sterol-deficient mutant steroid  methyltransferase 1 displays defects in immune responses induced by the  flagellin-derived peptide flg22. Variable-angle total internal  reflection fluorescence microscopy (VA-TIRFM) coupled with   single-particle tracking showed that the spatiotemporal dynamics of   FLS2-GFP changed on a millisecond time scale and that the FLS2-GFP dwell  time at the plasma membrane increased in cells treated with a   sterol-extracting reagent when compared with untreated counterparts.We  further demonstrate that flg22-induced FLS2 clustering and endocytosis  involves the sterol-associated endocytic pathway, which is distinct from  the clathrin-mediated pathway. Moreover, flg22 enhanced the  colocalization of FLS2-GFP with the membrane microdomain marker Flot  1-mCherry and FLS2 endocytosis via the sterol-associated pathway. This  indicates that plants may respond to pathogen attacks by regulating two  different endocytic pathways.Taken together, our results suggest the key role of sterol homeostasis in flg22-induced plant defense responses.中文摘要（谷歌机翻）植物跨膜受体激酶FLAGELLIN SENSING 2（FLS2）对于先天免疫至关重要。尽管以前的研究已经报道了通过网格蛋白介导的途径介导的FLS2介导的信号转导和胞吞作用，但尚不清楚其他内吞途径是否会影响FLS2介导的防御反应。在这里，我们显示拟南芥的甾醇缺乏突变体类固醇甲基转移酶1在鞭毛蛋白衍生的肽flg22诱导的免疫反应中显示缺陷。可变角度全内反射荧光显微镜（VA-TIRFM）结合单粒子跟踪显示，在处理的细胞中，FLS2-GFP的时空动态在毫秒级变化，质膜上的FLS2-GFP停留时间增加与未经处理的对应物相比，含固醇提取剂。我们进一步证明，flg22诱导的FLS2聚集和内吞涉及与固醇相关的内吞途径，这不同于网格蛋白介导的途径。此外，flg22通过固醇相关途径增强了FLS2-GFP与膜微区标记Flot  1-mCherry和FLS2内吞的共定位。这表明植物可以通过调节两种不同的内吞途径来响应病原体侵袭。两者合计，我们的结果表明固醇稳态在flg22诱导的植物防御反应中的关键作用。结果表明：相比野生型，smt1（甾醇甲基转移酶1）突变体表现出迅速且强烈的Ca2+吸收速率高于根冠（R0和R1），低于根伸长区（R2）。增加，说明突变体对于flg22更为敏感。smt1突变没有改变FLS2的同源寡聚状态，但影响FLS2簇形成。smt1突变体中FLS2的内吞功能受到损伤。由上述结果得到一个假设，即甾醇相关的内吞途径对于flg22诱导的FLS2动力学和植物防御至关重要。

NMT是基因功能的活体检测技术，已被31位诺贝尔奖得主所在单位，及北大、清华、中科院使用。期刊：Chemosphere主题：浮萍超富集Cd的分子机制研究标题：Comparative transcriptome analysis of duckweed   (Landoltia punctata) in response to cadmium provides insights into   molecular mechanisms underlying hyperaccumulation影响因子：4.427检测指标：Cd2+流速检测样品：浮萍根（根尖、距离根尖400μm，1000μm）Cd2+流速流实验处理方法：50 μM CdCl2处理30minCd2+流速流实验测试液成份：0.1 mM KCl，0.1 mM MgCl2、0.05 mM CaCl2、0.3 mM MES、50μM CdCl2，pH5.5作者：中科院青岛生物能源与过程研究所周功克、徐华英文摘要Cadmium (Cd) is a detrimental environmental   pollutant. Duckweeds have been considered promising candidates for Cd   phytoremediation. Although many physiological studies have been   conducted, the molecular mechanisms underlying Cd hyperaccumulation in   duckweeds are largely unknown.In this study, clone 6001 of  Landoltia punctata, which showed high Cd tolerance, was obtained by  large-scale screening of over 200 duckweed clones. Subsequently, its  growth, Cd flux, Cd accumulation, and Cd distribution characteristics  were investigated. To further explore the global molecular mechanism, a  comprehensive transcriptome analysis was performed. For RNA-Seq, samples  were treated with 20 μM CdCl2 for 0, 1, 3, and 6 days.In total,  9,461, 9,847, and 9615 differentially expressed unigenes (DEGs) were  discovered between Cd-treated and control (0 day) samples. DEG  clustering and enrichment analysis identified several biological   processes for coping with Cd stress. Genes involved in DNA repair acted as an early response to Cd, while RNA and protein metabolism would be   likely to respond as well.Furthermore, the carbohydrate  metabolic flux tended to be modulated in response to Cd stress, and  upregulated genes involved in sulfur and ROS metabolism might cause high  Cd tolerance. Vacuolar sequestration most likely played an important  role in Cd detoxification in L. punctata 6001. These novel findings  provided important clues for molecular assisted screening and breeding  of Cd hyperaccumulating cultivars for phytoremediation.中文摘要（谷歌机翻）镉（Cd）是有害的环境污染物。浮萍被认为是镉植物修复的有前途的候选人。尽管已进行了许多生理研究，但浮萍中Cd过度富集的分子机制尚不清楚。在这项研究中，通过大规模筛选200多个浮萍克隆，获得了对镉具有高耐受性的Landoltia   punctata克隆6001。随后，研究了其生长，Cd通量，Cd积累和Cd分布特征。为了进一步探索全局分子机制，进行了全面的转录组分析。对于RNA-Seq，样品用20μMCdCl2处理0、1、3和6天。在经镉处理的样品和对照（0天）样品之间总共发现了9,461、9,847和9615个差异表达的单基因（DEG）。 DEG聚类和富集分析确定了应对Cd胁迫的几种生物学过程。参与DNA修复的基因是对Cd的早期反应，而RNA和蛋白质代谢也可能也有响应。此外，碳水化合物的代谢通量倾向于响应Cd胁迫而被调节，而参与硫和ROS代谢的基因上调可能导致对Cd的高耐受性。液泡螯合最有可能在点球藻6001的Cd解毒中发挥重要作用。这些新发现为分子辅助筛选和育种Cd超富集植物进行植物修复提供了重要线索。结果表明：叶片（F）的Cd2+吸收速率高于根冠（R0和R1），低于根伸长区（R2）。叶片中，从节点（F2）到顶点（F0）的Cd2+吸收速率逐渐下降。根和叶片的Cd2+吸收速率显著较低,从而更容易适应Cd胁迫。