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Genes :北京林业大学丨盐胁迫下钾转运体LrKUP8抑制K+外排(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: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12h
K+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(谷歌机翻译)

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。

文章链接:https://www.mdpi.com/2073-4425/10/8/600

 

 

PBJ IAA流新成果:农科院棉花所、浙江农科院丨AKR2A协调IAA和H2O2积累调控棉纤维伸长

转自中关村旭月非损伤微测技术产业联盟


期刊:Plant Biotechnology Journal
主题:AKR2A协调IAA和H2O2积累调控棉纤维伸长
标题:AKR2A participates in the regulation of cotton fibre development by modulating biosynthesis of verylong- chain fatty acids
影响因子:6.840
检测指标:IAA、H2O2流速
检测样品:棉纤维尖端
IAA、H2O2流实验处理方法:15 DPA
IAA、H2O2流实验测试液成份:0.1 mM KCl, 0.1 mM CaCl2, 0.1 mM MgCl2, 0.5 mM NaCl,0.3 mM MES, 0.2 mM Na2SO4 and 0.1% sucrose, pH 6.0
作者:中国农科院棉花所李付广、浙江省农科院沈国新

英文摘要

The biosynthesis of very‐long‐chain fatty acids (VLCFAs) and their transport are required for fibre development. However, whether other regulatory factors are involved in this process is unknown. We report here that overexpression of an Arabidopsis gene ankyrin repeat‐containing protein 2A (AKR2A) in cotton promotes fibre elongation.

RNA‐Seq analysis was employed to elucidate the mechanisms of AKR2A in regulating cotton fibre development. The VLCFA content and the ratio of VLCFAs to short‐chain fatty acids increased in AKR2A transgenic lines. In addition, AKR2A promotes fibre elongation by regulating ethylene and synergizing with the accumulation of auxin and hydrogen peroxide. Analysis of RNA‐Seq data indicates that AKR2A up‐regulates transcript levels of genes involved in VLCFAs’ biosynthesis, ethylene biosynthesis, auxin and hydrogen peroxide signalling, cell wall and cytoskeletal organization. Furthermore, AKR2A interacted with KCS1 in Arabidopsis both in vitro and in vivo. Moreover, the VLCFA content and the ratio of VLCFAs to short‐chain fatty acids increased significantly in seeds of AKR2A‐overexpressing lines and AKR2A/KCS1 co‐overexpressing lines, while AKR2A mutants are the opposite trend.

Our results uncover a novel cotton fibre growth mechanism by which the critical regulator AKR2A promotes fibre development via activating hormone signalling cascade by mediating VLCFA biosynthesis. This study provides a potential candidate gene for improving fibre yield and quality through genetic engineering.

 

中文摘要(谷歌机翻译)

超长链脂肪酸(VLCFA)的生物合成及其运输是纤维发展所必需的。但是,此过程是否涉及其他监管因素尚不清楚。我们在此报告,棉花中拟南芥基因锚蛋白重复序列蛋白2A(AKR2A)的过表达促进了纤维的伸长。

RNA‐Seq分析用于阐明AKR2A调节棉纤维发育的机制。在AKR2A转基因品系中,VLCFA含量和VLCFA与短链脂肪酸的比率增加。另外,AKR2A通过调节乙烯并与植物生长素和过氧化氢的积累协同作用来促进纤维伸长。RNA-Seq数据分析表明,AKR2A上调参与VLCFAs生物合成,乙烯生物合成,生长素和过氧化氢信号,细胞壁和细胞骨架组织的基因的转录水平。此外,在体外和体内,AKR2A与拟南芥中的KCS1相互作用。此外,AKR2A过表达品系和AKR2A / KCS1共过表达品系的种子中VLCFA含量和VLCFA与短链脂肪酸的比率显着增加,而AKR2A突变体则相反。

我们的结果揭示了一种新颖的棉纤维生长机制,关键调节剂AKR2A通过介导VLCFA生物合成,通过激活激素信号传导级联来促进纤维发育。这项研究为通过基因工程提高纤维产量和质量提供了潜在的候选基因。

 

结果表明:WT中IAA的平均外排速率为4262.30 fmol·cm-2·s-1;而AKR2A-2和AKR2A-57的外排速率平均值分别是14298.01 fmol·cm-2·s-1和13685.83 fmol·cm-2·s-1,显然AKR2A-OE的IAA外排速率明显高于WT。
WT中H2O2的平均外排速率为0.02 pmol·cm-2·s-1;而AKR2A-2和AKR2A-57的吸收速率平均值分别是0.49 pmol·cm-2·s-1和0.80 pmol·cm-2·s-1
相比于WT,AKR2A-OE的IAA外排速率和H2O2吸收速率在纤维伸长过程中明显增加。

文章链接:https://onlinelibrary.wiley.com/doi/full/10.1111/pbi.13221

 

 

J PLANT NUTR SOIL SC :中科院成都生物所丨低氮提高云杉氮利用率新机制(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: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(谷歌机翻译)

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流入: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。

文章链接:https://onlinelibrary.wiley.com/doi/10.1002/jpln.201900103

 

 

Environ Exp Bot:河南农业大学丨水稻根部响应缺氧的分子机制研究(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:Environmental and Experimental Botany
主题:水稻根部响应缺氧的分子机制研究
标题:Comparative morphological and transcriptomic responses of lowland and upland rice to root-zone hypoxia
影响因子:3.712
检测指标:IAA、O2流速
检测样品:水稻(低地和高地两种)距根尖0,200, 300, 600, 900, 1200, 1500, 2500 μm
IAA、O2流速流实验处理方法:10日龄幼苗分别在充足氧(4.5-6.0 mg·L−1)和缺氧(0.9-2.1 mg·L−1)条件下培养5d
IAA、O2流速流实验测试液成份:0.1mM KCl, 0.1mM CaCl2, 0.1mM MgCl2, 0.3mM MES, pH 5.5
作者:河南农业大学赵全志、辛泽毓、刘娟

英文摘要

Lowland and upland rice, as for two ecotypes, that have been exhibited different tolerance levels under hypoxia conditions. However, the molecular mechanisms underlying rice root hypoxia tolerance between them are not fully understood.

This study was performed to assess the morphological, physiological and transcriptional changes of roots in one lowland rice YueFu (YF) and one upland rice IRAT109 (IR) genotype. A morpho-physiological analysis revealed that compared to IR, YF showed less reduction of root length, root and shoot biomass, formed more aerenchyma in the root, and kept more oxygen influxes in root under hypoxia conditions. Indole-3-acetic acid (IAA) fluxes patterns exhibited a different response to hypoxia in YF and IR.

The contents of IAA, ethylene and hydrogen peroxide were significantly increased in YF and IR, but nitric oxide significantly increased only in YF under hypoxic conditions. Subsequently, transcriptome analysis revealed that more differentially expressed genes (DEGs) were identified to respond to hypoxia in YF than IR. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses showed that DEGs in both genotypes enriched in energy metabolism, aerenchyma formation, reactive oxygen species, and cell wall modification, whereas more related DEGs in YF significantly enriched in these pathways than IR. The specific DEGs in YF especially enriched in phytohormone metabolism and signaling, such as auxin, jasmonic acid, and ethylene, but the specific DEGs in IR especially enriched in photosynthesis. All these results demonstrate that YF is more tolerant to hypoxia than IR, and elucidate some specific mechanisms underlying the differential hypoxia tolerance in lowland and upland rice.

This study has provided valuable candidate genes for genetic improvement of rice in adapting to hypoxia stress.

 

中文摘要(谷歌机翻译)

就两种生态型而言,低陆和旱稻在低氧条件下表现出不同的耐受水平。然而,它们之间的水稻根缺氧耐受性的分子机制尚不完全清楚。

这项研究的目的是评估一种低陆稻悦富(YF)和一种陆稻IRAT109(IR)基因型的根的形态,生理和转录变化。形态生理学分析表明,与IR相比,YF在低氧条件下显示的根长,根和茎生物量减少减少,在根中形成更多的气孔,并保持更多的氧流入。吲哚-3-乙酸(IAA)通量模式在YF和IR中显示出对缺氧的不同响应。

在低氧条件下,IAF,乙烯和过氧化氢的含量在YF和IR中显着增加,而一氧化氮仅在YF中显着增加。随后,转录组分析显示,与IR相比,鉴定出更多的差异表达基因(DEG)对YF缺氧有反应。基因本体论(GO)和《京都基因与基因组百科全书》(KEGG)分析表明,两种基因型的DEG均富含能量代谢,通气组织形成,活性氧种类和细胞壁修饰,而YF中更多相关的DEG则显着丰富了这些途径比红外线YF中的特定DEGs尤其富含植物激素代谢和信号传导,例如生长素,茉莉酸和乙烯,而IR中的特定DEGs尤其富含光合作用。所有这些结果表明,YF比IR更耐缺氧,并阐明了低陆稻和旱稻差异耐缺氧性的某些特定机制。

该研究为水稻遗传适应低氧胁迫提供了有价值的候选基因。

 

结果表明:在氧气充足(Aer)的条件下,分生区吸收O2比较剧烈。YF(低地品种)和IR(高地品种)在300μm处吸收的O2最多,其流速值分别达到79.14 pmol·cm−2·s−1和83.72 pmol·cm−2·s−1,而在缺氧(Hyp)条件下分别为44.77 pmol·cm−2·s−1和40.08 pmol·cm−2·s−1;IR明显降低的更多一些。这说明,分生区是水稻根部感受氧的关键区域,IR对缺氧条件更加敏感。
根分生区同时也是IAA较为敏感的区域。缺氧条件下,除IR的300μm处之外,两种水稻均表现为吸收IAA。氧气充足时,YF和IR在600μm处的IAA吸收速率分别为29,740 fmol·cm−2·s-1和35,092 fmol·cm−2·s-1;而在缺氧条件下分别为8864 fmol·cm−2·s-1和5441 fmol·cm−2·s-1。结合IAA含量测定结果,发现缺氧促使根部积累IAA。这说明生长素参与了缺氧条件下根系发育的调控过程。

文章链接:https://www.sciencedirect.com/science/article/pii/S0098847219315138?via%3Dihub

 

Plant Cell Environ:北京林业大学丨沙冬青虫害/盐害交互抗性机制研究(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 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(谷歌机翻译)

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.

文章链接:https://onlinelibrary.wiley.com/doi/10.1111/pce.13662