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NMT历史上的今天丨J Proteome Res:胞外Ca2+内流对松果花粉管发育影响的蛋白质组学和细胞学综合分析

2008年04月01日,中科院植物研究所林金星、吴小琴用NMT在Journal of Proteome Research上发表了标题为Integrative proteomic and cytological analysis of the effects of extracellular Ca2+ influx on Pinus bungeana pollen tube development的研究成果。

  • 期刊:J Proteome Res
  • 主题:胞外Ca2+内流对松果花粉管发育影响的蛋白质组学和细胞学综合分析
  • 标题:Integrative proteomic and cytological analysis of the effects of extracellular Ca2+ influx on Pinus bungeana pollen tube development
  • 影响因子:5.684
  • 检测指标:Ca2+流速
  • 通讯作者:中科院植物研究所林金星吴小琴

英文摘要

Ca2+ is an essential ion in the control of pollen germination and tube growth. However, the control of pollen tube development by Ca2+ signaling and its interactions with cytoskeletal components, energy-providing pathways, and cell-expansion machinery remain elusive. Here, we used nifedipine (Nif) to study Ca2+ functions in differential protein expression and other cellular processes in Pinus bungeana pollen tube growth.

Proteomics analysis indicated that 50 proteins showed differential expression with varying doses of Nif. Thirty-four of these were homologous to previously reported proteins and were classified into different functional categories closely related to tip-growth machinery.

Blocking the L-type Ca2+ channel with Nif in the pollen tube membrane induced several early alterations within a short time, including a reduction of extracellular Ca2+ influx and a subsequently dramatic decrease in cytosolic free Ca2+ concentration ([Ca2+]c), concomitant with ultrastructural abnormalities and changes in the abundance of proteins involved in energy production and signaling. Secondary alterations included actin filament depolymerization, disrupted patterns of endocytosis/exocytosis, and cell wall remodeling, along with changes in the proteins involved in these processes.

These results suggested that extracellular Ca2+ influx was necessary for the maintenance of the typical tip-focused [Ca2+]c gradient in the P. bungeana pollen tube, and that reduced adenosine triphosphate production (ATP), depolymerization of the cytoskeleton, and abnormal endocytosis/exocytosis, together with enhanced rigidity of cell walls, were responsible for the growth arrest observed in pollen tubes treated with Nif.

 

中文摘要(谷歌机翻)

Ca2+是控制花粉萌发和管生长的必需离子。但是,通过Ca2+信号传导控制花粉管发育及其与细胞骨架成分,能量提供途径和细胞扩增机制的相互作用仍然难以捉摸。在这里,我们使用硝苯地平(Nifdipine)研究松果花粉花粉管生长中Ca2+在差异蛋白表达和其他细胞过程中的功能。

蛋白质组学分析表明,随着Nif剂量的变化,50种蛋白质表现出差异表达。其中的34个与先前报道的蛋白质同源,并被归类为与尖端生长机制密切相关的不同功能类别。

在花粉管膜中用Nif阻断L型Ca2+通道可在短时间内引起数个早期变化,包括细胞外Ca2+内流减少和随后胞浆游离Ca2+浓度([Ca2+] c)急剧下降,并伴有超微结构能量产生和信号传导中涉及的蛋白质异常和丰度变化。次要的变化包括肌动蛋白丝解聚,内吞/胞吐作用的破坏模式,细胞壁重塑以及这些过程中涉及的蛋白质变化。

这些结果表明,胞外Ca2+内流对于维持P. bungeana花粉管中典型的尖端集中[Ca2+] c梯度是必要的,并且减少了三磷酸腺苷的产生(ATP),细胞骨架的解聚以及异常的内吞/胞吐作用以及细胞壁的刚性增强是造成在用Nif处理的花粉管中观察到的生长停滞的原因。

Figure 1. Dose-dependent inhibitory effects of exogenous Nif on extracellular Ca2+ influx at the pollen tube tips. A typical extracellular Ca2+ influx can be detected in the very tip of control pollen tube, and 100 µM Nif treatment decreased the Ca2+ influx, while 250 and 500 µM Nif treatment greatly reduced this Ca2+ influx.

新冠肺炎干细胞治疗NMT创新平台

 

植物重金属创新科研平台成果:Cu-Cd、Zn-Cd、Cu-Zn复合污染对水稻毒性和重金属吸收的影响

转自中关村旭月非损伤微测技术产业联盟

 


期刊:环境污染与防治
主题:Cu-Cd、Zn-Cd、Cu-Zn复合污染对水稻毒性和重金属吸收的影响
样品:水稻
检测指标:Cu2+、Cd2+
作者:首都师范大学生命科学学院王学东、季冬雪

摘要

以"中花11号"水稻品种为研究对象,通过溶液培养探讨了不同浓度配比的Cu-Cd、Zn-Cd、Cu-Zn对水稻根相对伸长毒性及元素吸收的影响。

结果表明,Cu、Cd、Zn浓度与水稻相对根伸长均符合剂量效应关系,3种金属离子对水稻毒性次序为Cu>Cd>Zn;在重金属总量为50%的毒性效应浓度(EC50)下,Cu-Cd对水稻毒性因浓度配比不同而表现不同的作用方式,而Zn-Cd、Cu-Zn对水稻毒性均表现出拮抗作用。在Cu-Cd复合处理下,Cd的加入促进了水稻对Cu的吸收,而Cu的加入对水稻幼苗的Cd吸收表现出不同作用;在Zn-Cd复合处理中,不同浓度Zn的加入均抑制Cd的吸收,使得水稻幼苗中Cd含量降低了9.1%~48.4%;Cu-Zn复合处理中,高浓度Cu的加入(EC40、EC30、EC25)抑制水稻对Zn的吸收。

Zn-Cd拮抗作用主要和Zn抑制了水稻对Cd的吸收有关,而Cu-Zn的拮抗作用可能和高浓度的Cu抑制了Zn的吸收有关。

 

文章链接:http://kns.cnki.net/KCMS/detail/detail.aspx?dbcode=CJFQ&dbname=CJFDLAST2018&filename=HJWR201810013&v=MjkzMTkxRnJDVVJMT2ZZT1JzRnl6bVZiekpMU2ZjZkxHNEg5bk5yNDlFWjRSOGVYMUx1eFlTN0RoMVQzcVRyV00=

 

活性氧创新科研平台成果:中农学者阐明提高籼稻愈伤组织培养力的分子机制

转自中关村旭月非损伤微测技术产业联盟

【Nature Commun】 中国农大付永彩组阐明提高籼稻愈伤组织培养力的分子机制


期刊:Nature Communications
主题:中农学者阐明提高籼稻愈伤组织培养力的分子机制
标题:A common wild rice-derived BOC1 allele reduces callus browning in indica rice transformation
影响因子:11.878
检测指标:H2O2流速
检测样品:水稻愈伤组织
H2O2流实验处理方法:不同水稻传代培养21天的愈伤组织
H2O2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,pH 6.5
作者:中国农业大学付永彩、张坤、苏晶晶、徐敏

英文摘要

Callus browning, a common trait derived from the indica rice cultivar (Oryza sativa L.), is a challenge to transformation regeneration.

Here, we report the map-based cloning of BROWNING OF CALLUS1 (BOC1) using a population derived from crossing Teqing, an elite indica subspecies exhibiting callus browning, and Yuanjiang, a common wild rice accession (Oryza rufipogon Griff.) that is less susceptible to callus browning.

We show that BOC1 encodes a SIMILAR TO RADICAL-INDUCED CELL DEATH ONE (SRO) protein. Callus browning can be reduced by appropriate upregulation of BOC1, which consequently improves the genetic transformation efficiency. The presence of a Tourist-like miniature inverted-repeat transposable element (Tourist MITE) specific to wild rice in the promoter of BOC1 increases the expression of BOC1 in callus.

BOC1 may decrease cell senescence and death caused by oxidative stress. Our study provides a gene target for improving tissue culturability and genetic transformation.

中文摘要(谷歌机翻)

愈伤组织褐变是源自rice稻品种(Oryza sativa L.)的常见性状,对转化再生提出了挑战。

在这里,我们报告了基于图谱的克隆CALLUS1(BOC1)的克隆,该群体来自穿越易感褐变的优良in稻亚种特清和较不易感的常见野生稻种(Oryza rufipogon Griff)杂交的jiang江。愈伤组织褐变。

我们显示,BOC1编码类似到自由基诱导的细胞死亡一(SRO)蛋白。通过适当上调BOC1可以减少愈伤组织的褐变,从而提高遗传转化效率。BOC1启动子中特有的野生稻特有的类似于游客的微型反向重复转座因子(Tourist MITE)的存在增加了BOC1在愈伤组织中的表达。

BOC1可以减少氧化应激引起的细胞衰老和死亡。我们的研究为改善组织可培养性和遗传转化提供了基因靶标。

 

文章链接:https://www.nature.com/articles/s41467-019-14265-0

 

MP西农:钙流在H2S调控ABA信号通路研究中充当关键生理证据

转自中关村旭月非损伤微测技术产业联盟


期刊:Molecular Plant
主题:硫化氢参与调节ABA调控气孔关闭的分子机制
标题:Hydrogen Sulfide Positively Regulates Abscisic Acid Signaling through Persulfidation of SnRK2.6 in Guard Cells
影响因子:10.812
检测指标:Ca2+流速
检测样品:拟南芥保卫细胞
Ca2+流实验处理方法:5周龄拟南芥幼苗,10μM ABA/100μM NaHS瞬时处理
Ca2+流实验测试液成份:Ca2+:0.1 mM KCl, 0.1 mM CaCl2, 0.1 mM MgCl2, 0.5 mM NaCl, 0.3 mM MES, 0.2 mM Na2SO4, pH 6.0
作者:西北农林科技大学李积胜

保卫细胞检测(转自旭月公司)

英文摘要

The phytohormone abscisic acid (ABA) plays pivotal roles in triggering stomatal closure and facilitating adaptation of plants to drought stress. Hydrogen Sulfide (H2S), a small signaling gas molecule, is involved in ABA-dependent stomatal closure. However, how H2S regulates ABA signaling remains largely unclear.

Here, we show that ABA induces the production of H2S catalyzed by L-CYSTEINE DESULFHYDRASE1 (DES1) in guard cells and H2S in turn positively regulates ABA signaling through persulfidation of Open Stomata 1 (OST1)/SNF1-RELATED PROTEIN KINASE2.6 (SnRK2.6). Two cysteine (Cys) sites, Cys131 and Cys137 that are exposed on the surface of SnRK2.6 and closed to the activation loop, were identified to be persulfidated, which promotes the activity of SnRK2.6 and its interaction with ABA response element-binding factor2 (ABF2), a transcription factor downstream of ABA signaling.

When Cys131, Cys137 or both in SnRK2.6 was substituted with Serine (S), H2S-induced SnRK2.6 activity and SnRK2.6-ABF2 interaction were partially (SnRK2.6C131S and SnRK2.6C137S) or completely (SnRK2.6C131SC137S) comprised. Introduction of SnRK2.6C131S, SnRK2.6C137S, or SnRK2.6C131SC137S into ost1-3 mutant could not rescue the mutant phenotype, showing less sensitive to ABA- and H2S-induced stomatal closure and Ca2+ influx as well as increased water loss and decreased drought tolerance.

Taken together, our study reveals a novel post-translational regulatory mechanism of ABA signaling in which H2S persulfidates SnRK2.6 to promote ABA signaling and ABA-induced stomatal closure.

中文摘要(谷歌机翻)

植物激素脱落酸(ABA)在触发气孔关闭和促进植物适应干旱胁迫方面起着关键作用。硫化氢(H2S)是一种小的信号气体分子,与ABA依赖的气孔关闭有关。但是,H2S如何调节ABA信号在很大程度上尚不清楚。

在这里,我们显示ABA诱导L-半胱氨酸脱硫酶1(DES1)在保卫细胞中催化H2S的产生,而H2S则通过开放气孔1(OST1)/ SNF1相关蛋白激酶2.6(SnRK2)的过硫化而积极调节ABA信号传导。.6)。暴露在SnRK2.6表面并靠近激活环的两个半胱氨酸(Cys)位点Cys131和Cys137被鉴定为过硫化的,这促进了SnRK2.6的活性及其与ABA反应元件结合的相互作用factor2(ABF2),ABA信号下游的转录因子。

当SnRK2.6中的Cys131,Cys137或两者都被丝氨酸(S)取代时,H2S诱导的SnRK2.6活性和SnRK2.6-ABF2相互作用部分(SnRK2.6C131S和SnRK2.6C137S)或完全(SnRK2.6C131SC137S)包括在内。将snRK2.6C131S,SnRK2.6C137S或SnRK2.6C131SC137S引入ost1-3突变体无法挽救突变体表型,显示出对ABA和H2S诱导的气孔关闭和Ca2 +流入的敏感性较低,以及水分流失和干旱减少公差。

综上所述,我们的研究揭示了一种新的ABA信号转导后调控机制,其中H2S过硫化SnRK2.6以促进ABA信号转导和ABA诱导的气孔关闭。

文章链接:https://www.sciencedirect.com/science/article/pii/S1674205220300046?via%3Dihub

 

NMT历史上的今天丨Plant J:拟南芥WRKY8与VQ9调节盐胁迫耐受性

转自中关村旭月非损伤微测技术产业联盟

NMT历史上的今天

2013年03月02日,中科院西双版纳热带植物园余迪求、胡彦如用NMT在Plant Journal上发表了标题为Arabidopsis transcription factor WRKY8 functions antagonistically with its interacting partner VQ9 to modulate salinity stress tolerance的研究成果。

 

  • 期刊:Plant Journal
  • 主题:拟南芥WRKY8与VQ9调节盐胁迫耐受性
  • 标题:Arabidopsis transcription factor WRKY8 functions antagonistically with its interacting partner VQ9 to modulate salinity stress tolerance
  • 影响因子:6.582
  • 检测指标:K+流速
  • 通讯作者:中科院西双版纳热带植物园余迪求、胡彦如

英文摘要

The WRKY transcription factors have been demonstrated to play crucial roles in regulating stress responses; however, the exact mechanisms underlying their involvement in stress responses are not fully understood.

Arabidopsis WRKY8 was predominantly expressed in roots and was highly upregulated by salt treatment. Disruption of WRKY8 rendered plants hypersensitive to salt, showing delayed germination, inhibited post‐germination development and accelerated chlorosis. Further investigation revealed that WRKY8 interacted with VQ9, and their interaction decreased the DNA‐binding activity of WRKY8.

The VQ9 protein was exclusively localized in the nucleus, and VQ9 expression was strongly responsive to NaCl treatment. Mutation of VQ9 enhanced tolerance to salt stress, indicating that VQ9 acts antagonistically with WRKY8 to mediate responses to salt stress. The antagonist functions of WRKY8 and VQ9 were consistent with an increased or reduced Na+/K+ concentration ratio, as well as contrasting expression patterns of downstream stress‐responsive genes in salt‐stressed wrky8 and vq9 mutants.

Moreover, chromatin immunoprecipitation (ChIP) assays showed that WRKY8 directly bound the promoter of RD29A under salt conditions. These results provided strong evidence that the VQ9 protein acts as a repressor of the WRKY8 factor to maintain an appropriate balance of WRKY8‐mediated signaling pathways to establish salinity stress tolerance.

 

中文摘要(谷歌机翻)

WRKY转录因子已被证明在调节压力反应中起关键作用。然而,它们参与应激反应的确切机制尚不完全清楚。

拟南芥WRKY8主要在根中表达,并通过盐处理高度上调。WRKY8的破坏使植物对盐高度敏感,表现出延迟的发芽,抑制了发芽后的发育并加速了萎黄病。进一步的研究表明,WRKY8与VQ9相互作用,并且它们的相互作用降低了WRKY8的DNA结合活性。

VQ9蛋白专门位于细胞核中,并且VQ9表达对NaCl处理有强烈反应。VQ9突变增强了对盐胁迫的耐受性,表明VQ9与WRKY8拮抗,介导对盐胁迫的响应。WRKY8和VQ9的拮抗剂功能与Na+ / K+浓度比的增加或降低,以及盐胁迫的wrky8和vq9突变体中下游胁迫反应基因的相反表达模式一致。

此外,染色质免疫沉淀(ChIP)分析表明,WRKY8在盐条件下直接结合RD29A的启动子。这些结果提供了有力的证据,表明VQ9蛋白可作为WRKY8因子的阻遏物,以维持WRKY8介导的信号通路的适当平衡,从而建立盐度胁迫耐受性。

(c) Net K+ efflux in root tips. Seeds were germinated on MS agar medium for 4 days in a vertical manner. The net immediate K+ efflux was measured using the non-injuring technique after the addition of salt. The insert shows the mean efflux rates within the measuring period of 0–20 min.

文章链接:https://onlinelibrary.wiley.com/doi/full/10.1111/tpj.12159