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Light at the end of the tunnel: FRAP assays combined with super resolution microscopy confirm the presence of a tubular vacuole network in meristematic plant cells 隧道尽头的曙光FRAP 检测与超分辨率显微镜相结合,证实分生植物细胞中存在管状液泡网络
Pub Date : 2024-09-21 DOI: 10.1093/plcell/koae243
David Scheuring, Elena A Minina, Falco Krueger, Upendo Lupanga, Melanie Krebs, Karin Schumacher
Plant vacuoles play key roles in cellular homeostasis, performing catabolic and storage functions, and regulating pH and ion balance. Despite their essential role, there is still no consensus on how vacuoles are established. A model proposing that the endoplasmic reticulum is the main contributor of membrane for growing vacuoles in meristematic cells has been challenged by a study proposing that plant vacuoles are formed de novo by homotypic fusion of multivesicular bodies (MVBs). Here, we use the Arabidopsis thaliana root as a model system to provide a systematic overview of successive vacuole biogenesis stages, starting from the youngest cells proximate to the quiescent center. We combine in vivo high- and super-resolution (STED) microscopy to demonstrate the presence of tubular and connected vacuolar structures in all meristematic cells. Using customized fluorescence recovery after photobleaching (FRAP) assays, we establish different modes of connectivity and demonstrate that thin, tubular vacuoles, as observed in cells near the quiescent center, form an interconnected network. Finally, we argue that a growing body of evidence indicates that vacuolar structures cannot originate from MVBs alone but receive membrane material from different sources simultaneously.
植物液泡在细胞平衡中发挥着关键作用,具有分解代谢和储存功能,并能调节 pH 值和离子平衡。尽管液泡发挥着重要作用,但人们对液泡是如何形成的仍未达成共识。有研究提出,植物液泡是通过多囊体(MVBs)的同型融合从新形成的,这对提出内质网是分生细胞中生长液泡的主要膜贡献者的模型提出了挑战。在这里,我们以拟南芥根为模型系统,从接近静止中心的最年轻细胞开始,系统地概述了液泡的连续生物发生阶段。我们结合体内高分辨和超分辨(STED)显微镜,证明了所有分生组织细胞中都存在管状和相连的液泡结构。利用定制的光漂白后荧光恢复(FRAP)测定,我们建立了不同的连接模式,并证明在静止中心附近的细胞中观察到的细管状液泡形成了一个相互连接的网络。最后,我们认为,越来越多的证据表明,液泡结构不可能仅源于 MVB,而是同时接收来自不同来源的膜材料。
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引用次数: 0
From the archives: On abiotic stress signaling: An ON/OFF switch for the heat stress response in wheat, connecting gibberellin signaling and salt stress, and calcium homeostasis and stress sensitivity. 来自档案:非生物胁迫信号:小麦热胁迫反应的开关、赤霉素信号传导与盐胁迫的联系、钙稳态与胁迫敏感性。
Pub Date : 2024-09-20 DOI: 10.1093/plcell/koae258
Leiyun Yang
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引用次数: 0
From histone acetylation to abundance: The role of GCN5-CAMTA2 interaction in wheat grain weight. 从组蛋白乙酰化到丰度:GCN5-CAMTA2 相互作用在小麦粒重中的作用。
Pub Date : 2024-09-20 DOI: 10.1093/plcell/koae257
Regina Mencia
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引用次数: 0
Seeing red: Light and temperature-dependent complex formation reveal new insights into phytochrome activity. 看见红色:与光和温度有关的复合物形成揭示了植物色素活性的新见解。
Pub Date : 2024-09-20 DOI: 10.1093/plcell/koae255
Rory Osborne
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引用次数: 0
Well prepared: How trichome polymorphism creates an early-warning system against herbivory. 准备充分:毛状体的多态性如何创造出一种防止食草动物侵害的预警系统。
Pub Date : 2024-09-18 DOI: 10.1093/plcell/koae253
Leonard Blaschek
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引用次数: 0
Shedding light on photosystem II components in the dark. 揭示黑暗中的光系统 II 成分。
Pub Date : 2024-09-18 DOI: 10.1093/plcell/koae254
Nora Flynn
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引用次数: 0
NUCLEAR FACTOR-Y–POLYCOMB REPRESSIVE COMPLEX2 dynamically orchestrates starch and seed storage protein biosynthesis in wheat 核因子 Y-POLYCOMB REPRESSIVE COMPLEX2 动态协调小麦淀粉和种子贮藏蛋白的生物合成
Pub Date : 2024-09-18 DOI: 10.1093/plcell/koae256
Jinchao Chen, Long Zhao, Haoran Li, Changfeng Yang, Xuelei Lin, Yujing Lin, Hao Zhang, Mengxia Zhang, Xiaomin Bie, Peng Zhao, Shengbao Xu, David Seung, Xiansheng Zhang, Xueyong Zhang, Yingyin Yao, Dongzhi Wang, Jun Xiao
The endosperm in cereal grains is instrumental in determining grain yield and seed quality, as it controls starch and seed storage protein (SSP) production. In this study, we identified a specific nuclear factor-Y (NF-Y) trimeric complex in wheat (Triticum aestivum L.), consisting of TaNF-YA3-D, TaNF-YB7-B, and TaNF-YC6-B, and exhibiting robust expression within the endosperm during grain filling. Knockdown of either TaNF-YA3 or TaNF-YC6 led to reduced starch but increased gluten protein levels. TaNF-Y indirectly boosted starch biosynthesis genes by repressing TaNAC019, a repressor of cytosolic small ADP-glucose pyrophosphorylase 1a (TacAGPS1a), sucrose synthase 2 (TaSuS2), and other genes involved in starch biosynthesis. Conversely, TaNF-Y directly inhibited the expression of Gliadin-γ-700 (TaGli-γ-700) and low molecular weight-400 (TaLMW-400). Furthermore, TaNF-Y components interacted with SWINGER (TaSWN), the histone methyltransferase subunit of Polycomb repressive complex 2 (PRC2), to repress TaNAC019, TaGli-γ-700, and TaLMW-400 expression through trimethylation of histone H3 at lysine 27 (H3K27me3) modification. Notably, weak mutation of FERTILIZATION INDEPENDENT ENDOSPERM (TaFIE), a core PRC2 subunit, reduced starch but elevated gliadin and LMW-GS contents. Intriguingly, sequence variation within the TaNF-YB7-B coding region was linked to differences in starch and SSP content. Distinct TaNF-YB7-B haplotypes affect its interaction with TaSWN-B, influencing the repression of targets like TaNAC019 and TaGli-γ-700. Our findings illuminate the intricate molecular mechanisms governing TaNF-Y–PRC2-mediated epigenetic regulation for wheat endosperm development. Manipulating the TaNF-Y complex holds potential for optimizing grain yield and enhancing grain quality.
谷物的胚乳控制着淀粉和种子贮藏蛋白(SSP)的产生,因此在决定谷物产量和种子质量方面起着重要作用。在这项研究中,我们在小麦(Triticum aestivum L.)中发现了一种特异的核因子-Y(NF-Y)三聚体复合物,由 TaNF-YA3-D、TaNF-YB7-B 和 TaNF-YC6-B 组成,在谷粒充实过程中在胚乳中表现出强劲的表达。敲除 TaNF-YA3 或 TaNF-YC6 会导致淀粉含量降低,但谷蛋白含量增加。TaNF-Y 通过抑制细胞膜小 ADP-葡萄糖焦磷酸化酶 1a(TacAGPS1a)的抑制因子 TaNAC019、蔗糖合成酶 2(TaSuS2)以及其他参与淀粉生物合成的基因,间接促进了淀粉生物合成基因的表达。相反,TaNF-Y 直接抑制麸质-γ-700(TaGli-γ-700)和低分子量-400(TaLMW-400)的表达。此外,TaNF-Y成分与多聚核糖抑制复合体2(PRC2)的组蛋白甲基转移酶亚基SWINGER(TaSWN)相互作用,通过组蛋白H3在赖氨酸27处的三甲基化(H3K27me3)修饰抑制TaNAC019、TaGli-γ-700和TaLMW-400的表达。值得注意的是,核心 PRC2 亚基 FERTILIZATION INDEPENDENT ENDOSPERM(TaFIE)的弱突变降低了淀粉含量,但提高了麦胶蛋白和 LMW-GS 的含量。有趣的是,TaNF-YB7-B 编码区内的序列变异与淀粉和 SSP 含量的差异有关。不同的 TaNF-YB7-B 单倍型会影响其与 TaSWN-B 的相互作用,从而影响对 TaNAC019 和 TaGli-γ-700 等靶标的抑制。我们的发现揭示了TaNF-Y-PRC2介导的小麦胚乳发育表观遗传调控的复杂分子机制。操纵 TaNF-Y 复合物有望优化谷物产量和提高谷物品质。
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引用次数: 0
The rice R2R3 MYB transcription factor FOUR LIPS connects brassinosteroid signaling to lignin deposition and leaf angle 水稻 R2R3 MYB 转录因子 FOUR LIPS 将黄铜类固醇信号与木质素沉积和叶片角度联系起来
Pub Date : 2024-09-11 DOI: 10.1093/plcell/koae251
Huichao Liu, Jie Zhang, Junxue Wang, Zhibin Fan, Xiaoxiao Qu, Min Yan, Chunxia Zhang, Kezhen Yang, Junjie Zou, Jie Le
Leaf angle is an important agronomic trait for crop architecture and yield. In rice (Oryza sativa), the lamina joint is a unique structure connecting the leaf blade and sheath that determines leaf angle. Brassinosteroid (BR) signaling involving GLYCOGEN SYNTHASE KINASE-3 (GSK3)/SHAGGY-like kinases and BRASSINAZOLE-RESISTANT1 (BZR1) has a central role in regulating leaf angle in rice. In this study, we identified the atypical R2R3-MYB transcription factor FOUR LIPS (OsFLP), the rice homolog of Arabidopsis (Arabidopsis thaliana) AtFLP, as a participant in BR-regulated leaf angle formation. The spatiotemporal specificity of OsFLP expression in the lamina joint was closely associated with lignin deposition in vascular bundles and sclerenchyma cells. OsFLP mutation caused loose plant architecture with droopy flag leaves and hypersensitivity to BRs. OsBZR1 directly targeted OsFLP, and OsFLP transduced BR signals to lignin deposition in the lamina joint. Moreover, OsFLP promoted the transcription of the phenylalanine ammonia-lyase family genes OsPAL4 and OsPAL6. Intriguingly, OsFLP feedback regulated OsGSK1 transcription and OsBZR1 phosphorylation status. In addition, an Ala-to-Thr substitution within the OsFLP R3 helix-turn-helix domain, an equivalent mutation to that in Osflp-1, affected the DNA-binding ability and transcriptional activity of OsFLP. Our results reveal that OsFLP functions with OsGSK1 and OsBZR1 in BR signaling to maintain optimal leaf angle by modulating the lignin deposition in mechanical tissues of the lamina joint.
叶片角度是影响作物结构和产量的重要农艺性状。在水稻(Oryza sativa)中,叶片关节是连接叶片和叶鞘的独特结构,它决定了叶片的角度。芸苔素类固醇(BR)信号传导涉及 GLYCOGEN SYNTHASE KINASE-3 (GSK3)/SHAGGY-like 激酶和 BRASSINAZOLE-RESISTANT1 (BZR1),在调节水稻叶片角度方面起着核心作用。在这项研究中,我们发现了非典型 R2R3-MYB 转录因子 FOUR LIPS(OsFLP),它是拟南芥(Arabidopsis thaliana)AtFLP 的水稻同源物,参与了 BR 调控的叶角形成。OsFLP在叶片连接处表达的时空特异性与维管束和叶脉细胞中木质素的沉积密切相关。OsFLP 突变导致植物结构松散,旗叶下垂,并对 BRs 过敏。OsBZR1 直接靶向 OsFLP,OsFLP 将 BR 信号转导至木质素在叶片连接处的沉积。此外,OsFLP 还促进了苯丙氨酸氨解酶家族基因 OsPAL4 和 OsPAL6 的转录。耐人寻味的是,OsFLP 反馈调节了 OsGSK1 的转录和 OsBZR1 的磷酸化状态。此外,OsFLP R3螺旋-转螺旋结构域中的Ala-Thr取代(与Osflp-1中的突变相当)影响了OsFLP的DNA结合能力和转录活性。我们的研究结果表明,OsFLP与OsGSK1和OsBZR1一起在BR信号转导中发挥作用,通过调节叶片关节机械组织中的木质素沉积来维持最佳叶片角度。
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引用次数: 0
Plant Phytochrome Interactions Decode Light and Temperature Signals 植物植物色素相互作用解码光和温度信号
Pub Date : 2024-09-11 DOI: 10.1093/plcell/koae249
Chengwei Yi, Uwe Gerken, Kun Tang, Michael Philipp, Matias D Zurbriggen, Jürgen Köhler, Andreas Möglich
Plant phytochromes perceive red and far-red light to elicit adaptations to the changing environment. Downstream physiological responses revolve around red-light-induced interactions with phytochrome-interacting factors (PIF). Phytochromes double as thermoreceptors, owing to the pronounced temperature dependence of thermal reversion from the light-adapted Pfr to the dark-adapted Pr state. Here, we assess whether thermoreception may extend to the phytochrome:PIF interactions. While the association between Arabidopsis (Arabidopsis thaliana) PHYTOCHROME B (PhyB) and several PHYTOCHROME-INTERACTING FACTOR (PIF) variants moderately accelerates with temperature, the dissociation does more so, thus causing net destabilization of the phytochrome:PIF complex. Markedly different temperature profiles of PIF3 and PIF6 might underlie stratified temperature responses in plants. Accidentally, we identify a photoreception mechanism under strong continuous light, where the extent of phytochrome:PIF complexation decreases with red-light intensity rather than increases. Mathematical modeling rationalizes this attenuation mechanism and ties it to rapid red-light-driven Pr⇄Pfr interconversion and complex dissociation out of Pr. Varying phytochrome abundance, e.g., during diurnal and developmental cycles, and interaction dynamics, e.g., across different PIFs, modify the nature and extent of attenuation, thus permitting light-response profiles more malleable than possible for the phytochrome Pr⇄Pfr interconversion alone. Our data and analyses reveal a photoreception mechanism with implications for plant physiology, optogenetics, and biotechnological applications.
植物的植物色素能感知红光和远红光,以适应不断变化的环境。下游生理反应围绕着红光诱导的与植物色素相互作用因子(PIF)的相互作用。由于从适应光的 Pfr 状态到适应暗的 Pr 状态的热还原具有明显的温度依赖性,因此植物色素兼具热敏感受器的双重作用。在这里,我们评估了温感知是否可能延伸到植物色素与 PIF 的相互作用。拟南芥 PHYTOCHROME B(PhyB)与几种 PHYTOCHROME-INTERACTING FACTOR(PIF)变体之间的结合会随着温度的升高而适度加快,而解离则会更快,从而导致植物色素:PIF 复合物的净不稳定性。PIF3 和 PIF6 明显不同的温度曲线可能是植物分层温度反应的基础。我们意外地发现了强连续光下的光接收机制,在这种机制下,植物色素:PIF 复合物的程度会随着红光强度的增加而降低,而不是增加。数学模型合理地解释了这种衰减机制,并将其与红光驱动的 Pr⇄Pfr 快速相互转化和从 Pr 中解离出复合物联系起来。不同的植物色素丰度(如在昼夜周期和发育周期中)和相互作用动态(如在不同的 PIF 之间)会改变衰减的性质和程度,从而使光响应曲线比单纯的植物色素 Pr⇄Pfr 相互转换更具延展性。我们的数据和分析揭示了一种光接收机制,对植物生理学、光遗传学和生物技术应用具有重要意义。
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引用次数: 0
Alternative splicing of a potato disease resistance gene maintains homeostasis between growth and immunity 马铃薯抗病基因的交替剪接可维持生长和免疫之间的平衡
Pub Date : 2024-06-28 DOI: 10.1093/plcell/koae189
Biying Sun, Jie Huang, Liang Kong, Chuyun Gao, Fei Zhao, Jiayong Shen, Tian Wang, Kangping Li, Luyao Wang, Yuanchao Wang, Dennis A Halterman, Suomeng Dong
Plants possess a robust and sophisticated innate immune system against pathogens and must balance growth with rapid pathogen detection and defense. The intracellular receptors with nucleotide-binding leucine-rich repeat (NLR) motifs recognize pathogen-derived effector proteins and thereby trigger the immune response. The expression of genes encoding NLR receptors is precisely controlled in multifaceted ways. The alternative splicing (AS) of introns in response to infection is recurrently observed but poorly understood. Here we report that the potato (Solanum tuberosum) NLR gene RB undergoes AS of its intron, resulting in two transcriptional isoforms, which coordinately regulate plant immunity and growth homeostasis. During normal growth, RB predominantly exists as intron-retained isoform RB_IR, encoding a truncated protein containing only the N-terminus of the NLR. Upon late blight infection, the pathogen induces intron splicing of RB, increasing the abundance of RB_CDS, which encodes a full-length and active R protein. By deploying the RB splicing isoforms fused with a luciferase reporter system, we identified IPI-O1 (also known as Avrblb1), the RB cognate effector, as a facilitator of RB AS. IPI-O1 directly interacts with potato splicing factor StCWC15, resulting in altered localization of StCWC15 from the nucleoplasm to the nucleolus and nuclear speckles. Mutations in IPI-O1 that eliminate StCWC15 binding also disrupt StCWC15 re-localization and RB intron splicing. Thus, our study reveals that StCWC15 serves as a surveillance facilitator that senses the pathogen-secreted effector and regulates the trade-off between RB-mediated plant immunity and growth, expanding our understanding of molecular plant–microbe interactions.
植物拥有强大而复杂的先天免疫系统来抵御病原体,必须在生长与快速检测和防御病原体之间取得平衡。细胞内具有核苷酸结合富亮氨酸重复(NLR)结构的受体能识别病原体衍生的效应蛋白,从而触发免疫反应。编码 NLR 受体的基因表达受到多方面的精确控制。内含子的替代剪接(AS)对感染的反应经常被观察到,但却鲜为人知。在这里,我们报告了马铃薯(Solanum tuberosum)NLR 基因 RB 的内含子发生了 AS 变化,产生了两种转录异构体,它们协调调节植物免疫和生长平衡。在正常生长过程中,RB 主要以内含子保留异构体 RB_IR 的形式存在,编码一种仅含有 NLR N 端的截短蛋白。晚疫病感染时,病原体会诱导 RB 的内含子剪接,增加 RB_CDS 的丰度,后者编码全长的活性 R 蛋白。通过将 RB 剪接异构体与荧光素酶报告系统融合,我们发现 RB 同源效应物 IPI-O1(又称 Avrblb1)是 RB AS 的促进因子。IPI-O1 与马铃薯剪接因子 StCWC15 直接相互作用,导致 StCWC15 从核质到核仁和核斑点的定位改变。消除 StCWC15 结合的 IPI-O1 基因突变也会破坏 StCWC15 的重新定位和 RB 内含子的剪接。因此,我们的研究揭示了 StCWC15 是一种监视促进因子,它能感知病原体分泌的效应物,并调节 RB 介导的植物免疫与生长之间的权衡,从而拓展了我们对植物与微生物分子相互作用的认识。
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引用次数: 0
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The Plant Cell
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