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Incomplete filling in the basal region of maize endosperm: timing of development of starch synthesis and cell vitality. 玉米胚乳基部的不完全填充:淀粉合成和细胞活力的发展时间。
IF 6.2 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2024-11-01 Epub Date: 2024-09-30 DOI: 10.1111/tpj.17043
Xian-Min Chen, Zhi-Wei Wang, Xiao-Gui Liang, Feng-Yuan Li, Bin-Bin Li, Gong Wu, Fei Yi, Tim L Setter, Si Shen, Shun-Li Zhou

Starch synthesis in maize endosperm adheres to the basipetal sequence from the apex downwards. However, the mechanism underlying nonuniformity among regions of the endosperm in starch accumulation and its significance is poorly understood. Here, we examined the spatiotemporal transcriptomes and starch accumulation dynamics in apical (AE), middle (ME), and basal (BE) regions of endosperm throughout the filling stage. Results demonstrated that the BE had lower levels of gene transcripts and enzymes facilitating starch synthesis, corresponding to incomplete starch storage at maturity, compared with AE and ME. Contrarily, the BE showed abundant gene expression for genetic processing and slow progress in physiological development (quantified by an index calculated from the expression values of development progress marker genes), revealing a sustained cell vitality of the BE. Further analysis demonstrated a significant parabolic correlation between starch synthesis and physiological development. An in-depth examination showed that the BE had more active signaling pathways of IAA and ABA than the AE throughout the filling stage, while ethylene showed the opposite pattern. Besides, SNF1-related protein kinase1 (SnRK1) activity, a regulator for starch synthesis modulated by trehalose-6-phosphate (T6P) signaling, was kept at a lower level in the BE than the AE and ME, corresponding to the distinct gene expression in the T6P pathway in starch synthesis regulation. Collectively, the findings support an improved understanding of the timing of starch synthesis and cell vitality in regions of the endosperm during development, and potential regulation from hormone signaling and T6P/SnRK1 signaling.

玉米胚乳中的淀粉合成遵循从顶点向下的基瓣序列。然而,人们对胚乳各区域淀粉积累不均匀的机制及其意义还知之甚少。在这里,我们研究了胚乳顶端(AE)、中间(ME)和基部(BE)区域在整个灌浆期的时空转录组和淀粉积累动态。结果表明,与 AE 和 ME 相比,BE 中促进淀粉合成的基因转录本和酶的水平较低,这与成熟时淀粉储存不完全有关。与此相反,BE 的基因处理基因表达量丰富,生理发育进度缓慢(根据发育进度标记基因的表达值计算出的指数进行量化),显示出 BE 细胞的持续活力。进一步分析表明,淀粉合成与生理发育之间存在明显的抛物线相关性。深入研究表明,在整个灌浆期,BE 的 IAA 和 ABA 信号通路比 AE 更活跃,而乙烯则相反。此外,SNF1相关蛋白激酶1(SnRK1)的活性(SNF1相关蛋白激酶1是淀粉合成的调控因子,受6-磷酸三卤淀粉(T6P)信号调控)在BE中保持在低于AE和ME的水平,这与淀粉合成调控中T6P通路的不同基因表达相对应。总之,这些发现有助于人们更好地了解胚乳发育过程中淀粉合成和细胞活力的时间以及激素信号和T6P/SnRK1信号的潜在调控。
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引用次数: 0
Integrated genomic, transcriptomic, and metabolomic analyses of Ilex hylonoma provide insights into the triterpenoid saponin biosynthesis. 对 Ilex hylonoma 的基因组、转录组和代谢组进行综合分析,有助于深入了解三萜类皂苷的生物合成。
IF 6.2 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2024-11-01 Epub Date: 2024-09-27 DOI: 10.1111/tpj.17046
Landi Feng, Yingjun Yao, Minghui Kang, Wengjie Yang, Yu Han, Wei Liu, Xiaonan Li, Na Li, Yongqi Hu, Jianquan Liu, Quanjun Hu

Ilex is known for its rich content of secondary metabolites, particularly triterpenoid saponins. These compounds hold significant value in natural remedies and herbal medicine. However, the molecular mechanisms responsible for triterpenoid biosynthesis in plants of this genus remain largely unexplored. In this study, we successfully generated the first chromosome-scale genome of Ilex hylonoma. The assembly, comprising 20 anchored chromosomes, has an N50 contig size of 2.13 Mb and a scaffold size of 33.68 Mb. Comparative genome analyses with two other congeners with available chromosome-level genomes suggested that an end-to-end chromosome fusion event likely contributed to the reduction in chromosome number from n = 20 to n = 19 within this genus. By integrating transcriptomic and metabolomic data, we identified the gene expression patterns and metabolite profiles of I. hylonoma across three commonly utilized medicinal tissues. We subsequently pinpointed candidate genes involved in the regulation of triterpenoid saponin biosynthesis, including CYP450 genes, UGT genes, and associated transcription factors. Furthermore, yeast heterologous expression analysis revealed that ihyl08363 catalyzed the conversion of β-amyrin into oleanolic acid, while ihyl04303 catalyzed the C-2α hydroxylation of oleanolic acid to produce maslinic acid. This integrated analysis provides valuable insights into the biosynthesis of important triterpenoid saponins in medicinal Ilex plants.

Ilex 以其丰富的次生代谢物含量而闻名,尤其是三萜类皂甙。这些化合物在天然疗法和草药方面具有重要价值。然而,该属植物中三萜类生物合成的分子机制在很大程度上仍未得到探索。在这项研究中,我们成功生成了 Ilex hylonoma 的首个染色体级基因组。该基因组由 20 条固定染色体组成,N50 等位基因大小为 2.13 Mb,支架大小为 33.68 Mb。与其他两个具有染色体级基因组的同属植物进行的基因组比较分析表明,端对端染色体融合事件很可能是导致该属植物染色体数从 n = 20 减少到 n = 19 的原因。通过整合转录组和代谢组数据,我们确定了 I. hylonoma 在三种常用药用组织中的基因表达模式和代谢物特征。随后,我们确定了参与调控三萜类皂苷生物合成的候选基因,包括 CYP450 基因、UGT 基因和相关转录因子。此外,酵母异源表达分析表明,ihyl08363 催化 β-amyrin 转化为齐墩果酸,而 ihyl04303 催化齐墩果酸的 C-2α 羟基化生成马斯林酸。这项综合分析为了解药用 Ilex 植物中重要的三萜类皂苷的生物合成提供了宝贵的信息。
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引用次数: 0
SlBTB19 interacts with SlWRKY2 to suppress cold tolerance in tomato via the CBF pathway. SlBTB19 与 SlWRKY2 相互作用,通过 CBF 途径抑制番茄的耐寒性。
IF 6.2 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2024-11-01 Epub Date: 2024-09-25 DOI: 10.1111/tpj.17040
Jin Xu, Sidi Liu, Jiachen Hong, Rui Lin, Xiaojian Xia, Jingquan Yu, Yanhong Zhou

Cold stress restricts the metabolic and physiological activities of plants, thereby affecting their growth and development. Although broad-complex, tramtrack, and bric-à-brac (BTB) proteins are essential for diverse biological processes and stress responses, the mechanisms underlying BTB-mediated cold responses remain not fully understood. Here, we characterize the function of the cold-induced SlBTB19 protein in tomato (Solanum lycopersicum). Overexpression of SlBTB19 resulted in increased plant sensitivity to cold stress, whereas SlBTB19 knockout mutants exhibited a cold-tolerance phenotype. Further analyses, including protein-protein interaction studies and cell-free degradation assays, revealed that SlBTB19 interacts with and destabilizes the transcription factor SlWRKY2. Using virus-induced gene silencing (VIGS) to silence SlWRKY2 in both wild-type and slbtb19 mutants, we provided genetic evidence that SlWRKY2 acts downstream of SlBTB19 in regulating cold tolerance. Importantly, we demonstrated that SlWRKY2 positively regulates cold tolerance in a CRT/DRE binding factor (CBF)-dependent manner. Under cold stress, SlWRKY2 binds to the W-box in the CBF1 and CBF3 promoters, directly activating their expression. In summary, our findings identify a SlBTB19-SlWRKY2 module that negatively regulates the CBF-dependent cold tolerance pathway in tomato.

冷胁迫会限制植物的代谢和生理活动,从而影响其生长和发育。虽然广谱复合蛋白、电车轨道蛋白和砖块蛋白(BTB)对多种生物过程和胁迫响应至关重要,但人们对 BTB 介导的冷响应机制仍不完全清楚。在这里,我们描述了番茄(Solanum lycopersicum)中冷诱导的 SlBTB19 蛋白的功能。过表达 SlBTB19 提高了植物对冷胁迫的敏感性,而 SlBTB19 基因敲除突变体则表现出耐寒表型。进一步的分析,包括蛋白质-蛋白质相互作用研究和无细胞降解测定,发现 SlBTB19 与转录因子 SlWRKY2 相互作用并破坏其稳定性。我们利用病毒诱导基因沉默(VIGS)技术沉默了野生型和 slbtb19 突变体中的 SlWRKY2,提供了 SlWRKY2 在调控耐寒性方面作用于 SlBTB19 下游的遗传学证据。重要的是,我们证明了 SlWRKY2 以一种依赖 CRT/DRE 结合因子(CBF)的方式正向调节耐寒性。在冷胁迫下,SlWRKY2 与 CBF1 和 CBF3 启动子中的 W-box 结合,直接激活它们的表达。总之,我们的研究结果发现了一个 SlBTB19-SlWRKY2 模块,它能负向调节番茄中依赖 CBF 的耐寒途径。
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引用次数: 0
Exit control: the role of Arabidopsis hydathodes in auxin storage and nutrient recovery. 出口控制:拟南芥水瘤在辅助素储存和养分恢复中的作用。
IF 6.2 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2024-11-01 DOI: 10.1111/tpj.17118
Gwendolyn Kirschner
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引用次数: 0
Phylogenomic insights into the diversity and evolution of RPW8-NLRs and their partners in plants. 植物中 RPW8-NLRs 及其伙伴的多样性和进化的系统发生学见解。
IF 6.2 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2024-11-01 Epub Date: 2024-09-23 DOI: 10.1111/tpj.17034
Huiyu Qin, Junyuan Cheng, Guan-Zhu Han, Zhen Gong

Plants use nucleotide-binding leucine-rich repeat receptors (NLRs) to sense pathogen effectors, initiating effector-triggered immunity (ETI). NLRs containing RESISTANCE TO POWDERY MILDEW 8 domain (RNLs) function as "helper" NLRs in flowering plants and support the immune responses mediated by "sensor" NLRs in cooperation with lipase-EP domain fused proteins (EP proteins). Despite their crucial roles in ETI, much remains unclear about the evolutionary trajectories of RNLs and their functional partners EP proteins. Here, we perform phylogenomic analyses of RNLs in 90 plants, covering the major diversity of plants, and identify the presence of RNLs in land plants and green algae, expanding the distribution of RNLs. We uncover a neglected major RNL group in gymnosperms, besides the canonical major group with NRG1s and ADR1s, and observe a drastic increase in RNL repertoire size in conifers. Phylogenetic analyses indicate that RNLs originated multiple times through domain shuffling, and the evolution of RNLs underwent a birth-and-death process. Moreover, we trace the origin of EP proteins back to the last common ancestor of vascular plants. We find that both RNLs and EP proteins evolve mainly under negative selection, revealing strong constraints on their function. Concerted losses and positive correlation in copy number are observed between RNL and EP sublineages, suggesting their cooperation in function. Together, our findings provide insights into the origin and evolution of plant helper NLRs, with implications for predicting novel innate immune signaling modules.

植物利用核苷酸结合富亮氨酸重复受体(NLRs)感知病原体效应因子,启动效应因子触发免疫(ETI)。在开花植物中,含有抗粉尘微粒 8 结构域(RNLs)的 NLRs 起着 "辅助 "NLRs 的作用,它们与脂肪酶-EP 结构域融合蛋白(EP 蛋白)合作,支持由 "传感器 "NLRs 介导的免疫反应。尽管 RNLs 及其功能伙伴 EP 蛋白在 ETI 中起着至关重要的作用,但它们的进化轨迹仍有许多不清楚之处。在这里,我们对 90 种植物中的 RNLs 进行了系统进化分析,涵盖了植物的主要多样性,并确定了陆生植物和绿藻中 RNLs 的存在,扩大了 RNLs 的分布范围。我们在裸子植物中发现了一个被忽视的主要 RNL 组,此外还有 NRG1s 和 ADR1s 的典型主要组,并观察到针叶树中 RNL 的剧增。系统发育分析表明,RNLs是通过结构域的洗牌多次起源的,RNLs的进化经历了一个出生-死亡的过程。此外,我们将 EP 蛋白的起源追溯到维管植物的最后一个共同祖先。我们发现,RNLs 和 EP 蛋白都主要是在负选择下进化的,这揭示了对其功能的强烈限制。在 RNL 和 EP 亚系之间观察到了拷贝数的协同损失和正相关性,这表明它们在功能上是相互合作的。总之,我们的发现为植物辅助 NLRs 的起源和进化提供了见解,对预测新型先天性免疫信号模块具有重要意义。
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引用次数: 0
Uracil phosphoribosyltransferase is required to establish a functional cytochrome b6f complex. 尿嘧啶磷酸核糖转移酶是建立功能性细胞色素 b6f 复合物的必要条件。
IF 6.2 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2024-11-01 Epub Date: 2024-09-25 DOI: 10.1111/tpj.17036
Vanessa Scherer, Leo Bellin, Serena Schwenkert, Martin Lehmann, Jannis Rinne, Claus-Peter Witte, Kathrin Jahnke, Andreas Richter, Tobias Pruss, Anne Lau, Lisa Waller, Sebastian Stein, Dario Leister, Torsten Möhlmann

Arabidopsis uracil phosphoribosyltransferase (UPP) is an essential enzyme and plants lacking this enzyme are strongly compromised in chloroplast function. Our analysis of UPP amiRNA mutants has confirmed that this vital function is crucial to establish a fully functional photosynthesis as the RIESKE iron sulfur protein (PetC) is almost absent, leading to a block in photosynthetic electron transport. Interestingly, this function appears to be unrelated to nucleotide homeostasis since nucleotide levels were not altered in the studied mutants. Transcriptomics and proteomic analysis showed that protein homeostasis but not gene expression is most likely responsible for this observation and high light provoked an upregulation of protease levels, including thylakoid filamentation temperature-sensitive 1, 5 (FtsH), caseinolytic protease proteolytic subunit 1 (ClpP1), and processing peptidases, as well as components of the chloroplast protein import machinery in UPP amiRNA lines. Strongly reduced PetC amounts were not only detected by immunoblot from mature plants but in addition in a de-etiolation experiment with young seedlings and are causing reduced high light-induced non-photochemical quenching Φ(NPQ) but increased unregulated energy dissipation Φ(NO). This impaired photosynthesis results in an inability to induce flavonoid biosynthesis. In addition, the levels of the osmoprotectants raffinose, proline, and fumarate were found to be reduced. In sum, our work suggests that UPP assists in stabilization PetC during import, processing or targeting to the thylakoid membrane, or protects it against proteolytic degradation.

拟南芥尿嘧啶磷酸核糖基转移酶(UPP)是一种重要的酶,缺乏这种酶的植物叶绿体功能严重受损。我们对 UPP amiRNA 突变体的分析证实,由于 RIESKE 铁硫蛋白(PetC)几乎缺失,导致光合电子传递受阻,因此这一重要功能对于建立功能完备的光合作用至关重要。有趣的是,这一功能似乎与核苷酸平衡无关,因为在所研究的突变体中,核苷酸水平没有发生变化。转录组学和蛋白质组学分析表明,蛋白质平衡而非基因表达很可能是导致这一观察结果的原因,强光引起了蛋白酶水平的上调,包括UPP amiRNA株系中的叶绿体丝状温敏1、5(FtsH)、酪蛋白溶解蛋白酶蛋白水解亚基1(ClpP1)、加工肽酶以及叶绿体蛋白质导入机制的组分。不仅通过免疫印迹从成熟植株中检测到 PetC 数量大幅减少,而且在幼苗去叶绿素实验中也检测到 PetC 数量大幅减少,这导致强光诱导的非光化学淬灭Φ(NPQ)减少,但非调控能量耗散Φ(NO)增加。光合作用受损导致无法诱导类黄酮的生物合成。此外,还发现渗透保护剂棉子糖、脯氨酸和富马酸盐的水平降低。总之,我们的研究结果表明,UPP 在 PetC 的导入、加工或定向到类木质膜的过程中起到了稳定 PetC 的作用,或保护 PetC 免受蛋白酶降解。
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引用次数: 0
Decrease in purifying selection pressures on wheat homoeologous genes: tetraploidization versus hexaploidization. 小麦同源基因纯化选择压力的降低:四倍体化与六倍化。
IF 6.2 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2024-11-01 Epub Date: 2024-10-20 DOI: 10.1111/tpj.17047
Akihiro Ezoe, Daisuke Todaka, Yoshinori Utsumi, Satoshi Takahashi, Kanako Kawaura, Motoaki Seki

A series of polyploidizations in higher-order polyploids is the main event affecting gene content in a genome. Each polyploidization event can lead to massive functional divergence because of the subsequent decrease in selection pressure on duplicated genes; however, the causal relationship between multiple rounds of polyploidization and the functional divergence of duplicated genes is poorly understood. We focused on the Triticum-Aegilops complex lineage and compared selection pressure before and after tetraploidization and hexaploidization events. Although both events led to decreased selection pressure on homoeologous gene pairs (compared with diploids and tetraploids), the initial tetraploidization had a greater impact on selection pressure on homoeologous gene pairs than did subsequent hexaploidization. Consistent with this, selection pressure on expression patterns for the initial event relaxed more than those for the subsequent event. Surprisingly, the decreased selection pressure on these homoeologous genes was independent of the existence of in-paralogs within the same subgenome. Wheat homoeologous pairs had different evolutionary consequences compared with orthologs related to other mechanisms (ancient allopolyploidization, ancient autopolyploidization, and small-scale duplication). Furthermore, tetraploidization and hexaploidization also seemed to have different evolutionary consequences. This suggests that homoeologous genes retain unique functions, including functions that are unlikely to be preserved in genes generated by the other duplication mechanisms. We found that their unique functions differed between tetraploidization and hexaploidization (e.g., reproductive and chromosome segregation processes). These findings imply that the substantial number of gene pairs resulting from multiple allopolyploidization events, especially initial tetraploidization, may have been a unique source of functional divergence.

高阶多倍体的一系列多倍体化是影响基因组中基因含量的主要事件。由于重复基因的选择压力随之降低,每次多倍体化都可能导致大规模的功能分化;然而,人们对多轮多倍体化与重复基因功能分化之间的因果关系知之甚少。我们重点研究了Triticum-Aegilops复合品系,并比较了四倍体化和六倍化前后的选择压力。虽然这两个事件都导致同源基因对的选择压力下降(与二倍体和四倍体相比),但最初的四倍体化对同源基因对选择压力的影响要大于随后的六倍化。与此相一致的是,初始事件对表达模式的选择压力比后续事件的选择压力更宽松。令人惊讶的是,这些同源基因选择压力的降低与同一亚基因组中是否存在内同源基因无关。小麦同源基因对与其他机制(古代异源多倍体化、古代自源多倍体化和小规模复制)相关的直向同源基因相比,具有不同的进化后果。此外,四倍体化和六倍化似乎也有不同的进化后果。这表明同源基因保留了独特的功能,包括其他复制机制产生的基因不太可能保留的功能。我们发现,它们的独特功能在四倍体化和六倍化过程中有所不同(如生殖和染色体分离过程)。这些发现意味着,由多次异源多倍体化(尤其是最初的四倍体化)产生的大量基因对可能是功能分化的独特来源。
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引用次数: 0
The histone chaperones ASF1 and HIRA are required for telomere length and 45S rDNA copy number homeostasis. 组蛋白伴侣 ASF1 和 HIRA 是端粒长度和 45S rDNA 拷贝数平衡所必需的。
IF 6.2 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2024-11-01 Epub Date: 2024-10-14 DOI: 10.1111/tpj.17041
Adéla Machelová, Martina Nešpor Dadejová, Michal Franek, Guillaume Mougeot, Lauriane Simon, Samuel Le Goff, Céline Duc, Jasmin Bassler, Martin Demko, Jana Schwarzerová, Sophie Desset, Aline V Probst, Martina Dvořáčková

Genome stability is significantly influenced by the precise coordination of chromatin complexes that facilitate the loading and eviction of histones from chromatin during replication, transcription, and DNA repair processes. In this study, we investigate the role of the Arabidopsis H3 histone chaperones ANTI-SILENCING FUNCTION 1 (ASF1) and HISTONE REGULATOR A (HIRA) in the maintenance of telomeres and 45S rDNA loci, genomic sites that are particularly susceptible to changes in the chromatin structure. We find that both ASF1 and HIRA are essential for telomere length regulation, as telomeres are significantly shorter in asf1a1b and hira mutants. However, these shorter telomeres remain localized around the nucleolus and exhibit a comparable relative H3 occupancy to the wild type. In addition to regulating telomere length, ASF1 and HIRA contribute to silencing 45S rRNA genes and affect their copy number. Besides, ASF1 supports global heterochromatin maintenance. Our findings also indicate that ASF1 transiently binds to the TELOMERE REPEAT BINDING 1 protein and the N terminus of telomerase in vivo, suggesting a physical link between the ASF1 histone chaperone and the telomere maintenance machinery.

染色质复合物在复制、转录和 DNA 修复过程中促进组蛋白从染色质中装载和排出,染色质复合物的精确协调对基因组稳定性有重大影响。在这项研究中,我们研究了拟南芥 H3 组蛋白伴侣素 ANTI-SILENCING FUNCTION 1(ASF1)和 HISTONE REGULATOR A(HIRA)在维护端粒和 45S rDNA 位点中的作用,这些基因组位点特别容易受到染色质结构变化的影响。我们发现 ASF1 和 HIRA 对端粒长度的调节都至关重要,因为 ASF1a1b 和 HIRA 突变体的端粒明显缩短。然而,这些较短的端粒仍然定位在核仁周围,并表现出与野生型相当的相对 H3 占有率。除了调节端粒长度,ASF1 和 HIRA 还有助于沉默 45S rRNA 基因并影响其拷贝数。此外,ASF1还支持全球异染色质的维持。我们的研究结果还表明,ASF1能瞬时结合体内的端粒重复结合1蛋白和端粒酶的N末端,这表明ASF1组蛋白伴侣与端粒维持机制之间存在物理联系。
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引用次数: 0
A novel RAV transcription factor from pear interacts with viral RNA-silencing suppressors to inhibit viral infection. 梨中的一种新型 RAV 转录因子与病毒 RNA 沉默抑制因子相互作用,抑制病毒感染。
IF 6.2 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2024-11-01 Epub Date: 2024-09-23 DOI: 10.1111/tpj.17037
Yin-Shuai Xie, Qi Zeng, Wen-Ting Huang, Jin-Ying Wang, Han-Wei Li, Shang-Zhen Yu, Can Liu, Xue-Qing Zhang, Chen-Lu Feng, Wen-Hao Zhang, Tian-Zhong Li, Yu-Qin Cheng

In plants, RNA silencing constitutes a strong defense against viral infection, which viruses counteract with RNA-silencing suppressors (RSSs). Understanding the interactions between viral RSSs and host factors is crucial for elucidating the molecular arms race between viruses and host plants. We report that the helicase motif (Hel) of the replicase encoded by apple stem grooving virus (ASGV)-the main virus affecting pear trees in China-is an RSS that can inhibit both local and systemic RNA silencing, possibly by binding double-stranded (ds) siRNA. The transcription factor related to ABSCISIC ACID INSENSITIVE3/VIVIPAROUS1 from pear (PbRAV1) enters the cytoplasm and binds Hel through its C terminus, thereby attenuating its RSS activity by reducing its binding affinity to 21- and 24-nt ds siRNA, and suppressing ASGV infection. PbRAV1 can also target p24, an RSS encoded by grapevine leafroll-associated virus 2 (GLRaV-2), with similar negative effects on p24's suppressive function and inhibition of GLRaV-2 infection. Moreover, like the positive role of the PbRAV1 homolog from grapevine (VvRAV1) in p24's previously reported RSS activity, ASGV Hel can also hijack VvRAV1 and employ the protein to sequester 21-nt ds siRNA, thereby enhancing its own RSS activity and promoting ASGV infection. Furthermore, PbRAV1 neither interacts with CP, an RSS encoded by grapevine inner necrosis virus, nor has any obvious effect on CP's RSS activity. Our results identify an RSS encoded by ASGV and demonstrate that PbRAV1, representing a novel type of RAV transcription factor, plays a defensive role against viral infection by targeting viral RSSs.

在植物体内,RNA 沉默是抵御病毒感染的一种强有力的防御手段,而病毒则利用 RNA 沉默抑制因子(RSSs)来抵消这种防御手段。了解病毒 RSS 与宿主因子之间的相互作用对于阐明病毒与宿主植物之间的分子军备竞赛至关重要。我们报告说,苹果茎沟病毒(ASGV)--影响中国梨树的主要病毒--编码的复制酶的螺旋酶基序(Hel)是一种RSS,可能通过结合双链(ds)siRNA抑制局部和系统RNA沉默。与梨中的 ABSCISIC ACID INSENSITIVE3/VIVIPAROUS1 有关的转录因子(PbRAV1)进入细胞质并通过其 C 末端与 Hel 结合,从而通过降低其与 21-nt 和 24-nt ds siRNA 的结合亲和力来削弱其 RSS 活性,并抑制 ASGV 感染。PbRAV1 还能靶向葡萄卷叶相关病毒 2(GLRaV-2)编码的 RSS p24,对 p24 的抑制功能和抑制 GLRaV-2 感染产生类似的负面影响。此外,与之前报道的葡萄中的 PbRAV1 同源物(VvRAV1)在 p24 的 RSS 活性中的积极作用一样,ASGV Hel 也能劫持 VvRAV1 并利用该蛋白来封存 21-nt ds siRNA,从而增强自身的 RSS 活性并促进 ASGV 感染。此外,PbRAV1 既不与葡萄内坏死病毒编码的 RSS CP 相互作用,也不对 CP 的 RSS 活性产生任何明显影响。我们的研究结果确定了一种由 ASGV 编码的 RSS,并证明了 PbRAV1 是一种新型的 RAV 转录因子,它通过靶向病毒 RSS 发挥防御病毒感染的作用。
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引用次数: 0
Integrating multiregulatory analysis reveals the negative regulatory function of miR482a in the response of poplar to canker pathogen infection. 多调控整合分析揭示了 miR482a 在杨树应对腐烂病病原体感染过程中的负调控功能。
IF 6.2 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2024-11-01 Epub Date: 2024-09-24 DOI: 10.1111/tpj.17039
Xiaoqian Yang, Ruen Yu, Jiahao Liu, Dandan Xiao, Chun Wang, Tiantian Fu, Yuzhang Yang, Kaijing Rong, Yanwei Wang

Canker disease caused by the bacterium Lonsdalea populi is one of the most destructive diseases affecting poplar stems. However, the detailed stress response mechanisms of poplar have not been widely characterized. To explore the diverse regulatory RNA landscape and the function of key regulators in poplar subjected to L. populi stress, we integrated time-course experiment with mock-inoculation (CK) and inoculation (IN) with L. populi at the first, third, and sixth day (IN1, IN3, IN6) on Populus × euramericana cv. '74/76' (107), small RNA-seq, whole transcriptome-wide analysis, degradome analysis and transgenic experiments. A total of 98 differentially expressed (DE) miRNA, 17 974 DEmRNA, and 807 DElncRNA were identified in poplar infected by L. populi, presenting dynamic changes over the infection course. Regulatory networks among RNAs were further constructed. Notably, a network centered on ptc-miR482a in CK-vs-IN3 contained most DEGs. We show that miR482a and miR1448 are located in one transcript as a polycistron. Overexpression of pre-miR482a-miR1448 (OX482-1448) and pre-miR482a (OX482) increased poplar susceptibility to canker pathogen with reduced accumulation of reactive oxygen species, while the suppression of miR482a (STTM482) conferred poplar disease resistance. PHA7 was validated as the target of miR482a with degradome sequencing and tobacco transient co-transformation, its expression being downregulated in OX482-1448 and OX482 lines. Additionally, a series of phasiRNAs were triggered by miR482a targeting PHA7, forming regulatory cascades with more RLP, NBS-LRR, and PK genes, further verifying the defense function of miR482a. These findings provide insights for understanding the roles of ncRNAs and regulatory networks involved in poplar immunity.

由白杨龙须菜细菌引起的枯萎病是影响杨树茎干的最具破坏性的病害之一。然而,杨树详细的应激反应机制尚未得到广泛表征。为了探索杨树在受到褐斑病菌胁迫时的多种调控 RNA 图谱以及关键调控因子的功能,我们在 Populus × euramericana cv. '74/76' (107) 上进行了模拟接种(CK)和在第一、第三和第六天接种(IN1、IN3、IN6)褐斑病菌的时程实验、小 RNA-seq、全转录组分析、降解组分析和转基因实验。结果发现,在被杨树褐斑病菌感染的杨树中,共有 98 个差异表达(DE)miRNA、17 974 个 DEmRNA 和 807 个 DElncRNA 在感染过程中呈现动态变化。进一步构建了 RNA 之间的调控网络。值得注意的是,CK-vs-IN3 中以 ptc-miR482a 为中心的网络包含了大多数 DEGs。我们发现,miR482a 和 miR1448 位于一个多聚体转录本中。过表达pre-miR482a-miR1448(OX482-1448)和pre-miR482a(OX482)会增加杨树对腐烂病病原体的易感性,减少活性氧的积累,而抑制miR482a(STM482)会赋予杨树抗病性。通过降解组测序和烟草瞬时共转化验证了 PHA7 是 miR482a 的靶标,其表达在 OX482-1448 和 OX482 株系中被下调。此外,以 PHA7 为靶标的 miR482a 触发了一系列 phasiRNAs,与更多的 RLP、NBS-LRR 和 PK 基因形成调控级联,进一步验证了 miR482a 的防御功能。这些发现为了解参与杨树免疫的 ncRNAs 的作用和调控网络提供了启示。
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