Plant Science最新文献_第10页

CRISPR/Cas9-mediated GhFT-targeted mutagenesis prolongs indeterminate growth and alters plant architecture in cotton CRISPR/ cas9介导的ghft靶向诱变延长了棉花的不确定生长并改变了植株结构。

IF 4.2 2区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Plant Science

Pub Date : 2024-12-24 DOI: 10.1016/j.plantsci.2024.112374

Na Sang , Bin Ma , Hui Liu , Tingting Feng , Xianzhong Huang

The shift from vegetative to reproductive growth is an important developmental transition that affects flowering and maturation, architecture, and ecological adaptability in plants. The florigen-antiflorigen system universally controls flowering and plant architecture, and changes to the ratio of these components alter this transition and disrupt growth. The genes FT (FLOWERING LOCUS T), encoding the florigen protein FT, and CETS [CENTRORADIALIS (CEN)/TERMINAL FLOWER1 (TFL1)/SELF-PRUNING (SP)], encoding antiflorigen proteins, have opposing roles. Upland cotton (Gossypium hirsutum) is one of the world’s most widely cultivated cotton varieties, and its complex allotetraploid genome contains only one homoeologous pair of FT genes (GhFT-A and GhFT-D). The functionally conserved gene GhFT promotes flowering and plays a role in plant architecture, although the molecular regulation of flowering and plant architecture in cotton remains unclear. In this study, CRISPR/Cas9 technology was used to induce mutations in the first and second exons of GhFT, respectively. G. hirsutum cv. YZ-1 was transformed with a CRISPR/Cas9-GhFT vector using Agrobacterium tumefaciens, and a diverse set of mutations was identified at the editing site. Compared with the wild type, mutant plants could not transition between vegetative and reproductive growth, and significant alterations to plant architecture were observed. Quantitative RT-PCR revealed downregulation of the homologous floral meristem identity genes APETALA1 (GhAP1) and OVEREXPRESSION OF CONSTANS 1 (GhSOC1) and upregulation of the TFL1 homologs GhTFL1–1 and GhTFL1–2. These results suggested that GhFT played a significant role in flowering time and plant architecture and that the ratio of florigen-antiflorigen components was critical to producing improved cotton varieties. This study provided a basis for future investigations of molecular breeding in cotton and guidance for the agricultural production of this crop.

植物从营养生长到生殖生长是一个重要的发育转变，影响着植物的开花和成熟、结构和生态适应性。致花剂-抗致花剂系统普遍控制着开花和植物结构，这些成分比例的变化改变了这种转变并破坏了生长。编码致花蛋白FT的基因FT（开花位点T）和编码抗致花蛋白的基因cts [CENTRORADIALIS (CEN)/TERMINAL FLOWER1 (TFL1)/SELF-PRUNING (SP)]具有相反的作用。陆地棉（Gossypium hirsutum）是世界上种植最广泛的棉花品种之一，其复杂的异源四倍体基因组仅包含一对同源的FT基因（GhFT-A和GhFT-D）。功能保守的基因GhFT促进开花并在植物结构中发挥作用，尽管棉花开花和植物结构的分子调控尚不清楚。在本研究中，利用CRISPR/Cas9技术分别诱导GhFT的第一外显子和第二外显子发生突变。毛囊草。YZ-1被使用农杆菌的CRISPR/Cas9-GhFT载体转化，在编辑位点发现了一系列不同的突变。与野生型相比，突变株不能在营养生长和生殖生长之间转换，植株结构发生了显著变化。定量RT-PCR显示，同源花分生组织识别基因APETALA1 （GhAP1）和CONSTANS 1 （GhSOC1）过表达下调，TFL1同源基因GhTFL1-1和GhTFL1-2上调。综上所述，GhFT对棉花开花时间和植株结构有显著影响，促花剂与抗花剂组分的比例对棉花品种的改良具有重要影响。本研究为今后棉花分子育种的研究奠定了基础，并为棉花的农业生产提供了指导。

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引用次数: 0

Nanomaterials impact in phytohormone signaling networks of plants − A critical review 纳米材料对植物激素信号网络的影响

IF 4.2 2区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Plant Science

Pub Date : 2024-12-24 DOI: 10.1016/j.plantsci.2024.112373

Garima Tripathi , Shrestha Dutta , Anamika Mishra , Soumyadeep Basu , Vishesh Gupta , Chinnaperumal Kamaraj

Nanotechnology offers a transformative approach to augment plant growth and crop productivity under abiotic and biotic stress conditions. Nanomaterials interact with key phytohormones, triggering the synthesis of stress-associated metabolites, activating antioxidant defense mechanisms, and modulating gene expression networks that regulate diverse physiological, biochemical, and molecular processes within plant systems. This review critically examines the impact of nanoparticles on both conventional and genetically modified crops, focusing on their role in nutrient delivery systems and the modulation of plant cellular machinery. Nanoparticle-induced reactive oxygen species (ROS) generation plays a central role in altering secondary metabolite biosynthesis, highlighting their function as potent elicitors and stimulants in plant systems. The review underscores the significant contribution of nanoparticles in enhancing stress resilience through the modulation of phytohormonal signaling pathways, offering novel insights into their potential for improving crop health and productivity under environmental stressors.

纳米技术提供了一种在非生物和生物胁迫条件下增强植物生长和作物生产力的变革性方法。纳米材料与关键的植物激素相互作用，触发应激相关代谢物的合成，激活抗氧化防御机制，并调节调节植物系统内各种生理、生化和分子过程的基因表达网络。这篇综述批判性地研究了纳米颗粒对传统和转基因作物的影响，重点关注它们在营养输送系统和植物细胞机制调节中的作用。纳米颗粒诱导的活性氧（ROS）的产生在改变次生代谢物的生物合成中起着核心作用，突出了它们在植物系统中作为有效的激发子和兴奋剂的功能。这篇综述强调了纳米颗粒通过调节植物激素信号通路在增强胁迫恢复力方面的重要贡献，为它们在环境胁迫下改善作物健康和生产力的潜力提供了新的见解。

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引用次数: 0

AtTRM11 as a tRNA 2-methylguanosine methyltransferase modulates flowering and bacterial resistance via translational regulation tRNA 2-甲基鸟苷甲基转移酶AtTRM11通过翻译调控调控开花和细菌抗性。

IF 4.2 2区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Plant Science

Pub Date : 2024-12-22 DOI: 10.1016/j.plantsci.2024.112368

Zhengyi Lv , Lun Guan , Ruixuan Yao , Hanchen Chen , Hailang Wang , Xukai Li , Xiaodong Xu , Liangcai Peng , Youmei Wang , Peng Chen

2-methylguanosine is an eukaryote-specific modified nucleoside in transfer RNAs, and m²G10 is catalyzed by Trm11-Trm112 protein complex in eukaryotic tRNAs. Here, we show that loss-of-function mutation of the Arabidopsis Trm11 homolog AtTRM11 resulted in m²G deficiency associated with disturbed ribosome assembly and overall transcriptome changes, including genes involved in flowering regulation and plant-pathogen interaction. The attrm11 mutant showed phenotypes of enlarged rosette leaves and early flowering, as well as enhanced resistance to Pseudomonas bacterial infection. AtTRM11 could partially rescue the m²G nucleoside level in yeast trm11 mutant, and AtTRM11 protein mostly resided in cytosol and physically interacted with AtTRM112b in planta. AtTRM11 was mostly expressed in shoot apex, root tip, and distal end of rosette leaves. KEGG enrichment analysis of differentially expressed genes between trm11 mutant and wild type indicated changes in pathways including phenopropanoid biosynthesis, plant-pathogen interaction, plant hormone signal transduction and MAPK signaling, suggesting that the pleiotropic phenotypes of the attrm11 mutant can be ascribed to translational and transcriptional changes.

2-甲基鸟苷是转移rna中真核特异性修饰核苷，m2G10在真核tRNAs中由Trm11-Trm112蛋白复合物催化。在这里，我们发现拟南芥Trm11同源物AtTRM11的功能缺失突变导致m2G缺乏，与核糖体组装紊乱和整体转录组变化有关，包括参与开花调节和植物-病原体相互作用的基因。attrm11突变体表现出莲座叶增大、开花提前、抗假单胞菌感染能力增强的表型。在酵母trm11突变体中，AtTRM11可以部分恢复m2G核苷水平，并且在植物中AtTRM11蛋白主要驻留在细胞质中，并与AtTRM112b发生物理相互作用。AtTRM11主要表达于莲座叶片的茎尖、根尖和远端。对trm11突变体与野生型差异表达基因的KEGG富集分析表明，trm11突变体在类表型生物合成、植物-病原体相互作用、植物激素信号转导和MAPK信号转导等途径上发生了变化，表明trm11突变体的多效表型可归因于翻译和转录的变化。

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引用次数: 0

The Arabidopsis heterotrimeric G protein α subunit binds to and inhibits the inward rectifying potassium channel KAT1 拟南芥异三聚体G蛋白α亚基结合并抑制向内整流钾通道KAT1。

IF 4.2 2区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Plant Science

Pub Date : 2024-12-20 DOI: 10.1016/j.plantsci.2024.112363

Jiang-Fan Guo , Hui Zhou , Zhuo-Ran Hu , Ya-Lan Yang , Wen-Bin Wang , Yan-Ru Zhang , Xue Li , Nuerkaimaier Mulati , Ying-Xin Li , Lu Wu , Yu Long , Jun-Min He

In animal cells, Gα subunit of the heterotrimeric G proteins can bind to both the N-terminal and C-terminal domains of G-protein-activated inwardly rectifying K⁺ channels (GIRKs) to inhibit their activities. In Arabidopsis guard cells, the Gα subunit GPA1 mediates multiple stimuli-regulated stomatal movements via inhibiting guard cell inward-rectifying K⁺ (K⁺_in) current, but it remains unclear whether GPA1 directly interacts with and inhibits the activities of K⁺_in channels. Here, we found that GPA1 interacted with the transmembrane domain rather than the intracellular domain of the Shaker family K⁺_in channel KAT1. Two-Electrode Voltage-Clamp experiments in Xenopus oocytes demonstrated that GPA1 significantly inhibited KAT1 channel activity. However, GPA1 could not inhibit the assembly of KAT1 as well as KAT2 as homo- and hetero-tetramers and alter the subcellular localization and protein stability of these channels. In conclusion, these findings reveal a novel regulatory mechanism for Gα inhibition of the Shaker family K⁺_in channel KAT1 via binding to its channel transmembrane domains but without affecting its subcellular localization, protein stability and the formation of functional homo- and hetero-tetramers. This suggests that in both animal and plant cells, Gα can regulate K⁺_in channels through physical interaction, albeit with differing mechanisms of interaction and regulation.

在动物细胞中，异三聚体G蛋白的Gα亚基可以结合到G蛋白激活的内向纠偏K+通道（GIRKs）的n端和c端结构域，从而抑制其活性。在拟南芥保护细胞中，Gα亚基GPA1通过抑制保护细胞内向纠偏K+ (K+ In)电流介导多种刺激调节的气孔运动，但GPA1是否直接与通道中K+的活性相互作用并抑制其活性尚不清楚。在这里，我们发现GPA1与通道KAT1中的Shaker家族K+的跨膜结构域而不是细胞内结构域相互作用。双电极电压钳实验表明，GPA1能显著抑制爪蟾卵母细胞中KAT1通道的活性。然而，GPA1不能抑制KAT1和KAT2作为同源和异源四聚体的组装，也不能改变这些通道的亚细胞定位和蛋白质稳定性。综上所述，这些发现揭示了一种新的调节机制，通过与通道跨膜结构域结合，而不影响其亚细胞定位、蛋白质稳定性和功能同源和异源四聚体的形成，从而抑制Gα在通道KAT1中的作用。这表明，在动物和植物细胞中，Gα都可以通过物理相互作用调节通道中的K+，尽管相互作用和调节机制不同。

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引用次数: 0

Review: Recent advances in unraveling the genetic architecture of kernel row number in maize 综述：揭示玉米核行数遗传结构的最新进展。

IF 4.2 2区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Plant Science

Pub Date : 2024-12-20 DOI: 10.1016/j.plantsci.2024.112366

Yizhu Wang, Ranjan K. Shaw, Xingming Fan

Kernel row number (KRN) is an important trait in maize that significantly impacts maize yield. The high heritability of KRN underscores its significance in maize breeding programs. In this review, we summarize recent advances in understanding the mechanisms underlying the formation, differentiation, and regulation of KRN in maize. Specifically, we have discussed gene mapping studies, functional validation of KRN-associated genes, and the application of gene editing techniques to KRN in maize. We summarized the various methods used to map and fine-map QTLs controlling KRN and provide an overview of the current status of cloned KRN-regulating genes. Despite the identification of many genes associated with KRN, the complexity of its regulation—arising from multiple loci and intricate gene interactions—remains a challenge. Balancing KRN with kernel number per row (KNR) and kernel weight is critical for optimizing yield while ensuring stability across different environments. Furthermore, we analyzed the influence of environmental factors on KRN, noting that despite its high heritability, environmental conditions can significantly affect this trait. Combining genotype-phenotype relationships with environmental data using big data and artificial intelligence could enhance maize breeding efficiency and accelerate genetic gains. This review emphasizes the importance of balancing traits, integrating environmental factors, and leveraging advanced technologies in maize breeding to achieve optimal yield and stress tolerance. Finally, we outlined future research perspectives aimed at developing high-yielding maize varieties through advances in KRN-related research.

粒行数是影响玉米产量的重要性状。KRN的高遗传力突出了其在玉米育种中的重要意义。本文综述了玉米KRN形成、分化和调控机制的最新研究进展。具体来说，我们讨论了基因定位研究，KRN相关基因的功能验证，以及基因编辑技术在玉米KRN中的应用。我们总结了用于定位和精细定位控制KRN的qtl的各种方法，并概述了克隆KRN调控基因的现状。尽管已经确定了许多与KRN相关的基因，但其调控的复杂性-由多个位点和复杂的基因相互作用引起-仍然是一个挑战。平衡KRN与每行核数（KNR）和核权对于优化产量，同时确保不同环境下的稳定性至关重要。此外，我们分析了环境因素对KRN的影响，发现尽管其具有高遗传力，但环境条件可以显著影响该性状。利用大数据和人工智能技术，将基因型-表型关系与环境数据相结合，可以提高玉米育种效率，加快遗传增益。本文强调了平衡性状、综合环境因素和利用先进技术在玉米育种中实现最佳产量和抗逆性的重要性。最后，通过krn相关研究的进展，展望了未来玉米高产品种的研究前景。

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引用次数: 0

An AP2/ERF transcription factor GhERF109 negatively regulates plant growth and development in cotton AP2/ERF转录因子GhERF109负向调控棉花的生长发育。

IF 4.2 2区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Plant Science

Pub Date : 2024-12-20 DOI: 10.1016/j.plantsci.2024.112365

Huiyun Shi , Ghulam Qanmber , Zuoren Yang , Yuling Guo , Shuya Ma , Sheng Shu , Yujun Li , Zhongxu Lin , Fuguang Li , Zhao Liu

Cotton is an important source of natural fibers. The AP2/ethylene response factor (ERF) family is one of the largest plant-specific transcription factors (TFs) groups, playing key roles in plant growth and development. However, the role of ERF TFs in cotton’s growth and development remains unclear. In this study, we identified GhERF109, a nuclear-localized ERF, which showed significant expression differences between ZM24 and pag1 cotton. Heterologous overexpression of GhERF109 in Arabidopsis resulted in reduced plant height, shortened root length, and reduced silique lengths compared to wild-type (WT) plants. In contrast, silencing GhERF109 in cotton led to a significant increase in plant height due to the elongation of stem cells. Overexpression of GhERF109 in cotton also produced a compact plant type with a notable reduction in height. RNA-seq analysis of GhERF109-silenced plants revealed 4123 differentially expressed genes (DEGs), with many upregulated genes involved in auxin response, polar transport, cell expansion, cell cycle regulation, brassinolide (BL) biosynthesis, and very long-chain fatty acid (VLCFA) pathways. These findings suggest that GhERF109 integrates auxin and other signaling pathways to suppress plant growth, providing valuable genetic material for breeding programs to improve mechanized cotton harvesting.

棉花是天然纤维的重要来源。AP2/乙烯响应因子（ERF）家族是最大的植物特异性转录因子（TFs）家族之一，在植物生长发育中起着关键作用。然而，ERF TFs在棉花生长发育中的作用尚不清楚。在本研究中，我们鉴定了GhERF109，这是一个核定位的ERF，在ZM24和pag1棉花中表达差异显著。与野生型（WT）植株相比，异源过表达GhERF109可导致植株高度降低、根长缩短和茎长缩短。相比之下，在棉花中沉默GhERF109，由于干细胞的伸长，导致植株高度显著增加。在棉花中过表达GhERF109也产生了株型紧凑且株高显著降低的株型。对gherf109沉默植物的RNA-seq分析发现了4123个差异表达基因（DEGs），其中许多上调基因涉及生长素反应、极性运输、细胞扩增、细胞周期调节、油菜素内酯（BL）生物合成和甚长链脂肪酸（VLCFA）途径。这些发现表明GhERF109整合了生长素和其他信号通路来抑制植物生长，为提高棉花机械化收获的育种计划提供了有价值的遗传物质。

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引用次数: 0

Regulation of PILS genes by bZIP transcription factor TGA7 in tomato plant growth bZIP转录因子TGA7在番茄植株生长中的调控作用

IF 4.2 2区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Plant Science

Pub Date : 2024-12-18 DOI: 10.1016/j.plantsci.2024.112359

Qixiang Zheng , Xiaole Meng , Xiaojing Fan , Shangyu Chen , Kangqi Sang , Jingquan Yu , Yanhong Zhou , Xiaojian Xia

Auxin plays a pivotal role in plant growth regulation. The PIN-FORMED (PIN) proteins facilitate long-distance polar auxin transport, whereas the recently identified PIN-LIKES (PILS) proteins regulate intracellular auxin homeostasis. However, the auxin transport mechanisms in horticultural crops remain largely unexplored. Here, we identified and characterized PILS genes in tomato (Solanum lycopersicum). Promoter analysis revealed enrichment in TGA[C/T]G motifs, suggesting transcriptional regulation by TGA factors in the bZIP family. Subcellular localization studies confirmed that all tomato PILS proteins localize in the endoplasmic reticulum. PILS2 exhibited the highest expression across examined tissues, and its close homologue PILS6 showed a similar but less pronounced expression pattern. Silencing PILS2 significantly inhibited shoot and root growth. Phylogenetic and expression analyses identified the homologs of Arabidopsis TGA1, TGA3, TGA4, and TGA7 in tomato genome, with tomato TGA7 showing higher expression in roots. Notably, silencing tomato TGA7, but not TGA1, TGA3, or TGA4, strongly impaired shoot and root growth. Molecular assays demonstrated that TGA7 directly binds to the PILS2 promoter to activate its transcription. These findings uncover a TGA7-PILS2 regulatory module that governs plant growth and offer new insights into the function and regulation of PILS genes in tomato.

生长素在植物生长调控中起着举足轻重的作用。PIN- formed （PIN）蛋白促进长距离极性生长素运输，而最近发现的PIN- like （PILS）蛋白调节细胞内生长素稳态。然而，生长素在园艺作物中的转运机制仍未得到充分研究。在此，我们对番茄（Solanum lycopersicum）中的PILS基因进行了鉴定和表征。启动子分析显示，bZIP家族中TGA[C/T]G基序富集，表明TGA因子参与了bZIP家族的转录调控。亚细胞定位研究证实，所有的番茄PILS蛋白都定位于内质网。PILS2在检查的组织中表现出最高的表达，其相近的同源物PILS6表现出相似但不太明显的表达模式。沉默PILS2显著抑制了茎和根的生长。系统发育和表达分析发现拟南芥TGA1、TGA3、TGA4和TGA7在番茄基因组中具有同源性，其中TGA7在番茄根中表达量较高。值得注意的是，沉默番茄TGA7，而不是TGA1， TGA3或TGA4，会严重损害茎和根的生长。分子分析表明，TGA7直接结合到PILS2启动子上，激活其转录。这些发现揭示了调控植物生长的TGA7-PILS2调控模块，并为番茄中PILS基因的功能和调控提供了新的见解。

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引用次数: 0

ATP-binding cassette G23 is required for Arabidopsis seed coat suberization 拟南芥种皮脱落需要atp结合盒G23。

IF 4.2 2区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Plant Science

Pub Date : 2024-12-17 DOI: 10.1016/j.plantsci.2024.112361

Ryeo Jin Kim, Yuyang Zhang, Mi Chung Suh

Suberin is an extracellular hydrophobic polymer deposited in seed coats that acts as a barrier to regulate the movement of ions, water, and gases, and protects seeds against pathogens. However, the molecular mechanisms underlying suberin deposition in the seed coat remain unknown. In this study, the in planta role of ATP-binding cassette G23 (ABCG23) was investigated in the Arabidopsis seed coat. ABCG23 transcripts were predominantly expressed in the outer integument1 (oi1) of seed coats and the endodermal cells of roots. The fluorescence of the eYFP:ABCG23 construct was observed in the plasma membranes of the tobacco epidermis, seed coat oi1, and root endodermal cells. Seed coats of abcg23–1 and abcg23–2 mutants exhibited reduced autofluorescence under UV light and increased permeability to tetrazolium salts. Total suberin loads and major suberin components, C24 ω-hydroxy fatty acids and 1, ω-dicarboxylic acids were significantly decreased in the mutant seed coats. The ratio of seed germination and seedling establishment of abcg23–1 and abcg23–2 was significantly reduced compared to the WT under salt and osmotic stress conditions. The bimolecular fluorescence complementation assay showed homodimeric interactions of ABCG-2, −6, −20, and −23 and heterodimeric interactions between ABCG23 and ABCG-2, −6, −11, or −20. Our findings indicate that ABCG23 contributes to the transport of suberin monomers in the Arabidopsis seed coats.

木素是一种沉积在种皮上的细胞外疏水性聚合物，作为调节离子、水和气体运动的屏障，保护种子免受病原体的侵害。然而，种皮中木质素沉积的分子机制尚不清楚。本研究研究了atp结合盒G23 （ABCG23）在拟南芥种皮中的作用。ABCG23转录本主要表达于种皮外被1 （oi1）和根的内胚层细胞。构建的eYFP:ABCG23在烟草表皮、种皮油1和根内胚层细胞的质膜上观察到荧光。abcg23-1和abcg23-2突变体种皮在紫外光下的自身荧光减弱，对四氮唑盐的渗透性增加。突变体种皮的亚色素总含量和主要亚色素成分C24 ω-羟基脂肪酸和1，ω-二羧酸含量显著降低。在盐胁迫和渗透胁迫条件下，abcg23-1和abcg23-2的种子发芽率和成苗率均显著低于WT。双分子荧光互补实验显示ABCG-2、-6、-20和-23的同二聚体相互作用以及ABCG23与ABCG-2、-6、-11和-20的异二聚体相互作用。我们的研究结果表明，ABCG23参与了拟南芥种皮中亚胺单体的运输。

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引用次数: 0

Enhancing starch levels, granule size and phosphate content in Chlamydomonas reinhardtii through overexpression of ChlreSEX4 通过过表达ChlreSEX4提高莱茵衣藻淀粉水平、颗粒大小和磷酸盐含量。

IF 4.2 2区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Plant Science

Pub Date : 2024-12-15 DOI: 10.1016/j.plantsci.2024.112360

Florencia Torresi , Julieta B. Carrillo , Diego F. Gomez-Casati , Maria V. Busi , Mariana Martín

Chlamydomonas reinhardtii is a green alga that has been widely used as a model organism for studying various cellular processes, including starch metabolism. In this alga, starch undergoes continuous phosphorylation during its synthesis and degradation. We recently identified and characterized ChlreSEX4 (starch excess 4), a glucan phosphatase from C. reinhardtii, orthologous to Arabidopsis thaliana SEX4, which is able to bind and dephosphorylate amylopectin in vitro. To explore the possibility of manipulating starch phosphorylation levels in C. reinhardtii, we overexpressed the ChlreSEX4 gene in Chlamydomonas and characterized the resulting lines. Results showed a high phosphatase activity in the overexpressing lines, accompanied by an increase in starch content, greater granule size and higher levels of granule-bound phosphate, without changes in triglyceride content. This work allowed us not only to discover a new method to enhance starch accumulation without affecting the lipid content of the alga, but also to obtain a more phosphorylated starch, which would have diverse applications in biotechnology.

莱茵衣藻（Chlamydomonas reinhardtii）是一种绿藻，被广泛用作研究包括淀粉代谢在内的各种细胞过程的模式生物。在这种藻类中，淀粉在合成和降解过程中经历了持续的磷酸化。我们最近鉴定并鉴定了一种来自拟南芥（Arabidopsis thaliana）的葡聚糖磷酸酶ChlreSEX4 (starch excess 4)，它与拟南芥的SEX4同源，能够在体外结合支链淀粉并使其去磷酸化。为了探索控制衣藻淀粉磷酸化水平的可能性，我们在衣藻中过表达了ChlreSEX4基因，并对所产生的品系进行了表征。结果表明，在过表达系中，磷酸酶活性高，淀粉含量增加，颗粒大小和颗粒结合磷酸盐水平较高，甘油三酯含量没有变化。这项工作不仅使我们发现了一种在不影响藻类脂质含量的情况下提高淀粉积累的新方法，而且还获得了一种磷酸化程度更高的淀粉，这将在生物技术中有多种应用。

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引用次数: 0

Effect analysis of S5-interacting genes on rice hybrid sterility using nontransgenic gamete killer 利用非转基因配子杀手分析 S5 相互作用基因对水稻杂种不育性的影响。

IF 4.2 2区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Plant Science

Pub Date : 2024-12-13 DOI: 10.1016/j.plantsci.2024.112357

Jie Li , Fu Huang , Yingxia Jiang, Jianglei Rao, Yourong Fan, Jiangyi Yang

While hybrids between japonica and indica rice exhibit strong heterosis, they often suffer from hybrid sterility (HS). Hybrid fertility of the embryo sac is predominantly regulated by a three-gene system (comprising closely linked ORF3, ORF4 and ORF5) at rice S5 locus. The cooperation of ORF5+ and ORF4+ can result in endoplasmic reticulum (ER) stress and sporophytically kill all embryo sacs, while ORF3+ can gametophytically protect the residing embryo sac. We previously identified four S5-interacting genes (SIGs) using a transgenic line BL^ORF5⁺ (Balilla carrying transgenic ORF5+) and a wide compatibility variety Dular (DL or D). Homozygote and hemizygote of ORF5+ transgene had significantly different spikelet fertility (SF), which disturbed the phenotypic effects of SIGs. However, HS effects of SIGs under the endogenous (nontransgenic) gamete killer remained unknown. We formerly constructed a semisterile near isogenic line (NIL) S5-BL/NJ by introgressing S5 fragment of indica rice Nanjing11 (NJ or N, carrying ORF3+ORF4-ORF5+ haplotypes) into the genome of japonica rice Balilla (BL or B, carrying ORF3-ORF4+ORF5- haplotypes). The gamete-protecting effect of ORF3+ in NJ may confuse SF effect of the SIGs, so we knocked out ORF3+ of S5-NJ/NJ and crossed it with BL to get gamete-killing S5-BL/NJ^ΔORF3+, which can kill all (KA) gametes (abbreviated as enS5KA). Compared with the exS5KA line (a NIL carrying ORF5+ transgenic, wihch can kill all gamete), the enS5KA line conferred SIGs a more pronounced SF effect. The enS5KA,SIG-DDDD (four SIGs carry homozygous DL alleles) genotype caused a SF of about 78 %, while SF of exS5KA,SIG-DDDD was only about 62 %. Moreover, all SIGs acted in a sporophytic manner without segregation distortion of genotype. Although enS5KA,SIG-DDDD plants had high SF, the ER stress still existed. The ovule section revealed that enS5KA,SIG-BBBB genotype (four SIGs carry homozygous BL allele, with ER stress and SF < 5 %) caused abnormal degradation of nucellar cells and functional megaspores. In contrast, enS5KA,SIG-DDDD genotype preserved most nucellar cells and functional megaspores. These results lay the foundation for further research on HS mechanism of S5 and SIGs and cloning of candidate genes.

虽然粳稻和籼稻的杂交种表现出很强的异交性，但它们经常会出现杂交不育症（HS）。胚囊的杂交育性主要受水稻 S5 基因座上的三基因系统（包括紧密相连的 ORF3、ORF4 和 ORF5）调控。ORF5+ 和 ORF4+ 的协同作用会导致内质网（ER）应激，孢子体会杀死所有胚囊，而 ORF3+ 则能配子体保护驻留的胚囊。此前，我们利用转基因品系 BLORF5+（携带转基因 ORF5+ 的 Balilla）和广相容性品种 Dular（DL 或 D）鉴定出了四个 S5 相互作用基因（SIG）。ORF5+转基因的同源杂合子和半杂合子的小穗生育力（SF）明显不同，这干扰了SIGs的表型效应。然而，SIGs 在内源（非转基因）配子杀手下的 HS 效应仍然未知。以前，我们通过将籼稻南京11（NJ或N，携带ORF3+ORF4-ORF5+单倍型）的S5片段导入粳稻Balilla（BL或B，携带ORF3-ORF4+ORF5-单倍型）的基因组，构建了半固定近等基因系（NIL）S5-BL/NJ。NJ中ORF3+的配子保护作用可能会混淆SIG的SF效应，因此我们敲除了S5-NJ/NJ的ORF3+，并将其与BL杂交，得到了能杀死所有（KA）配子的配子杀伤型S5-BL/NJΔORF3+（简称enS5KA）。与exS5KA品系（携带ORF5+的转基因NIL，可杀死所有配子）相比，enS5KA品系赋予SIG的SF效应更明显。enS5KA,SIG-DDDD（四个 SIG 携带同源 DL 等位基因）基因型的 SF 值约为 78%，而 exS5KA,SIG-DDDD 的 SF 值仅为 62%。此外，所有 SIG 都以孢子体方式发挥作用，基因型没有发生分离失真。虽然 enS5KA,SIG-DDDD 植株的 SF 值很高，但仍然存在 ER 胁迫。胚珠切片显示，enS5KA,SIG-BBBB 基因型（4 个 SIG 携带同源 BL 等位基因，具有 ER 胁迫，SF < 5%）导致核细胞异常退化和功能性巨孢子。相反，enS5KA,SIG-DDDD 基因型保留了大部分核细胞和功能性巨孢子。这些结果为进一步研究 S5 和 SIG 的 HS 机制以及克隆候选基因奠定了基础。

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