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IPTG- and estradiol-inducible gene expression systems in the unicellular red alga Cyanidioschyzon merolae. 单细胞红藻中IPTG和雌二醇诱导的基因表达系统。
IF 6.9 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2026-01-06 DOI: 10.1093/plphys/kiaf575
Takayuki Fujiwara, Shunsuke Hirooka, Shota Yamashita, Shin-Ya Miyagishima

The genetically tractable unicellular red alga Cyanidioschyzon merolae has a remarkably simple genome (4,775 nucleus-encoded proteins) and cellular architecture. It contains only a single set of most membranous organelles, making it a valuable tool for elucidating the fundamental mechanisms of photosynthetic eukaryotes. However, as in other genetically tractable eukaryotic algae, previously developed systems for inducible gene expression rely on environmental stimuli such as heat shock or ammonium depletion, which impact cellular physiology and thus limit their usage. To overcome this issue, we developed IPTG- and estradiol-inducible gene expression systems in C. merolae in which the addition of these chemicals itself has no impact on cellular growth or the transcriptome. Additionally, we established IPTG- and estradiol-inducible protein knockdown systems and successfully degraded the endogenous chloroplast division protein DRP5B using the estradiol-inducible system. These systems facilitate functional genomic analyses in C. merolae, especially for understanding physiological mechanisms and their interactions in photosynthetic eukaryotes.

单细胞红藻merolae具有非常简单的基因组(4775个核编码蛋白)和细胞结构。它只包含一组大多数膜细胞器,使其成为阐明光合作用真核生物基本机制的有价值的工具。然而,与其他遗传易感的真核藻类一样,以前开发的诱导基因表达系统依赖于环境刺激,如热休克或铵耗竭,这会影响细胞生理,从而限制其使用。为了克服这一问题,我们在C. merolae中开发了IPTG和雌二醇诱导的基因表达系统,其中添加这些化学物质本身对细胞生长或转录组没有影响。此外,我们建立了IPTG和雌二醇诱导的蛋白敲低系统,并成功地利用雌二醇诱导系统降解了内源性叶绿体分裂蛋白DRP5B。这些系统促进了merolae的功能基因组分析,特别是了解光合真核生物的生理机制及其相互作用。
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引用次数: 0
BnaMYB52 negatively regulates drought resistance by controlling stomatal and non-stomatal water loss in Brassica napus BnaMYB52通过控制甘蓝型油菜气孔和非气孔水分损失负调控抗旱性
IF 7.4 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2026-01-04 DOI: 10.1093/plphys/kiaf681
Xiaowei Wu, Jiang Ye, Xiang Li, Lintang Xu, Qian Qu, Yuyan Xiang, Jinglu Zhou, Shuai Fang, Liangqian Yu, Xu Han, Liang Guo, Xuan Yao
Brassica napus is one of the most important oil crops worldwide, and its production is severely threatened by drought stress. Breeding drought-resistant cultivars is needed to cope with global climate change. However, few drought resistance regulators have been identified, and the molecular mechanism of drought resistance is largely unknown in B. napus. Here, we characterized a typical R2R3-MYB transcription factor, BnaMYB52, as a negative regulator of drought resistance in B. napus. The disruption of BnaMYB52 resulted in enhanced drought resistance, whereas overexpression of BnaA09.MYB52 reduced drought resistance. Further analyses showed that BnaMYB52 affects leaf water loss from both stomata and the cuticle to negatively regulate drought resistance. We demonstrated that BnaMYB52 acts as a transcription repressor regulating the expression of the downstream targets BnaMYB96 and BnaMYB30, controlling both ABA signaling and wax biosynthesis, and BnaMYC2, controlling stomatal density. This study uncovers the molecular mechanism of BnaMYB52-regulated drought resistance and provides a genetic resource for the molecular breeding of drought-resistant B. napus cultivars.
甘蓝型油菜是世界上最重要的油料作物之一,其生产受到干旱胁迫的严重威胁。培育抗旱品种是应对全球气候变化的需要。然而,目前发现的抗旱性调控因子很少,甘蓝型油菜抗旱性的分子机制在很大程度上是未知的。本研究发现,典型的R2R3-MYB转录因子BnaMYB52是甘蓝型油菜抗旱性的负调控因子。BnaMYB52基因的破坏导致了抗旱性的增强,而BnaA09基因的过表达导致了抗旱性的增强。MYB52降低了抗旱性。进一步分析表明,BnaMYB52同时影响叶片气孔和角质层的水分流失,负调控叶片抗旱性。我们发现BnaMYB52作为一个转录抑制因子,调控下游靶点BnaMYB96和BnaMYB30的表达,控制ABA信号和蜡的生物合成,BnaMYC2则控制气孔密度。本研究揭示了bnamyb52调控甘蓝型油菜抗旱性的分子机制,为甘蓝型油菜抗旱性的分子育种提供了遗传资源。
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引用次数: 0
The multifunctional protein OsAIM1 regulates floret opening and closure timing via jasmonic acid-mediated lodicule dynamics in rice. 多功能蛋白OsAIM1通过茉莉酸介导的水稻小泡动力学调节小花的开放和关闭时间。
IF 6.9 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2026-01-02 DOI: 10.1093/plphys/kiaf690
Yuan Hu, Haishen Li, Haigang Hou, Song Cui, Zhuang Xu, Benyuan Hao, Liang Cai, Liang Zhu, Jie Wang, Kongyou Chang, Wen Li, Weihao Shao, Shanshan Zhu, Chao Li, Zhigang Zhao, Ling Jiang, Yunlu Tian, Xi Liu, Shijia Liu, Liangming Chen, Shirong Zhou, Jianmin Wan

Rice (Oryza sativa) indica and japonica inter-subspecific hybrids hold significant potential for increasing yields. However, differences in diurnal flower-opening time (DFOT) between the two subspecies limit the effective exploitation of this heterosis. Additionally, the timing of post-anthesis glume closure (PAGC) affects both hybrid seed yield and quality. Despite their importance, the molecular mechanisms underlying these processes, particularly glume closure, remain poorly understood. In this study, we identify OsAIM1 as a pivotal regulator of both DFOT and PAGC in rice. The aim1-2 mutant exhibits delayed DFOT and impaired PAGC while maintaining normal floret structure, emphasizing its crucial role in floret dynamics. OsAIM1 is highly expressed in lodicules and encodes a peroxisome-localized multifunctional protein. Functional analyses reveal that OsAIM1 regulates lodicule swelling during floret opening and withering post-anthesis, processes essential for glume movement. We further demonstrate that OsAIM1-dependent jasmonic acid (JA) biosynthesis is indispensable for coordinating floret opening and closure and influences sugar transport to ensure proper lodicule dynamics. Importantly, natural variation in the OsAIM1 coding region contributes to DFOT divergence between japonica and indica subspecies, providing a molecular basis for their asynchronous flowering. These findings establish OsAIM1 as a key regulator of floret dynamics and a promising molecular target for synchronizing flowering in hybrid rice production.

水稻(Oryza sativa)籼稻和粳稻亚种间杂交种具有显著的增产潜力。然而,两个亚种之间日开花时间的差异限制了这一杂种优势的有效利用。此外,花后颖片闭合(PAGC)的时间对杂交种子的产量和品质都有影响。尽管它们很重要,但这些过程的分子机制,特别是颖片闭合机制,仍然知之甚少。在这项研究中,我们发现OsAIM1是水稻DFOT和PAGC的关键调节因子。在维持正常小花结构的同时,aim1-2突变体表现出延迟的DFOT和受损的PAGC,强调了其在小花动力学中的重要作用。OsAIM1在小泡中高度表达,编码过氧化物酶体定位的多功能蛋白。功能分析表明,OsAIM1调控小花开放和花后凋谢过程中的小叶膨胀,这是颖花运动所必需的过程。我们进一步证明,osaim1依赖性茉莉酸(JA)的生物合成对于协调小花的开放和关闭是必不可少的,并影响糖的运输以确保适当的小叶动力学。重要的是,OsAIM1编码区的自然变异有助于粳稻和籼稻亚种之间的DFOT差异,为它们的非同步开花提供了分子基础。这些发现表明OsAIM1是水稻小花动态的关键调控因子,是杂交水稻同步开花的一个有前景的分子靶点。
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引用次数: 0
Evolution of phenolic conjugate incorporation in plant sporopollenin. 植物孢粉中酚类偶联物结合的演化。
IF 6.9 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2025-12-31 DOI: 10.1093/plphys/kiaf680
Lei Xu, An-Mei Zhu, Yi-Fan Ju, Gui Yan, Yu-Chi He

Sporopollenin, a structurally complex and chemically recalcitrant biopolymer, forms the outer exine layer of plant spores and pollen, protecting male gametes from environmental stresses. Varying levels of phenolic constituents are incorporated as building blocks into sporopollenin in many species, but how this process evolved remains unclear. Using an optimized alkaline hydrolysis method, along with NMR spectroscopy and GC-MS, we determined that plants have evolved the ability to incorporate phenolic compounds into sporopollenin in a phylogenetically ordered manner. Covalently linked phenolic constituents, including p-coumarate (p-CA), ferulate, p-hydroxybenzoate, naringenin, the canonical monolignol p-hydroxyphenyl unit and guaiacyl unit, occur in sporopollenin of vascular but not non-vascular plants. Evolutionary analyses showed that the metabolic scaffold for phenolic precursors evolved before the integration of phenolics into sporopollenin in vascular plants. The conserved multicopper oxidase SCULP1, which incorporates p-CA into sporopollenin, co-occurred with p-coumaroylation of sporopollenin in vascular plants, likely contributing to the prevalence of p-coumaroylated sporopollenin. These findings provide an evolutionary framework for understanding genetic associations with sporopollenin chemical diversification and plant adaptation.

孢粉素是一种结构复杂、化学性质顽固的生物聚合物,形成植物孢子和花粉的外外外表皮层,保护雄性配子免受环境胁迫。在许多物种中,不同水平的酚类成分作为构建块被纳入孢子孢粉,但这一过程是如何进化的尚不清楚。利用优化的碱性水解方法,以及核磁共振光谱和气相色谱-质谱分析,我们确定植物已经进化出了将酚类化合物整合到孢粉素中的能力。共价连接的酚类成分,包括对香豆酸酯(p-CA)、阿魏酸酯、对羟基苯甲酸酯、柚皮素、典型单脂醇对羟基苯基单位和愈创木酰单位,存在于维管植物的孢子花粉中,而不存在于非维管植物中。进化分析表明,在维管植物中,酚类前体的代谢支架在酚类物质整合到孢子花粉素之前就已经进化了。保守的多铜氧化酶SCULP1将p-CA整合到孢子粉中,与维管植物孢子粉的对香豆素化共同发生,可能导致了对香豆素化孢子粉的流行。这些发现为理解孢粉素化学多样化和植物适应的遗传关联提供了一个进化框架。
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引用次数: 0
mTACT: A cell type-specific transportome-scale amiRNA toolbox to overcome functional redundancy in Arabidopsis. mTACT:一个细胞类型特异性转运体规模的amiRNA工具箱,以克服拟南芥的功能冗余。
IF 6.9 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2025-12-31 DOI: 10.1093/plphys/kiaf682
Moran Anfang, Shir Ben Yaakov, Ning Su, Anat Shafir, Jenia Binenbaum, Reem Haj Yahya, Xikai Yu, Carl Procko, Hamtual Bar, Joanne Chory, Julian I Schroeder, Yosef Fichman, Itay Mayrose, Eilon Shani, Yuqin Zhang

In plants, both developmental processes and environmental responses are spatiotemporally regulated by an assembly of signaling molecules such as hormones, secondary metabolites, and ions. The ability of these signaling molecules to move within and across plant tissues is essential for various developmental cues. However, the characterization of transported signaling molecules and their translocation mechanisms is difficult due to the functional redundancy of plant genomes and shortcomings in methodologies. Here, we report our development of the Multi Targeted AmiRNA Cell type-specific Transportome-scale (mTACT) toolbox, which can be used to reveal phenotypic plasticity in plants. mTACT is based on a large set of artificial microRNAs (amiRNAs), each designed to optimally target multiple members of a particular gene family encoding transporter proteins. In total, the mTACT toolbox includes 5,565 amiRNAs, targeting 81.7% of the Arabidopsis (Arabidopsis thaliana) transportome. The amiRNA library can be driven under 12 cell type-specific promoters, allowing the design of spatial-specific genetic screens. mTACT is further divided into eight sub-libraries of amiRNAs targeting a functionally defined protein class. A proof-of-concept screen validated the mTACT approach by identifying phenotypes linked to both known and unidentified genes. With the ability to overcome functional redundancy in a transportome-scale, cell type-specific manner, the mTACT toolbox will allow the plant research community to study previously hidden genetic factors required for long- and short-distance translocation of signaling molecules.

在植物中,发育过程和环境反应都是由一系列信号分子(如激素、次生代谢物和离子)在时空上进行调节的。这些信号分子在植物组织内和组织间移动的能力对于各种发育线索至关重要。然而,由于植物基因组的功能冗余和方法上的缺陷,表征转运信号分子及其易位机制是困难的。在这里,我们报告了我们开发的多靶向AmiRNA细胞类型特异性转运体规模(mTACT)工具箱,该工具箱可用于揭示植物的表型可塑性。mTACT是基于大量的人工微rna (amiRNAs),每个微rna都被设计成最优地靶向编码转运蛋白的特定基因家族的多个成员。总的来说,mTACT工具箱包括5565个amirna,靶向81.7%的拟南芥(拟南芥)转运组。amiRNA文库可以在12个细胞类型特异性启动子下驱动,从而允许设计空间特异性遗传筛选。mTACT进一步分为八个靶向功能定义蛋白类的amirna亚库。通过识别与已知和未知基因相关的表型,概念验证筛选验证了mTACT方法。mTACT工具箱具有以运输体规模和细胞类型特异性的方式克服功能冗余的能力,将使植物研究界能够研究以前隐藏的信号分子长距离和短距离易位所需的遗传因素。
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引用次数: 0
No More Fishes: Plants as a Bio-Factory for Omega-3 Fatty Acid. 不再有鱼:植物是Omega-3脂肪酸的生物工厂。
IF 7.4 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2025-12-26 DOI: 10.1093/plphys/kiaf679
Prateek Jain,Maneesh Lingwan
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引用次数: 0
Bigger is not always better: Optimizing leaf area index with narrow leaf shape in soybean 不是越大越好:窄叶型优化大豆叶面积指数
IF 7.4 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2025-12-24 DOI: 10.1093/plphys/kiaf663
Bishal G Tamang, Gregory Bernard, Carl J Bernacchi, Brian W Diers, Elizabeth A Ainsworth
Modern soybean (Glycine max) varieties have higher than optimal leaf area index (LAI), which could divert resources from reproductive growth. Altering leaf shape could be a simple strategy to reduce LAI. To test this, we developed 204 near-isogenic lines differing in leaf morphology by introgressing the ln allele that confers the narrow-leaf trait from two donor parents (PI 612713A and PI 547745) into the elite, broad-leaf cultivar LD11-2170. We evaluated the lines across two locations and two row spacings (38-cm and 76-cm) to assess how reduced investment in leaf area influences canopy architecture, crop physiology, and yield. Narrow-leaf lines showed 13% lower peak LAI and 3% lower digital biomass compared to broad-leaf counterparts yet maintained yield parity (5,756 vs. 5.801 kg ha-1, p = 0.43) across environmental conditions. Photosynthetic capacity remained largely unchanged, with narrow-leaf lines showing modest increases in electron transport rate and leaf mass per area. Narrow-leaf lines achieved similar canopy closure timing despite lower LAI, suggesting architectural compensation mechanisms. The most striking difference appeared in seed packaging, with 34% of pods containing four seeds in narrow-leaf lines compared to only 1.8% in broad-leaf lines. There was a nonlinear relationship between peak LAI and yield, with optimal LAI values of 9-11 varying by environment. These findings show that the single-gene GmJAG1-controlled narrow-leaf trait offers a tractable strategy for reducing LAI and maintaining high productivity. This could reduce the metabolic costs associated with excessive canopy development and support sustainable agriculture under increasing climate variability.
现代大豆(Glycine max)品种具有较高的最优叶面积指数(LAI),可将资源从生殖生长中分流出来。改变叶片形状可能是降低LAI的一个简单策略。为了验证这一点,我们通过将来自两个供体亲本(PI 612713A和PI 547745)的ln等位基因渗入到精英阔叶品种LD11-2170中,获得了204个叶片形态不同的近等基因系。我们评估了两个位置和两行间距(38 cm和76 cm)的线,以评估叶面积投资减少如何影响冠层结构、作物生理和产量。与阔叶系相比,窄叶系的LAI峰值低13%,数字生物量低3%,但在各种环境条件下均保持产量持平(5,756对5.801 kg ha-1, p = 0.43)。光合能力基本保持不变,窄叶线显示出电子传递速率和每面积叶质量的适度增加。尽管LAI较低,但窄叶线的冠层闭合时间相似,这表明存在建筑补偿机制。最显著的差异出现在种子包装上,窄叶系34%的荚果含有4颗种子,而阔叶系只有1.8%。LAI峰值与产量呈非线性关系,9 ~ 11的最佳LAI值随环境变化而变化。这些结果表明,单基因gmjag1控制的窄叶性状为降低LAI和保持高产提供了一种可处理的策略。这可以减少与冠层过度开发相关的代谢成本,并支持气候变率日益增加的可持续农业。
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引用次数: 0
From Glossy to Glaucous: How TaMYB96-2D Controls Wax Deposition and Drought Resilience. 从光滑到白霜:TaMYB96-2D如何控制蜡沉积和抗旱能力。
IF 7.4 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2025-12-24 DOI: 10.1093/plphys/kiaf675
Ritu Singh,Erin Cullen
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引用次数: 0
Eco-physiological responses of Hieracium pilosella and Trifolium pratense to reduced air pressure 毛茛和三叶草对低气压的生态生理反应
IF 7.4 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2025-12-24 DOI: 10.1093/plphys/kiaf631
Bouchra El Omari, Silvia Lembo, Matteo Dainese, Paul Illmer, Nadine Praeg, Andreas Meul, Dolores Asensio, Georg Niedrist
Climate change is a major factor shaping the distribution of plant species. A well-documented response consequence is the upward shift of plant species to higher elevations as they track their thermal niches. However, plants migrating upward face complex environmental changes shaped by multiple interacting factors. Among these, reduced air pressure remains relatively understudied, its effects are often confounded with other covarying parameters. This study investigated the direct impact of reduced air pressure on the eco-physiological responses of two plant species (Hieracium pilosella L. and Trifolium pratensis L.). The plants were grown for four weeks in controlled climatic chambers under different air pressures (85, 75, and 62 kPa), while all other environmental parameters were kept constant. At the end of the experiment, photosynthesis, chlorophyll fluorescence, growth, carbohydrate content, carbon stable isotopes, and plant nitrogen concentrations were determined. Reduced air pressure decreased growth, carbon isotopic discrimination and chlorophyll content, but increased CO2 fixation efficiency and carbohydrate accumulation in the leaves. These results suggest that reduced air pressure impacts plant performance during upslope migration and may, in turn, contribute to shaping future distribution patterns in alpine ecosystems.
气候变化是影响植物物种分布的一个主要因素。一个有充分证据的响应结果是植物物种在追踪其热生态位时向上迁移到更高的海拔。然而,植物向上迁移面临着多种相互作用因素形成的复杂环境变化。其中,气压降低的研究相对较少,其影响往往与其他共变参数相混淆。研究了气压降低对两种植物(Hieracium pilosella L.和Trifolium pratensis L.)生态生理反应的直接影响。这些植物在不同气压(85、75和62 kPa)的可控气候室中生长四周,而所有其他环境参数保持不变。实验结束时,测定光合作用、叶绿素荧光、生长、碳水化合物含量、碳稳定同位素和植物氮浓度。气压降低降低了叶片的生长、碳同位素识别和叶绿素含量,但增加了叶片的CO2固定效率和碳水化合物积累。这些结果表明,在上坡迁移过程中,气压降低会影响植物的表现,并可能反过来影响未来高山生态系统的分布格局。
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引用次数: 0
Deciphering the regulatory network underlying auxin depletion-induced fruitlet abscission in litchi ( Litchi chinensis Sonn.) 生长素耗竭诱导荔枝果实脱落的调控网络解析
IF 7.4 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2025-12-23 DOI: 10.1093/plphys/kiaf676
Zidi He, Xingshuai Ma, Hang Zhang, Zishang Kong, Fei Wang, Ye Yuan, Minglei Zhao, Jianguo Li
Auxin transport through the abscission zone (AZ) is crucial for preventing organ abscission in plants; however, the regulatory mechanisms involved remain poorly understood. Here, we demonstrate that exogenous auxin application alone fully inhibits abscission triggered by litchi (Litchi chinensis Sonn.) fruitlet removal, which depletes auxin in the AZ. Following fruitlet removal, we observed sequential and significant alterations in seven biological processes within the AZ, including the transient suppression of auxin signaling and activation of ethylene and abscisic acid (ABA) signaling, followed by the increased production of reactive oxygen species (ROS), reduced carbohydrate content, and ultimately, the induction of programmed cell death (PCD) and cell wall remodeling (CWR). Moreover, we identified 34 transcription factors as potential key regulators and constructed transcriptional regulatory networks involved in auxin depletion-induced abscission. Notably, we characterized LcMYB62 as a positive regulator of abscission, likely functioning by transactivating genes associated with cell wall remodeling. We further showed that the transcription factor AUXIN RESPONSE FACTOR 5 (LcARF5) binds to and activates LcMYB62, suggesting a LcARF5-LcMYB62-CWR transcriptional regulatory cascade in litchi fruitlet abscission. Overall, our findings provide a comprehensive overview of the gene regulatory network governing auxin-mediated fruitlet abscission in litchi, providing insights into the mechanisms by which auxin depletion in the AZ triggers this process.
植物生长素通过脱落区运输是防止器官脱落的关键。然而,所涉及的监管机制仍然知之甚少。在这里,我们证明了外源生长素单独施用完全抑制了荔枝(litchi chinensis Sonn.)去果引发的脱落,这消耗了荔枝体内的生长素。在去果之后,我们观察到七个生物过程的顺序和显著变化,包括生长素信号的短暂抑制和乙烯和脱落酸(ABA)信号的激活,随后活性氧(ROS)的产生增加。减少碳水化合物含量,并最终诱导程序性细胞死亡(PCD)和细胞壁重塑(CWR)。此外,我们确定了34个转录因子作为潜在的关键调控因子,并构建了参与生长素消耗诱导脱落的转录调控网络。值得注意的是,我们认为LcMYB62是脱落的积极调节因子,可能通过反激活与细胞壁重塑相关的基因来发挥作用。我们进一步发现,转录因子AUXIN RESPONSE factor 5 (LcARF5)结合并激活LcMYB62,表明LcARF5-LcMYB62- cwr转录调控级联在荔枝果实脱落中起作用。总的来说,我们的研究结果提供了生长素介导的荔枝果实脱落的基因调控网络的全面概述,提供了对AZ中生长素耗尽触发这一过程的机制的见解。
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引用次数: 0
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Plant Physiology
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