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Unique structural attributes of the PSI-NDH supercomplex in Physcomitrium patens. 专利植物中 PSI-NDH 超级复合物的独特结构属性。
IF 6.2 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2024-11-03 DOI: 10.1111/tpj.17116
Monika Opatíková, Roman Kouřil

Cyclic electron transport around photosystem I (PSI) is essential for the protection of the photosynthetic apparatus in plants under diverse light conditions. This process is primarily mediated by Proton Gradient Regulation 5 protein/Proton Gradient Regulation 5-like photosynthetic phenotype 1 protein (PGR5/PGRL1) and NADH dehydrogenase-like complex (NDH). In angiosperms, NDH interacts with two PSI complexes through distinct monomeric antennae, LHCA5 and LHCA6, which is crucial for its higher stability under variable light conditions. This interaction represents an advanced evolutionary stage and offers limited insight into the origin of the PSI-NDH supercomplex in evolutionarily older organisms. In contrast, the moss Physcomitrium patens (Pp), which retains the lhca5 gene but lacks the lhca6, offers a glimpse into an earlier evolutionary stage of the PSI-NDH supercomplex. Here we present structural evidence of the Pp PSI-NDH supercomplex formation by single particle electron microscopy, demonstrating the unique ability of Pp to bind a single PSI in two different configurations. One configuration closely resembles the angiosperm model, whereas the other exhibits a novel PSI orientation, rotated clockwise. This structural flexibility in Pp is presumably enabled by the variable incorporation of LHCA5 within PSI and is indicative of an early evolutionary adaptation that allowed for greater diversity at the PSI-NDH interface. Our findings suggest that this variability was reduced as the structural complexity of the NDH complex increased in vascular plants, primarily angiosperms. This study not only clarifies the evolutionary development of PSI-NDH supercomplexes but also highlights the dynamic nature of the adaptive mechanisms of plant photosynthesis.

在不同的光照条件下,围绕光系统 I(PSI)的循环电子传递对于保护植物的光合装置至关重要。这一过程主要由质子梯度调节 5 蛋白/质子梯度调节 5 样光合表型 1 蛋白(PGR5/PGRL1)和 NADH 脱氢酶样复合物(NDH)介导。在被子植物中,NDH 通过不同的单体触角 LHCA5 和 LHCA6 与两个 PSI 复合物相互作用,这对其在多变光照条件下保持较高的稳定性至关重要。这种相互作用代表了一个高级进化阶段,对更古老生物体中 PSI-NDH 超级复合物的起源提供了有限的启示。相比之下,保留了 lhca5 基因但缺乏 lhca6 基因的藓类 Physcomitrium patens(Pp)则为 PSI-NDH 超级复合物的早期进化阶段提供了一瞥。在这里,我们通过单颗粒电子显微镜展示了 Pp PSI-NDH 超级复合物形成的结构证据,证明了 Pp 以两种不同构型结合单个 PSI 的独特能力。一种构型与被子植物模型非常相似,而另一种构型则表现出一种顺时针旋转的新型 PSI 取向。Pp的这种结构灵活性可能是通过LHCA5在PSI中的可变结合实现的,并表明早期的进化适应使PSI-NDH界面具有更大的多样性。我们的研究结果表明,随着维管植物(主要是被子植物)中 NDH 复合物结构复杂性的增加,这种可变性有所降低。这项研究不仅阐明了 PSI-NDH 超级复合体的进化发展,还突出了植物光合作用适应机制的动态性质。
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引用次数: 0
Effects of phosphorylation on CsTT12 transport function: A comparative phosphoproteomic analysis of flavonoid biosynthesis in tea plants (Camellia sinensis). 磷酸化对 CsTT12 转运功能的影响:茶树(Camellia sinensis)黄酮类化合物生物合成的磷酸化蛋白质组比较分析
IF 6.2 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2024-11-03 DOI: 10.1111/tpj.17120
Na-Na Wang, Ke-Yan Xiu, Min Deng, Qi-Yun Liu, Di-Di Jin, Qiao-Mei Zhao, Huang-Qiang Su, Ting-Ting Qiu, Hai-Yan Wang, Ya-Jun Liu, Xiao-Lan Jiang, Tao Xia, Li-Ping Gao

Monomeric flavan-3-ols and their oligomeric forms, proanthocyanidins (PAs), are closely related to the bitterness of tea beverages. Monomeric flavan-3-ols are characteristic flavor compounds in tea. Increasing the content of PAs and anthocyanins enhances the resistance of tea plants to pathogen invasion but decreases the quality of tea beverages. MATE family transporters play a critical role in transferring monomeric flavan-3-ols and anthocyanins into vacuoles for storage or subsequent condensation into PAs. Their activities modulate the ratio of monomeric flavan-3-ols to PAs and increase anthocyanin content in tea plants. In this study, it was observed that the gene expression and protein phosphorylation level of the MATE transporter CsTT12, a vacuole-localized flavonoid transporter, were notably upregulated following exogenous sucrose treatment, promoting PA synthesis in tea plants. Further analysis revealed that overexpression of CsTT12 and CsTT12S17D significantly increased the content of anthocyanins and PAs in plants, whereas CsTT12S17A did not. In CsTT12 knockdown plants, PA's accumulation decreased significantly, while monomeric catechin content increased. Moreover, phosphorylation modification enhanced the vacuolar membrane localization of CsTT12, whereas dephosphorylation weakened its vacuolar membrane localization. This study uncovers the crucial role of phosphorylation in flavonoid biosynthesis and provides insights into balancing quality improvements and resistance enhancement.

单体黄烷-3-醇及其低聚体形式原花青素(PA)与茶饮料的苦味密切相关。单体黄烷-3-醇是茶叶中特有的风味化合物。增加 PAs 和花青素的含量可增强茶树抵抗病原体入侵的能力,但会降低茶饮料的品质。MATE 家族转运体在将单体黄烷-3-醇和花青素转移到液泡中储存或随后缩合成 PA 方面发挥着关键作用。它们的活动调节了单体黄烷-3-醇与 PA 的比例,增加了茶树中的花青素含量。本研究观察到,外源蔗糖处理后,液泡定位的黄酮类转运体MATE转运体CsTT12的基因表达和蛋白磷酸化水平显著上调,促进了茶树中PA的合成。进一步的分析表明,过表达 CsTT12 和 CsTT12S17D 能显著增加植物中花青素和 PA 的含量,而 CsTT12S17A 则不能。在 CsTT12 基因敲除的植株中,PA 的积累明显减少,而单体儿茶素的含量增加。此外,磷酸化修饰增强了 CsTT12 的液泡膜定位,而去磷酸化则削弱了其液泡膜定位。这项研究揭示了磷酸化在黄酮类化合物生物合成中的关键作用,并为平衡品质改善和提高抗性提供了启示。
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引用次数: 0
Genome-wide associated study identifies FtPMEI13 gene conferring drought resistance in Tartary buckwheat. 全基因组关联研究发现 FtPMEI13 基因赋予鞑靼荞麦抗旱性。
IF 6.2 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2024-11-03 DOI: 10.1111/tpj.17119
Jiayue He, Yanrong Hao, Yuqi He, Wei Li, Yaliang Shi, Muhammad Khurshid, Dili Lai, Chongzhong Ma, Xiangru Wang, Jinbo Li, Jianping Cheng, Alisdair R Fernie, Jingjun Ruan, Kaixuan Zhang, Meiliang Zhou

Tartary buckwheat is known for its ability to adapt to intricate growth conditions and to possess robust stress-resistant properties. Nevertheless, it remains vulnerable to drought stress, which can lead to reduced crop yield. To identify potential genes involved in drought resistance, a genome-wide association study on drought tolerance in Tartary buckwheat germplasm was conducted. A gene encoding pectin methylesterase inhibitors protein (FtPMEI13) was identified, which is not only associated with drought tolerance but also showed induction during drought stress and abscisic acid (ABA) treatment. Further analysis revealed that overexpression of FtPMEI13 leads to improved drought tolerance by altering the activities of antioxidant enzymes and the levels of osmotically active metabolites. Additionally, FtPMEI13 interacts with pectin methylesterase (PME) and inhibits PME activity in response to drought stress. Our results suggest that FtPMEI13 may inhibit the activity of FtPME44/FtPME61, thereby affecting pectin methylesterification in the cell wall and modulating stomatal closure in response to drought stress. Yeast one-hybrid, dual-luciferase assays, and electrophoretic mobility shift assays demonstrated that an ABA-responsive transcription factor FtbZIP46, could bind to the FtPMEI13 promoter, enhancing FtPMEI13 expression. Further analysis indicated that Tartary buckwheat accessions with the genotype resulting in higher FtPMEI13 and FtbZIP46 expression exhibited higher drought tolerance compared to the others. This suggests that this genotype has potential for application in Tartary buckwheat breeding. Furthermore, the natural variation of FtPMEI13 was responsible for decreased drought tolerance during Tartary buckwheat domestication. Taken together, these results provide basic support for Tartary buckwheat breeding for drought tolerance.

鞑靼荞麦以其适应复杂生长条件的能力和强大的抗逆性而著称。然而,它仍然很容易受到干旱胁迫的影响,从而导致作物减产。为了确定参与抗旱的潜在基因,研究人员对鞑靼荞麦种质的抗旱性进行了全基因组关联研究。研究发现了一个编码果胶甲基酯酶抑制蛋白(FtPMEI13)的基因,该基因不仅与抗旱性有关,而且在干旱胁迫和脱落酸(ABA)处理过程中显示出诱导作用。进一步分析表明,过表达 FtPMEI13 可通过改变抗氧化酶的活性和渗透活性代谢物的水平来提高耐旱性。此外,FtPMEI13 与果胶甲基酯酶(PME)相互作用,抑制 PME 在干旱胁迫下的活性。我们的研究结果表明,FtPMEI13 可能会抑制 FtPME44/FtPME61 的活性,从而影响细胞壁中果胶的甲基化,并调节干旱胁迫下的气孔关闭。酵母单杂交、双荧光素酶测定和电泳迁移测定表明,ABA响应转录因子FtbZIP46能与FtPMEI13启动子结合,从而增强FtPMEI13的表达。进一步的分析表明,FtPMEI13 和 FtbZIP46 表达量较高的基因型的鞑靼荞麦品种与其他品种相比具有更高的耐旱性。这表明该基因型在鞑靼荞麦育种中具有应用潜力。此外,在鞑靼荞麦驯化过程中,FtPMEI13的自然变异是导致耐旱性降低的原因。综上所述,这些结果为鞑靼荞麦的抗旱育种提供了基本支持。
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引用次数: 0
Diurnal control of H3K27me1 deposition shapes expression of a subset of cell cycle and DNA damage response genes. H3K27me1 沉积的昼夜控制影响着细胞周期和 DNA 损伤反应基因子集的表达。
IF 6.2 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2024-11-01 DOI: 10.1111/tpj.17114
Jorge Fung-Uceda, María Sol Gómez, Laura Rodríguez-Casillas, Anna González-Gil, Crisanto Gutierrez

Rhythmic oscillation of biological processes helps organisms adapt their physiological responses to the most appropriate time of the day. Chromatin remodeling has been described as one of the molecular mechanisms controlling these oscillations. The importance of these changes in transcriptional activation as well as in the maintenance of heterochromatic regions has been widely demonstrated. However, little is still known on how diurnal changes can impact the global status of chromatin modifications and, hence, control gene expression. In plants, the repressive mark H3K27me1, deposited by ARABIDOPSIS TRITHORAX-RELATED PROTEIN 5 and 6 (ATXR5 and 6) methyltransferases, is largely associated with transposable elements but also covers lowly expressed genes. Here we show that this histone modification is preferentially deposited during the night. In euchromatic regions, it is found along the bodies of DNA damage response genes (DDR), where it is needed for their proper expression. The absence of H3K27me1 translates into an enhanced expression of DDR genes that follows a rhythmic oscillation pattern. This evidences a link between chromatin modifications and their synchronization with the diurnal cycle in order to accurately modulate the activation of biological processes to the most appropriate time of the day.

生物过程的节律振荡有助于生物体根据一天中最合适的时间调整其生理反应。染色质重塑被描述为控制这些振荡的分子机制之一。这些变化在转录激活和维持异染色质区域方面的重要性已得到广泛证实。然而,人们对昼夜变化如何影响染色质修饰的整体状态并进而控制基因表达仍然知之甚少。在植物中,抑制性标记 H3K27me1 由 ARABIDOPSIS TRITHORAX-RELATED PROTEIN 5 和 6(ATXR5 和 6)甲基转移酶沉积,主要与转座元件相关,但也覆盖低表达基因。我们在这里发现,这种组蛋白修饰在夜间优先沉积。在外显子区域,它沿着 DNA 损伤应答基因(DDR)的主体被发现,这些基因的正常表达需要它。H3K27me1 的缺失会导致 DDR 基因的表达增强,并呈现出有节奏的振荡模式。这证明了染色质修饰与昼夜周期同步之间的联系,以便在一天中最合适的时间准确调节生物过程的激活。
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引用次数: 0
A MYB transcription factor underlying plant height in sorghum qHT7.1 and maize Brachytic 1 loci. 在高粱 qHT7.1 和玉米 Brachytic 1 基因座中,一个 MYB 转录因子是植株高度的基础。
IF 6.2 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2024-11-01 DOI: 10.1111/tpj.17111
Qi Mu, Jialu Wei, Hallie K Longest, Hua Liu, Si Nian Char, Jacob T Hinrichsen, Laura E Tibbs-Cortes, Gregory R Schoenbaum, Bing Yang, Xianran Li, Jianming Yu

Manipulating plant height is an essential component of crop improvement. Plant height was generally reduced through breeding in wheat, rice, and sorghum to resist lodging and increase grain yield but kept high for bioenergy crops. Here, we positionally cloned a plant height quantitative trait locus (QTL) qHT7.1 as a MYB transcription factor controlling internode elongation, cell proliferation, and cell morphology in sorghum. A 740 bp transposable element insertion in the intronic region caused a partial mis-splicing event, generating a novel transcript that included an additional exon and a premature stop codon, leading to short plant height. The dominant allele had an overall higher expression than the recessive allele across development and internode position, while both alleles' expressions peaked at 46 days after planting and progressively decreased from the top to lower internodes. The orthologue of qHT7.1 was identified to underlie the brachytic1 (br1) locus in maize. A large insertion in exon 3 and a 160 bp insertion at the promoter region were identified in the br1 mutant, while an 18 bp promoter insertion was found to be associated with reduced plant height in a natural recessive allele. CRISPR/Cas9-induced gene knockout of br1 in two maize inbred lines showed significant plant height reduction. These findings revealed functional connections across natural, mutant, and edited alleles of this MYB transcription factor in sorghum and maize. This enriched our understanding of plant height regulation and enhanced our toolbox for fine-tuning plant height for crop improvement.

控制株高是作物改良的重要组成部分。小麦、水稻和高粱的育种通常会降低株高以抗倒伏和增加谷物产量,但生物能源作物的育种则会保持较高的株高。在这里,我们定位克隆了一个株高数量性状位点(QTL)qHT7.1,它是一个控制高粱节间伸长、细胞增殖和细胞形态的 MYB 转录因子。内含子区域的 740 bp 转座元件插入引起了部分错误剪接事件,产生了一个包含额外外显子和过早终止密码子的新转录本,导致植株矮小。显性等位基因在整个发育过程和节间位置上的表达量总体高于隐性等位基因,而两个等位基因的表达量都在播种后 46 天达到峰值,并从节间顶部到下部逐渐降低。qHT7.1的直向同源物被确定为玉米brachytic1(br1)基因座的基础。在 br1 突变体中发现了外显子 3 中的大插入和启动子区域的 160 bp 插入,而在自然隐性等位基因中发现 18 bp 启动子插入与植株高度降低有关。在两个玉米近交系中,CRISPR/Cas9诱导的br1基因敲除显示植株高度显著降低。这些发现揭示了高粱和玉米中这种 MYB 转录因子的天然、突变和编辑等位基因之间的功能联系。这丰富了我们对植株高度调控的理解,增强了我们微调植株高度以改良作物的工具箱。
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引用次数: 0
Exploring an economic and highly efficient genetic transformation and genome-editing system for radish through developmental regulators and visible reporter. 通过发育调节剂和可见报告基因,探索经济高效的萝卜遗传转化和基因组编辑系统。
IF 5.3 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2024-11-01 Epub Date: 2024-10-10 DOI: 10.1111/tpj.17068
Xiaofang Yi, Congcong Wang, Xiaoqi Yuan, Mi Zhang, Changwei Zhang, Tiaojiao Qin, Haiyun Wang, Liang Xu, Liwang Liu, Yan Wang

Radish (Raphanus sativus L.) is one of the most important root vegetable crops worldwide. However, gene function exploration and germplasm innovation still face tremendous challenges due to its extremely low transformation efficiency. Here, an economic and highly efficient genetic transformation method for radish was explored by Agrobacterium rhizogenes-mediated transformation with the help of combining special developmental regulator (DR) genes and the visual identification reporter. Firstly, the RUBY gene, a betalain biosynthesis system, could result in a visual red-violet color used as a convenient and effective reporter for monitoring transgenic hairy roots screening of radish. However, the hairy roots-to-shoots conversion system of radish still stands as a barrier to the obtainment of whole transgenic plants, although different hormone combinations and various culture conditions were tried. Following, two DR genes including Wuschel2 (Wus2) and isopentenyl transferase (ipt), as well as their combination Wus2-ipt were introduced for the shoot regeneration capacity improvement. The results showed that the transgenic shoots could be directly generated without externally supplying any hormones in the presence of a Wus2-ipt combination. Then, Wus2-ipt along with the RUBY reporter was employed to establish an efficient genetic transformation system of radish. Moreover, this system was applied in generating gene-edited radish plants and the phytoene desaturase (RsPDS) gene was effectively knockout through albino phenotype observation and sequencing analysis. These findings have the potential to be widely applied in genetic transformation and genome-editing genetic improvement of other vegetable species.

萝卜(Raphanus sativus L.)是世界上最重要的根茎蔬菜作物之一。然而,由于其转化效率极低,基因功能探索和种质创新仍面临巨大挑战。本文通过根瘤农杆菌介导的转化,结合特殊的发育调节因子(DR)基因和视觉识别报告基因,探索了一种经济、高效的萝卜遗传转化方法。首先,RUBY 基因是一种甜菜碱生物合成系统,可产生视觉上的红紫色,是监测萝卜转基因毛细根筛选的一种方便有效的报告基因。然而,尽管尝试了不同的激素组合和各种培养条件,萝卜毛细根到根的转化系统仍然是获得完整转基因植株的障碍。为了提高萝卜芽的再生能力,研究人员引入了两个 DR 基因,包括 Wuschel2(Wus2)和异戊烯基转移酶(ipt),以及它们的组合 Wus2-ipt。结果表明,在 Wus2-ipt组合存在的情况下,无需外部提供任何激素就能直接产生转基因芽。随后,Wus2-ipt 与 RUBY 报告基因一起被用于建立一个高效的萝卜遗传转化系统。此外,该系统还被应用于生成基因编辑萝卜植株,通过白化表型观察和测序分析,植二烯去饱和酶(RsPDS)基因被有效敲除。这些发现有望广泛应用于其他蔬菜物种的遗传转化和基因组编辑遗传改良。
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引用次数: 0
Deep learning to capture leaf shape in plant images: Validation by geometric morphometrics. 深度学习捕捉植物图像中的叶形:几何形态计量学验证。
IF 5.3 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2024-11-01 Epub Date: 2024-10-09 DOI: 10.1111/tpj.17053
Ladislav Hodač, Kevin Karbstein, Lara Kösters, Michael Rzanny, Hans Christian Wittich, David Boho, David Šubrt, Patrick Mäder, Jana Wäldchen

Plant leaves play a pivotal role in automated species identification using deep learning (DL). However, achieving reproducible capture of leaf variation remains challenging due to the inherent "black box" problem of DL models. To evaluate the effectiveness of DL in capturing leaf shape, we used geometric morphometrics (GM), an emerging component of eXplainable Artificial Intelligence (XAI) toolkits. We photographed Ranunculus auricomus leaves directly in situ and after herbarization. From these corresponding leaf images, we automatically extracted DL features using a neural network and digitized leaf shapes using GM. The association between the extracted DL features and GM shapes was then evaluated using dimension reduction and covariation models. DL features facilitated the clustering of leaf images by source populations in both in situ and herbarized leaf image datasets, and certain DL features were significantly associated with biological leaf shape variation as inferred by GM. DL features also enabled leaf classification into morpho-phylogenomic groups within the intricate R. auricomus species complex. We demonstrated that simple in situ leaf imaging and DL reproducibly captured leaf shape variation at the population level, while combining this approach with GM provided key insights into the shape information extracted from images by computer vision, a necessary prerequisite for reliable automated plant phenotyping.

植物叶片在使用深度学习(DL)进行物种自动识别中发挥着关键作用。然而,由于深度学习模型固有的 "黑箱 "问题,实现对叶片变异的可重现捕捉仍具有挑战性。为了评估深度学习在捕捉叶片形状方面的有效性,我们使用了几何形态计量学(GM),它是可扩展人工智能(XAI)工具包的一个新兴组件。我们直接在原地和草本化后拍摄了 Ranunculus auricomus 的叶片。从这些相应的叶片图像中,我们使用神经网络自动提取了DL特征,并使用GM对叶片形状进行了数字化处理。然后,我们使用降维和协变模型评估了提取的 DL 特征与 GM 形状之间的关联。在原位和草本化叶片图像数据集中,DL特征有助于按来源种群对叶片图像进行聚类,而且某些DL特征与GM推断的生物叶片形状变化有显著关联。DL 特征还能在错综复杂的 R. auricomus 种群中将叶片分类为形态-系统发生组。我们证明,简单的原位叶片成像和 DL 可以在种群水平上重复捕捉叶片形状的变异,而将这种方法与基因改造相结合,则可以深入了解计算机视觉从图像中提取的形状信息,这是可靠的自动化植物表型的必要前提。
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引用次数: 0
Fast and simple fluorometric measurement of phloem loading exposes auxin-dependent regulation of Arabidopsis sucrose transporter AtSUC2. 对韧皮部负荷进行快速、简单的荧光测量,揭示拟南芥蔗糖转运体 AtSUC2 受植物生长素调控的情况。
IF 6.2 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2024-11-01 DOI: 10.1111/tpj.17110
Yunjuan Ren, Ziyu Zhang, Diana Zhanakhmetova, Wenhui Li, Shaolin Chen, Tomáš Werner, Johannes Liesche

The rate of sucrose export from leaves is a major factor in balancing whole-plant carbon and energy partitioning. A comprehensive study of its dynamics and relationship to photosynthesis, sink demand, and other relevant processes is hampered by the shortcomings of current methods for measuring sucrose phloem loading. We utilize the ability of sucrose transporter proteins, known as SUCs or SUTs, to specifically transport the fluorescent molecule esculin in a novel assay to measure phloem loading rates. Esculin was administered to source leaves and its fluorescence in the leaf extract was measured after 1 or 2 h. Dicot plants with an active phloem loading strategy showed an export-dependent reduction of esculin fluorescence. Relative leaf esculin export rates correlated with leaf export rates of isotopic carbon and phloem exudate sucrose levels. We used esculin experiments to examine the effects of phytohormones on phloem loading in Arabidopsis, showing, for example, that auxin induces phloem loading while cytokinin reduces it. Transcriptional regulation of AtSUC2 by AUXIN RESPONSE FACTOR1 (ARF1) corroborated the link between auxin signaling and phloem loading. Unlike established methods, the esculin assay is rapid and does not require specialized equipment. Potential applications and limitations of the esculin assay are discussed.

叶片的蔗糖输出率是平衡整个植物碳和能量分配的一个主要因素。由于目前测量蔗糖韧皮部负载的方法存在缺陷,因此无法全面研究蔗糖韧皮部负载的动态及其与光合作用、吸收汇需求和其他相关过程的关系。我们利用蔗糖转运蛋白(又称 SUC 或 SUT)特异性转运荧光分子 esculin 的能力,采用一种新的测定方法来测量韧皮部负载率。双子叶植物具有活跃的韧皮部负载策略,表现出依赖于输出的埃斯库林荧光减少。叶片的相对esculin输出率与叶片的同位素碳输出率和韧皮部渗出的蔗糖水平相关。我们利用esculin实验研究了植物激素对拟南芥韧皮部负载的影响,结果显示,例如,辅助素会诱导韧皮部负载,而细胞分裂素则会降低韧皮部负载。AUXIN RESPONSE FACTOR1(ARF1)对 AtSUC2 的转录调控证实了叶绿素信号与韧皮部负载之间的联系。与既有方法不同的是,esculin 分析法快速且不需要专门设备。本文讨论了埃斯库灵测定法的潜在应用和局限性。
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引用次数: 0
Natural variation at the cotton HIC locus increases trichome density and enhances resistance to aphids. 棉花 HIC 基因座的自然变异可增加毛状体密度,增强对蚜虫的抗性。
IF 5.3 1区 生物学 Q1 PLANT SCIENCES Pub Date : 2024-11-01 Epub Date: 2024-10-04 DOI: 10.1111/tpj.17050
Yanan Wang, Qi Zhou, Jilong Zhang, Haiyan He, Zhigang Meng, Yuan Wang, Sandui Guo, Rui Zhang, Chengzhen Liang

Plant trichomes are an excellent model for studying cell differentiation and development, providing crucial defenses against biotic and abiotic stresses. There is a well-established inverse relationship between trichome density and aphid prevalence, indicating that higher trichome density leads to reduced aphid infestations. Here we present the cloning and characterization of a dominant quantitative trait locus, HIC (hirsute cotton), which significantly enhances cotton trichome density. This enhancement leads to markedly improved resistance against cotton aphids. The HIC encodes an HD-ZIP IV transcriptional activator, crucial for trichome initiation. Overexpression of HIC leads to a substantial increase in trichome density, while knockdown of HIC results in a marked decrease in density, confirming its role in trichome regulation. We identified a variant in the HIC promoter (-810 bp A to C) that increases transcription of HIC and trichome density in hirsute cotton compared with Gossypium hirsutum cultivars with fewer or no trichomes. Interestingly, although the -810 variant in the HIC promoter is the same in G. barbadense and hirsute cotton, the presence of a copia-like retrotransposon insertion in the coding region of HIC in G. barbadense causes premature transcription termination. Further analysis revealed that HIC positively regulates trichome density by directly targeting the EXPANSIN A2 gene, which is involved in cell wall development. Taken together, our results underscore the pivotal function of HIC as a primary regulator during the initial phases of trichome formation, and its prospective utility in enhancing aphid resistance in superior cotton cultivars via selective breeding.

植物毛状体是研究细胞分化和发育的绝佳模型,可提供抵御生物和非生物胁迫的重要防御能力。毛状体密度与蚜虫发生率之间存在着公认的反比关系,表明毛状体密度越高,蚜虫发生率越低。在这里,我们展示了一个显性数量性状基因座 HIC(多毛棉花)的克隆和特征描述,该基因座能显著提高棉花的毛状体密度。这种增强显著提高了对棉蚜的抗性。HIC 编码一个 HD-ZIP IV 转录激活子,对毛状体的启动至关重要。过量表达 HIC 会导致毛状体密度大幅增加,而敲除 HIC 则会导致密度明显降低,这证实了它在毛状体调控中的作用。我们在 HIC 启动子中发现了一个变体(-810 bp A 到 C),与毛状体较少或没有毛状体的长硬毛棉花栽培品种相比,它能增加 HIC 的转录和毛状体密度。有趣的是,虽然 HIC 启动子中的 -810 变体在长硬毛棉花和长硬毛棉花中是相同的,但在长硬毛棉花中,HIC 编码区中 copia 样逆转录质子插入会导致转录过早终止。进一步的分析表明,HIC 通过直接靶向参与细胞壁发育的 EXPANSIN A2 基因对毛状体密度进行正向调节。综上所述,我们的研究结果强调了 HIC 作为毛状体形成初期的主要调控因子的关键功能,以及它在通过选择性育种提高优良棉花品种抗蚜虫能力方面的应用前景。
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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 : 2024-10-30 DOI: 10.1111/tpj.17118
Gwendolyn Kirschner
<p>Hydathodes are organs on the leaves of all vascular plants. They regulate the secretion of fluids derived from the xylem sap (Bellenot et al., <span>2022</span>; Cerutti et al., <span>2019</span>). When stomata are closed at night and the humidity level levels are too high, the xylem delivers excess water from the roots, which is secreted at the hydathodes in a process called guttation (Figure 1a) (Singh, <span>2020</span>). Hydathodes are composed of an epidermal surface layer with water pores, and an inner parenchyma, called the epithem, which is highly vascularized and constitutes a direct connection between leaf surface and xylem vessels (Figure 1b) (Bellenot et al., <span>2022</span>). Hydathodes were first described by the German botanist Anton de Bary in 1877, and named by the Austrian botanist Gottlieb Haberlandt in 1897, from the Greek ‘hyda’ (water) and ‘hodos’ (way) (Bellenot et al., <span>2022</span>). When Jean-Marc Routaboul, the corresponding author of the highlighted publication, joined Laurent Noël's team at INRAE, France, in 2018, he was surprised to find that hydathodes and the process of guttation were not well understood at the molecular level. Therefore, Routaboul and his colleagues set out to test two long-standing hypotheses about hydathodes: that hydathodes are sites of auxin accumulation, and that they facilitate the withholding of nutrients from guttation fluids (Routaboul et al., <span>2024</span>).</p><p>These hypotheses are based on genes expressed in hydathodes, including those for auxin biosynthesis, transport, and signalling. Moreover, the presence of auxin in hydathodes was detected by antibodies and by using the auxin signalling reporter <i>DR5</i> (Aloni et al., <span>2003</span>). Other hydathode-specific genes encode membrane transporters for amino acids, sugar or ions (Nagai et al., <span>2013</span>), potentially preventing nutrient loss through guttation. For their study, Routaboul <i>et al</i>. combined RNAseq of hydathode-enriched tissue by deep sequencing with a detailed metabolomic analysis of guttation fluids.</p><p>First, the authors compared the transcriptome of macro-dissected leaf margins containing hydathodes with the transcriptome of leaf blade tissue of mature Arabidopsis leaves. They found higher expression of genes related to auxin metabolism, stress, DNA, plant cell wall, transport, RNA and lipids in the hydathode-enriched tissue. Genes related to glucosinolate synthesis and transport, the sulfation pathway, metal handling or photosynthesis were more highly expressed in the leaf blade. Because many genes related to auxin biosynthesis were expressed in hydathodes, the authors measured the accumulation of free auxin in hydathode-enriched tissue and leaf blades with liquid chromatography/mass spectrometry (LC/MS) and found nearly 40% more free auxin in hydathode-enriched tissue than in leaf blades. Reporter gene expression confirmed that genes encoding the key auxin biosynthetic enzymes Tryp
水瘤是所有维管植物叶片上的器官。它们调节木质部汁液的分泌(Bellenot 等人,2022 年;Cerutti 等人,2019 年)。当夜间气孔关闭、湿度水平过高时,木质部会从根部输送多余的水分,这些水分在称为 "内渗化 "的过程中分泌到水瘤(图 1a)(Singh,2020 年)。水瘤由带有水孔的表皮表层和称为表皮的内部实质组成,后者血管高度发达,是叶片表面和木质部血管之间的直接连接(图 1b)(Bellenot 等人,2022 年)。水瘤由德国植物学家 Anton de Bary 于 1877 年首次描述,1897 年由奥地利植物学家 Gottlieb Haberlandt 命名,源自希腊语 "hyda"(水)和 "hodos"(路)(Bellenot 等人,2022 年)。该论文的通讯作者让-马克-劳塔布尔(Jean-Marc Routaboul)于 2018 年加入法国国家高等农艺研究所劳伦特-诺埃尔(Laurent Noël)的团队,他惊讶地发现,人们对水合作用和肠化过程在分子水平上并不十分了解。因此,Routaboul和他的同事们开始验证有关水瘤的两个长期存在的假说:水瘤是助长素积累的场所,以及水瘤有助于从内蜕液中扣留养分(Routaboul等人,2024年)。这些假说基于水瘤中表达的基因,包括那些用于助长素生物合成、运输和信号传递的基因。此外,通过抗体和使用辅酶信号报告基因 DR5(Aloni 等人,2003 年),可以检测到水瘤中存在辅酶。其他水瘤特异性基因编码氨基酸、糖或离子的膜转运体(Nagai 等人,2013 年),有可能防止营养物质通过内脏流失。在他们的研究中,Routaboul 等人通过深度测序对富含水瘤的组织进行了 RNAseq 测序,并对开沟液进行了详细的代谢组学分析。首先,作者比较了含有水瘤的大面积解剖叶缘的转录组和成熟拟南芥叶片组织的转录组。他们发现,在富含水瘤的组织中,与辅酶代谢、胁迫、DNA、植物细胞壁、运输、RNA 和脂质有关的基因表达量较高。与葡萄糖苷酸合成和运输、硫酸化途径、金属处理或光合作用有关的基因在叶片中的表达量更高。由于许多与辅酶生物合成有关的基因在水叶中表达,作者用液相色谱/质谱法(LC/MS)测量了水叶富集组织和叶片中游离辅酶的积累,发现水叶富集组织中的游离辅酶比叶片中多近 40%。报告基因表达证实,拟南芥色氨酸氨基转移酶 1 (TAA1)、YUCCA2、YUCCA5、YUCCA8 和 YUCCA9 等关键辅素生物合成酶的编码基因在水瘤中特异性表达。这表明,由于局部辅助素的生物合成,水瘤中的辅助素浓度很高。两种 GH3 IAA-氨基合成酶通过将过量的 IAA 连接成氨基酸共轭物来维持辅素平衡,它们在水合作用富集部分的表达量也更高。因此,作者测量了该途径的下游产物氧化吲哚-3-AIA。水合叶中较高的氧化吲哚-3-AIA 表明水合叶具有较高的辅助素储存能力。辅助素信号和响应基因,如辅助素响应因子(ARFs)和辅助素/吲哚乙酸(Aux/IAAs),在水瘤中也有较高的表达,这共同表明水瘤是辅助素合成和信号传导的活性位点、如前所示(Krouk 等人,2010 年;Misson 等人,2004 年),作者发现许多编码水、离子(硝酸盐、磷酸盐、硫酸盐、钙、锌、铁、铜、氯化物和砷酸硼)转运体、激素转运体(ABA、GA、辅助素和细胞分裂素)以及糖、肽、蜡和其他有机化合物转运体的基因在水瘤中高表达,这表明水瘤可能是积极改变内渗液的部位。因此,作者对叶柄中渗出的木质部液体进行了采样,并利用气相色谱/质谱法将其成分与从切除水瘤的叶片中采样的水瘤前液体和从水瘤中采集的内渗液进行了比较。在代谢物到达水合器之前,叶片组织捕获了 91% 的代谢物。在水合作用前的液体中,78%的剩余代谢物在水合作用释放之前被捕获。肠液中 23 种代谢物(包括氨基酸、有机酸、糖类和肌醇)的浓度低于前阴极液。比色测定显示,水合阳极吸收了无机磷酸盐(Pi)和硝酸盐。
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