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Overexpression of plasma membrane SUT1 in poplar alters lateral sucrose partitioning in stem and promotes leaf necrosis. 杨树质膜SUT1过表达改变茎侧蔗糖分配,促进叶片坏死。
IF 2.3 3区 生物学 Q2 PLANT SCIENCES Pub Date : 2025-03-12 eCollection Date: 2025-03-01 DOI: 10.1002/pld3.70023
Liang-Jiao Xue, Moh'd I Hozain, Christopher J Frost, Afraz Talebi, Batbayar Nyamdari, Kavita B Aulakh, Ran Zhou, Scott A Harding, Chung-Jui Tsai

In Populus and many other tree species, photoassimilate sucrose diffuses down a concentration gradient via symplastically connected mesophyll cells to minor vein phloem for long-distance transport. There is no evidence for apoplastic phloem-loading in Populus. However, plasma membrane sucrose transporters (SUT1 and SUT3) orthologous to those associated with apoplastic phloem loading are expressed in vascular tissues of poplar. While SUT3 functions in sucrose import into developing xylem, the role of SUT1 remains unclear. Here, we overexpressed PtaSUT1 in Populus tremula x P. alba to examine the effects on sucrose partitioning in transgenic plants. Overall leaf sucrose levels were similar between wild type and transgenic lines. Stem sucrose levels were not changed in bark but were significantly reduced in the adjacent xylem, suggesting hindered intercellular sucrose trafficking from the phloem to the developing xylem. Fully expanded leaves of transgenic plants deteriorated prematurely with declining photosynthesis prior to severe necrotic spotting. Necrotic spotting advanced most rapidly in the distal portion of mature leaves and was accompanied by sharp hexose increases and sharp sucrose decreases there. Leaf transcriptome profiling and network inference revealed the down-regulation of copper proteins and elevated expression of copper microRNAs prior to noticeable leaf injury. Our results suggest ectopic expression of PtaSUT1 altered sucrose partitioning in stems with systemic effects on leaf health and copper homeostasis mediated in part by sucrose-sensitive copper miRNAs.

在杨树和许多其他树种中,光同化蔗糖通过叶肉细胞的交感连接沿浓度梯度向下扩散到小叶脉韧皮部进行长距离运输。没有证据表明杨树有外质体韧皮部负荷。然而,与外质体韧皮部负荷相关的质膜蔗糖转运蛋白(SUT1和SUT3)在杨树维管组织中表达。虽然SUT3在发育木质部的蔗糖输入中起作用,但SUT1的作用尚不清楚。在此,我们在白杨中过表达PtaSUT1,以研究其对转基因植物中蔗糖分配的影响。野生型和转基因系叶片总蔗糖水平相似。树皮的茎蔗糖水平没有变化,但相邻木质部的蔗糖水平显著降低,表明韧皮部向发育中的木质部的细胞间蔗糖运输受到阻碍。完全展开的转基因植株叶片在发生严重的坏死斑染前,光合作用下降,叶片过早退化。坏死点斑在成熟叶的远端发展最快,并伴有己糖急剧增加和蔗糖急剧减少。叶片转录组分析和网络推断显示,在叶片明显损伤之前,铜蛋白下调,铜microrna表达升高。我们的研究结果表明,PtaSUT1的异位表达改变了茎中的蔗糖分配,并在一定程度上通过对蔗糖敏感的铜mirna介导对叶片健康和铜稳态的系统性影响。
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引用次数: 0
Genomic Markers Associated With Soybean Resistance to the Stem Borer, Dectes texanus (Coleoptera: Cerambycidae). 大豆对茎螟虫(Dectes texanus)抗性的基因组标记。
IF 2.3 3区 生物学 Q2 PLANT SCIENCES Pub Date : 2025-03-12 eCollection Date: 2025-03-01 DOI: 10.1002/pld3.70040
Sarah Johnson, David Hyten

The Dectes stem borer, Dectes texanus LeConte (Coleoptera: Cerambycidae), can significantly reduce yields by causing significant lodging in soybean. While this stem borer has not been considered a major pest of soybean, damage from it is increasing in the United States Midwest region with no current elite cultivars found resistant. Our objective was to map quantitative trait loci (QTL) that reduce girdled stems caused by Dectes stem borer infection and infestation of Dectes stem borer. A genome-wide association study (GWAS) using 50,000 single nucleotide polymorphisms was used to analyze data from a population of maturity group (MG) V to VII soybean accessions grown in North Carolina, which had been scored for Dectes stem borer larvae infestation and girdled stems caused by Dectes stem borer infestation. The GWAS identified 3 QTL with reduced larvae infestation and 4 QTL for reduced girdled stems. Allele effects ranged from 1% to 9% reduced larvae infestation or girdled stems. The QTL identified and germplasm containing the beneficial alleles can be used for improving resistance to the damage caused by the Dectes stem borer in elite soybean cultivars.

牛虻(Dectes texanus LeConte)(鞘翅目:天牛科)通过在大豆上造成显著的倒伏而显著降低产量。虽然这种茎蛀虫不被认为是大豆的主要害虫,但在美国中西部地区,它的危害正在增加,目前还没有发现具有抗性的优良品种。我们的目的是绘制减少黄斑干螟虫感染和侵染引起的环茎数量性状位点(QTL)。利用5万个单核苷酸多态性进行全基因组关联研究(GWAS),分析了北卡罗莱纳州成熟组(MG) V ~ VII大豆群体的数据,并对该群体进行了Dectes茎螟虫幼虫侵染和Dectes茎螟虫侵染引起的环茎评分。GWAS鉴定出3个减少幼虫侵染的QTL和4个减少环茎的QTL。等位基因效应范围为1%至9%,减少幼虫侵染或缠绕茎。所鉴定的QTL和含有有益等位基因的种质资源可用于提高大豆优良品种对Dectes茎螟虫危害的抗性。
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引用次数: 0
Suppression of Chorismate Mutase 1 in Hybrid Poplar to Investigate Potential Redundancy in the Supply of Lignin Precursors. 抑制杂交杨木枝酸突变酶1研究木质素前体供应的潜在冗余。
IF 2.3 3区 生物学 Q2 PLANT SCIENCES Pub Date : 2025-03-12 eCollection Date: 2025-03-01 DOI: 10.1002/pld3.70053
Yaseen Mottiar, Timothy J Tschaplinski, John Ralph, Shawn D Mansfield

Chorismate is an important branchpoint metabolite in the biosynthesis of lignin and a wide array of metabolites in plants. Chorismate mutase (CM), the enzyme responsible for transforming chorismate into prephenate, is a key regulator of metabolic flux towards the synthesis of aromatic amino acids and onwards to lignin. We examined three CM genes in hybrid poplar (Populus alba × grandidentata; P39, abbreviated as Pa×g) and used RNA interference (RNAi) to suppress the expression of Pa×gCM1, the most highly expressed isoform found in xylem tissue. Although this strategy was successful in disrupting Pa×gCM1 transcripts, there was also an unanticipated increase in lignin content, a shift towards guaiacyl lignin units, and more xylem vessels with smaller lumen areas, at least in the most severely affected transgenic line. This was accompanied by compensatory expression of the other two CM isoforms, Pa×gCM2 and Pa×gCM3, as well as widespread changes in gene expression and metabolism. This study investigates potential redundancy within the CM gene family in the developing xylem of poplar and highlights the pivotal role of chorismate in plant metabolism, development, and physiology.

木质素酸盐是木质素生物合成中重要的分枝点代谢物,也是植物中广泛存在的代谢物。Chorismate mutase (CM)是将choris酸转化为预苯酸盐的酶,是芳香氨基酸合成和木质素代谢通量的关键调节因子。对杂交杨杨(Populus alba × granddentata;P39,缩写为Pa×g),并使用RNA干扰(RNAi)抑制木质部组织中表达最高的亚型Pa×gCM1的表达。尽管这种策略成功地破坏了Pa×gCM1转录本,但至少在受影响最严重的转基因品系中,木质素含量也出现了意想不到的增加,向愈创木酰基木质素单位转变,木质部血管增多,管腔面积减小。这伴随着另外两种CM亚型Pa×gCM2和Pa×gCM3的代偿性表达,以及基因表达和代谢的广泛变化。本研究探讨了杨树木质部发育过程中CM基因家族的潜在冗余,并强调了choris酸盐在植物代谢、发育和生理中的关键作用。
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引用次数: 0
Loss of Pleiotropic Regulatory Functions in Tannin1, the Sorghum Ortholog of Arabidopsis Master Regulator TTG1. 拟南芥主调控物TTG1的高粱同源物单宁1多效调控功能缺失
IF 2.3 3区 生物学 Q2 PLANT SCIENCES Pub Date : 2025-03-12 eCollection Date: 2025-03-01 DOI: 10.1002/pld3.70055
Anthony Schuh, Geoffrey P Morris

Transcriptional master regulators are often targeted to improve plant traits, but antagonistic pleiotropic effects of these regulators can hamper this approach. The Myb-bHLH-WDR (MBW) complex is a broadly conserved transcriptional regulator affecting pigmentation, biotic stress resistance, and abiotic stress tolerance. We investigated the function of sorghum grain pigmentation regulator Tannin1, the ortholog of Arabidopsis pleiotropic WD40 regulator TTG1, to test for conserved pleiotropic regulatory effects and to better understand the evolution of the MBW complex in Poaceae. We characterized genome-wide differential expression of leaf tissue using RNA sequencing in near-isogenic lines (NILs) that contrasted wildtype Tan1 and loss-of-function tan1-b alleles, under optimal temperature and chilling stress. Notably, Gene Ontology analyses revealed no pathways with differential expression between Tan1 and tan1-b NILs, suggesting that, in contrast to Arabidopsis TTG1, Tannin1 has no pleiotropic regulatory role in leaves. Further, NILs had no visible difference in anthocyanin pigmentation, and no genes with known or expected function in flavonoid synthesis were differentially expressed. Genome-wide, only 18 total genes were differentially expressed between NILs, with six of these genes located inside the NIL introgression region, an observation most parsimoniously explained by cis-regulatory effects unrelated to Tannin1 regulation. Comparing our findings with known function of TTG1 orthologs in maize, rice, and Arabidopsis, we conclude that pleiotropic regulatory function in leaf tissue was likely lost in panicoid grass evolution before the sorghum-maize split. These findings inform future molecular breeding of MBW regulated traits and highlight the benefit of subfunctionalization to relieve pleiotropic constraints.

转录主调控因子通常是改善植物性状的目标,但这些调控因子的拮抗多效效应会阻碍这种方法的实现。Myb-bHLH-WDR (MBW)复合体是一种广泛保守的转录调控因子,影响色素沉着、生物抗逆性和非生物抗逆性。我们研究了拟南芥多效性WD40调节因子TTG1的同源物——高粱籽粒色素调控因子Tannin1的功能,以检验其保守的多效性调控作用,并更好地了解禾科植物MBW复合物的进化。在最佳温度和低温胁迫下,研究人员利用RNA测序对近等基因系(NILs)叶片组织的全基因组差异表达进行了表征,这些系对比了野生型Tan1和失去功能的Tan1 -b等位基因。值得注意的是,基因本体分析显示Tan1和Tan1 -b NILs之间没有差异表达的途径,这表明与拟南芥TTG1相比,Tannin1在叶片中没有多效性调节作用。此外,NILs在花青素色素沉着方面没有明显差异,在类黄酮合成中没有已知或预期功能的基因差异表达。在全基因组范围内,NILs之间总共只有18个基因存在差异表达,其中6个基因位于NIL渗入区,这一观察结果最简洁地解释为与单宁1调控无关的顺式调控效应。将我们的发现与已知的TTG1同源物在玉米、水稻和拟南芥中的功能进行比较,我们得出结论,在高粱-玉米分裂之前,穗草的叶片组织中可能失去了多效性调节功能。这些发现为未来MBW调控性状的分子育种提供了信息,并强调了亚功能化在缓解多效性限制方面的益处。
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引用次数: 0
ALPHA: A High Throughput System for Quantifying Growth in Aquatic Plants. ALPHA:水生植物生长定量的高通量系统。
IF 2.3 3区 生物学 Q2 PLANT SCIENCES Pub Date : 2025-03-12 eCollection Date: 2025-03-01 DOI: 10.1002/pld3.70048
Kassidy A Robinson, Victoria Augoustides, Tanaka Madenyika, Ryan C Sartor

The need for more sustainable agricultural systems is becoming increasingly apparent. The global demand for agricultural products-food, feed, fuel and fiber-will continue to increase as the global population continues to grow. This challenge is compounded by climate change. Not only does a changing climate make it difficult to maintain stable yields but current agricultural systems are a major source of greenhouse gas emissions and continue to drive the problem further. Therefore, future agricultural systems must not only increase production but also significantly decrease negative environmental impacts. One approach to addressing this is to begin breeding and cultivating new plant species that have fundamental sustainability advantages over our existing crops. The Lemnaceae, commonly known as duckweeds, are one family of plants that have potential to increase output and reduce the negative environmental impacts of agricultural production. Herein we describe the Automated Lab-scale PHenotyping Apparatus, ALPHA, for high-throughput phenotyping of Lemnaceae. ALPHA is being used for selective breeding of one species, Lemna gibba, toward the goal of creating a new crop for use in sustainable agricultural systems. ALPHA can be used on many small aquatic plant species to assess growth rates in different environmental conditions. A proof of principle use case is demonstrated where ALPHA is used to determine saltwater tolerance of six different clones of L. gibba.

对更加可持续的农业系统的需求日益明显。随着全球人口的持续增长,全球对农产品(食品、饲料、燃料和纤维)的需求将继续增加。气候变化加剧了这一挑战。气候变化不仅使保持稳定的产量变得困难,而且目前的农业系统是温室气体排放的主要来源,并将继续推动这一问题的进一步发展。因此,未来的农业系统不仅要提高产量,还要显著减少对环境的负面影响。解决这个问题的一个方法是开始培育新的植物物种,这些物种比我们现有的作物具有根本的可持续性优势。扁豆科,俗称浮萍,是一个有潜力增加产量和减少农业生产对环境的负面影响的植物家族。在这里,我们描述了自动化实验室规模的表型仪,ALPHA,用于高通量表型的lemaceae。ALPHA被用于一种物种的选择性育种,即长臂猿,目的是创造一种用于可持续农业系统的新作物。ALPHA可用于许多小型水生植物物种,以评估不同环境条件下的生长速率。一个原理证明用例被证明,其中ALPHA被用来确定六个不同的长臂猿克隆的盐水耐受性。
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引用次数: 0
Unequal Genetic Redundancies Among MYC bHLH Transcription Factors Underlie Seedling Photomorphogenesis in Arabidopsis. MYC bHLH转录因子不平等遗传冗余是拟南芥幼苗光形态发生的基础。
IF 2.3 3区 生物学 Q2 PLANT SCIENCES Pub Date : 2025-02-13 eCollection Date: 2025-02-01 DOI: 10.1002/pld3.70042
Vikas Garhwal, Sreya Das, Sreeramaiah N Gangappa

Light is one of the most critical ecological cues controlling plant growth and development. Plants have evolved complex mechanisms to cope with fluctuating light signals. In Arabidopsis, bHLH transcription factors MYC2, MYC3, and MYC4 have been shown to play a vital role in protecting plants against herbivory and necrotrophic pathogens. While the role of MYC2 in light-mediated seedling development has been studied in some detail, the role of MYC3 and MYC4 still needs to be discovered. Here, we show that MYC4 negatively regulates seedling photomorphogenesis, while the MYC3 function seems redundant. However, the genetic analysis reveals that MYC3/MYC4 together act as positive regulators of seedling photomorphogenic growth as the myc3myc4 double mutants showed exaggerated hypocotyl growth compared to the myc3 and myc4 single mutants and Col-0. Intriguingly, the loss of MYC2 function in the myc3myc4 double mutant background (myc2myc3myc4) resulted in further enhancement in the hypocotyl growth than myc3myc4 double mutants in WL, BL and FRL, suggesting that MYC2/3/4 together play an essential and positive role in meditating optimal seedling photomorphogenesis. Besides, MYC3/MYC4 genetically and physically interact with HY5 to partially inhibit its function in controlling hypocotyl and photo-pigment accumulation. Moreover, our results suggest that COP1 physically interacts and degrades MYC3 and MYC4 through the 26S proteasomal pathway and controls their response to dark and light for fine-tuning HY5 function and seedling photomorphogenesis.

光是控制植物生长发育最重要的生态信号之一。植物已经进化出复杂的机制来应对波动的光信号。在拟南芥中,bHLH转录因子MYC2, MYC3和MYC4已被证明在保护植物免受草食性和坏死性病原体的侵害中发挥重要作用。虽然MYC2在光介导的幼苗发育中的作用已经有了一些详细的研究,但MYC3和MYC4的作用仍有待发现。在这里,我们发现MYC4负调控幼苗光形态发生,而MYC3的功能似乎是多余的。然而,遗传分析表明,MYC3/MYC4共同对幼苗光形态发育起积极调节作用,因为myc3myc4双突变体比MYC3和MYC4单突变体和Col-0表现出更大的下胚轴生长。有趣的是,MYC2在myc3myc4双突变背景(myc2myc3myc4)中的功能缺失导致下胚轴生长比在WL、BL和FRL中的myc3myc4双突变体进一步增强,这表明MYC2/3/4共同在优化幼苗光形态发生中发挥了重要的积极作用。此外,MYC3/MYC4在遗传和物理上与HY5相互作用,部分抑制其控制下胚轴和光色素积累的功能。此外,我们的研究结果表明,COP1通过26S蛋白酶体途径与MYC3和MYC4进行物理相互作用和降解,并控制它们对黑暗和光线的反应,从而微调HY5功能和幼苗光形态发生。
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引用次数: 0
LeafDNet: Transforming Leaf Disease Diagnosis Through Deep Transfer Learning. LeafDNet:通过深度迁移学习转化叶片疾病诊断。
IF 2.3 3区 生物学 Q2 PLANT SCIENCES Pub Date : 2025-02-12 eCollection Date: 2025-02-01 DOI: 10.1002/pld3.70047
Tofayet Sultan, Mohammad Sayem Chowdhury, Nusrat Jahan, M F Mridha, Sultan Alfarhood, Mejdl Safran, Dunren Che

The health and productivity of plants, particularly those in agricultural and horticultural industries, are significantly affected by timely and accurate disease detection. Traditional manual inspection methods are labor-intensive, subjective, and often inaccurate, failing to meet the precision required by modern agricultural practices. This research introduces an innovative deep transfer learning method utilizing an advanced version of the Xception architecture, specifically designed for identifying plant diseases in roses, mangoes, and tomatoes. The proposed model introduces additional convolutional layers following the base Xception architecture, combined with multiple trainable dense layers, incorporating advanced regularization and dropout techniques to optimize feature extraction and classification. This architectural enhancement enables the model to capture complex, subtle patterns within plant leaf images, contributing to more robust disease identification. A comprehensive dataset comprising 5491 images across four distinct disease categories was employed for the training, validation, and testing of the model. The experimental results showcased outstanding performance, achieving 98% accuracy, 99% precision, 98% recall, and a 98% F1-score. The model outperformed traditional techniques as well as other deep learning-based methods. These results emphasize the potential of this advanced deep learning framework as a scalable, efficient, and highly accurate solution for early plant disease detection, providing substantial benefits for plant health management and supporting sustainable agricultural practices.

及时和准确的疾病检测对植物的健康和生产力,特别是农业和园艺工业中的植物的健康和生产力产生重大影响。传统的人工检测方法劳动强度大,主观,而且往往不准确,无法满足现代农业实践所要求的精度。本研究引入了一种创新的深度迁移学习方法,利用Xception架构的高级版本,专门用于识别玫瑰,芒果和西红柿的植物病害。该模型在基本异常架构的基础上引入了额外的卷积层,结合多个可训练的密集层,结合先进的正则化和dropout技术来优化特征提取和分类。这种结构上的增强使模型能够捕捉植物叶片图像中复杂、微妙的模式,有助于更可靠的疾病识别。一个包含5491张不同疾病类别图像的综合数据集被用于模型的训练、验证和测试。实验结果显示了出色的性能,达到98%的正确率,99%的精密度,98%的召回率和98%的f1分数。该模型优于传统技术以及其他基于深度学习的方法。这些结果强调了这种先进的深度学习框架作为早期植物病害检测的可扩展、高效和高度准确的解决方案的潜力,为植物健康管理和支持可持续农业实践提供了实质性的好处。
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引用次数: 0
A Promoter Collection for Cell-Targeted Analysis Within the Stomatal Complex. 气孔复合体细胞靶向分析的启动子集合。
IF 2.3 3区 生物学 Q2 PLANT SCIENCES Pub Date : 2025-02-12 eCollection Date: 2025-02-01 DOI: 10.1002/pld3.70045
Thanh-Hao Nguyen, Jovaras Krasauskas, Thu Binh-Anh Nguyen, Azka Noureen, Mark Smedley, John M Christie, Wendy Harwood, Michael R Blatt, Penny Hundleby

Stomatal aperture is driven by changes in turgor of the guard cells that surround the stomatal pore. Epidermal cells immediately surrounding the guard cells are thought to contribute to the kinetics of aperture changes through changes in their turgor that opposes the guard cells and thought their putative roles in solute storage for use by the guard cells. Nonetheless, our knowledge remains fragmentary of surrounding cell mechanics, in large part because the tools and strategies needed to target the surrounding cells independent of the guard cells are limited. Here, we have analyzed sets of promoters for Arabidopsis, Brassica, and barley that will allow physiological studies of the roles of epidermal cells and also surrounding cells in the case of barley in stomatal behavior. These tissue-specific promoters offer distinct advantages over widely used, constitutive promoters by enabling precise and targeted gene expression within guard cells and the adjacent epidermal cells. As genetic tools, the promoters will have applications in strategies centered on physiological analyses and differential comparisons following expression targeted between the guard cells and the foliar epidermis as a whole. As such, they are well suited to questions around the mechanics of solute and water flux that will advance an understanding of the stomatal complex in these model species.

气孔开度是由气孔周围保卫细胞的膨胀变化所驱动的。保护细胞周围的表皮细胞被认为通过其与保护细胞对立的膨胀变化而促进了孔径变化的动力学,并认为它们在为保护细胞使用的溶质储存中起着假定的作用。尽管如此,我们对周围细胞力学的知识仍然是零碎的,很大程度上是因为独立于保护细胞靶向周围细胞所需的工具和策略是有限的。在这里,我们分析了拟南芥、芸苔和大麦的启动子,这些启动子将允许对大麦的表皮细胞和周围细胞在气孔行为中的作用进行生理学研究。这些组织特异性启动子与广泛使用的组成型启动子相比,具有明显的优势,可以在保护细胞和邻近表皮细胞中精确和靶向地表达基因。作为遗传工具,启动子将在保护细胞与叶表皮之间的生理分析和表达差异比较策略中得到应用。因此,它们非常适合解决溶质和水通量的机制问题,这将促进对这些模式物种气孔复合体的理解。
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引用次数: 0
Delivery of Marker-Free DNA to Plant Genome by the Transgenic Selection-Associated Fragment Elimination (T-SAFE) System. 利用转基因选择相关片段消除(T-SAFE)系统将无标记DNA传递到植物基因组。
IF 2.3 3区 生物学 Q2 PLANT SCIENCES Pub Date : 2025-02-05 eCollection Date: 2025-02-01 DOI: 10.1002/pld3.70046
Yi Yang, Huan Chang, Leiwen Pan, Dongbei Guo, Shun Peng, Ting Mao, Yuehui Zhang, Shui Wang

The presence of a selection marker in transgenic plants has raised public concerns regarding health safety. We have developed a CRISPR/Cas9-based DNA delivery system termed transgenic selection-associated fragment elimination (T-SAFE). The T-SAFE system comprises four cassettes: the selection marker, CRISPR/Cas9, spacer-plus-protospacer adjacent motif (SP), and the cargo. The first two cassettes, the selection marker and CRISPR/Cas9, are collectively referred to as SCC. The SCC is flanked by two identical SPs derived from the fruit fly Ebony gene, which efficiently facilitate the SCC cleavage and subsequently lead to self-elimination of the selection marker upon integration of exogenous DNA into the plant genome. To inhibit the production of a functional Cas9 protein in bacteria, the IV2 intron of the potato ST-LS1 gene has been incorporated into the Cas9 gene. Additionally, the Cas9 gene is driven by a reproductive cell-specific or inducible promoter to avoid SCC cleavage in nonreproductive plant cells. These innovative features allow the T-SAFE system to achieve an elimination efficiency of the selection marker ranging from 10%-30% in Arabidopsis and 5%-8% in rice, with a DNA delivery capacity of approximately 10 kb. This approach offers a safe means for genetically modifying plants.

转基因植物中选择标记的存在引起了公众对健康安全的关注。我们开发了一种基于CRISPR/ cas9的DNA传递系统,称为转基因选择相关片段消除(T-SAFE)。T-SAFE系统包括四个盒:选择标记、CRISPR/Cas9、间隔器加原间隔器相邻基序(SP)和货物。前两个盒,选择标记和CRISPR/Cas9,统称为SCC。SCC两侧有两个来自果蝇乌木基因的相同SPs,这有效地促进了SCC的切割,并在外源DNA整合到植物基因组后导致选择标记的自我消除。为了抑制细菌中功能性Cas9蛋白的产生,马铃薯ST-LS1基因的IV2内含子被并入Cas9基因中。此外,Cas9基因由生殖细胞特异性或诱导启动子驱动,以避免非生殖植物细胞中的SCC切割。这些创新特性使T-SAFE系统能够实现选择标记的消除效率,在拟南芥中为10%-30%,在水稻中为5%-8%,DNA传递能力约为10 kb。这种方法为转基因植物提供了一种安全的方法。
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引用次数: 0
Chromosome Numbers and Reproductive Life Cycles in Green Plants: A Phylotranscriptomic Perspective. 绿色植物的染色体数目和生殖生命周期:系统转录组学的视角。
IF 2.3 3区 生物学 Q2 PLANT SCIENCES Pub Date : 2025-01-24 eCollection Date: 2025-01-01 DOI: 10.1002/pld3.70044
Rijan R Dhakal, Alex Harkess, Paul G Wolf

The strong correlation between reproductive life cycle type and chromosome numbers in green plants has been a long-standing mystery in evolutionary biology. Within green plants, the derived condition of heterosporous reproduction has emerged from the ancestral condition of homospory in disparate locations on the phylogenetic tree at least 11 times, of which three lineages are extant. In all green plant lineages where heterospory has emerged, there has been a significant downsizing in chromosome numbers. This dynamic has been investigated without clear answers for many decades. In this study, we combine known ideas from existing literature with novel methods, tools, and data to generate fresh insights into an old question. Using gene family evolution models and selection analyses, we identified gene families that have undergone significant expansion, contraction, or selection in heterosporous lineages. Alongside lineage-specific genomic changes, our results revealed shared genomic changes/trends among heterosporous lineages. We found expansions in gene families related to developmental regulation, signaling pathways, and stress responses across heterosporous groups. Notably, the MATE efflux family showed consistent expansion and evidence of selection in heterosporous lineages, suggesting a potentially conserved role in heterospory evolution. These findings could provide novel avenues to investigate and probe the underlying mechanism that may underpin the association between heterospory and genomic changes. The general importance of chromosome numbers, structure, and sizes in cellular biology notwithstanding, the association between the emergence of heterosporous reproduction and chromosome number reduction/genome downsizing is not fully understood. It remains unclear why there exists an association between aspects of biology at such disparate levels as reproductive life cycles and chromosome numbers/genome size. Exploring and answering this conundrum of evolutionary biology can add to our broader understanding of life sciences and of biology at different levels. Applying the novel tools and methods emerging from ongoing progress in biotechnology and computational sciences presents an opportunity to make new inroads into this long-standing question.

在进化生物学中,绿色植物的生殖生命周期类型与染色体数目之间的密切相关性一直是一个谜。在绿色植物中,异孢子生殖的衍生条件在系统发育树上的不同位置至少出现了11次,其中有3个谱系是现存的。在所有出现异孢子的绿色植物谱系中,染色体数目都有显著的缩小。几十年来,人们一直在研究这种动态,但没有明确的答案。在这项研究中,我们将现有文献中的已知观点与新颖的方法、工具和数据结合起来,对一个老问题产生新的见解。利用基因家族进化模型和选择分析,我们确定了在异孢子谱系中经历了显著扩展、收缩或选择的基因家族。除了谱系特异性基因组变化外,我们的研究结果还揭示了异孢子谱系之间共有的基因组变化/趋势。我们发现在异孢子群体中,与发育调控、信号通路和应激反应相关的基因家族有所扩展。值得注意的是,MATE外排家族在异孢子谱系中表现出持续的扩张和选择证据,表明其在异孢子进化中可能具有保守作用。这些发现为研究异孢子与基因组变化之间的关系提供了新的途径。尽管染色体数目、结构和大小在细胞生物学中具有普遍的重要性,但异孢子生殖的出现与染色体数目减少/基因组缩小之间的关系尚不完全清楚。目前尚不清楚为什么在生殖生命周期和染色体数目/基因组大小等不同水平的生物学方面存在关联。探索和回答进化生物学的这个难题可以增加我们对生命科学和不同层次的生物学的更广泛的理解。应用从生物技术和计算科学的持续进展中出现的新工具和方法,为解决这个长期存在的问题提供了新的机会。
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