An Enhancer Trap system to track developmental dynamics in Marchantia polymorpha
An Enhancer Trap system to track developmental dynamics in Marchantia polymorpha
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追踪地钱发育动态的增强陷阱系统
DOI：
10.1101/2023.07.06.547788
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Marron A
中科院分区：
文献类型：
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作者：
Marron A
与维管植物相比，流线型遗传学、实验易处理性和相对形态学简单性的组合使得地钱成为研究植物生物学许多方面的理想模型系统。在这里，我们描述了一个转化载体相结合的组成型荧光膜标记与核标记，这是由附近的增强子元件调节，并使用此产生的增强子陷阱线地钱库筛选芽从这些线允许新的标记线，包括标记假根和油细胞的鉴定和表征。该图书馆允许识别的边缘组织运行的叶状体边缘，突出显示在叶状体发育。该标记的表达与生长素水平相关。我们产生了多个标记的分生组织顶端缺口区域，具有不同的空间表达模式，在顶端缺口切除后的分生组织再生过程中的不同时间重新出现，并有不同的反应生长素补充或抑制。这表明，有proximodistal亚结构内的顶切迹，否则无法观察到。我们使用我们的标记来研究地钱孢子的发育，观察分生组织的出现，定义了原丝体到原叶体阶段的过渡，以及随后在原叶体阶段产生边缘组织。外源生长素处理在原丝体阶段阻止分生组织出现，但不抑制细胞分裂，导致具有许多假根的愈伤组织样孢子，而抑制生长素合成和运输的外源生长素处理不阻止分生组织出现。这个增强子陷阱系统为社区提供了有用的资源，并将有助于未来的地钱研究。
A combination of streamlined genetics, experimental tractability and relative morphological simplicity compared to vascular plants makes the liverwortMarchantia polymorphaan ideal model system for studying many aspects of plant biology. Here we describe a transformation vector combining a constitutive fluorescent membrane marker with a nuclear marker that is regulated by nearby enhancer elements and use this to produce a library of enhancer trap lines for Marchantia.Screening gemmae from these lines allowed the identification and characterization of novel marker lines, including markers for rhizoids and oil cells. The library allowed the identification of a margin tissue running around the thallus edge, highlighted during thallus development. The expression of this marker is correlated with auxin levels. We generated multiple markers for the meristematic apical notch region, which have different spatial expression patterns, reappear at different times during meristem regeneration following apical notch excision and have varying responses to auxin supplementation or inhibition. This reveals that there are proximodistal substructures within the apical notch that could not be observed otherwise. We employed our markers to study Marchantia sporeling development, observing meristem emergence as defining the protonema‐to‐prothallus stage transition, and subsequent production of margin tissue during the prothallus stage. Exogenous auxin treatment stalls meristem emergence at the protonema stage but does not inhibit cell division, resulting in callus‐like sporelings with many rhizoids, whereas pharmacologically inhibiting auxin synthesis and transport does not prevent meristem emergence. This enhancer trap system presents a useful resource for the community and will contribute to future Marchantia research.