Efficient Agrobacterium-mediated Transformation of the Elite–Indica Rice Variety Komboka
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Author:
Van T. Luu, Melissa Stiebner, Paula Emmerich Maldonado, Sandra Valdés, Didier Marín, Gerardo Delgado, Virginia Laluz, Lin-Bo Wu, Paul Chavarriaga, Joe Tohme, Inez H. Slamet-Loedin and Wolf B. Frommer,
Date:
2020-09-05
[Abstract] Genetic transformation is crucial for both investigating gene functions and for engineering of crops to introduce new traits. Rice (Oryza sativa L.) is an important model in plant research, since it is the staple food for more than half of the world’s population. As a result, numerous transformation methods have been developed for both indica and japonica rice. Since breeders continuously develop new rice varieties, transformation protocols have to be adapted for each new variety. Here we provide an optimized transformation protocol with detailed tips and tricks for a new African variety Komboka using immature embryos. In Komboka, we obtained an apparent transformation rate of up to 48% for GUS/GFP reporter gene constructs using this optimized protocol. This ...
[摘要] [摘要 ] 遗传转化对于研究基因功能和农作物工程引入新性状均至关重要。水稻(Oryza sativa L.)是植物研究中的重要模型,因为它是世界一半以上人口的主食。其结果是,大量的转化方法已经开发了两个籼稻和粳稻米。由于育种者不断开发新的水稻品种,因此必须针对每个新品种适应转化方案。在这里,我们为使用未成熟胚的非洲新品种Komboka 提供了一种优化的转化协议,包括详细的技巧和窍门。在Komboka中,我们使用此优化方案对GUS / GFP报告基因构建体的表观转化率高达48%。该协议也适用于其他优质lite 稻品种。
[背景 ] 为植物的遗传转化的各种方法公顷一直在开发,例如穿孔(Shimamoto PEG介导的原生质体转染。等人,1989;达塔。等人,1992),生物射弹转化(Christou的等人,1991)和农杆菌-介导的转化(Slamet-Loedin 等,2014)。农杆菌介导的转化是将DNA引入植物的最广泛使用的方法之一(van Wordragen and Dons,1992)。该方法已被广泛用于研究,并已成为生物技术的关键先决条件。自从开发新的育种技术(如基因组编辑)以来,它就变得越来越重要(Char ...
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Electron Tomography to Study the Three-dimensional Structure of Plasmodesmata in Plant Tissues–from High Pressure Freezing Preparation to Ultrathin Section Collection
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Author:
Date:
2018-01-05
[Abstract] Plasmodesmata (PD) are nanometric (~20 nm wide) membrane lined pores encased in the cell walls of the adjacent plant cells. They allow the cells to exchange all types of molecules ranging from nutrients like sugar, hormones, to RNAs and various proteins. Unfortunately, they are also hijacked by phyto-viruses, enabling them to spread from cell-to-cell and then systematically throughout the whole plant. Their central position in plant biology makes it crucial to understand their physiology and especially link their function to their structure. Over the past 50 years, electron microscopists have observed them and attempted to ultrastructurally characterize them. They laid the foundation of what is known about these pores (Tilney et al., 1991; Ding et al., 1992; Oparka and ...
[摘要] Plasmodesmata(PD)是包裹在相邻植物细胞的细胞壁中的纳米(〜20nm宽)膜衬里的孔。它们允许细胞交换从糖,激素,RNA到各种蛋白质等营养物质的所有类型的分子。不幸的是,它们也被植物病毒劫持,使它们从细胞间传播,然后在整个植物体系中传播。它们在植物生物学中的核心地位使得理解其生理机制,尤其是将其功能与其结构联系起来至关重要。在过去的50年中,电子显微镜观察家们已经观察到了这些现象,并试图用超微结构来表征它们。他们为已知的这些毛孔奠定了基础(Tilney等人,1991; Ding等人,1992; Oparka和Roberts,2001; Nicolas等人, et al。,2017a)。
尽管三维电子显微镜(3D-EM)爆炸,PD超微结构仍然不支持这种技术。第一个技术难点是在尽可能接近原生状态的情况下处理它们。其次,由于染色/固定试剂穿透率差,其体积增大,含水量高以及存在酸性液泡,植物样品显示自己难以加工。最重要的是,它们在细胞壁上的独特位置和它们的纳米尺寸使得难以方便地染色以便看到这些孔隙的内部运作。
这里我们详细描述Nicolas et al。(2017b)中使用的协议,对PD进行细节化处理,并生成高分辨率的X线断层图。
【背景】高压冻结(HPF)依赖于样品中存在的水的玻璃化。通过以足够高的冷冻速度(10 ...
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In vitro Floral Induction of Cuscuta reflexa
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Author:
Date:
2017-01-20
[Abstract] Floral initiation and development in the angiosperms is the essential step on which the yield of the plant depends. Sometimes external climate or any abiotic stress hinders the floral initiation and ultimately affect the plant yield. Hence, in vitro floral induction and development can help to overcome the external climatic factor. Furthermore, the protocol for in vitro floral induction in the parasitic angiosperm like Cuscuta reflexa has not been reported yet. We have standardized the protocol for floral induction in the parasitic plant Cuscuta reflexa. In this study it is established that MMS (modified Murashige Skoog) media supplemented with 2 mg L-1 NAA (naphthalene acetic acid, plant growth regulator) and 2 mg L-1 2,4-D ...
[摘要] 被子植物的花卉引发和发育是植物产量依赖的关键步骤。有时,外部气候或任何非生物胁迫阻碍花卉开始,最终影响植物产量。因此,体外花卉诱导和发育可以帮助克服外部气候因素。此外,尚未报道如在睾丸反射中的寄生被子植物体外花诱导的方案。我们已经规定了寄生植物茧丝反射中花诱导的方案。在本研究中,确定补充有2mg L NAA(萘乙酸,植物生长调节剂)和2mg L -1的MMS(改良的Murashige Skoog)培养基> 2,4-D(2,4-二氯苯氧基丙酸,植物生长调节剂)在体外拍摄后支持花诱导。此外,我们发现补充有2mg L-sup-2,4-D的MMS培养基从淋巴结外植体直接快速诱导花蕾,而没有任何芽伸长。该协议将有助于研究人员在其他被子植物植物中诱导花along与镰刀菌反射。背景 睾丸反射是寄生在大量被子植物上寄生的被子植物(Kuijt,1969)。在宿主植物中造成严重损害和产量损失的能力使得该物种对科学研究非常重要(Nun and Mayer,1999)。除了少数具有功能性叶绿体(Hibberd等人,1998)之外,大多数Cuscuta 物种是非叶绿体。从较大的胚胎开发幼苗。已经报道了白色介质中的反射(Maheshweri和Baldev,1961)。花卉诱导。反射炎在短时间内以及黑暗条件已经在体外中看到(Baldev,1962)。 ...
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