| Generation and Selection of Transgenic Olive Plants
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Author:
Date:
2017-11-20
[Abstract] Olive (Olea europaea L.) is one of the most important oil crops in the Mediterranean basin. Biotechnological improvement of this species is hampered by the recalcitrant nature of olive tissue to regenerate in vitro. In previous investigations, our group has developed a reliable Agrobacterium-mediated transformation protocol using olive somatic embryos as explants (Torreblanca et al., 2010). Embryogenic cultures derived from radicles of matured zygotic embryos are infected with Agrobacterium tumefaciens, AGL1 strain, containing a binary plasmid with the gene of interest and the nptII selection gene. After a meticulous selection procedure, carried out using solid and liquid media supplemented with paromomycin, the putative transformed ...
[摘要] 橄榄油（Olea europaea L.）是地中海盆地最重要的石油作物之一。生物技术的改善受到橄榄组织在体外再生的顽固性质的阻碍。在以前的研究中，我们小组开发了一个可靠的土壤杆菌介导的转化方案，使用橄榄体细胞胚作为外植体（Torreblanca et al。，2010）。含有具有感兴趣基因和nptII选择基因的二元质粒的根瘤农杆菌（Agrobacterium tumefaciens）AGL1菌株感染源自成熟合子胚胎胚根的胚性培养物。经过细致的选择程序，使用补充巴龙霉素的固体和液体培养基进行，建立推定的转化系。其转基因性质的初步确认是通过PCR扩增进行的。之后，可以通过有效的再生方案获得植物，其主要特征是使用低离子强度的矿物制剂，在液体培养基中用于培养物同步的阶段以及使用半渗透性乙酸纤维素膜进行胚胎成熟（Cerezo et al。 ，2011）。转基因插入的最终确认通过Southern或Northern分析使用再生植物的叶样品进行。
【背景】由Torreblanca等人开发的协议。 （2010）与Rugini等人（2000）开发的先前的橄榄转化协议有所不同，主要是外植体的种类，土壤杆菌菌株及其选择方法。 Rugini等人（2000）使用胚性质量作为外植体，将其在LBA4404农杆菌菌株的细菌悬浮液中温育48小时。感染后，将外植体在水中漂洗并在补充有250mg ...
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| Quantifying Auxin Metabolites in Young Root Tissue of Medicago truncatula by Liquid Chromatography Electrospray-ionisation Quadrupole Time-of-flight (LC-ESI-QTOF) Tandem Mass Spectrometry
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Author:
Date:
2016-06-20
[Abstract] Auxins represent a major group of phytohormones controlling plant development. The spatio-temporal regulation of auxin gradients is essential for the initiation, growth and correct development of plant organs. Because auxins and their metabolites occur at trace levels in plant tissue, experiments requiring identification plus their selective and specific quantification can be most conveniently achieved using mass spectrometry (MS) and the associated chromatographic methods. With the advent of appropriate liquid-based ionisation techniques, emphasis has moved from the use of gas chromatography as the sample interface to the MS (GC/MS), with its concomitant need for derivatisation, to the more sensitive liquid chromatography tandem mass spectrometry (LC-MS/MS). We describe an optimized ...
[摘要] 生长素运输能力的测量提供了涉及植物中生长素运输的生理机制的定量数据。 这种技术很容易执行，并给出快速的结果。 将放射性标记的生长素（吲哚-3-乙酸）通过琼脂块加入到Medic藜苜蓿的根部。 使用液体闪烁计数器测量根中放射性标记的生长素摄取的最终放射性。 在这里，我们描述了生长素运输能力周围的结瘤敏感区在年轻幼苗根的测量。 truncatula 响应根瘤菌接种。 类似的测定可以适用于其他植物物种并回答其他生物学问题。...
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| Water Deficit Treatment and Measurement in Apple Trees
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Author:
Date:
2015-02-05
[Abstract] Water is considered perhaps the most limiting factor for plant growth and productivity (Boyer, 1982), and climate change predicts more frequent, more severe and longer drought periods for a significant portion of the world in coming years. Unfortunately, drought resistance is particularly difficult to measure due in part to the complexity of the underlying biology that contributes to a plant’s ability to cope with water limitations. For example, water deficit is frequently examined by detaching leaves or withholding water for a set period of time prior to tissue collection. Such approaches may elucidate the early stages of drought response but are generally not physiologically relevant for maintenance of drought resistance over a longer period. A more realistic approach is to impose a ...
[摘要] 水被认为可能是植物生长和生产力的最大限制因素（Boyer，1982），气候变化预测未来几年世界上相当大一部分地区的更频繁，更严重和更长的干旱期。不幸的是，抗旱性特别难以测量，部分原因是基础生物学的复杂性，有助于植物应对水分限制的能力。例如，在组织收集之前，通常通过将叶子或保留水分离一段设定的时间来检查水缺乏。这些方法可以阐明干旱反应的早期阶段，但是通常在较长时期内维持抗旱性不是生理学相关的。更现实的方法是用持续的土壤水分水平强加逐步的水限制，特别是在多年生木本的情况下。我们在这里描述一个协议，在受控的实验室条件下施加长期缺水，允许分子生物学方法来了解木本植物如何生存的严重水限制。代表性数据可以在Artlip等人（1997）和Bassett等人（2014）中找到。
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