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首页> 外文期刊>In Vitro Cellular and Development Biology. Plant: Journal of the Tissue Culture Association >Optimization of methods for using polyethylene glycol as a non-permeating osmoticum for the induction of microspore embryogenesis in the Brassicaceae
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Optimization of methods for using polyethylene glycol as a non-permeating osmoticum for the induction of microspore embryogenesis in the Brassicaceae

机译：优化使用聚乙二醇作为非渗透性渗透液来诱导十字花科小孢子胚胎发生的方法

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摘要

The objective of this work was to enhance the quality and quantity of microspore-derived embryos of cruciferous species by using polyethylene glycol (PEG) to replace sucrose in the culture medium. The main advantage in using PEG is that it produces embryos that are morphologically more similar to zygotic embryos and have enhanced germination capabilities. When microspores were cultured in full strength NLN medium supplemented with 25% (w/v) PEG, the addition of 3 ml of full strength NLN with 13% (w/v) sucrose at 14 d was beneficial for embryo quality and quantity. Experiments showed that this PEG system could be used for a number of Brassica napus cultivars, as well as a number of other cruciferous species. PEG enhanced microspore embryogenesis in B. nigra, Crambe abyssinica, and Raphanus oleifera. Microspore-derived embryos were obtained from all cruciferous species evaluated (B. alboglabra, B. carinata, B. juncea, B. rapa, B. nigra, R. oleifera, Crambe abyssinica, Sinapis alba) using either sucrose or PEG as the osmoticum. Microspore embryogenesis was induced in B. napus in PEG-based cultures without a 32°C heat shock (i.e., 4, 15, 18, and 24°C). These temperature conditions were non-inductive when sucrose was used as the osmoticum. Spontaneous chromosome doubling occurred in 64–92% of the regenerated plants when PEG was used in the NLN culture medium, whereas in culture medium containing sucrose, the spontaneous doubling rate was 2–18%.
机译：这项工作的目的是通过使用聚乙二醇（PEG）替代培养基中的蔗糖来提高十字花科物种的小孢子来源胚的质量和数量。使用PEG的主要优点是它产生的胚胎在形态上与合子胚更相似，并且具有增强的发芽能力。当在补充有25％（w / v）PEG的全强度NLN培养基中培养小孢子时，在14 d加入3 ml含13％（w / v）蔗糖的全强度NLN有利于胚胎的质量和数量。实验表明，该PEG系统可用于许多甘蓝型油菜品种，以及许多其他十字花科物种。 PEG增强了黑芽孢杆菌，深层蛤ram和油菜萝卜中的小孢子胚胎发生。使用蔗糖或PEG作为渗透液，从所有评估的十字花科物种（小白芥，卡氏小芥，芥菜，小白菜，黑麦，小油菜，克拉姆比阿比西尼卡，白芥子）中获得小孢子来源的胚。 。在32°C无热休克（即4、15、18和24°C）的PEG基培养物中，甘蓝型油菜诱导小孢子胚胎发生。当使用蔗糖作为渗透剂时，这些温度条件是非感应的。当在NLN培养基中使用PEG时，再生植物的自发染色体倍增发生在64–92％，而在含蔗糖的培养基中，自发倍增率为2–18％。

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