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曼陀罗和黑种草二倍体及四倍体植株与离体培养物的流式细胞术研究

Flow cytometric investigations of diploid and tetraploid plants and in vitro cultures of Datura stramonium and Hyoscyamus niger.

作者信息

Weber Jost, Georgiev Vasil, Pavlov Atanas, Bley Thomas

机构信息

Partec GmbH, Am Flugplatz 13, 02828 Görlitz, Germany.

出版信息

Cytometry A. 2008 Oct;73(10):931-9. doi: 10.1002/cyto.a.20628.

Abstract

Plant in vitro systems are valuable sources for the production of biological active substances. However, changed profiles of secondary metabolites, and low, variable yields possibly caused by genetic instabilities complicate their industrial implementation. DNA profiling of plant in vitro cultures may provide data for the selection of highly producing in vitro cultures. Diploid and tetraploid Datura stramonium and Hyoscyamus niger plant as well as calli, and hairy root lines derived from them were analyzed by flow cytometry. Plant in vitro cultures undergo several cycles of endoreduplication more than the explants from which they were obtained. The highest cycle values were observed in calli (e.g. 1.19 for diploid H. niger) possibly induced by the growth factors. However, hairy roots cultivated without growth factor exhibited significant degrees of endoreduplication (cycle value 0.88 for diploid H. niger). Sets of five hairy root lines from each plant and ploidy level showed consistent within-set ploidy patterns. The ploidy profiles of investigated plant in vitro and in vivo differ. For the first time we report that hairy roots of two Solanaceae species undergo endoreduplication. Theploidy profiles of in vitro cultures (hairy roots and calli) seem to be influenced by the genome size, the growth factors applied, and the type of in vitro culture. The transformation of several hairy root lines showed no differences in the ploidy patterns.

摘要

植物体外系统是生物活性物质生产的宝贵来源。然而,次生代谢产物谱的改变以及可能由遗传不稳定性导致的低产量和产量波动,使其工业化应用变得复杂。植物体外培养物的DNA分析可为高产体外培养物的选择提供数据。通过流式细胞术分析了二倍体和四倍体曼陀罗、黑种草植株以及由它们衍生的愈伤组织和毛状根系。植物体外培养物比获得它们的外植体经历更多轮的核内复制。在愈伤组织中观察到最高的复制轮值(例如,二倍体黑种草为1.19),这可能是由生长因子诱导的。然而,在没有生长因子的情况下培养的毛状根表现出显著程度的核内复制(二倍体黑种草的复制轮值为0.88)。来自每种植物和倍性水平的五组毛状根系显示组内倍性模式一致。所研究的植物体外和体内的倍性谱不同。我们首次报道两种茄科植物的毛状根会发生核内复制。体外培养物(毛状根和愈伤组织)的倍性谱似乎受基因组大小、所应用的生长因子以及体外培养类型的影响。几个毛状根系的转化在倍性模式上没有差异。

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