Plant Breeding and Acclimatization Institute - National Research Institute, Radzików, 05-870, Błonie, Poland.
J Appl Genet. 2022 Dec;63(4):663-675. doi: 10.1007/s13353-022-00717-9. Epub 2022 Aug 19.
Somatic embryogenesis is a plant regeneration method that can be exploited in tissue culture systems for a variety of tasks, such as genetic modification or the selection of somaclones with advantageous characteristics. Therefore, it is crucial to create efficient regeneration procedures and comprehend how medium components affect regeneration effectiveness or the degree of variation created in plant tissue cultures. The level of tissue culture-induced variation in triticale regenerants was examined in the current study in relation to the concentration of copper and silver ions in the induction media as well as the length of time immature zygotic embryo explants were incubated on these media. The high degree of variation (45%) revealed by the methylation-sensitive amplified fragment length polymorphism approach for estimating variation included 38% DNA sequence alterations, 6% DNA demethylation, and 1% de novo DNA methylation. Different levels of variance were found in relation to various DNA sequence settings. The CHG context had the most alterations, whereas CG experienced the fewest; sequence variation predominated in each sequence context. Lower copper ion concentrations showed the most variance. However, it could not be connected to the duration of in vitro culture or the effect of silver ions. Accordingly, we think that altering the concentration of copper ions in the induction medium may throw off the equilibrium of the metabolic processes in which copper is involved, resulting in tissue culture-induced variation.
体细胞胚胎发生是一种植物再生方法,可在组织培养系统中用于各种任务,例如遗传修饰或选择具有有利特征的体细胞无性系。因此,创建有效的再生程序并了解培养基成分如何影响再生效率或在植物组织培养中产生的变异程度至关重要。本研究探讨了组织培养诱导的黑小麦再生体的变异程度与诱导培养基中铜和银离子的浓度以及未成熟的合子胚外植体在这些培养基上培养的时间长短之间的关系。通过甲基化敏感扩增片段长度多态性方法估算变异,发现变异程度很高(45%),包括 38%的 DNA 序列改变、6%的 DNA 去甲基化和 1%的从头 DNA 甲基化。不同的 DNA 序列设置会发现不同的变异程度。CHG 上下文的改变最多,而 CG 则最少;在每个序列上下文中,序列变异都占主导地位。较低的铜离子浓度显示出更多的变异。但是,这与体外培养时间或银离子的作用无关。因此,我们认为改变诱导培养基中铜离子的浓度可能会破坏涉及铜的代谢过程的平衡,从而导致组织培养诱导的变异。