Plant Breeding and Acclimatization Institute-National Research Institute, Błonie, Radzików, 05-870, Poland.
J Appl Genet. 2022 Dec;63(4):677-690. doi: 10.1007/s13353-022-00719-7. Epub 2022 Aug 26.
Triticale regeneration via anther culture faces many difficulties, e.g., a low percentage of regenerated plants and the presence of albinos. Plant regeneration may be affected by abiotic stresses and by ingredients added to the induction medium. The latter influences biochemical pathways and plant regeneration efficiency. Among such ingredients, copper and silver ions acting as cofactors for enzymatic reactions are of interest. However, their role in plant tissue cultures and relationships with biochemical pathways has not been studied yet.The study evaluated relationships between DNA methylation, changes in DNA sequence variation, and green plant regeneration efficiency influenced by copper and silver ions during triticale plant regeneration. For this purpose, a biological model based on donor plants and their regenerants, a methylation-sensitive amplified fragment length polymorphism, and structural equation modeling were employed.The green plant regeneration efficiency varied from 0.71 to 6.06 green plants per 100 plated anthers. The values for the components of tissue culture-induced variation related to cytosine methylation in a CHH sequence context (where H is A, C, or T) were 8.65% for sequence variation, 0.76% for DNA demethylation, and 0.58% for de novo methylation. The proposed model states that copper ions affect the regeneration efficiency through cytosine methylation and may induce mutations through, e.g., oxidative processes, which may interfere with the green plant regeneration efficiency. The linear regression confirms that the plant regeneration efficiency rises with increasing copper ion concentration in the absence of Ag ions in the induction medium. The least absolute shrinkage and selection operator regression shows that de novo methylation, demethylation, and copper ions may be involved in the green plant regeneration efficiency. According to structural equation modeling, copper ions play a central role in the model determining the regeneration efficiency.
通过花药培养进行黑小麦再生面临许多困难，例如再生植株的比例低和出现白化苗。植物再生可能受到非生物胁迫和添加到诱导培养基中的成分的影响。后者会影响生化途径和植物再生效率。在这些成分中，作为酶促反应辅助因子的铜和银离子引起了人们的兴趣。然而，它们在植物组织培养中的作用及其与生化途径的关系尚未得到研究。本研究评估了铜和银离子对黑小麦植物再生过程中 DNA 甲基化、DNA 序列变异变化和绿苗再生效率的影响之间的关系。为此，采用了基于供体植物及其再生体的生物模型、甲基化敏感扩增片段长度多态性和结构方程模型。绿苗再生效率在每 100 个接种的花药中有 0.71 到 6.06 株。与 CHH 序列背景（其中 H 是 A、C 或 T）中胞嘧啶甲基化相关的组织培养诱导变异的组成部分的值为序列变异 8.65%、DNA 去甲基化 0.76%和从头甲基化 0.58%。所提出的模型表明，铜离子通过胞嘧啶甲基化影响再生效率，并可能通过氧化过程诱导突变，这可能会干扰绿苗再生效率。线性回归证实，在诱导培养基中没有 Ag 离子的情况下，随着铜离子浓度的增加，植物再生效率会提高。最小绝对收缩和选择算子回归表明，从头甲基化、去甲基化和铜离子可能与绿苗再生效率有关。根据结构方程模型，铜离子在决定再生效率的模型中起着核心作用。
