背景:植物体外技术的发展带来了在再生体中鉴定的变异,称为体细胞克隆或组织培养诱导变异(TCIV)。S-腺苷-L-蛋氨酸(SAM),谷胱甘肽(GSH),低甲基化果胶(LMP),和Cu(II)离子可能与绿色植物再生效率(GPRE)和TCIV有关,根据对大麦(大麦)和部分小黑麦(×Triticosecalespp。威特马克前A.加缪1927年)。使用结构方程模型(SEM),这些代谢物与代谢途径(克雷布斯和杨循环,糖酵解,转硫化),但不适合小黑麦.利用代谢组学和(epi)遗传数据,该研究试图建立小黑麦再生效率统计模型。培养物的诱导培养基补充有各种量的Cu(II)和Ag(I)离子用于再生。植物再生的时期也发生了变化。供体植物,花药衍生的再生剂,和metAFLP用于分析关于对称DNA的TCIV(CG,CHG)和不对称(CHH)序列上下文。衰减全反射-傅立叶转移红外(ATR-FTIR)光谱用于收集LMP的代谢组学信息,SAM,GSH。要构建数据,采用结构方程模型。
结果:根据metAFLP分析,CHH上下文中的平均序列变化为8.65%,0.58%为从头甲基化。特定于LMP区域的FTIR光谱的吸光度,SAM,和GSH被用作引入SEM模型的变量值。每100个接种花药的绿色再生剂的平均数量为2.55。
结论:果胶去甲基化的量,SAM,从头甲基化,和GSH在模型中连接以解释GPRE。通过改变培养基中Cu(II)离子的浓度,影响果胶的含量,小黑麦的GPRE可以增加。
BACKGROUND: The development of the plant in vitro techniques has brought about the variation identified in regenerants known as somaclonal or tissue culture-induced variation (TCIV). S-adenosyl-L-methionine (SAM), glutathione (GSH), low methylated pectins (LMP), and Cu(II) ions may be implicated in green plant regeneration efficiency (GPRE) and TCIV, according to studies in barley (Hordeum vulgare L.) and partially in triticale (× Triticosecale spp. Wittmack ex A. Camus 1927). Using structural equation models (SEM), these metabolites have been connected to the metabolic pathways (Krebs and Yang cycles, glycolysis, transsulfuration), but not for triticale. Using metabolomic and (epi)genetic data, the study sought to develop a triticale regeneration efficiency statistical model. The culture\'s induction medium was supplemented with various quantities of Cu(II) and Ag(I) ions for regeneration. The period of plant regeneration has also changed. The donor plant, anther-derived regenerants, and metAFLP were utilized to analyze TCIV concerning DNA in symmetric (CG, CHG) and asymmetric (CHH) sequence contexts. Attenuated Total Reflectance-Fourier Transfer Infrared (ATR-FTIR) spectroscopy was used to gather the metabolomic information on LMP, SAM, and GSH. To frame the data, a structural equation model was employed.
RESULTS: According to metAFLP analysis, the average sequence change in the CHH context was 8.65%, and 0.58% was de novo methylation. Absorbances of FTIR spectra in regions specific for LMP, SAM, and GSH were used as variables values introduced to the SEM model. The average number of green regenerants per 100 plated anthers was 2.55.
CONCLUSIONS: The amounts of pectin demethylation, SAM, de novo methylation, and GSH are connected in the model to explain GPRE. By altering the concentration of Cu(II) ions in the medium, which influences the amount of pectin, triticale\'s GPRE can be increased.