Itziar Aurora Montalbán, Ander Castander-Olarieta, Antonia Maiara Marques do Nascimento, Sonia Suárez-Álvarez, Ana Herrán, Luiza Giacomolli Polesi, Franklin Back, Neusa Steiner, Miguel Pedro Guerra, Paloma Moncaleán
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Abstract
Applying stress factors such as high temperatures during the different stages of somatic embryogenesis is important to uncover the molecular mechanisms of stress response and adaptation, and as a strategy to produce plants adapted to harsh environmental conditions derived from climate changes. In this sense, the present work aims to study the effect of high temperatures applied during the maturation of somatic embryogenesis in the ploidy stability, the amino acid and polyamine profiles of the somatic embryos obtained, and the morphological characteristics of the somatic plantlets. The results revealed that the maturation temperature did not affect the morphology of the resulting somatic plantlets, nor the ploidy and genome size of phenotypically normal somatic embryos, whose ploidy and DNA content levels were similar to those found in mature zygotic embryos. Nonetheless, a slight but significant reduction of the genome size of aberrant somatic embryos was observed. The maturation temperature changed the levels of glycine, arginine, lysine, and ornithine. These last three amino acids are precursors of the polyamines detected. Regarding this, putrescine levels were higher in somatic embryos from the highest maturation temperature (5 min pulse at 60 ºC), however, the amount of this polyamine in all samples was much lower than spermidine, spermine and cadaverine. In conclusion, the different temperatures applied did not lead to substantial changes in the ploidy level, endogenous PAs of the somatic embryos developed, or in the morphology of the somatic plantlets. Significant changes in the endogenous amino acids were observed, which may be linked to PAs metabolism and other metabolic pathways involved in stress response.
摘要 在体细胞胚胎发生的不同阶段应用高温等胁迫因素，对于揭示胁迫响应和适应的分子机制，以及作为培育适应气候变化带来的恶劣环境条件的植物的一种策略非常重要。因此，本研究旨在研究体细胞胚胎发生成熟过程中的高温对体细胞胚胎倍性稳定性、氨基酸和多胺谱以及体细胞小植株形态特征的影响。结果表明，成熟温度并不影响所产生的体细胞小植株的形态，也不影响表型正常的体细胞胚胎的倍性和基因组大小，其倍性和 DNA 含量水平与成熟的合子胚胎相似。不过，异常体细胞胚胎的基因组大小有轻微但显著的减少。成熟温度改变了甘氨酸、精氨酸、赖氨酸和鸟氨酸的含量。后三种氨基酸是检测到的多胺的前体。关于这一点，最高成熟温度（60 ºC 下 5 分钟脉冲）下的体细胞胚胎中的腐胺含量较高，但所有样本中的这种多胺含量都远低于精胺、精胺和尸胺。总之，不同的温度不会导致体细胞倍性水平、发育的体细胞胚胎内源性多胺或体细胞小植株的形态发生重大变化。观察到内源氨基酸发生了显著变化，这可能与 PAs 代谢和其他参与胁迫反应的代谢途径有关。
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