Pérez Marta, Viejo Marcos, LaCuesta Maite, Toorop Peter, Cañal María Jesús
Lab. Fisiología Vegetal, Dpto. B.O.S., Facultad de Biología, Oviedo University, C/Catedrático Rodrigo Uría, s/n, 33071 Oviedo, Asturias, Spain; Intituto Universitario de Biotecnología de Asturias (IUBA asociado con CSIC), Spain.
Departamento de Biología Vegetal y Ecología, Facultad de Farmacia, UPV/EHU, Vitoria-Gasteiz E-01006, Spain.
J Plant Physiol. 2015 Jan 15;173:51-61. doi: 10.1016/j.jplph.2014.07.028. Epub 2014 Sep 17.
Somatic embryogenesis is a powerful alternative to conventional mass propagation of Quercus suber L. However, poor quality and incomplete maturation of somatic embryos restrict any application. Given that epigenetic and hormonal control govern many developmental stages, including maturation of zygotic embryos, global DNA methylation and abscisic acid (ABA) were analyzed during development and maturation of cork oak somatic embryos. Our results indicated that development of somatic embryos concurred with a decrease in 5-mdC. In contrast, endogenous ABA content showed a transient increase with a peak in immature E2 embryos denoting the onset of the maturation phase. A cold stratification phase was necessary for embryos to acquire germination ability, which coincided with a significant decrease in 5-mdC and ABA content. Immunohistochemical analyses showed that there was a specific spatial-temporal regulation during embryogenesis, particularly after the cold treatment. The acquisition of germination capacity concurred with a general low 5-mdC signal in the root meristem, while retention of the 5-mdC signal was mainly located in the shoot meristem and provascular tissues. Conversely, ABA immunolocalization was mainly located in the root and shoot apical meristems. Furthermore, a strong decrease in the ABA signal was observed in the root cap after the stratification treatment suggesting a role for the root cap during development of somatic embryos. These results suggest that, in addition to ABA, epigenetic control appears to play an important role for the correct maturation and subsequent germination of cork oak somatic embryos.
体细胞胚胎发生是栓皮栎传统大规模繁殖的一种有效替代方法。然而,体细胞胚胎的质量差和成熟不完全限制了其任何应用。鉴于表观遗传和激素调控控制着许多发育阶段,包括合子胚的成熟,因此在栓皮栎体细胞胚胎的发育和成熟过程中分析了全球DNA甲基化和脱落酸(ABA)。我们的结果表明,体细胞胚胎的发育与5-甲基胞嘧啶(5-mdC)的减少同时发生。相反,内源ABA含量显示出短暂增加,在未成熟的E2胚胎中达到峰值,表明成熟阶段开始。冷分层阶段对于胚胎获得萌发能力是必要的,并与5-mdC和ABA含量的显著降低同时发生。免疫组织化学分析表明,在胚胎发生过程中存在特定的时空调节,特别是在冷处理后。萌发能力的获得与根分生组织中普遍较低的5-mdC信号同时发生,而5-mdC信号的保留主要位于茎分生组织和原维管组织中。相反,ABA免疫定位主要位于根和茎顶端分生组织中。此外,分层处理后在根冠中观察到ABA信号强烈降低,表明根冠在体细胞胚胎发育过程中起作用。这些结果表明,除了ABA外,表观遗传控制似乎对栓皮栎体细胞胚胎的正确成熟和随后的萌发起重要作用。