Plana Dagmara, Fuentes Alejandro, Alvarez Marta, Lara Regla M, Alvarez Félix, Pujol Merardo
National Institute of Agricultural Sciences, San Joséde las Lajas, Havana, Cuba.
Biotechnol J. 2006 Oct;1(10):1153-7. doi: 10.1002/biot.200500042.
Many available methodologies for in vitro regeneration of commercial tomato varieties promote not only the production of normal shoots but also individual leaves, shoots without apical meristems and vitrified structures. All these abnormal formations influence and diminish the regeneration efficiency. At the basis of this phenomenon lies callus development. We optimized an alternative procedure by which the regeneration occurs without abnormal shoot formation. The portion including the proximal part of hypocotyls and the radicle was cultured on medium consisting of Murashige and Skoog salts, 4 mg/L thiamine, 100 mg/L mio-inositol and 3% sucrose. After two-three weeks, 60% explants showed adventitious shoot formation. No changes in the morphological characteristics of regenerated plants and fruits were observed as compared with parents. Karyotypic analysis of regenerated plants showed no variations in chromosome number. The optimized procedure offers the advantage of tomato plant regeneration avoiding callus formation, which enables normal plant recovery with an efficiency ranging from 1.45 +/- 0.05 to 2.57 +/- 0.06 shoots per explant in Campbell-28, Amalia, Lignon, and Floradel cultivars.
许多用于商业番茄品种离体再生的现有方法不仅能促进正常芽的产生,还会促进单个叶片、无顶端分生组织的芽以及玻璃化结构的形成。所有这些异常结构都会影响并降低再生效率。这种现象的根源在于愈伤组织的发育。我们优化了一种替代方法,通过该方法再生过程中不会形成异常芽。将包括下胚轴近端部分和胚根的部分接种在由Murashige和Skoog盐、4 mg/L硫胺素、100 mg/L肌醇和3%蔗糖组成的培养基上。两到三周后,60%的外植体出现不定芽形成。与亲本相比,再生植株和果实的形态特征没有观察到变化。再生植株的核型分析表明染色体数目没有变化。该优化方法具有番茄植株再生的优势,避免了愈伤组织的形成,在Campbell - 28、Amalia、Lignon和Floradel品种中,每外植体可实现1.45±0.05至2.57±0.06个芽的正常植株再生效率。