Bernard Françoise, Bazarnov Hossein Shaker, Khatab Leila Javadi, Darabi Ahmad Shafiei, Sheidai Massoud
Department of Biology, Faculty of Sciences, Plant Physiology and Biotechnology Laboratory, Shahid Beheshti University, Tehran, Iran.
Pak J Biol Sci. 2007 Jun 15;10(12):1977-83. doi: 10.3923/pjbs.2007.1977.1983.
Ferula gummosa Boiss. a highly valuable medicinal plant which naturally propagates in very limited areas of the Middle East with specific environmental conditions. The production of Ferula gummosa somatic embryos and the karyological analysis of somatic seedlings were the purpose of this study. High frequency indirect embryogenesis was induced in callus derived from zygotic embryonic axes. Embryogenesis was obtained when callus tissues were placed onto an agar induction Murashige and Skoog medium with 1-naphthalene acetic acid and after the transfer of the cultures in a thermoperiod regime of 16 h, 19 degrees C/8 h, 7 degrees C under photoperiod of 16 h light/8h dark. Embryogenic callus tissues were maintained by subculture on induction medium. Globular proliferation was achieved with suspension culture in the Murashige and Skoog medium added with 1-naphthalene acetic acid or 2,4-dichlorophenoxyacetic acid for two weeks. Maturation of embryos and development of plantlets arose on the induction agar medium, but was better after transfer into the hormone free Murashige and Skoog medium. However, the level of abnormal embryos was high. Direct embryogenesis was obtained from somatic seedlings. The best results were obtained from hypocotyl explants. Embryo induction was achieved by two week culture of the explants in 2,4-dichlorophenoxyacetic acid liquid medium; somatic embryo growth and maturation was recovered on the hormone free medium. High level of abnormalities was recorded in the culture. Karyological analysis showed a high incidence level of cytochimerism in somatic seedlings with chromosome stickiness, polypoidy and aneuploidy in metaphase cells of the same root tip. The frequency of these karyological changes varied with the type of somatic embryos with regard to morphological abnormalities. Normal and abnormal rooted somatic seedlings were able to grow until production of the first leaf and then entered dormancy in the same manner as zygotic plantlets.
胶阿魏（Ferula gummosa Boiss.）是一种极具价值的药用植物，自然繁殖区域非常有限，分布于中东具有特定环境条件的地区。本研究旨在进行胶阿魏体细胞胚胎的生产以及体细胞幼苗的染色体分析。从合子胚轴衍生的愈伤组织中诱导出了高频间接胚胎发生。当将愈伤组织置于添加了1-萘乙酸的琼脂诱导型Murashige和Skoog培养基上，并在16小时光照/8小时黑暗的光周期下，于16小时19℃/8小时7℃的变温条件下转移培养物后，获得了胚胎发生。通过在诱导培养基上继代培养来维持胚性愈伤组织。在添加了1-萘乙酸或2,4-二氯苯氧乙酸的Murashige和Skoog培养基中进行两周的悬浮培养，实现了球形胚增殖。胚胎的成熟和幼苗的发育在诱导琼脂培养基上出现，但转移到无激素的Murashige和Skoog培养基后效果更好。然而，异常胚胎的比例较高。从体细胞幼苗中获得了直接胚胎发生。下胚轴外植体取得了最佳结果。通过将外植体在2,4-二氯苯氧乙酸液体培养基中培养两周实现胚胎诱导；体细胞胚胎的生长和成熟在无激素培养基上恢复。培养中记录到高水平的异常情况。染色体分析表明，体细胞幼苗中细胞嵌合体的发生率很高，同一根尖中期细胞存在染色体粘连、多倍体和非整倍体现象。这些染色体变化的频率因体细胞胚胎的形态异常类型而异。正常和异常生根的体细胞幼苗能够生长至第一片叶子产生，然后以与合子幼苗相同的方式进入休眠。
