Zayed Eman M M, Abdelbar Ola H
Central Laboratory of Date Palm Research and Development, Agriculture Research Center, 9 Gamma Street, Giza, 12622, Egypt.
Department of Agricultural Botany, Faculty of Agriculture, Ain Shams University, Cairo, Egypt.
Methods Mol Biol. 2017;1637:129-144. doi: 10.1007/978-1-4939-7156-5_12.
Rapid production of somatic embryogenesis and date palm regeneration is achieved by culturing immature female inflorescence explants. Inflorescence explants are soft, creamy in color, average 6-7 cm in length, and cultured on Murashige and Skoog (MS) medium containing 1 mg/L thidiazuron (TDZ). Callus induction occurs after 4-5 weeks of culture on the callus induction medium. Subsequently, callus develops embryogenic calli on MS medium supplemented with 0.1 mg/L naphthalene acetic acid (NAA). Histological samples were collected successively at the culturing time and during morphogenetic changes throughout the developmental stages of somatic embryos. Initiation of callus and different successive developmental stages for somatic embryos including two-celled, four-celled, globular, bipolar, and fully developed cotyledonary somatic embryos were observed. Mature somatic embryos develop within 10-12 weeks after culture establishment.
通过培养未成熟雌花序外植体可实现体细胞胚胎发生和枣椰树再生的快速生产。花序外植体质地柔软，颜色呈奶油色，平均长度为6 - 7厘米，并在含有1毫克/升噻苯隆（TDZ）的Murashige和Skoog（MS）培养基上培养。在愈伤组织诱导培养基上培养4 - 5周后会发生愈伤组织诱导。随后，愈伤组织在添加有0.1毫克/升萘乙酸（NAA）的MS培养基上发育成胚性愈伤组织。在培养期间以及体细胞胚胎发育的整个形态发生变化过程中，连续收集组织学样本。观察到愈伤组织的起始以及体细胞胚胎的不同连续发育阶段，包括二细胞、四细胞、球形、双极和完全发育的子叶体细胞胚胎。成熟的体细胞胚胎在培养建立后的10 - 12周内发育形成。
