Fki Lotfi, Kriaa Walid, Nasri Ameni, Baklouti Emna, Chkir Olfa, Masmoudi Raja B, Rival Alain, Drira Noureddine
Laboratory of Plant Biotechnology, Faculty of Sciences of Sfax, University of Sfax, Route de Sokra, Sfax, Tunisia.
UMR DIADE, Cirad BioS, IRD, Montpellier Cedex 5, France.
Methods Mol Biol. 2017;1637:99-106. doi: 10.1007/978-1-4939-7156-5_9.
This chapter describes an efficient protocol for large-scale micropropagation of date palm. Somatic embryo-derived plants are regenerated from highly proliferating suspension cultures. Friable embryogenic callus is initiated from juvenile leaves using slightly modified Murashige and Skoog (MS) medium supplemented with 0.1 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D). Suspension cultures consisting of proembryonic masses are established from highly competent callus for somatic embryogenesis using half-strength MS medium enriched with 0.1 mg/L 2,4-D and 300 mg/L activated charcoal. The productivity of cultures increased 20-fold when embryogenic cell suspensions were used instead of standard protocols on solidified media. The overall production of somatic embryos mostly exceeds 10,000 units per liter per month. Partial desiccation of mature somatic embryos, corresponding to a decrease in water content from 90 down to 75%, significantly improved germination rates.
本章介绍了一种高效的枣椰树大规模微繁殖方案。体细胞胚衍生植株从高度增殖的悬浮培养物中再生。使用添加了0.1毫克/升2,4-二氯苯氧乙酸（2,4-D）的略微改良的Murashige和Skoog（MS）培养基，从幼叶诱导出易碎胚性愈伤组织。使用富含0.1毫克/升2,4-D和300毫克/升活性炭的半强度MS培养基，从用于体细胞胚胎发生的高活性愈伤组织中建立由原胚团组成的悬浮培养物。当使用胚性细胞悬浮液而非固化培养基上的标准方案时，培养物的生产力提高了20倍。体细胞胚的总体产量大多超过每月每升10,000个单位。成熟体细胞胚的部分干燥，对应于含水量从90%降至75%，显著提高了发芽率。
