Mazri Mouaad Amine, Meziani Reda, Belkoura Ilham, Mokhless Boutaïna, Nour Souad
1Institut National de la Recherche Agronomique, CRRA-Marrakech, UR Agro-Biotechnologie, Laboratoire de Biotechnologie Végétale, BP 533, Marrakech, Morocco.
2Institut National de la Recherche Agronomique, CRRA-Errachidia, UR Systèmes Oasiens, Laboratoire National de Culture des Tissus du Palmier Dattier, BP 2, Errachidia, Morocco.
3 Biotech. 2018 Apr;8(4):215. doi: 10.1007/s13205-018-1235-x. Epub 2018 Apr 9.
An efficient regeneration system via a combined pathway of organogenesis and somatic embryogenesis was developed for date palm ( L.) cv. Mejhoul. Adventitious buds were obtained from shoot-tip explants with a frequency of 53.3% after 9 months of culture: 6 months on half-strength Murashige and Skoog (MS/2) medium containing 14.2 µM indole-3-acetic acid (IAA), 13.4 µM 1-naphthaleneacetic acid (NAA) and 0.5 µM 6-(dimethylallylamino) purine (2iP), and 3 months on MS/2 medium supplemented with 1.1 µM IAA, 1.1 µM NAA, 0.5 µM 2iP, 2.2 µM 6-benzyladenine (BA) and 0.4 µM kinetin. Adventitious bud segments were used as explants to induce somatic embryogenesis, and the effects of different concentrations (22.5, 45, 90, 225 or 450 µM) of 3,6-dichloro-o-anisic acid (dicamba) and 4-amino-3,5,6-trichloropicolinic acid (picloram) were evaluated. The optimal medium for somatic embryogenesis induction was MS medium supplemented with 45 µM picloram and 5 µM 2iP, in which the somatic embryogenesis rate was 70%. For somatic embryo maturation, the effects of sorbitol, mannitol, polyethylene glycol (PEG) and abscisic acid (ABA) were tested. The highest maturation rate (88.6 mature somatic embryos per 100 mg fresh weight callus) was observed on liquid MS medium supplemented with 20 g L PEG. Subsequent somatic embryo germination was achieved with up to 52.0% in MS medium containing 2.5 µM NAA and 2.5 µM BA. The regenerated plantlets were transferred to the glasshouse where 76.0% of them survived.
通过器官发生和体细胞胚胎发生的联合途径，为海枣（Phoenix dactylifera L.）品种“梅久尔”建立了一种高效的再生系统。在培养9个月后，从茎尖外植体获得不定芽，频率为53.3%：在含有14.2µM吲哚-3-乙酸（IAA）、13.4µM 1-萘乙酸（NAA）和0.5µM 6-（二甲基烯丙基氨基）嘌呤（2iP）的1/2强度Murashige和Skoog（MS/2）培养基上培养6个月，然后在添加了1.1µM IAA、1.1µM NAA、0.5µM 2iP、2.2µM 6-苄基腺嘌呤（BA）和0.4µM激动素的MS/2培养基上培养3个月。将不定芽切段用作外植体诱导体细胞胚胎发生，并评估了不同浓度（22.5、45、90、225或450µM）的3,6-二氯邻茴香酸（麦草畏）和4-氨基-3,5,6-三氯吡啶甲酸（毒莠定）的影响。诱导体细胞胚胎发生的最佳培养基是添加了45µM毒莠定和5µM 2iP的MS培养基，其中体细胞胚胎发生率为70%。对于体细胞胚胎成熟，测试了山梨醇、甘露醇、聚乙二醇（PEG）和脱落酸（ABA）的影响。在添加20g/L PEG的液体MS培养基上观察到最高的成熟率（每100mg鲜重愈伤组织有88.6个成熟体细胞胚胎）。随后，在含有2.5µM NAA和2.5µM BA的MS培养基中，体细胞胚胎的萌发率高达52.0%。再生植株被转移到温室中，其中76.0%存活。
