El Abidine Triqui Zine, Guédira Abdelkarim, Chlyah Averil, Chlyah Hassane, Souvannavong Vongthip, Haïcour Robert, Sihachakr Darasinh
Laboratoire de physiologie végétale, département de biologie, faculté des sciences, BP 1014, Rabat, Maroc.
C R Biol. 2008 Mar;331(3):198-205. doi: 10.1016/j.crvi.2007.11.009. Epub 2007 Dec 21.
Lateral buds of six cultivars of sweet potato were induced to form embryogenic callus in a culture medium solidified with two types of gelling agents, Agar or Gelrite, and supplemented with various concentrations of auxins, 2,4-D, 2,4,5-T and Picloram. Of the six cultivars screened, only three gave an embryogenic response. Best results with an average of 3.53% embryogenic response were obtained with the medium solidified with Agar, while in Gelrite only 0.45% of lateral buds gave rise to embryogenic callus. The interaction between the genotype and auxins was highly significant; particularly the optimal response was obtained with cv. Zho and 865 yielding 10.7 and 14.7% somatic embryogenesis, respectively, in the medium containing 2,4,5-T or Picloram. The plant conversion was dramatically improved by subculture of the embryogenic callus on the medium with the combination of 1 microM 2,4-D and 1 microM Kinetin or 5 microM ABA alone before transfer of mature embryos onto hormone-free medium. The embryogenic callus of sweet potato and its sustained ability to further regenerate plants have regularly been maintained for several years by frequent subculture in 5 microM 2,4,5-T or the combination of 10 microM 2,4-D and 1 microM BAP or kinetin. The embryo-derived plants seemed apparently genetically stable and similar to the hexaploid parental plants, based on morphological analysis and their ploidy level determined by using flow cytometry.
在以两种凝胶剂(琼脂或吉兰糖胶)固化并添加不同浓度生长素(2,4-二氯苯氧乙酸、2,4,5-三氯苯氧乙酸和毒莠定)的培养基中,诱导六个甘薯品种的侧芽形成胚性愈伤组织。在筛选的六个品种中,只有三个产生了胚性反应。以琼脂固化的培养基获得了最佳结果,平均胚性反应率为3.53%,而在吉兰糖胶中,只有0.45%的侧芽产生了胚性愈伤组织。基因型与生长素之间的相互作用非常显著;特别是在含有2,4,5-三氯苯氧乙酸或毒莠定的培养基中,品种Zho和865分别产生了10.7%和14.7%的体细胞胚胎发生,获得了最佳反应。在将成熟胚转移到无激素培养基之前,将胚性愈伤组织继代培养在含有1微摩尔2,4-二氯苯氧乙酸和1微摩尔激动素或单独5微摩尔脱落酸的培养基上,显著提高了植株转化率。通过在5微摩尔2,4,5-三氯苯氧乙酸或10微摩尔2,4-二氯苯氧乙酸与1微摩尔苄氨基嘌呤或激动素的组合中频繁继代培养,甘薯的胚性愈伤组织及其进一步再生植株的持续能力已定期保持数年。基于形态分析和使用流式细胞仪测定的倍性水平,胚性植株似乎在遗传上是稳定的,并且与六倍体亲本植株相似。