Kamo K, Jones B, Castillon J, Bolar J, Smith F
U.S. Department of Agriculture, U.S. National Arboretum, Floral and Nursery Plants Research Unit, B-010A Room 238, BARC West, Beltsville, MD 20705, USA.
Plant Cell Rep. 2004 Jun;22(11):787-92. doi: 10.1007/s00299-003-0723-5. Epub 2004 Mar 12.
Plant regeneration from embryogenic cells of two Rosa hybrida cultivars, Kardinal and Classy, was increased by dispersing embryogenic callus in liquid medium for 3 h followed by size-fractionation to isolate proembryogenic masses that were smaller than 530 microm. Dispersed callus of three cultivars, Kardinal, Classy, and Tineke, produced 61-135 cotyledonary-stage embryos/100 mg fresh weight (FW) as compared to intact callus that had not been dispersed, which produced only zero to three cotyledonary-stage embryos/100 mg FW. Over 500 cotyledonary-stage embryos/100 mg FW callus developed from proembryogenic masses of Kardinal, Classy, and Tineke following 2 months of culture on solidified Murashige and Skoog's basal salts medium supplemented with 0.25% activated charcoal. Cotyledonary-stage embryos of Classy that developed from both dispersed callus and fractionated cells of various sizes showed a significantly higher conversion frequency to plants (28%) than cotyledonary-stage embryos isolated from intact callus (9%). The highest conversion frequencies for Kardinal (50-58%) occurred from cotyledonary-stage embryos that developed from dispersed callus and from the fraction of cells smaller than 850 microm.
通过将两种杂交茶香月季品种（红衣主教和优雅）的胚性细胞在液体培养基中分散3小时，然后进行大小分级以分离小于530微米的原胚团，可提高其植株再生率。与未分散的完整愈伤组织相比，红衣主教、优雅和廷内克这三个品种的分散愈伤组织每100毫克鲜重（FW）可产生61 - 135个子叶期胚，而未分散的完整愈伤组织每100毫克FW仅产生0至3个子叶期胚。在添加了0.25%活性炭的固化Murashige和Skoog基础盐培养基上培养2个月后，红衣主教、优雅和廷内克的原胚团发育出超过500个子叶期胚/100毫克FW愈伤组织。从分散愈伤组织和各种大小分级细胞发育而来的优雅子叶期胚向植株的转化频率（28%）显著高于从未分散愈伤组织分离得到的子叶期胚（9%）。红衣主教的最高转化频率（50 - 58%）出现在从分散愈伤组织和小于850微米的细胞分级发育而来的子叶期胚中。
