以带1 mm叶柄的月季叶片为外植体诱导出胚性愈伤和体细胞胚胎,对月季体细胞胚胎植株再生进行研究。结果表明,月季体细胞胚胎再生与培养基种类、培养基坡度和体细胞胚胎自身类型等密切相关。月季体细胞胚胎在SP/R培养基(含1.0 mg/L 6-BA+0.05 mg/L NAA+3.0mg/L GA3)上比在EM培养基(含1.0 mg/L 2,4–D+0.1 mg/L TDZ)上能再生出更多小植株,在EP培养基(含3.0 mg/L 2,4–D+0.5 mg/L TDZ)上则逐步褐化死亡;将培养基放置成坡面,有利于体细胞胚胎分化出芽和根,提高植株再生率;在体细胞胚胎的子叶尚未完全开张(呈90°~150°角)前转入倾斜的SP/R培养基可以再生出正常植株,体细胞胚胎子叶充分伸展甚至向外翻转时(大于150°角)再转入倾斜的SP/R培养基,体细胞胚胎则会回到愈伤组织形态,丧失植株再生能力。
Leaves with a l mm petiole of China Rose (R. chinensis Jacq.) were used as explant to induce embryogenic callus and embryos for study of plant regeneration of rose. The results showed that the regeneration of rose somatic embryos was closely related to types of culture media, slope of medium and type of embryo itself. The higher regenerated frequency was obtain on SP/R medium containing 1.0 mg/L 6-BA + 0.05 mg/L NAA+ 3.0 mg/L GA3 than on EM medium with 1.0 mg/L 2,4-D + 0.1 mg/L TDZ. Somatic embryos on EP media with 3.0 mg/L 2,4-D + 0.5 mg/L TDZ would be browned and subsequently necrosed. The sloping and even vertical medium, with probably different water and nutrition level in different medium height, was benefit for the proliferation of shoot and root of somatic embryos and showed higher regenerated frequency. Somatic embryos should be transferred to SP/R medium for normal plant regeneration before the cotyledons stretch thoroughly. The somatic embryo with cotyledons stretch thoroughly would go back to calli and lose capacity of regeneration on SP/R medium.