以混培法离体诱导油茶多倍体细胞的初步研究

摘要

为了研究秋水仙素诱导油茶多倍体细胞的效果,结合植物组织培养方法,以'华硕'嫩叶为材料,用不同浓度的秋水仙素处理进行不同时间长度的培养,利用流式细胞仪测定培养获得的愈伤组织相对DNA含量,石蜡切片显微观察愈伤组织细胞形态.结果表明:在一定的浓度范围内,随着秋水仙素浓度的升高和处理时间的延长,叶盘的死亡率逐渐升高,愈伤率逐渐降低;相对于对照组而言,各处理组均可诱导获得多倍体细胞,其中以40 mg·L-1的秋水仙素处理20 d和以20 mg·L-1的秋水仙素处理30 d培养获得的油茶愈伤组织细胞其相对DNA含量倍数均在2.0以上,其核质比为(0.130±0.023),且在显微镜下还能观察到细胞分生团.%In order to study effects of colchicine on inducing polyploid cells in Camellia oleifera,using tender leaves of 'Huashuo' as materials, the materials were treated by different concentrations of colchicine for different durations, combining with plant tissue culture method.Relative DNA content in polyploid cells was determined by flow cytometry, and paraffin sections were used for microscopic observation of callus cell.The results show that:at a certain concentration range, with increase of colchicine concentration and extension of treatment time, mortality of leaf disc was increased gradually, and callus rate was decreased gradually. Compared with the control group, polyploid cells could be induced in each treatment group. Among them, multiples of relative DNA contents in obtained callus were both above 2.0 in the treatments of 40 mg·L-1colchicine for 20 days and 20 mg·L-1colchicine for 30 days, and nuclear-cytoplasmic ratio was (0.130±0.023). Moreover, some cell meristems could be found under microscope.