野生唐古特白刺悬浮细胞系的建立及生长特性

以野生唐古特白刺的茎段、叶片为外植体,研究了NaClO对外植体的消毒效果和激素2,4-D、NAA和6-BA对愈伤组织诱导及愈伤组织形态的影响,并利用疏松型愈伤组织建立了白刺悬浮细胞系,对其生长特征进行了分析。结果表明:对茎段和叶片用2%的NaClO消毒10 min最适宜,时间过长或过短均会影响成活率;3种激素对愈伤组织的相对生长量和形态均有影响,其中,2,4-D是主要影响因子,且茎段诱导出的愈伤组织相对生长量比叶片的高;根据愈伤组织的生长速度和形态,适宜用浅黄色疏松型愈伤组织构建唐古特白刺悬浮细胞系,最佳试验组合为MS+2,4-D 1.5 mg·L^-1+NAA 0.2 mg·L^-1+6-BA 0.4 mg·L^-1,最佳继代接种量为7.5 mL母液,生长曲线呈"S"型,培养第7天细胞分裂指数达最大值(5.1%),培养第3天细胞活力最强(吸光值为0.69),存活率在细胞生长延迟期和稳定期较对数期下降略快,但均保持在84% 93%间。

Establishment of Cell Suspension Culture and Growth Characteristics of Wild Nitraria tangutorum

Taking stems and leaves of wild Nitraria tangutorum Bobr. as explants to study the disinfectant effects by NaClO on explants and the effects of 2,4-D, NAA and 6-BA on callus induction and morphology. The callus morphology was analyzed and cell suspension systems were established with the loose type callus, its growth characteristics were observed. The results show that the best way of disinfection of explants was 10 minutes with NaClO, the longer or shorter the sterilization time, the lower the survival rate. Three hormones had effects on relative growth and morphology of callus and the 2,4-D was a major factor, the relative growth rate of callus from stem was higher than that of leaves. To be confirmed by callus growth rate and morphological characteristics, cultured on MS medium containing 2,4-D 1.5 mg·L^-1, NAA 0.2 mg·L^-1 and 6-BA 0.4 mg·L^-1 was the optimum treatment combination for the establishment of N. tangutorum suspension cell lines. The calluses were loose type with light yellow, and the best inoculation dosage was 7.5 mL mother liquor. Under such condition, the cells grew in an 'S' curve. The cell mitotic index reached the maximum value of 5.1% at the seventh day, at the third day of cultured the cell got the strongest activity and the light absorption value was 0.69. The survival rate in the lag phase and stationary phase decreased slightly faster than logarithmic phase, but maintained between 84%-93%.