烟草胚状体诱导的影响因素

Factors Affecting Induction in Tobacco Embryoid by in vitro Culture

【目的】为了烟草胚状体诱导培养体系能得到优化。【方法】以2个烟草杂交品种的花药与叶片分别作为试验材料,对影响胚状体诱导的因子:基因型、外源激素浓度及比例、培养条件、接种密度等进行对比研究。【结果】所有的供试材料均可被诱导出烟草胚状体,但不同基因型间诱导率差异显著,且花药被诱导出胚状体的能力要强于叶片;在IAA与6-BA浓度不超过0.5 mg/L时,胚状体诱导率较不加激素能得到显著提升,超过该浓度后,随着激素浓度的提升反而会使胚状体诱导率下降,愈伤组织诱导率上升;进行10 d的暗处理能显著提升胚状体的诱导率。【结论】诱导胚状体产生的最佳培养条件为采用叶片为诱导材料,每培养皿接种35颗,使用1/2 MS+0.125 mg/L IAA+0.5 mg/L 6-BA培养基,在黑暗条件下诱导10 d后,再转移至光照下培养。

【Objective】The study aimed to establish the anther culture system.【Method】The anther and leaves from two kind of tobacco were used to study the effects of genotype,exogenous hormones,external culture factors,inoculum density etc.on the anther culture.【Result】The facts showed that all experimental material could induce tobacco seeding,and different material had different rate of haploid embryo.And the ability of the anthers to induce embryoid was stronger than that of leaves;The exogenous hormone treatment showed that lower IAA and 6-BA tend to form embryoid.Higher combination would increase the rate of callus.10 days without light before the lighting culture would promote efficiency.【Conclusion】1/2 MS+0.125 mg/L IAA+0.5 mg/L 6-BA was the best medium to improve the tobacco leaves embryoid culture.35 pollens per dish and culturing without light for 10 days and then transferred to light.