以筛选得到的四倍体稗草成熟胚作为外植体, 进行了组织培养与快速繁殖技术的研究。结果表明: 最佳外植体消毒方式为70%乙醇浸泡30 s+0.1%(w/v)的升汞消毒15 min。最佳诱导培养基为MS盐、DL维生素+2.5 mg/L 2,4-D+300 mg/L谷氨酰胺+500 mg/L脯氨酸+3%麦芽糖。将愈伤组织转入分化培养基(MS+50 mg/L肌醇+1.0 mg/L 6-BA+0.2 mg/L KT+0.5 mg/L NAA+0.25 mg/L IAA+3%麦芽糖)上, 培养20 d后超过70%的愈伤组织分化成苗。将3-5 cm长的分化苗转入1/2 MS生根培养基进行生根培养。炼苗后, 移入营养土与珍珠岩(2∶1)的基质中, 移栽成活率高达90%以上。该体系的建立为稗草抗除草剂基因的功能验证提供了前提条件。

We report a successful and efficient plant regeneration system using mature embryo of barnyardgrass (Echinochloa crus). The effect of different sterilization procedures on explant culture and the effect of plant growth regulators on regeneration process and acclimatization of transplanting were assessed. It was found that EtOH+0.1%HgCl2 was optimal for explant sterilization. Essentially, dehulled and sterilized seeds were plated on callus induction and subculture medium composed of MS basal salts and DL vitamins plus 2.5 mg/L 2,4-D, 300 mg/L glutamine, 500 mg/L proline, 3% maltose, 0.8% (w/v) agar at pH 5.8. Differentiation was carried out on a differentiation medium MS with 50 mg/L inositol plus 1.0 mg/L 6-BA, 0.2 mg/L kinetin KT, 0.5 mg/L NAA, 0.25 mg/L IAA, 3% maltose, 0.8% (w/v) agar at pH 5.8. Plantlets ranging from 3 to 5 cm in length were transferred to the rooting media comprising 1/2MS basal salts. In vitro regenerated plantlets were washed to remove the traces of agar and then were transferred to pots containing nutritious soil and perlite (2∶1, v/v), and the survival rate was over 90%. The protocol developed in this study will facilitate barnyardgrass generation and is useful for the production of herbicide-resistant transgenic plants.