‘章姬’草莓茎段愈伤组织诱导及高频植株再生体系的建立

Efficient callus induction and high-frequency regeneration system establishment of strawberry ‘Akihime'

该研究以‘章姬’脱毒苗带节匍匐茎段为外植体,以MS为基本培养基,进行单因素预实验,选取适合‘章姬’草莓生长的植物激素种类和质量浓度范围,进而通过L_9(3~4)正交实验研究不同植物激素种类及其质量浓度对愈伤组织诱导、丛芽发生及植株再生的影响。结果表明:采用的3种基本培养基中,B_5和1/2MS对抑制培养过程中的褐化现象明显优于MS,而附加20 g·L^(-1)Na_2S_2O_3在保证材料存活的前提下,大大降低了褐化率;去除褐化的材料在MS+0.1 mg·L^(-1)6-BA+0.05 mg·L^(-1)2,4-D+0.1 mg·L^(-1)NAA中可同步进行愈伤组织诱导和丛芽发生;丛芽增殖培养基为MS+0.1 mg·L^(-1)6-BA+0.1 mg·L^(-1)NAA,30 d后繁殖系数可达12.86;试管苗生根则在1/2MS+1.0 g·L^(-1)AC中进行,35 d后可获得生长健壮的再生植株,生根率92.50%。再生苗移栽成活率在95%以上。该研究建立了‘章姬’草莓体外高效快速繁殖体系,对短期内为‘章姬’草莓栽培提供大量种性稳定、质量优良的种苗具有重要意义,同时为其他草莓品种的体外快繁提供了技术参考。

In order to cultivate an improved variety with higher potential yield and establish an efficient in vitro regeneration system of strawberry‘Akihim’（Fragaria×ananassa）,callus induction and plant regeneration protocol was designed for solve browning problem.A formal L_9（3~4）orthogonal experiment was designed to investigate the browning research in primary culture using explants of creeping stems excised from aseptic seedlings.Based on the improved medium above,singlefactor experiments were conducted to select effective plant growth regulators and appropriate concentrations on blank MS medium.A L_9（3~4）orthogonal experiment was designed to study effects of types and concentrations of plant growth regulators on callus induction,adventitious shoot formation,and plant regeneration.The results indicated that MS medium was inferior to B_5and 1/2MS in inhibiting browning condition.The browning rate was dramatically reduced while the callus still survived on the medium with the addition of 20 g·L^（-1）Na_2S_2O_3.The callus induction and shoot formation of the explants were observed on MS＋0.1 mg·L^（-1）6-BA＋0.05 mg·L^（-1）2,4-D＋0.1 mg·L^（-1）NAA with effective inhibiting browning.The optimal medium protocol for multiple shoots proliferation was MS＋0.1 mg·L^（-1）6-BA＋0.1 mg·L^（-1）NAA where the proliferation coefficient was 12.86 after 30 d.Healthily regenerated plants were yielded on culture medium 1/2MS＋1.0 g·L^（-1）AC after 35 d,with a rooting rate of 92.50%.More than 95%of plantlets survived after transplanting into field.The rapid propagation system is helpful to provide homogeneous progeny and high quality seedlings for cultivation of strawberry‘Akihime’as well as a technical reference for other strawberry species in vitro regeneration.