药用植物白及组培苗瓶内复壮研究

The Rejuvenation of Medicinal Plant Bletilla striata in Vitro

对白及(Bletilla striata)试管苗进行复壮研究,以期解决其羸弱纤细、不利于生根培养及生根后成活率低等问题,同时优化复壮培养条件。在课题组已有的白及增殖及生根培养基配方中,分别添加不同质量浓度植物生长延缓剂多效唑(PP333)和矮壮素(CCC)并改变培养基碳源浓度,以不同继代试管苗为材料,通过单因素和L9(34)正交实验研究不同材料和不同植物生长延缓剂种类及其质量浓度对试管苗复壮、繁殖生长、生根培养及驯化移栽的影响。实验表明,4代苗在MS+活性炭1.0 g/L+香蕉泥50 g/L+6-BA 0.5 mg/L+NAA 0.01 mg/L+PP3330.5 mg/L+CCC 0.1 mg/L+蔗糖40 g/L中获得最佳的复壮效果,培养70天后,试管苗基茎直径平均达(0.26±0.06) cm,繁殖系数2.13,假鳞茎发生率100%;复壮后的试管苗在1/2MS+活性炭1.0 g/L+香蕉泥80 g/L+6-BA 0.1 mg/L+NAA 1.0 mg/L+蔗糖15 g/L中培养60天后,可获得具假鳞茎、根粗苗壮的再生植株,生根率100%;再生苗移栽成活率亦达100%。本研究完善了白及体外快繁及复壮条件,建立了高效的植株再生体系。

Since the transplanting seedlings are difficult to adapt to rooting culture and domestication due to the tenuous stems in medium, the rejuvenation research was conducted using test-tube plantlets of Bletilla striata by optimizing the conditions of the medium. Based on the proliferative and rhizogenic formulas established in our previous study, the single factor and L9(34) orthogonal experiment were carried out using different subculture plantlets, different concentrations of plant growth retardant(PP333and CCC) and carbon source, aiming to investigate the impact of different types and concentrations of plant growth retardant and explants on rejuvenation, multiplication, rooting culture and transplanting. L9(34) orthogonal test exhibited that the optimal rejuvenation medium was MS+ 1.0 g/L AC+ 50 g/L banana puree+ 0.5 mg/L 6-BA+ 0.01 mg/L NAA+ 0.5 mg/L PP333+ 0.1 mg/L CCC+ 40 g/L sucrose for the fourth generation. After 70 d, the diameter of the plantlets’ basal part of stem, the growth coefficient and the occurrence rate was(0.26±0.06) cm, 2.13 and 100%,respectively. The pseudobulbs and roots were sent forth from rejuvenation plantlets after 60 d on the 1/2 MS medium containing 1.0 g/L AC, 80 g/L banana puree, 0.1 mg/L 6-BA, 1.0 mg/L NAA and 15 g/L sucrose. The survival rate of transplant seedlings was 100%. High-efficiency regeneration system of B. striata with optimal rejuvenation condition was established.