花叶玉簪试管苗限制生长保存研究

Study on in vitro Preservation of Hosta plantaginea by Slow Growth

玉簪属植物在园林景观及药理学方面具有很高的应用价值。野生玉簪属极易遭受自然灾害、病毒及病虫害等侵袭,组织培养继代也面临着维护成本高、容易污染等挑战。针对新品种培育及品种遗传稳定等需求,建立高安全、低维护、低消耗的保存技术具有重要意义。本研究以花叶玉簪无菌苗为材料,采用限制生长保存技术,运用正交设计,研究不同浓度的蔗糖、甘露醇、脱落酸、矮壮素对花叶玉簪限制生长试管苗的存活影响及生理生化响应。结果表明:在20~25℃条件下,玉簪试管苗在培养基中添加蔗糖50 g/L+甘露醇20 g/L+脱落酸(ABA,abscisic acid)1 mg/L+矮壮素(CCC,chlormequat chloride)20 mg/L,保存6个月后存活率仍为100%,无继代保存期可达对照2倍以上且维持性状稳定;保存后材料的超氧化物歧化酶(SOD,superoxide dismutase)、过氧化物酶(POD,peroxidase)、过氧化氢酶(CAT,catalase)酶活性均高于对照组,丙二醛(MDA,malondialdehyde)含量均低于对照组,保存材料的抗氧化酶活性增强,细胞膜脂氧化程度降低,延缓衰老进程,延长保存继代周期。

Hosta Tratt.plants have high value in landscape and pharmacology.Hosta Tratt.plants in the wild are vulnerable to natural disasters,viruses,diseases and insect pests,and the normal tissue culture is faced with high maintenance costs,and chances of pollution.Considering the need for breeding new varieties and maintaining the genetic stability,it is of great significance to establish a preservation technique with safety,low maintenance and low consumption.This research was conducted to obtain the optimal in vitro preservation protocol for Hosta plantaginea(Lam.)Asch.by using the orthogonal design and to study the effects of different concentrations of sucrose,mannitol,abscisic acid and CCC(chlormequat chloride)in the culture medium on the survival percentages and stability during the slow growth of in vitro plantlets of Hosta plantaginea(Lam.)Asch..The results showed that 100%survival of in vitro Hosta plantaginea(Lam.)Asch.plantlets was obtained after six-month storage at 20-25℃,which were cultured in the MS medium supplemented with 50 g/L sucrose,20 g/L mannitol,1 mg/L ABA,20 mg/L CCC and 0.7%agar(pH 5.8),and the storage life without subculture was twice as much as that of the control of 3 months on normal medium.The activities of SOD,POD and CAT in preserved in vitro plantlets were higher than those of the control group,and the content of MDA was lower than that of the control group,which indicated that the activity of antioxidant enzymes increased,the degree of lipid oxidation decreased,the senescence process was delayed,and the storage life was prolonged.