背蛇生体外再生体系的建立

Establishment of In Vitro Regeneration System for Aristolochia tuberosa

目的:筛选适宜的植株再生途径,建立背蛇生人工高效繁殖技术体系。方法:以背蛇生带芽茎段为外植体、以MS为基本培养基,分别考察了不同的植物激素种类和质量浓度对带芽茎段基部丛生芽的发生、增殖及生根的影响。结果:带芽茎段在MS+6-BA 1.5 mg/L+IBA 1.5 mg/L+KT 1.5 mg/L中培养10 d后节上腋芽开始萌发生长;30 d后基茎节处出现大量丛生芽,发生率为86%;45 d后在既有基部不定丛芽又有上部腋芽生长的簇状丛生系统,增殖系数达10.0以上。不定根的诱导则在1/2 MS+6-BA 0.1 mg/L+NAA 1.5 mg/L中进行,45 d后可获得生长健壮的再生植株,生根率100%;生根苗经过炼苗,移栽30 d后成活率达100%。结论:在背蛇生的体外快繁中,带芽茎段基茎丛生芽发生为最有效途径,该方式较愈伤组织间接发生不定芽效率及质量更高。该研究所建立的高效繁殖技术体系为濒危野生物种蛇背生的种质资源保护和规模化生产奠定了实验基础。

Objective:To screen the appropriate plant regeneration way of Aristolochia tuberosa and establish the artificial efficient propagation technology system.Methods:The effects of different kinds of plant hormones and concentrations on the occurrence,proliferation and plant regeneration of stem clusters with bud segments were studied with Aristolochia tuberosa stem segments with buds as explants and MS as basic medium.Results:The stem segments with buds was cultured in MS with 1.5 mg/L of 6-BA,1.5 mg/L of IBA and 1.5 mg/L of KT for 10 days,and the axillary buds began to germinate and grow.After 30 days,a large number of clumped buds appeared at the basal ganglia,with an incidence of 86%.After 45 days,a cluster system was formed with both basal adventitious plexus buds and upper axillary buds growing,with a proliferation coefficient of over 10.0.The adventitious roots were induced in 1/2 MS with 0.1 mg/L of 6-BA and 1.5 mg/L of NAA,and the regenerated plants with robust growth and rooting rate were obtained after 45 days,with 100%rooting rate.The survival rate of rooting seedlings was 100%after refining plants and 30 days of transplanting.Conclusion:The most effective way for rapid propagation of Aristolochia tuberosa in vitro is the occurrence of cluster buds from the base of stem segments with buds,which is more efficient and better than the indirect occurrence of adventitious buds from callus.The established efficient breeding technology system lays an experimental foundation for the conservation and large-scale production of the germplasm resources of the endangered wild species.