多头切花菊节间横切薄层不定芽再生的研究

Adventitous shoot regeneration from internode transverse thin cell layers of cut spray chrysanthemum.

利用节间横切薄层(transverse thin cell layers,tTCLs)培养技术建立不定芽再生体系,可以极大提高植物组织培养的再生频率,并有效控制不定芽玻璃化的发生率。该研究以4个多头切花菊品种:‘Repluse’、‘Regan EliteImproved’、‘Regan Elite Sunny’、‘Regan Elite White’的节间tTCLs为外植体,筛选出不定芽分化所需的最佳生长素种类和浓度、最佳的外植体来源和取材部位,并通过调控琼脂浓度和培养温度来有效降低不定芽玻璃化的发生率。结果表明:试验中的4个品种都能高频再生,不定芽再生率都在70%以上,Ⅰ级节间的tTCLs更易于分化不定芽;不同品种对生长素的种类要求不同,0.1 mg/L的生长素浓度为最适浓度;琼脂浓度过高或过低以及培养温度过高都会增加不定芽玻璃化的发生率。该研究建立了4个品种的高效不定芽再生体系,为菊花的良种繁育以及转基因研究提供了一条有效途径。

Internode transverse thin-cell-layers（tTCLs） is a high efficiency shoot regeneration system with low rate of hyperhydricity.In this study,internode tTCLs of four cut spray chrysanthemum cultivars,i.e.,＇Repluse＇,＇Regan Elite Improved＇,＇Regan Elite Sunny＇ and ＇Regan Elite White＇,were cultured on the medium supplemented with auxin.The goal is to select the optimum concentration of auxin and the optimum location of explants for shoot regeneration.Agar concentration and culture temperature were also explored to minimize the occurance of shoot hyperhydricity.Shoots emerged from all four cultivars＇ internode tTCLs,with the regenerating rate over 70%.Internode tTCLs of the first internode were able to regenerate more easily than other locations.The regeneration rate varied among cultivars applied with different concentrations of auxin,with 0.1 mg/L being the optimum.The frequency of hyperhydricity increased with agar concentration being too high or too low,or with high temperature.This study establishes an efficient system for adventitious shoot regeneration from cut spray chrysanthemum internode tTCLs of four cultivars,which may be further applied to the propagation of other improved varieties in transgenic research.