张洁 郑益平 杨成龙 林觅 林智敏

摘 要：为建立矮牵牛梅林的高频再生体系，以叶器官为材料，研究不同生长调节剂配比、叶片不同部位和不同形态愈伤对不定芽再生的影响。结果表明：6BA和NAA及其配比极显著影响矮牵牛梅林不定芽再生率，其中NAA 0.3 mg·L-1 极显著促进了矮牵牛梅林不定芽的再生，6BA和NAA浓度超过1.0 mg·L-1和0.5 mg·L-1时，刺激玻璃化的发生。叶片不同部位极显著影响不定芽的再生，芽分化率由高到低依次是叶柄>带主叶脉>不带主叶脉，愈伤组织形态决定芽分化能力和质量，叶柄直接分化丛芽是梅林再生的优良外植体，分化率达98.3%。芽诱导最佳培养基为MS+6BA 1.0 mg·L-1+NAA 0.3 mg·L-1，其产生不定芽无玻璃化且分化率高，为68.3%。叶片愈伤组织诱导最佳培养基为MS+6BA 1. 0 mg· L-1+NAA 0. 3 mg·L-1，其不定芽分化率为68.3%且未产生玻璃化苗;最佳生根培养基为1/2 MS培养基，生根率达100.0%;继代增殖培养最佳培养基为MS+6BA 0.5 mg·L-1+NAA 0.1 mg·L-1，丛生芽多且大小均匀。

关键词：矮牵牛梅林;组织培养;愈伤;植株再生

中圖分类号：S 682.2   文献标志码：A   文章编号：0253-2301（2021）05-0022-06

DOI： 10.13651/j.cnki.fjnykj.2021.05.005

Callus Induction and Plant Regeneration of Petunia Hybrida Meilin

ZHANG Jie， ZHENG Yi-ping， YANG Cheng-long， LIN Mi， LIN Zhi-min*

（Biotechnology Institute， Fujian Academy of Agricultural Sciences， Fuzhou， Fujian 350003， China）

Abstracts： In order to establish a high frequency regeneration system of Petunia hybrida Meilin， the effects of different ratios of growth regulators， different parts of leaves and different forms of callus on the regeneration of adventitious buds were studied. The results showed that 6BA， NAA and their ratios had significant effects on the regeneration rate of adventitious buds of Petunia hybrida. Among which，

0.3 mg·L-1 NAA significantly promoted the regeneration of adventitious buds of Meilin. When the concentration of 6BA and NAA respectively exceeded 1.0 mg·L-1 and 0.5

mg·L-1， the occurrence of vitrification was stimulated. Different parts of leaves significantly affected the regeneration of adventitious buds. The bud differentiation rate from high to low was petiole， with main vein， and without main vein. The ability and quality of bud differentiation were determined by the shape of callus. The direct differentiation of cluster buds from petioles was an excellent explant for the regeneration of Meilin， with the differentiation rate of 98.3%. The optimal medium for the induction of bud was MS+6BA 1.0 mg·L-1+NAA 0.3 mg·L-1， the adventitious buds were produced without vitrification and the differentiation rate was high， reaching 68.3%. The optimal medium for the induction of leaf callus was MS+6BA 1.0 mg·L-1+NAA 0.3 mg·L-1， and the differentiation rate of adventitious bud was 68.3% without vitrification. The best rooting medium was 1/2 MS medium， in which the rooting rate could reach 100.0%. The optimal medium for the multiplication culture was MS+6BA 0.5 mg·L-1+NAA 0.1 mg·L-1， and the cluster buds were

abundant and uniform in size.

Key words： Petunia hybrida Meilin;Tissue culture;Callus;Plant regeneration

矮牵牛Petunia hybrida为茄科碧冬茄属，一年或多年生草本花卉。1985年Horsch等[1]首次获得矮牵牛转基因植株，此后基因工程技术在矮牵牛的应用不断展开[2-4]，矮牵牛也因其生活周期短，遗传背景清晰且易于进行根癌农杆菌介导，逐渐成为花卉转基因的模式植物[5]。植物再生系统的建立是进行基因工程研究的实验基础，目前国内外对矮牵牛植株再生的研究较为深入，但矮牵牛再生方式受基因型限制，不同学者得出的结论也不尽相同[6-9]。矮牵牛梅林……