牡丹子叶离体再生研究

卫俨 张紫娟 李梅兰 胡永红 朱木兰

摘要：以牡丹（Paeonia suffruticosa）品种凤丹的子叶片为起始外植体，研究了不同种类、浓度及其配比的植物生长调节剂对牡丹愈伤组织诱导、不定芽分化及不定根诱导等离体再生过程的影响。结果表明，0.1%升汞浸泡7 min，再用10%次氯酸钠溶液浸泡5 min消毒效果最佳，胚的无菌率和萌发率分别为82.96%、56.03%；诱导愈伤组织的最佳NAA浓度为3.0 mg/L，出愈率达85.03%；最适宜的不定芽诱导培养基为WPM+1.0 mg/L 6-BA+0.3 mg/L NAA，不定芽诱导率可达77.35%，平均不定芽数为3.05个；最适宜的不定芽伸长培养基为WPM+0.3 mg/L 6-BA+2.0 mg/L GA3，不定芽伸长率为81.12%，芽的平均伸长长度可达1.93 cm；最适宜的生根培养基为1/2MS+2.0 mg/L NAA+1.5 mg/L IBA+1.0 mg/L AgNO3，生根率达54.31%。

关键词：牡丹（Paeonia suffruticosa）；子叶叶片；再生体系

中图分类号：S567.1+5；Q813.1+2 文献标识码：A 文章编号：0439-8114（2018）07-0117-05

DOI：10.14088/j.cnki.issn0439-8114.2018.07.028

Study on Regeneration System for Cotyledons in Peonyin Vitro

WEI Yan1，2，3，ZHANG Zi-juan1，2，3，LI Mei-Lan1，HU Yong-hong2，ZHU Mu-lan2，3

（1.College of Horticulture，Shanxi Agricultural University，Taigu 030801，Shanxi，China；

2.Institute of Physiology & Ecology，Shanghai Chen Shan Botanical Research Center，Shanghai 201602，China；

3.Institute of Physiology & Ecology，Shanghai Institutes for Biological Sciences，CAS，Shanghai 200032，China）

Abstract： The cotyledon leaves of peony（Paeonia suffruticosa） cultivars Fengdan was used as the initial explants，the effects of different species，concentration and compatibility of plant hormones on the regeneration process of peony cotyledon leaves were studied，including callus induction，adventitious bud differentiation and adventitious root induction. The results showed that the use of 0.1% mercury immersion for 7 minutes，then 10% sodium hypochlorite solution soak for 5 minutes is the best disinfection method，aseptic rate and germination rate of embryo were 82.96% and 56.03%. The optimum naphthylacetic acid （NAA） concentration in callus occurrence was 3.0 mg/L and the callus induction rate was 85.03%. The optimum medium for the adventitious buds occurrence was WPM+1.0 mg/L 6-benzylamino purine（6-BA）+0.3 mg/L NAA along with the frequency of differentiation was 77.35% and average number of adventitious buds was 3.05. The suitable medium for adventitious bud elongation was WPM+0.3 mg/L 6-BA+2.0 mg/L gibberellin（GA3）. The elongation efficiency was about 81.12%，and the average shoot length was 1.93 cm. The optimum rooting medium was 1/2MS+2.0 mg/L NAA+1.5 mg/L indole butyric acid（IBA）+1.0 mg/L AgNO3，the rooting rate reached 54.31%.

Key words： peony（Paeonia suffruticosa）；cotyledon leaves；regeneration system

牡丹（Paeonia suffruticosa）是芍藥科芍药属植物，为多年生落叶小灌木。花大色艳、富丽端庄，素有“花中之王”的美誉[1]。牡丹作为中国传统名花，自古以来颇受人们喜爱，常用作园林观赏花卉；牡丹子富含人体所需的氨基酸、维生素、多糖和多种不饱和脂肪酸[2，3]，以牡丹子仁为原料榨制而成的食用油，不饱和脂肪酸含量高达92%以上，其中α-亚麻酸占42%，多项指标超过被称为“液体黄金”的橄榄油[4，5]；牡丹根皮成分丹皮酚，有抗炎、降温、解热、解痉等多种药理[6]，为常用的镇痛、镇静之良药[7]。

虽然牡丹市场前景广阔，但是该物种的常规育苗方式耗时长、成本高、操作难度大。因此，通過组织培养技术建立牡丹离体再生体系具有重要意义。目前，国内外关于牡丹离体再生的研究尚未完全成熟，特别是再生过程中褐化严重、生根困难等问题阻碍了该物种的科研与生产进程[8-13]。……