摘要
目的优化黑果枸杞Lycium ruthenicum离体培养方法及条件,探索有效增殖方式,筛选适宜的植株再生途径,建立其人工高效繁殖技术体系。方法以无菌实生苗带1~2个节的茎段为材料,采用MS、改良的MS1以及改良的MS2为基本培养基,通过单因素、完全组合及L9(34)正交试验研究不同植物激素种类及其质量浓度对愈伤组织诱导、腋芽萌发、基茎不定丛芽诱导及植株再生的影响。结果在MS+NAA 1.0 mg/L+6-BA 0.1 mg/L培养基中,可诱导出大量愈伤组织,但其再分化能力较弱,培养35 d后最高增殖系数仅为4.36;而在MS+NAA 0.1 mg/L+6-BA 0.05 mg/L+KT 0.5 mg/L中培养,随着腋芽萌发,茎段与培养基接触的节处开始膨大并出现不定芽点,萌发出基茎丛生芽,发生率达100%,培养45 d增殖系数最高可达到42.84;试管苗生根的适宜培养基为改良的MS1+NAA 1.0 mg/L,培养40 d后生根率达98.9%;试管苗经炼苗后移栽成活率90%以上。结论研究通过基茎丛生芽这一全新的增殖途径,成功建立了黑果枸杞体外高效再生体系,不仅大大提高了试管苗的产量及品质,也为枸杞属其他植物的体外快繁提供了另一思路。
Objective To optimize the culture methods and conditions for the rapid propagation of Lycium ruthenicum in vitro, explore effective proliferation methods, and screen suitable plant regeneration pathways, so as to establish an artificial high-efficiency breeding technology system. Methods Stem segments with one or two nodes of aseptic seedlings were used as materials. MS, modified MS1 and modified MS2 were used as basic media. The effects of different plant hormones and their concentrations on callus induction, axillary bud germination, basal stem adventitious cluster bud induction and plant regeneration were studied by single factor, complete combination and L9(34) orthogonal experiments. Results A large number of callus were induced in MS + NAA 1.0 mg/L + 6-BA 0.1 mg/L medium, but the ability of redifferentiation was weak, and the highest multiplication coefficient was only 4.36 after 35 d of culture;While in MS + NAA 0.1 mg/L + 6-BA 0.05 mg/L + KT 0.5 mg/L medium, with axillary buds starting to germinate, the node of stem segment contacted with culture medium began to swell and appeared adventitious bud points, which sprouted basal stem cluster budswith the incidence rate of 100%: And the highest multiplication coefficient could up to 42.84 after 45 d of culture. The suitable medium(MS1 + NAA 1.0 mg/L) for rooting of test-tube seedlings was modified. After 40 d of culture, the rooting rate reached 98.9%;And the survival rate of transplanted tube seedlings after refining was over 90%. Conclusion In this study, the basal stem cluster buds were used as a new way for proliferation, and an efficient propagation system of L. barbarum in vitro was successfully established, which not only greatly improved the yield and quality of test-tube seedlings, but also provided another idea for the rapid propagation of other Lycium plants in vitro.
作者
李娜
黄衡宇
曾彪
LI Na;HUANG Heng-yu;ZENG Biao(Engineering Research Center for Reproducing Fine Varieties of Chinese Medicinal Plants,Yunnan University of Chinese Traditional Medicine,Kunming 650500,China;College of Traditional Chinese Medicine of Guangzhou University of Traditional Chinese Medicine,Guangzhou 510006,China;Lijiang Qiucheng Breeding Co.,Ltd.,Lijiang 674100,China;Yunnan Academy of Agricultural Sciences,Kunming 650200,China)
出处
《中草药》
CAS
CSCD
北大核心
2020年第13期3545-3553,共9页
Chinese Traditional and Herbal Drugs
基金
国家自然科学基金资助项目(81760695)
国家自然科学基金资助项目(81860674)
云南省科学技术厅-云南中医学院应用基础联合专项(2017FF117-033)联合资助。
关键词
黑果枸杞
茎段
基茎丛生芽
愈伤组织
体外快繁
Lycium ruthenicum Murr.
stem segment
basal stem cluster buds
callus
rapid propagation in vitro
