摘要
Baby Boom(BBM)基因是植物细胞全能性的关键调节因子,在胚胎发育再生、细胞增殖、愈伤组织快速生长、促进分化等方面起着重要的作用。由于西洋参遗传转化体系存在转化不稳定、转化效率低、转化周期长等问题,该研究拟利用基因枪法将玉米BBM基因转入到西洋参愈伤组织中,以研究其对愈伤组织的生长以及人参皂苷含量的影响,为建立高效的西洋参遗传转化体系奠定基础。经草铵膦抗性筛选及PCR分子鉴定共获得4个不同转化事件的西洋参转基因愈伤组织,对野生型和转基因愈伤组织在相同时间内的生长状态、生长速率进行比较;利用超高效液相色谱-三重四极杆质谱(UPLC-MS/MS)对转基因愈伤组织中人参皂苷成分含量进行测定。结果表明,转BBM基因西洋参愈伤组织的生长速率显著高于野生型,人参皂苷Rb1、Rg1、Ro及Re的含量均显著高于野生型。该研究初步明确了BBM基因具有提高西洋参愈伤组织的生长速率及人参皂苷含量的作用,为进一步建立稳定、快速、高效的人参属物药用植物遗传转化体系提供依据。
Baby Boom(BBM)gene is a key regulatory factor in embryonic development and regeneration,cell proliferation,callus growth,and differentiation promotion.Since the genetic transformation system of Panax quinquefolius is unstable with low efficiency and long period,this study attempted to transfer BBM gene of Zea mays to P.quinquefolius callus by gene gunship to investigate its effect on the callus growth and ginsenoside content,laying a foundation for establishing efficient genetic transformation system of P.quinquefolius.Four transgenic callus of P.quinquefolius with different transformation events were obtained by screening for glufosinate ammonium resistance and molecular identification by PCR.The growth state and growth rate of wild-type and transgenic callus were compared in the same growth period.The content of ginsenoside in transgenic callus was determined by ultra-high performance liquid chromatography-triple quadrupole mass spectrometry(UPLC-MS/MS).The results showed that transgenic callus growth rate was significantly higher than that of wild-type callus.In addition,the content of ginsenoside Rbi,Rgi,Ro,and Re was significantly higher than that in wild-type callus.The paper preliminarily proved the function of BBM gene in promoting growth rate and increasing ginsenoside content,which provided a scientific basis to establish a stable and efficient genetic transformation system for Panax plants in the future.
作者
王腾腾
胡进
焦红红
刘艳
周骏辉
赵玉洋
袁媛
WANG Teng-teng;HU Jin;JIAO Hong-hong;LIU Yan;ZHOU Jun-hui;ZHAO Yu-yang;YUAN Yuan(School of Traditional Chinese Medicine,Guangdong Pharmaceutical University,Guangzhou 510006,China;School of Chinese Medicinal Materials,Jilin Agricultural University,Changchun 130000,China;State Key Laboratory Breeding Base of Dao-di Herbs,National Resource Center for Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China;Institute of Crop Sciences,Chinese Academy of Agricultural Sciences,Beijing 100081,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2023年第12期3156-3161,共6页
China Journal of Chinese Materia Medica
基金
国家自然科学基金项目(82173926,81891013)
中国中医科学院科技创新工程重大攻关项目(CI2021A041,CI2021B014)
中央本级重大增减支项目(2060302)
国家中医药管理局青年岐黄学者培养项目
国家中医药管理局中医药创新团队及人才支持计划项目(ZYYCXTD-D-202005)。
