摘要
发根农杆菌(Agrobacterium rhizogenes)侵染植物后可诱导植物产生毛状根。菠菜(Spinacia oleracea)是常见的食用蔬菜,目前尚未见菠菜毛状根的研究报道。经筛选得到适合诱导菠菜毛状根的发根农杆菌菌株LBA9402,LBA9402侵染菠菜外植体茎后,毛状根的诱导率最高可达16%。菠菜毛状根呈白色,具有丰富的根毛,能在无外源激素的固体培养基上快速增殖生长。通过诱导菠菜毛状根产生愈伤组织并进行分化,获得了菠菜毛状根的再生植株,再生率为8%。此外,LBA9402可将含有Ri质粒的T-DNA和携带外源GFP基因的Ti质粒T-DNA共同导入外植体中。PCR检测和荧光显微观察结果显示, rol B及GFP基因在菠菜毛状根基因组中稳定表达,共转化频率为50%。
The plant can produce hairy roots after infection by Agrobacterium rhizogenes harboring Ri plasmids.Spinach(Spinacia oleracea)is a common edible vegetable,and its hairy root system has yet to be reported.In this study,A.rhizogenes LBA9402 was screened to be suitable for hairy root induction of spinach.The highest induction efficiency,16%,was reached when the spinach stem was infected by LBA9402.Hairy roots grew vigorously on SH solid medium without exogenous hormones.The hairy roots of spinach were white and had enormous root hairs.The regeneration plant obtained from the callus redifferentiated from spinach hair roots;the rate of regeneration was 8%.In addition,Ri T-DNA and Ti T-DNA carrying rol B genes and GFP gene,respectively,were co-transferred into cells of spinach and produced transgenic hairy roots.PCR analysis and fluorescence microscopy assays showed rol B gene and GFP gene in the hairy roots of spinach and expressed stably;the rate of co-transformation was 50%.
作者
徐悦
曹英萍
王玉
付春祥
戴绍军
Yue Xu;Yingping Cao;Yu Wang;Chunxiang Fu;Shaojun Dai(Key Laboratory of Saline-alkali Vegetation Ecology Restoration,Ministry of Education,Alkali Soil Natural EnvironmentalScience Center,Northeast Forestry University,Harbin150040,China;Shandong Technology Innovation Center of SyntheticBiology,Shandong Provincial Key Laboratory of Energy Genetics,Key Laboratory of Biofuels,Qingdao Institute of Bioenergyand Bioprocess Technology,Chinese Academy of Sciences,Qingdao266000,China;Development Center of Plant GermplasmResources,College of Life Sciences,Shanghai Normal University,Shanghai200234,China)
出处
《植物学报》
CAS
CSCD
北大核心
2019年第4期515-521,共7页
Chinese Bulletin of Botany
基金
青岛市创业创新领军人才(No.16-8-3-15-zhc)
中央高校基本科研业务费专项资金(No.2572017ET01)
上海市科学技术委员会科研计划(No.17391900600)
关键词
共转化
毛状根
植物再生体系
菠菜
co-transformation
hairy root
plant regeneration system
Spinacia oleracea
