In vitro regeneration and Agrobacterium-mediated genetic transformation of Euonymus alatus.
An in vitro plant regeneration method and an Agrobacterium tumefaciens-mediated genetic transformation protocol were developed for Euonymus alatus. More than 60% of cotyledon and 70% of hypocotyl sections from 10-day-old seedlings of E. alatus produced 2-4 shoots on woody plant medium (WPM) supplemented with 5.0 mg/l 6-benzylaminopurine (BA) plus 0.2 mg/l alpha-naphthalene acetic acid (NAA), and 77% of shoots produced roots on WPM medium with 0.3 mg/l NAA and 0.5 mg/l Indole-3-butyricacid (IBA). On infection with Agrobacterium tumefaciens strain EHA105 harboring a gusplus gene that contained a plant recognizable intron from the castor bean catalase gene to ensure plant-specific beta-glucuronidase (GUS) expression, 16% of cotyledon and 15% of hypocotyl explants produced transgenic shoots using kanamycin as a selection agent, and 67% of these shoots rooted. Stable insertion of T-DNA into the host genome was determined with organ- and tissue-specific expression of the gusplus gene and further confirmed with a PCR-based molecular analysis.
Chen
Yongqin
Y
Department of Plant Science, University of Connecticut, Storrs, CT 06269, USA. yi.li@uconn.edu
Lu
Litang
L
Deng
Wei
W
Yang
Xingyu
X
McAvoy
Richard
R
Zhao
Degang
D
Pei
Yan
Y
Luo
Keming
K
Duan
Hui
H
Smith
William
W
Thammina
Chandra
C
Zheng
Xuelian
X
Ellis
Donna
D
Li
Yi
Y
D016960:Agrobacterium tumefaciens / Q000378:metabolism*; D018548:Cotyledon / Q000502:physiology; D004269:DNA, Bacterial / Q000378:metabolism; D029756:Euonymus / Q000235:genetics / Q000502:physiology*; D018745:Genome, Plant / Q000235:genetics; D018546:Hypocotyl / Q000502:physiology; D018515:Plant Leaves / Q000166:cytology; D018517:Plant Roots / Q000166:cytology; D030821:Plants, Genetically Modified; D012038:Regeneration; D014170:Transformation, Genetic
D004269:DNA, Bacterial; C036483:T-DNA