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Record Details

Record 86 of 1454
Factors enhancing Agrobacterium tumefaciens-mediated gene transfer in peanut (Arachis hypogaea L.)
Author and Affiliation:
Egnin, M.(College of Agricultural, Tuskegee University, Center for Plant Biotechnology Research, Environmental and Natural Sciences, Tuskegee, Alabama 36088, United States)
Mora, A.
Prakash, C. S.
Mortley, D. G. [Principal Investigator]
Abstract: Parameters enhancing Agrobacterium-mediated transfer of foreign genes to peanut (Arachis hypogaea L.) cells were investigated. An intron-containing beta-glucuronidase uidA (gusA) gene under the transcriptional control of CaMV 35S promoter served as a reporter. Transformation frequency was evaluated by scoring the number of sectors expressing GUS activity on leaf and epicotyl explants. The 'Valencia Select' market type cv. New Mexico was more amenable to Agrobacterium transformation than the 'runner' market type cultivars tested (Florunner, Georgia Runner, Sunrunner, or South Runner). The disarmed Agrobacterium tumefaciens strain EHA101 was superior in facilitating the transfer of uidA gene to peanut cells compared to the disarmed strain C58. Rinsing of explants in half-strength Murashige-Skoog (MS) media prior to infection by Agrobacterium significantly increased the transformation efficiency. The use of cocultivation media containing high auxin [1.0 or 2.5 mg/l (4.53 micromolar or 11.31 micromolar) 2,4-D] and low cytokinin [0.25 or 0.5 mg/l (1.0 micromolar or 2.0 micromolar) BA] promoted higher transformation than either hormone-free or thidiazuron-containing medium. The polarity of the epicotyl during cocultivation was important; explants incubated in an inverted (vertically) manner followed by a vertically upright position resulted in improved transformation and shoot regeneration frequencies. Preculture of explants in MS basal medium or with 2.5 mg thidiazuron per l prior to infection drastically decreased the number of transformed zones. The optimized protocol was used to obtain transient transformation frequencies ranging from 12% to 36% for leaf explants, 15% to 42% for epicotyls. Initial evidence of transformation was obtained by polymerase chain reaction and subsequently confirmed by Southern analysis of regenerated plants.
Publication Date: Oct 01, 1998
Document ID:
20040088589
(Acquired Sep 07, 2004)
Subject Category: LIFE SCIENCES (GENERAL)
Document Type: Journal Article
Publication Information: In vitro cellular & developmental biology. Plant : journal of the Tissue Culture Association (ISSN 1054-5476); Volume 34; 4; 310-8
Publisher Information: United States
Description: In English
Distribution Limits: Unclassified; Publicly available; Unlimited
Rights: Copyright
NASA Terms: BACTERIA; GENE EXPRESSION; GENETICALLY MODIFIED PLANTS; GENETICS; LEGUMINOUS PLANTS; CULTURE MEDIA; CULTURE TECHNIQUES; CYTOLOGY; GENES; LEAVES; LIFE SUPPORT SYSTEMS; PLASMIDS; STEMS; TISSUE CULTURING; TISSUES (BIOLOGY)
Other Descriptors: ARACHIS HYPOGAEA/CYTOLOGY/GENETICS/MICROBIOLOGY; GENE EXPRESSION; PLANTS, GENETICALLY MODIFIED; RHIZOBIUM RADIOBACTER/GENETICS; TRANSFORMATION, GENETIC; COCULTURE; COMPARATIVE STUDY; CULTURE MEDIA; GENE TRANSFER TECHNIQUES; GLUCURONIDASE/GENETICS; INTRONS/GENETICS; PLANT LEAVES/CYTOLOGY/GENETICS/MICROBIOLOGY; PLANT SHOOTS/CYTOLOGY/GENETICS/MICROBIOLOGY; PLASMIDS/GENETICS; SUPPORT, U.S. GOV'T, NON-P.H.S; TISSUE CULTURE; NASA DISCIPLINE LIFE SUPPORT SYSTEMS; NON-NASA CENTER
Availability Source: Other Sources
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