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Title: A study of genetic transformation systems for groundnut (Arachis hypogaea L.) with the objective to genetically engineering resistance to Indian peanut clump virus
Authors: Cheung, Mam-Kim.
Award date: 1998
Presented at: University of Leicester
Abstract: Indian peanut clump virus (IPCV) causes serious yield losses in groundnut producing areas in India. The lack of natural resistance to IPCV has focused research into the development of novel sources of resistance. Coat protein-mediated resistance (CP-MR) strategy has been widely used to generate virus resistant plants. This approach was evaluated as a potential method for generating resistance to IPCV. In order to produce transgenic groundnut expressing the IPCV CP, an efficient plant regeneration system and gene transfer method were required. To establish a reliable and reproducible plant regeneration system, five previously reported groundnut tissue culture methods were evaluated with four Indian groundnut cultivars: JL24, Plover, Robert-21 and TMV-2. Groundnut regeneration could not be obtained using these five in vitro culture methods. Instead a genotype-independent method for rapidly producing fertile groundnut plants from half of a zygotic embryo with single cotyledon (HESC) explants was developed for cultivars JL24, Plover and TMV-2 A. tumefaciens- and microprojectile bombardment-mediated gene transfer techniques were evaluated for groundnut transformation. In addition, high-level plant expression vectors containing marker genes such luc, uidA-intron and sGFP were developed to detect transformation events. An in planta transformation technique was developed and evaluated using HESC explants. However, no transgenic groundnut plants were recovered. In the absence of a groundnut transformation system, the CP-MR strategy was evaluated in N. benthamiana. Transgenic plants containing the H-IPCV CP gene were found to be highly resistant to H-IPCV. Resistance levels ranged from completely susceptible CP+ plants with virus levels equivalent to H-IPCV infected CP- plants, to resistant CP+ plants where the presence of the infecting virus could not be detected. Transgenic H-IPCV CP+ plants were also shown to be highly resistant to D-IPCV, and partially resistant to L-IPCV.
Type: Thesis
Level: Doctoral
Qualification: PhD
Rights: Copyright © the author. All rights reserved.
Appears in Collections:Theses, Dept. of Biology
Leicester Theses

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