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A highly efficient Agrobacterium mediated transformation system of chickpea wilt pathogen Fusarium oxysporum f. sp. ciceri using DsRed-Express to follow root colonization

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Title A highly efficient Agrobacterium mediated transformation system of chickpea wilt pathogen Fusarium oxysporum f. sp. ciceri using DsRed-Express to follow root colonization
 
Creator Islam, Md. Nazrul
Nizam, Shadab
Verma, Praveen K.
 
Subject Fusarium oxysporum
Fusarium wilt
Chickpea
ATMT
DsRed-Express
 
Description The soil-borne fungus Fusarium oxysporum f. sp. ciceri (Foc) causes vascular wilt of chickpea (Cicer arietinum L.), resulting in substantial yield losses worldwide. Agrobacterium tumefaciens mediated transformation (ATMT) has served as a resourceful tool for plant-pathogen interaction studies and offers a number of advantages over conventional transformation systems. Here, we developed a highly efficient A. tumefaciens mediated transformation system for Foc. In addition, a binary vector for constitutive expression of red fluorescent protein (DsRed-Express) was used to study developmental stages and host-pathogen interactions. Southern hybridisation was performed to confirm the transformation event and the presence of T-DNA in selected hygromycin resistant transformants. Most of the transformants showed single copy integrations at random positions. Microscopic studies revealed significant levels of fluorescent protein, both in conidia and mycelia. Confocal microscopy of chickpea roots infected with the transformed Foc showed rapid colonisation. These studies will allow us to develop strategies to determine the mechanisms of Foc-chickpea interaction in greater detail and to apply functional genomics for the characterisation of involved genes at the molecular level either by insertional mutagenesis or gene knock-out.
This work is supported partially by research grant
(BT/AB/01/01/2008) provided by the Department of Biotechnology,
Government of India and a core grant from the National Institute
of Plant Genome Research, New Delhi.
 
Date 2014-05-13T08:17:33Z
2014-05-13T08:17:33Z
2012
6 February 2012
 
Type Article
 
Identifier Microbiol. Res., 167(6): 332-338
http://hdl.handle.net/123456789/237
 
Language en
 
Publisher Elsevier