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

Islam, Md.N. and Nizam, S and Verma, P.K. (2012) A highly efficient Agrobacterium mediated transformation system for chickpea wilt pathogen Fusarium oxysporum f. sp. ciceri using DsRed-Express to follow root colonisation. Microbiological Research. pp. 1-7.

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Abstract

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.

Item Type: Article
Additional Information: 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. The Foc Delhi isolate was provided by Dr. S.C. Dube, Division of Plant Pathology, Indian Agricultural Research Institute, New Delhi. S.N. acknowledges the University Grant commission for the fellowships.
Uncontrolled Keywords: Fusarium oxysporum; Fusarium wilt; Chickpea; ATMT; DsRed-Express
Author Affiliation: National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi 110 067, India
Subjects: Crop Improvement
Divisions: Chickpea
Depositing User: Mr. SanatKumar Behera
Date Deposited: 17 Mar 2012 12:39
Last Modified: 25 Apr 2012 04:55
Official URL: http://dx.doi.org/10.1016/j.micres.2012.02.001
URI: http://eprints.icrisat.ac.in/id/eprint/3934

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