|MUSTAFA, ROMA - National Institute Of Biotechnology And Genetic Engineering (NIBGE)
|HAMZA, MUHAMMAD - National Institute Of Biotechnology And Genetic Engineering (NIBGE)
|KAMAL, HIRA - National Institute Of Biotechnology And Genetic Engineering (NIBGE)
|MANSOOR, SHAHID - National Institute Of Biotechnology And Genetic Engineering (NIBGE)
|AMIN, IMRAN - National Institute Of Biotechnology And Genetic Engineering (NIBGE)
Submitted to: Molecular Biotechnology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 5/25/2017
Publication Date: 5/31/2017
Citation: Mustafa, R., Hamza, M., Kamal, H., Mansoor, S., Scheffler, J.A., Amin, I. 2017. Tobacco rattle virus (TRV) based silencing of cotton enoyl-CoA reductase (ECR) gene and the role of very long chain fatty acids in normal leaf development and resistance to wilt disease. Molecular Biotechnology. doi:10.1007/s12033-017-0014-y.
Interpretive Summary: Fungal pathogens can cause significant yield losses in cotton and currently there is no viable fungicide control. Modifying genes already present in plants in order to enhance their self- protection mechanisms is a possible solution. Using a method developed from a process that occurs naturally in Tobacco rattle virus (TRV) and causes targeted genes to be turned off, this study targeted and turned off the Enoyl-CoA reductase (ECR), a gene possibly involved in pathogen defense. Plants with an inactivated ECR gene were treated with two different fungi that cause wilt diseases (Verticillium and Fusarium). Plants previously resistant to wilt were susceptible and plants previously rated as susceptible exhibited even more extreme symptoms. The results indicate that this gene plays a role in the ability of the two wilt fungi to cause disease symptoms in cotton. Increasing the expression levels of ECR may enhance cotton’s self defense mechanisms and maintain yields in fungal infected fields.
Technical Abstract: A Tobacco rattle virus (TRV) based virus-induced gene silencing (VIGS) assay was employed as a reverse genetic approach to study gene function in cotton (Gossypium hirsutum). This approach was used to investigate the function of Enoyl-CoA reductase (GhECR) in pathogen defense. Amino acid sequence alignment of Arabidopsis ECR with homologous sequences from G. hirsutum, G. arboreum, G. herbaceum, and G. barbadense showed that ECR is evolutionarily conserved. TRV based silencing of GhECR in G. hirsutum induced a cell death / necrotic lesion-like phenotype. Reverse transcription polymerase chain reaction (RT-PCR) and real-time quantitative PCR showed reduced GhECR mRNA levels in TRV inoculated plants. Three isolates each of Verticillium dahliae and Fusarium oxysporum f. sp. vasinfectum (FOV) were used to infect GhECR silenced plants. Isolates of V. dahliae, King and VdlS.17 are defoliating types, and induced 74% and 57% foliar damage respectively, while isolate JR2, a non-defoliating type of V. dahliae, induced 68% foliar damage. In comparison to V. dahliae, FOV strains FOV5 and CA10 induced 60% and 57% foliar damage while RBHI induced only 13% foliar damage similar to the control plants. This is the first report showing that cotton GhECR is involved in resistance to different strains of V. dahliae and FOV.