Functional characterisation and evaluation of Erwinia mallotivora bifunctional chorismate mutase via TargeTron® gene knockout system


Citation

Abu Bakar, N., . and Juri, N. M, . and Sohaime, M. Z., . and Samsuddin, A. F., . (2024) Functional characterisation and evaluation of Erwinia mallotivora bifunctional chorismate mutase via TargeTron® gene knockout system. Journal of Tropical Agriculture and Food Science (Malaysia), 52 (1). 1 -10. ISSN 1394-9829

Abstract

Erwinia mallotivora is the causal agent of papaya dieback disease, a devastating diseases affecting papaya production in Malaysia. Preliminary omics studies had successfully uncovered the key effectors of Erwinia mallotivora’s (Em’s) pathogenicity, where chorismate mutase (CM) was among the most prominent effectors in E. mallotivora. An in-depth in silico characterisation and functional analysis of the CM was carried out. Sequence analysis revealed that E. mallotivora’s CM protein contained two catalytic motifs; chorismate mutase type II (CM 2) and prephenate dehydratase (PDT) and a regulatory domain (ACT), where no signal peptide was detected in its sequence. Based on this characterisation, it is confirmed that our targeted CM is a non-secreted bifunctional chorismate mutase/prephenate dehydratase (Cmp). Sequence homology search showed that EmCmp is highly conserved (>80% identity) with various bifunctional Cmp from several Erwinia and Pantoea species (n ≥60). To confirm its role in Em virulency, a Cmp knockout mutant of Em (EmΔCmp) was generated via TargeTron® Gene Knockout system. The EmΔCmp strain significantly loss its virulency to papaya seedlings as compared to the wild E. mallotivora strain in the inoculation assay, revealing the evidence of EmCmp function and importance as the target gene for future research towards disease management strategy in papaya.


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Abstract

Erwinia mallotivora is the causal agent of papaya dieback disease, a devastating diseases affecting papaya production in Malaysia. Preliminary omics studies had successfully uncovered the key effectors of Erwinia mallotivora’s (Em’s) pathogenicity, where chorismate mutase (CM) was among the most prominent effectors in E. mallotivora. An in-depth in silico characterisation and functional analysis of the CM was carried out. Sequence analysis revealed that E. mallotivora’s CM protein contained two catalytic motifs; chorismate mutase type II (CM 2) and prephenate dehydratase (PDT) and a regulatory domain (ACT), where no signal peptide was detected in its sequence. Based on this characterisation, it is confirmed that our targeted CM is a non-secreted bifunctional chorismate mutase/prephenate dehydratase (Cmp). Sequence homology search showed that EmCmp is highly conserved (>80% identity) with various bifunctional Cmp from several Erwinia and Pantoea species (n ≥60). To confirm its role in Em virulency, a Cmp knockout mutant of Em (EmΔCmp) was generated via TargeTron® Gene Knockout system. The EmΔCmp strain significantly loss its virulency to papaya seedlings as compared to the wild E. mallotivora strain in the inoculation assay, revealing the evidence of EmCmp function and importance as the target gene for future research towards disease management strategy in papaya.

Additional Metadata

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Item Type: Article
AGROVOC Term: Erwinia
AGROVOC Term: Carica papaya
AGROVOC Term: dieback
AGROVOC Term: pathogenicity
AGROVOC Term: virulence
AGROVOC Term: genetic transformation
AGROVOC Term: bioinformatics
AGROVOC Term: inoculation
AGROVOC Term: disease control
Geographical Term: Malaysia
Uncontrolled Keywords: chorismate mutase, knock out
Depositing User: Ms. Azariah Hashim
Date Deposited: 21 Nov 2025 02:33
Last Modified: 21 Nov 2025 02:33
URI: http://webagris.upm.edu.my/id/eprint/2697

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