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Identification and expression analysis of phytophthora cinnamomi CRN effector genes during infection of persea americana (Mill.)

Dissertation (MSc (Genetics))--University of Pretoria, 2023.

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Other Authors: Swart, Velushka
Format: Thesis
Language:English
Published: University of Pretoria 2024
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access_status_str Open Access
author2 Swart, Velushka
author_browse Swart, Velushka
author_facet Swart, Velushka
collection Thesis
dc_rights_str_mv © 2023 University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria.
description Dissertation (MSc (Genetics))--University of Pretoria, 2023.
format Thesis
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institution University of Pretoria (South Africa)
language English
last_indexed 2026-06-10T12:36:45.136Z
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provenance_str_mv Harvested via OAI-PMH from UPSpace — University of Pretoria Institutional Repository
publishDate 2024
publishDateRange 2024
publishDateSort 2024
publisher University of Pretoria
publisherStr University of Pretoria
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spelling oai:repository.up.ac.za:2263/94299 Identification and expression analysis of phytophthora cinnamomi CRN effector genes during infection of persea americana (Mill.) Swart, Velushka kayla.midgley@up.ac.za Van den Berg, Noelani Midgley, Kayla Alexis UCTD Plant-pathogen interactions Crinkler effectors molecular characterization in silico protein prediction Phytophthora Plant cell death Hemibiotroph Sustainable Development Goals (SDGs) SDG-02: Zero hunger Natural and agricultural sciences theses SDG-02 SDG-12: Responsible consumption and production Natural and agricultural sciences theses SDG-12 Dissertation (MSc (Genetics))--University of Pretoria, 2023. Phytophthora cinnamomi is a hemi-biotrophic plant pathogen impacting the South African avocado industry. Like other Phytophthora spp., this pathogen employs a repertoire of effectors to suppress host defence mechanisms and facilitate successful infection in host plants. Because P. cinnamomi is a hemi-biotroph, the pathogen would benefit from the suppression of cell death during the biotrophic phase early in infection and the subsequent induction of cell death later in infection when the pathogen switches to a necrotrophic phase. P. cinnamomi achieves this beneficial pattern of cell death through the differential expression and delivery of effectors at different stages of infection. Multiple effectors have been shown to play a role in host cell death manipulation in Phytophthora, but CRNs are of ongoing interest due to their ability to both supress and induce cell death during infection. The research in this thesis is a continuation of honours research conducted by me in 2020, where a pipeline was created to identify and validate P. cinnamomi CRN (PcinCRN) effector protein sequences from the P. cinnamomi GKB4 transcriptome. In the work conducted for this MSc degree, P. cinnamomi CRN expression profiles were analysed using dual RNA-seq data from P. cinnamomi infected avocado rootstocks. Candidate CRNs were selected based on their expression profile and their coding sequences were confirmed via cloning and Sanger sequencing. Amino acid sequences of CRN candidates and their variants were extracted and compared to other Phytophthora CRN sequences that have been functionally characterised as cell death inducers or suppressors during infection of host plant species. Tertiary protein structures for each CRN were predicted and their domains were assessed. Putative functions in cell death manipulation during infection of avocado were assigned to P. cinnamomi CRNs based on expression profiles, relatedness to other Phytophthora CRNs, domain analyses and protein folding predictions. From a list of 25 full-length P. cinnamomi CRNs, 13 CRN genes demonstrated expression profiles corresponding to a potential role in cell death manipulation during infection of avocado. The coding sequences of 10 candidate CRNs were confirmed, of which six were found to have two alleles. Additionally, one was found to have the potential to undergo alternative splicing. PcinCRN52 was molecular characterised as a cell death inducer during late infection and PcinCRN30, PcinCRN77, PcinCRN81 and PcinCRN86 were molecularly characterised as cell death suppressors during early infection. PcinCRN11, PcinCRN53, PcinCRN73, PcinCRN75 and PcinCRN95 were found to have variants that potentially play contradicting roles in cell death or where one variant may function in a different host plant species. The findings from this study suggest that P. cinnamomi CRNs may function by regulating each other, to either suppress or induce cell death at appropriate stages to promote successful colonisation of avocado. This study highlights the complex and dynamic interactions between CRN effectors and their potential impact on the outcome of P. cinnamomi infection of the host plant. The Hans Merensky Foundation Genetics MSc (Genetics) Unrestricted Faculty of Natural and Agricultural Sciences 2024-02-05T09:48:55Z 2024-02-05T09:48:55Z 2024-05-08 2023-11-15 Dissertation * A2024 http://hdl.handle.net/2263/94299 10.25403/UPresearchdata.25062875 en © 2023 University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria. application/pdf University of Pretoria
spellingShingle UCTD
Plant-pathogen interactions
Crinkler effectors
molecular characterization
in silico
protein prediction
Phytophthora
Plant cell death
Hemibiotroph
Sustainable Development Goals (SDGs)
SDG-02: Zero hunger
Natural and agricultural sciences theses SDG-02
SDG-12: Responsible consumption and production
Natural and agricultural sciences theses SDG-12
Identification and expression analysis of phytophthora cinnamomi CRN effector genes during infection of persea americana (Mill.)
title Identification and expression analysis of phytophthora cinnamomi CRN effector genes during infection of persea americana (Mill.)
title_full Identification and expression analysis of phytophthora cinnamomi CRN effector genes during infection of persea americana (Mill.)
title_fullStr Identification and expression analysis of phytophthora cinnamomi CRN effector genes during infection of persea americana (Mill.)
title_full_unstemmed Identification and expression analysis of phytophthora cinnamomi CRN effector genes during infection of persea americana (Mill.)
title_short Identification and expression analysis of phytophthora cinnamomi CRN effector genes during infection of persea americana (Mill.)
title_sort identification and expression analysis of phytophthora cinnamomi crn effector genes during infection of persea americana mill
topic UCTD
Plant-pathogen interactions
Crinkler effectors
molecular characterization
in silico
protein prediction
Phytophthora
Plant cell death
Hemibiotroph
Sustainable Development Goals (SDGs)
SDG-02: Zero hunger
Natural and agricultural sciences theses SDG-02
SDG-12: Responsible consumption and production
Natural and agricultural sciences theses SDG-12
url http://hdl.handle.net/2263/94299