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Candidatus liberibacterafricanus in non-rutaceous alternate host species from South Africa

Dissertation (MSc)--University of Pretoria, 2019.

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Other Authors: Pietersen, Gerhard
Format: Thesis
Language:English
Published: University of Pretoria 2020
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access_status_str Open Access
author2 Pietersen, Gerhard
author_browse Pietersen, Gerhard
author_facet Pietersen, Gerhard
collection Thesis
dc_rights_str_mv © 2020 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)--University of Pretoria, 2019.
format Thesis
id oai:repository.up.ac.za:2263/77829
institution University of Pretoria (South Africa)
language English
last_indexed 2026-06-10T12:38:26.101Z
license_str Other — see source repository
provenance_str_mv Harvested via OAI-PMH from UPSpace — University of Pretoria Institutional Repository
publishDate 2020
publishDateRange 2020
publishDateSort 2020
publisher University of Pretoria
publisherStr University of Pretoria
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source_str UPSpace — University of Pretoria Institutional Repository
spelling oai:repository.up.ac.za:2263/77829 Candidatus liberibacterafricanus in non-rutaceous alternate host species from South Africa Pietersen, Gerhard rochelle.jansevanrensburg@tuks.co.za Kruger, Kerstin Janse van Rensburg, Rochelle UCTD Dissertation (MSc)--University of Pretoria, 2019. Non-rutaceous plant species, potentially hosts to ‘Candidatus Liberibacter africanus’ (Laf) sensu lato, were sampled throughout the Cape Floristic Region from the Fynbos and Succulent Karoo biomes in South Africa, and tested for the presence of the insect-transmitted bacterial pathogen associated with ‘Citrus Greening disease’ (CG) in Citrus species (Rutaceae). Laf is considered a persistent problem to the production of citrus in South Africa as fruits produced from CG infected citrus are smaller in size, lopsided or misshaped and have a characteristic bitter taste. The information on the potential host range in indigenous and other plant species in South Africa is limited. In the current study three surveys were carried out during September 2017 (spring), January 2018 (summer), and August 2018 (winter) in the natural vegetation in Robertson, Worcester, Slanghoek, Vredendal, Lutzville and Klawer. Potential psyllid vectors were collected with vacuum sampling from approximately 20 randomly selected plant samples per plant species at each site. Branches and flowers, when available, of the same plants were collected for morphological identification. Leaf and petiole samples of 989 plant specimens, representing 19 plant families and 42 species, were collected. No typical galls induced on leaves by psyllid nymphs were observed on the plants. Psyllids were only collected from two Roepera foetida (Zygophyllaceae) plants. Of the 989 plant specimens of alternate host species tested for the presence of Liberibacters by real-time polymerase chain reaction (real-time PCR) assays 142 yielded a Ct value below the selected positive/negative threshold of 31 following the Liberibacter ‘Universal’ real-time PCR assay. Conventional PCR tests, including the amplification of the 16S rRNA, omp and rplJ genes of Laf, were conducted on these 142 plant specimens. Seven of these yielded very faint bands after gel electrophoresis analysis of the 16S rRNA conventional PCR test. Sanger sequencing of these suggested they were non-target amplicons. Therefore, none of the 42 plant species tested positive for ‘Ca. Liberibacter africanus’, the causal agent for CG in South African citrus. As a number of Atriplex semibaccata plants had yielded real-time PCR values <31 and yielded some amplicons with the 16S rRNA PCR, a sample with a DNA concentration above 250 ng/μl was selected for next-generation sequence (NGS) analysis using an Illumina HiSeq 2000 platform at Life Sequencing (Spain) to attempt to identify the presence of a potentially divergent Liberibacter. NGS data indicated that the bacterial entity amplified by the Liberibacter ‘Universal’ real-time PCR was not Liberibacter spp., but an, as yet, unidentified member of the Gammaproteobacteria. This bacterium may also be present in a number of other Atriplex samples tested during this study, including A. lindleyi and A. nummularia, which had also yielded amplicons in the real-time PCR assays. It may also be present in some of the other plant species testing positive in the real-time PCR test. The data generated from this study, and from the studies done in conjunction with this one, will be used for biological and epidemiological studies and the development of management strategies. Microbiology and Plant Pathology MSc Unrestricted 2020-12-29T11:50:49Z 2020-12-29T11:50:49Z 2020/04/30 2019 Dissertation Janse van Rensburg, R 2019, Candidatus liberibacterafricanus in non-rutaceous alternate host species from South Africa, MSc Dissertation, University of Pretoria, Pretoria, viewed yymmdd <http://hdl.handle.net/2263/77829> A2020 http://hdl.handle.net/2263/77829 en © 2020 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
Candidatus liberibacterafricanus in non-rutaceous alternate host species from South Africa
title Candidatus liberibacterafricanus in non-rutaceous alternate host species from South Africa
title_full Candidatus liberibacterafricanus in non-rutaceous alternate host species from South Africa
title_fullStr Candidatus liberibacterafricanus in non-rutaceous alternate host species from South Africa
title_full_unstemmed Candidatus liberibacterafricanus in non-rutaceous alternate host species from South Africa
title_short Candidatus liberibacterafricanus in non-rutaceous alternate host species from South Africa
title_sort candidatus liberibacterafricanus in non rutaceous alternate host species from south africa
topic UCTD
url http://hdl.handle.net/2263/77829