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Taxonomy and symbiotic effectiveness of Mesorhizobium associated with chickpea

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

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Other Authors: Muema, Esther K.
Format: Thesis
Language:English
Published: University of Pretoria 2023
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access_status_str Open Access
author2 Muema, Esther K.
author_browse Muema, Esther K.
author_facet Muema, Esther K.
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 (Biotechnology))--University of Pretoria, 2023.
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institution University of Pretoria (South Africa)
language English
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provenance_str_mv Harvested via OAI-PMH from UPSpace — University of Pretoria Institutional Repository
publishDate 2023
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publisher University of Pretoria
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spelling oai:repository.up.ac.za:2263/90580 Taxonomy and symbiotic effectiveness of Mesorhizobium associated with chickpea Muema, Esther K. u21568830@tuks.co.za Venter, S.N. (Stephanus Nicolaas) Steenkamp, Emma Theodora Beukes, Chrizelle W. Wanjofu, Edwin Isava Mesorhizobium Chickpea Plant growth promotion Symbiosis Phylogenetics UCTD Dissertation (MSc (Biotechnology))--University of Pretoria, 2023. Chickpea (Cicer arietinum L.) is one of the most produced pulses worldwide, with India being the biggest producer. The production of chickpea is spread throughout the world from its native range in Southeast Turkey to Australia and North and Central America. It comprises two major types, Desi and Kabuli, of which the former is adapted to winter conditions, while the latter is adapted to summer conditions. The genetic differences among each type have been used as part of breeding and improvement efforts. Chickpea production in Africa is mostly limited to Northern and Eastern Africa. Sub-Saharan Africa, which has enough arable land for the potential production of chickpea, remains unexplored. In addition, to overcome the overreliance on synthetic fertilizers, the application of nitrogen fixers collectively called legume nodulating bacteria (LNB) is encouraged. For chickpea, LNB have been found to belong to the genus Mesorhizobium. Two identified species (Mesorhizobium ciceri, and Mesorhizobium mediterraneum) sharing similar symbiosis loci are mainly associated with this interaction. The first research chapter of this study focused on screening a geographically diverse collection of chickpea symbionts in terms of nodulation efficacy and extra plant growth promotion traits. This was achieved by performing a glasshouse pot experiment where 21 Mesorhizobium strains (i.e., LMG14989, CC1192, USDA3392, SEMIA396, DSM30133, DSM1978, XAP4, XAP10, XAP11, LMG15046, LMG17147, LMG17149, IC59, ICCV3110.1, P16.1, P13.3, P17.1, P17.2, P20.1, P1.2, and IC3110.2) were tested on six genotypes (i.e., Dark variety, Light variety, ICCV3110, ICCV3111, ICCV3203, and ICCV4105) of the Desi variety. Several vegetative growth parameters (i.e., nodule fresh weights, shoot dry weights, average plant heights and symbiotic effectiveness) were measured and subjected to statistical analyses of variance using R software. This was followed by determining the plant growth promotion traits of the Mesorhizobium strains. These included siderophores production, ACC deaminase production, indole acetic acid (IAA) production and phosphate solubilization. Statistically, the factors, ‘chickpea genotype’ and ‘Mesorhizobium strain,’ had significant effects on the measured parameters but lacked significant interactions based on the analysis of variance. The Light variety examined outperformed the other chickpea genotypes on all tested parameters. In general, inoculation with strains LMG15046, CC1192, XAP4, XAP10, and LMG14989 resulted in the best performance for all the tested parameters. Furthermore, all the strains were able to produce IAA and solubilize phosphate except the South African field isolates, which could not solubilize phosphate. However, none of the 21 strains produced siderophores or showed ACC deaminase activity. These findings showed that nodulation and growth promotion of the strains varied broadly based on the six genotypes used, while their ability to produce IAA and solubilize phosphate supports previous findings for Mesorhizobium species. Interestingly, however, this study revealed the differential ability to solubilize phosphate by chickpea Mesorhizobium since the South African isolates failed to solubilize phosphate, a trait that has always been associated with the Mesorhizobium genus. The work presented in this chapter was published in the journal Microorganisms [10(12):2467] under the title “Nodulation and growth promotion of chickpea by Mesorhizobium isolates from diverse sources”. The second research chapter focussed on the taxonomic affiliations of the LNB examined. For this purpose, five housekeeping genes were sequenced to identify known species as well as to delineate new species. These analyses revealed five clusters, of which three represented novel species. They were B (XAP11, LMG17147 and IC59), C (SEMIA396, and LMG17149) and D (ICCV3110.1, P16.1, P13.3, P17.1, P17.2, P20.1, P1.2, and IC3110.2). The existence of these taxa were supported by genome-based Average Nucleotide Identity (ANI) and digital DNA-DNA hybridization (dDDH) values. Although the evaluated phenotypic data contained little information to conclusively differentiate between them, the strains in cluster D had a unique and consistent pattern for some traits (e.g., growth temperatures, pH, salinity, heavy metals sensitivity and antibiotic resistance). Investigation of the origin of the strains' symbiotic loci (i.e., nodA, nodC and nifH) revealed that they shared high sequence similarity with those from known chickpea Mesorhizobium nodulators, thus corroborating previous work on this legume. In conclusion, this study revealed varied performance in nodulation and growth promotion among different Mesorhizobium isolates on different genotypes of chickpea used. Since this study was conducted under controlled conditions, field experiments are recommended to test the promising Mesorhizobium strains under South African environmental conditions. FLAIR Microbiology and Plant Pathology MSc (Biotechnology) Unrestricted 2023-05-08T09:10:33Z 2023-05-08T09:10:33Z 2023-09 2023 Dissertation * S2023 http://hdl.handle.net/2263/90580 DOI: https://doi.org/10.25403/UPresearchdata.22060430.v1 https://doi.org/10.25403/UPresearchdata.22060430 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 Mesorhizobium
Chickpea
Plant growth promotion
Symbiosis
Phylogenetics
UCTD
Taxonomy and symbiotic effectiveness of Mesorhizobium associated with chickpea
title Taxonomy and symbiotic effectiveness of Mesorhizobium associated with chickpea
title_full Taxonomy and symbiotic effectiveness of Mesorhizobium associated with chickpea
title_fullStr Taxonomy and symbiotic effectiveness of Mesorhizobium associated with chickpea
title_full_unstemmed Taxonomy and symbiotic effectiveness of Mesorhizobium associated with chickpea
title_short Taxonomy and symbiotic effectiveness of Mesorhizobium associated with chickpea
title_sort taxonomy and symbiotic effectiveness of mesorhizobium associated with chickpea
topic Mesorhizobium
Chickpea
Plant growth promotion
Symbiosis
Phylogenetics
UCTD
url http://hdl.handle.net/2263/90580
https://doi.org/10.25403/UPresearchdata.22060430