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In vitro propagation of Haemanthus pumilio and H. albiflos (Amaryllidaceae) and the population genetics of H. pumilio
Thesis (MScAgric)--Stellenbosch University, 2021.
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| Format: | Thesis |
| Language: | en_ZA |
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Stellenbosch : Stellenbosch University
2021
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| _version_ | 1867613767850786816 |
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| access_status_str | Open Access |
| author | Madzivire, Panashe Kundai |
| author2 | Hills, Paul N. |
| author_browse | Hills, Paul N. Madzivire, Panashe Kundai |
| author_facet | Hills, Paul N. Madzivire, Panashe Kundai |
| author_sort | Madzivire, Panashe Kundai |
| collection | Thesis |
| dc_rights_str_mv | Stellenbosch University |
| description | Thesis (MScAgric)--Stellenbosch University, 2021. |
| format | Thesis |
| id | oai:scholar.sun.ac.za:10019.1/110019 |
| institution | Stellenbosch University (South Africa) |
| language | en_ZA |
| last_indexed | 2026-06-10T12:41:23.238Z |
| license_str | Other — see source repository |
| provenance_str_mv | Harvested via OAI-PMH from SUNScholar — Stellenbosch University Repository |
| publishDate | 2021 |
| publishDateRange | 2021 |
| publishDateSort | 2021 |
| publisher | Stellenbosch : Stellenbosch University |
| publisherStr | Stellenbosch : Stellenbosch University |
| record_format | dspace |
| source_str | SUNScholar — Stellenbosch University Repository |
| spelling | oai:scholar.sun.ac.za:10019.1/110019 In vitro propagation of Haemanthus pumilio and H. albiflos (Amaryllidaceae) and the population genetics of H. pumilio Madzivire, Panashe Kundai Hills, Paul N. Dreyer, L. L. Stellenbosch University. Faculty of AgriSciences. Dept. of Genetics. Institute for Plant Biotechnology. Haemanthus pumilio Haemanthus albiflos Plant micropropagation Plant population genetics Phylogenetics Amaryllidaceae -- Africa, Southern -- Classification UCTD Thesis (MScAgric)--Stellenbosch University, 2021. ENGLISH ABSTRACT: Haemanthus albiflos and H. pumilio are members of the Amaryllidaceae. H. albiflos is a widespread evergreen plant, while H. pumilio is an endangered narrow endemic species with only two known populations remaining. These populations, remnants of the one recently transferred from Wellington to the Stellenbosch University Botanical gardens and another in the Duthie Reserve in Stellenbosch, present with vastly different morphologies. It is therefore vital to understand the phylogenetics and population genetics of H. pumilio as well as to design a method of in vitro propagation to increase the numbers of these plants. This study analysed the phylogenetics of H. pumilio using non-coding nuclear (Internal transcribed spacer), plastid (trnL-trnF intergenic spacer) and mitochondrial (nadi1477) gene regions and the population genetics using inter-simple sequence repeats (ISSR) and start codon targeted (SCoT) polymorphisms. The resulting phylogenetic trees revealed that H. pumilio is closely related to H. canaliculatus and H. sanguineus. The dendrograms from the DNA fingerprinting separated the two populations into distinct groups, while also revealing higher within population diversity than between them. While this is unexpected in rare narrow endemics, it might be explained by recent divergence from a more widespread species, H. sanguineus. These levels of genetic diversity are encouraging for species conservation as they suggest that intrapopulation breeding will not be detrimental to population survival. However, the two populations may become more homogenous as there exists a high level of gene flow between them. A micropropagation protocol designed using H. albiflos plants was able to successfully multiply individuals from leaf explants within 8 weeks, likely due to its ability to readily multiply by offsets. This protocol proved difficult to replicate on H. pumilio and only a single treatment with 0.1 mg/L BAP and 2 mg/L NAA differentiated after 13 weeks. However, the study was able to identify two fungal species contaminating H. pumilio cultures that could be endophytes. Future population genetics studies should aim to ascertain whether the translocation of plants from the Wellington population has increased the gene flow, thus subsequently reducing the interpopulation diversity. Additionally, this study provides crucial preliminary results on an optimum H. pumilio tissue culture method and is the first to identify possible endophytes of this species. Future micropropagation research should be conducted into providing an understanding of the plant-symbiont relationship to ascertain whether the symbionts confer necessary advantages to the plant that can be exploited to improve H. pumilio tissue culture. AFRKAANSE OPSOMMING: Haemanthus albiflos en H. pumilio is lede van die Amaryllidaceae. H. albiflos is ‘n wydverspreide, immergroen plant, terwyl H. pumilio ‘n bedreigde, nou-endemiese spesie is waarvan daar slegs twee populasies oor is waarvan ons weet. Hierdie populasies, waarvan oorblywende lede van een onlangs vanaf Wellington oorgeplaas is na die Stellenbosch Universiteit botaniese tuin en die ander in die Duthie reservaat in Stellenbosch, toon sterk morfologiese verskille. Dit is daarom van kardinale belang om die filogenie en populasie genetika van H. pumilio te verstaan, en om ‘n metode van in vitro vermeerdering te ontwikkel om die aantal individue van hierdie plante te vermeerder. Hierdie studie het die filogenie van H. pumilio bestudeer deur van die nie-koderende kern (interne transkriberende plekhouer (spacer)) en plastiede (trnL-trnF intergeen plekhouer en nadi1477) geenareas gebruik te maak, en die populasie genetika deur van inter-eenvoudige basisvolgorde herhalings (ISSR) en beginkodon geteikende polimorfismes (SCoT) gebruik te maak. Die resulterende filogenetiese boom het getoon dat H. pumilio naverwant is aan H. canaliculatus en H. sanguineus. Die dendrogramme van die DNS vingerafdrukke het die twee populasies as twee aparte groepe geskei, en ook aangedui dat daar groter interne variasie is as variasie tussen die twee. Alhoewel dit onverwag is vir skaars, nou-endemiese spesies, mag dit verduidelik word as onlangse diversifikasie vanaf ‘n eens meer wydverspreide spesie, H. sanguineus. Hierdie vlakke van genetiese diversiteit is bemoedigend vir die bewaring van die spesie, aangesien dit suggereer dat intra-populasie teling nie nadelig sal wees vir populasie oorlewing nie. Die twee populasies mag egter meer homogeen raak, aangesien daar ‘n hoë vlak van geenvloei tussen hulle is. ʼn Mikrovermeerdering protokol is ontwerp deur van H. albiflos plante gebruik te maak, en kon suksesvol binne 8 weke individue deur blaar-eksplante vermeerder, waarskynlik weens die vermoë van die plant om geredelik deur plantdele te vermeerder. Dit was moeilik om hierdie protokol op H. pumilio te herhaal, en slegs ʼn enkele behandeling met 0.1 mg/L BAP en 2 mg/L NAA het eers na 13 weke gedifferensieer. Hierdie studie kon egter twee fungi spesies identifiseer wat H. pumilio kulture kontamineer, en hulle mag endofiete verteenwoordig. Toekomstige populasie genetiese studies moet mik om te bepaal of die translokasie van die Wellington populasie geenvloei vermeerder het, en dus daarna die interpopulasie diversiteit verminder het. Verder verskaf hierdie studie noodsaaklike voorlopige resultate van ʼn optimum weefselkultuur metode, en dit is die eerste studie wat moontlike endofiete binne hierdie spesie geïdentifiseer het. Toekomstige verdere mikro-vermeerdering navorsing moet mik om ʼn beter begrip te verkry van die plant-simbiont verwantskap, om te bepaal of die simbiont nodige voordele aan die plant verskaf wat gebruik kan word om weefselkutuur tegnieke vir H. pumilio te verbeter. Masters 2021-03-07T13:07:00Z 2021-04-21T14:36:40Z 2021-03-07T13:07:00Z 2021-04-21T14:36:40Z 2021-04 Thesis http://hdl.handle.net/10019.1/110019 en_ZA Stellenbosch University xi, 88 pages : illustrations application/pdf Stellenbosch : Stellenbosch University |
| spellingShingle | Haemanthus pumilio Haemanthus albiflos Plant micropropagation Plant population genetics Phylogenetics Amaryllidaceae -- Africa, Southern -- Classification UCTD Madzivire, Panashe Kundai In vitro propagation of Haemanthus pumilio and H. albiflos (Amaryllidaceae) and the population genetics of H. pumilio |
| title | In vitro propagation of Haemanthus pumilio and H. albiflos (Amaryllidaceae) and the population genetics of H. pumilio |
| title_full | In vitro propagation of Haemanthus pumilio and H. albiflos (Amaryllidaceae) and the population genetics of H. pumilio |
| title_fullStr | In vitro propagation of Haemanthus pumilio and H. albiflos (Amaryllidaceae) and the population genetics of H. pumilio |
| title_full_unstemmed | In vitro propagation of Haemanthus pumilio and H. albiflos (Amaryllidaceae) and the population genetics of H. pumilio |
| title_short | In vitro propagation of Haemanthus pumilio and H. albiflos (Amaryllidaceae) and the population genetics of H. pumilio |
| title_sort | in vitro propagation of haemanthus pumilio and h albiflos amaryllidaceae and the population genetics of h pumilio |
| topic | Haemanthus pumilio Haemanthus albiflos Plant micropropagation Plant population genetics Phylogenetics Amaryllidaceae -- Africa, Southern -- Classification UCTD |
| url | http://hdl.handle.net/10019.1/110019 |
| work_keys_str_mv | AT madzivirepanashekundai invitropropagationofhaemanthuspumilioandhalbiflosamaryllidaceaeandthepopulationgeneticsofhpumilio |