Genetic variation in Clivia miniata var. citrina

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Date
2006-06
Authors
Gagiano, Anthia
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Publisher
University of the Free State
Abstract
English: The genus Clivia is from the African tribe Haemanthaceae and a member of the family Amaryllidaceae. Clivia is endemic to southern Africa. Yellow Clivia are mutations of the orange-red standard forms that have appeared spontaneously in both wild and garden populations. Yellow Clivia plants are rare and desirable and were described as Clivia miniata var. citrina. Hobbyists from around the world trade in these ornamental plants initiating entire enterprises. Although the yellow form occurs naturally, many yellow clones have arisen through cultivation. Clones passed on from breeder to breeder have acquired different names. For directed breeding purposes in a thriving industry it is important to identify genetically similar plants. The aims of this study were to evaluate existing microsatellites for Clivia miniata var. citrina, to determine if AFLP analysis can distinguish among different plants within the genus Clivia and to determine genetic relatedness between different plants of ‘Vico’, ‘Giddy’ and ‘Natal Yellow’ cultivars. Previous studies done on Clivia include RAPD analysis and SSR analysis for Clivia. Work done in this study presents a first report of AFLP and SSR fingerprint analyses on C. miniata var. citrina. SSR fingerprint analysis revealed that the existing four SSR primer combinations were not applicable for studies on C. miniata var. citrina. AFLP analysis was optimised using a total of 28 EcoRI / MseI primer combinations. Primer combinations were evaluated using six randomly selected Clivia plants based on number of generated fragments, ability to score generated fragments, ability to detect polymorphism and level of polymorphic fragments. Fragments generated using EcoRI+3 primers in combination with Mse+4 primer combinations conformed to the chosen criteria. Primer combinations E-ACC with M-CATC, E-AGC with M-CATC and E-AGC with M-CTGG were selected for further studies on Clivia. AFLP analysis using three preselected primer combinations on 72 Clivia plants was successful in detecting genetic diversity and determining genetic relationships within closely related cultivated Clivia plants. Relatively high levels of genetic diversity (35%), as expected from known pedigree and species data, existed among Clivia plants. Genetic diversity within C. miniata and C. miniata var. citrina plants was high at 27%. Plants available for scrutiny were all genetically distinct. However, based on known pedigree data, names allocated to plants might not be truly representative of the true origin of the plants (e.g. Vico Meristem plants). Material obtained from different breeders could be distinguished at DNA level (e.g. ‘Giddy’ and ‘Natal Yellow’ cultivars). AFLP analysis revealed that different flower coloured plants (Apricot, Blush, Peach, Orange and Yellow) as well as plants from the same geographic areas were distributed together throughout the dendrogram with only a few of a certain colour grouping together. Known Group 1 Yellow and Group 2 Yellow were also present throughout the entire dendrogram, although the majority of known Group 1 Yellow plants grouped together. Clustering of the different species of the genus Clivia agreed with known pedigree data and hybrids included with their parents clustered according to known pedigree data. The phylogenetic relationships of natural populations of C. miniata indicated that all C. miniata plants shared a common ancestor. Clivia miniata from the same geographical area grouped together in the cladogram. More data would be required to prove these observations for all Clivia. Taxonomic status of the C. miniata var. citrina would depend on the monophyly of yellow Clivia plants. Orange flowered forms should be included to determine the validity of the current taxonomic status of these groups.
Afrikaans: Die genus Clivia is uit die tribes Haemanthaceae, deel van die familie Amaryllidaceae, en is endemies tot suider-Afrika. Geel Clivia blomme is ‘n kleurmutasie van die wilde oranje-rooi blomtipe wat spontaan binne beide natuurlike en gekultiveerde Clivia populasies verskyn. Die geel blomkleur is skaars en gesog en word as Clivia miniata var. citrina beskryf. Geel Clivia is ‘n gesogte rariteit wat deur geesdriftiges in ‘n handelsbedryf omskep is. Alhoewel die geel kleur in die natuur voorkom, is baie geelkleurige klone deur plantveredeling geskep en naamsverwarring het ontstaan aangesien elke teler ‘n plant na ‘n bekende persoon vernoem. Om spesifiek-gedrewe teling toe te pas is dit noodsaaklik om geneties identiese plante te identifiseer. Hierdie studie het ten doel gehad om bestaande mikrosatelliete op C. miniata var. citrina te toets, te bepaal of AFLP-analises tussen verskillende plante binne die genus Clivia kan onderskei en om te bepaal wat die genetiese verwantskappe tussen die verskillende ‘Vico’, ‘Giddy’ en ‘Natal Yellow’ kultivars is. Vorige werk wat op Clivia uitgevoer is, sluit RAPD- en SSR-analises in. Die huidige studie verteenwoordig die eerste verslag rakende die gebruik van AFLP- en SSRanalises op Clivia miniata var. citrina. SSR-vingerafdrukke het getoon dat die bestaande vier SSR-inleier kombinasies nie op C. miniata var. citrina werk nie. AFLP-analise is gestandaardiseer deur ‘n total van 28 EcoRI / MseI-voorvoerder kombinasies te gebruik. Die inleier kombinasies is op ses willekeurig geselekteerde Clivia plante getoets en is geëvalueer op grond van die aantal gegenereerde fragmente, vermoë om fragmente te dokumenteer, potensiaal om polimorfismes te erken asook aantal polimorfismes. Fragmente verkry deur van EcoRI+3 inleier in kombinasie met Mse+4 inleier gebruik te maak het bogenoemde kriteria die beste gepas. Inleier kombinasies E-ACC met M-CATC, E-AGC met M-CATC en E-AGC met M-CTGG is gekies vir verdere studies op Clivia. AFLP-analise met behulp van die bogenoemde inleier kombinasies is suksesvol op die hele steekproef van 72 Clivia plante toegepas en is suksesvol aangewend vir die bepaling van genetiese diversiteit asook genetiese verwantskappe tussen naverwante veredelde Clivia plante. Relatief hoë vlakke van genetiese diversiteit (35%), soos verwag gebaseer op bekende stamboom en spesiedata, is tussen lede van C. miniata en die ander Clivia species en tussen C. miniata en C. miniata var. citrina (27%) waargeneem. Al die plante binne die steekproef kon geneties van mekaar onderskei word. Gebaseer op bekende stamboomdata is gevind dat alle naam toevoegings nie in lyn met beskikbare stamboom inligting was nie, byvoorbeeld die Vico Meristeem plante. Materiaal wat van verskillende telers ontvang is kon op DNA-vlak onderskei word (bv. die ‘Giddy’ en ‘Natal Yellow’ kultivars). AFLP-analises het aangetoon dat verskillende blomkleur plante (Appelkoos, Ligpienk, Perskekleurig, Oranje en Geel) asook plante afkomstig van dieselfde geografiese area nie afsonderlik binne die dendrogramme gegroepeer het nie. Slegs ‘n paar plante het volgens kleur gegroepeer. Bekende Groep 1 Geel en Groep 2 Geel plante was deurgaans binne die dendrogram versprei, alhoewel die meeste Groep 1 Geel plante saam gegroepeer het. Groepering van die verskillende spesies in die genus Clivia het met beskikbare stamboomdata ooreengestem en basters met hul ouers het volgens stamboomdata saamgegroepeer. Die filogenetiese verwantskappe tussen natuurlike populasies van C. miniata het getoon dat alle C. miniata plante van ‘n gemeenskaplike voorouer afkomstig is. Clivia miniata plante afkomstig van dieselfde geografiese areas het saam gegroepeer in die cladogram. Meer inligting word benodig om hierdie bevinding toepaslik tot alle Clivia te kan maak. Die taksonomiese stand van C. miniata var. citrina berus op die monofelie van geel blomkleurige Clivia plante. Plante met oranje blomme behoort in toekomstige studies by geelkleuriges ingesluit te word om die taksonomiese stand van C. miniata var. citrina beter te verklaar.
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Keywords
Dissertation (M.Sc. (Plant Sciences (Genetics))--University of the Free State, 2006, Clivia -- South Africa -- Genetics, Clivia -- Classification, AFLP, Fingerprinting, Clivia miniata var
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