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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
161

Fingerprinting of full and half-sib black wattle (Acacia mearnsii) progenies using Random Amplified Polymorphic DNA (RAPD).

Naguran, Riann. January 2005 (has links)
Black wattle (Acacia mearnsii), which belongs to the genus Acacia, is one of the many species of trees or hardwoods grown commercially in South Africa. Black wattle is a species indigenous to Australia and was introduced into South Africa by the van der Plank brothers in 1864. These trees are grown in South Africa because of its tannin-rich bark, the extract of which is used by the leather tanning industry. Black wattle is also grown for its timber, timber products and pulp. The introduction and cultivation history of black wattle suggests that the South African plantations contain limited genetic variation with relatedness amongst groups estimated to be high, thus implying a narrow genetic base in the South African black wattle population. In this investigation, Random Amplified Polymorphic DNA (RAPD) was used to estimate the genetic variation between seven different black wattle groups. A total number of 34 individuals obtained from different areas in South Africa were examined; Piet Retief (group 47 and 50: half-sibs), Kumbula (group 85: unrelated individuals), Howick (group 400: unrelated individuals) and an unknown area (groups 88, 89, 91: full-sibs). As this investigation was the first of its kind, a DNA isolation method as well as a PCR-RAPD protocol had to be modified. Total genomic DNA was successfully extracted using the CTAB DNA extraction method. This method removed large amounts of tannin present in the cells of the black wattle leaves and extracted high quality DNA to conduct between 50-100 RAPD reactions. The DNA purities ranged from 0.1 to 1.8, with an average of 1.46. A total of fourteen 10-mer RAPD primer sequences were randomly selected from the Operon Technologies primer list A, and tested in this investigation. Of the 14 primers used, only nine primers produced clear, single and repeatable bands. Therefore nine primers were selected for subsequent analyses. Ninety one loci that generated bands ranging from 300-3050 base pairs were produced. Seven to 13 loci per primer were generated. A total of 95.6 % of the loci were polymorphic. The overall expected mean heterozygosity (H = 0.3) obtained in this study was high in comparison to other studies conducted on acacias. The high levels of genetic variation were attributed to mating systems, dissortative mating and geographic distribution. The statistical packages POPGENE and ARLEQUIN were used to analyse the RAPD fingerprints. The genetic measures, Nei's diversity and Shannon's Information Index, showed that there was greater diversity exhibited (Nei's gene diversity = 32.09 % and Shannon's = 48.31 %), in the whole population than in each of the groups (with average of Nei's gene diversity = 20.33 % and Shannon's = 34.64 %). With regards to individual group analyses, low levels of genetic variation was obtained in group 400 (unrelated), from the Howick region, and group 85 (unrelated), from the Kumbula region, (mean 0.14 and 0.17 respectively). The low genetic values were attributed to limited gene exchange occurring in these two areas, bottlenecks and selection pressures. Groups 88, 89 and 91, from the unknown region (full-sib groups), were the most variable in comparison to the other groups, with means of (0.27,0.24 and 0.18 respectively). These high genetic variation values could be due to the fact that gene migration could have occurred between these groups and others in the area. It is thought that most acacias are insect-pollinated and this could have lead to gene migration between groups or populations, thereby explaining the high mean values. The gene flow obtained for the seven groups (FST = 0.174) indicated that great genetic differentiation existed in this population of black wattle studied. This value is higher in comparison to other woody species; however it is similar to other acacia species. UPGMA cluster analysis using Nei's unbiased genetic distance, revealed four distinct clusters of groups corresponding to the distribution areas represented in this study. The Howick (group 400: unrelated) and Kumbula (group 85: unrelated) were more closely related to each other than to the other groups, since both these groups are from Natal. The Piet Retief groups (groups 47 and 50: half-sibs), branched-off together, indicating that they are distinct from the other groups. The pairwise analysis of identity showed that the relationship between the group from Howick (group 400: unrelated) and all the other groups from the other regions was the lowest, ranging from 64 % to 79 %. The relationship between all the groups beside the group from Howick (group 400: unrelated) was reasonably high, ranging from 78 % to 90 %. This distance displayed by group 400 (unrelated) from Howick in relation to the groups, is attributed to the fact that it is frost resistant and the other groups not. Genetic variation was also detected and partitioned, between and within groups, by Analysis of Molecular Variance (AMQVA). Majority of the variation existed within groups (82.65 %) but significant differentiation was recorded between groups (17.44 %). This high level of within group differentiation may be explained by many aspects, such as the species breeding system, genetic drift or genetic isolation of groups or populations. The application of RAPD fingerprinting in black wattle has provided a more in depth understanding of the genetic variation residing in the South African population. The results achieved implementing this technique has shown that significant genetic variation exists within the black wattle population in South Africa. The results obtained in this study are also important since it is contrary to the expectation that the black wattle population in South Africa has low genetic variation. This knowledge is of great value to genetically discriminate between individuals or groups, to improve the selection of superior genotypes and allowing improved quality control in breeding programmes and seed orchard management. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2005.
162

A quantitative genetic analysis of the effect of crossbreeding on the growth rate of the South African abalone, Haliotis midae

Vorster, Gysbert 04 1900 (has links)
Thesis (MSc)--University of Stellenbosch, 2003. / ENGLISH ABSTRACT: The genetic status of H. midae broodstock in the South African aquaculture industry reflects that of random samples originating from undomesticated wild populations. The nature of growth in abalone is very slow, taking between three and four years to reach a marketable size of between 60 to 100 grams. It is therefore of paramount importance to improve this trait in order to ensure global competitiveness and economic viability within the industry. Improving this negative characteristic through conventional selection methods is a long-term venture and alternative means that would yield instantaneous results had to be considered. Crossbreeding was identified as an alternative, short-term strategy to improve growth rate. A crossbreeding experiment was performed between two populations of the abalone, Haliotis midae, from the East (E) and West (W) Coast of South Africa. This was done to investigate the occurrence of heterosis for growth among the crossbred genotypes (East x West, West x East). Fifteen males and females from both the East and the West Coast populations were mated in a complete dialelle crossbreeding experiment to produce four progeny groups (WW, EE, EW and WE). Progeny groups were evaluated for weight (bW) and length gain (bL) over a specific growth period of 9 months. The results provided no evidence of significant differences in weight (P = 0.085) or length gain (P = 0.244) between the four progeny groups, giving no indication of significant heterosis for weight and length gain among the crossbred progenies of these East and West Coast populations. It is recommended that further efforts to obtain improved growth rate in the abalone, Haliotis midae, through crossbreeding only be considered in light of clear evidence of substantial genetic differentiation between such populations. / AFRIKAANSE OPSOMMING: Die huidige status van perlemoen, soos dit voorkom in akwakultuur bedrywighede in Suid Afrika, weerspieël dié van ‘n ewekansige monster vanuit wilde, natuurlike populasies. Perlemoen is inherent ‘n stadig groeiende organisme wat tussen drie en vier jaar neem om tot ‘n bemarkbare grote van 60 tot 100 gram te groei. Dit is dus uiters noodsaaklik om hierdie eienskap te verbeter ten einde die bedryf ekonomies lewensvatbaar en mededingend op wêreld markte te maak. Konvensionele seleksie as ‘n metode om hierdie negatiewe eienskap te verbeter is ‘n langtermyn onderneming wat die identifisering van ‘n korttermyn metode, wat ondmiddellike resultate lewer, noodsaak. Kruisteelt is geïdentifiseer as geskikte korttermyn oplossing aangesien dit onmiddellike resultate lewer. ‘n Kruisteel eksperiment is uitgevoer tussen twee populasies van die perlemoen, Haliotis midae, van die Ooskus (E = East) en die Weskus (W = West) van Suid Afrika. Dit is gedoen om die omvang van heterose vir groeitempo in die gekruisde nageslag (East x West, West x East) te bepaal. Fyftien mannetjies en wyfies van beide die Oos- en Weskus populasies is met mekaar gepaar in ‘n volledige dialleel kruising om vier nageslag groepe (WW, EE, EW en WE) te vorm. Die nageslag is geëvalueer ten opsigte van massa (bW) en lengte (bL) toename oor ‘n spesifieke groei tydperk van 9 maande. Die eksperimentele resultate dui daarop dat die vier nageslag groepe nie betekenisvol van mekaar verskil het ten opsigte van massa (P = 0.085) en lengte (P = 0.244) toename nie en dat daar dus geen aanduiding van heterose vir massa en lengte toename in die nageslag van kruisings tussen die Ooskus en Weskus populasies bestaan nie. Daar word aanbeveel dat kruisteling as ‘n metode van genetiese verbetering van groeitempo in Haliotis midae slegs oorweeg word in die lig van nuwe molekulêre bewyse van genoegsame genetiese differensiasie tussen sulke populasies.
163

The development of an enzyme linked immunosorbent assay for the detection of the South African strain(s) of grapevine fanleaf nepovirus

Liebenberg, Annerie 12 1900 (has links)
Thesis (MSc (Genetics))--Stellenbosch University, 2008. / South Africa is one of the top ten wine producing countries in the world. The South African wine industry contributes approximately R16.3 billion to South Africa’s annual gross domestic product with 42.8% of wine being exported. To compete with the top wine producing countries and to ensure a viable export market, South Africa needs to ensure that healthy, virus free propagation material is produced and sold. One of the viruses that need to be tested for is Grapevine fanleaf virus (GFLV). Grapevine fanleaf virus causes degeneration and malformation of berries, leaves and canes and is responsible for significant economic losses by reducing crop yields by as much as 80%, reducing the longevity of the vines and affecting fruit quality. It is widespread in the Breede River Valley of the Western Cape where the nematode vector, Xiphinema index, is prevalent. The Breede River Valley contributes approximately 30% of the total production of the local wine industry, and severe losses in this region could threaten the viticulture. The Plant Improvement Act states that all propagation material sold must be tested for GFLV by a reputable scientific technique. The technique commonly used in South Africa is the Double Antibody Sandwich - Enzyme-linked Immunosorbent Assay (DAS-ELISA) and the kits are imported from Europe at a significant cost to the South African viticulture industry. The objective of this study was to produce a reliable and sensitive diagnostic assay specific for the South African strains of GFLV. This project aimed to develop and optimize a DAS-ELISA, by using recombinant DNA technology to produce antibodies against bacterially expressed viral coat protein. Total RNA was extracted from GFLV infected grapevine material and the viral coat protein (CP) amplified. The CP was cloned into the pGex-6P-2 expression vector, fusing a Glutathione STransferase (GST) partner to the viral coat protein enhancing solubility and protein purification. Insufficient amounts of the soluble protein were expressed and purified, preventing the production of antibodies and thus the development of the DAS-ELISA. An alternative diagnostic rapid-direct-one-tube-RT-PCR assay was developed. This rapid-directone- tube-RT-PCR assay was compared to commercially available DAS-ELISA and ImmunoStrip tests (Agdia) to assess the reliability, sensitivity and specificity of the rapid-direct-one-tube-RTPCR assay. Twelve GFLV isolates from South Africa were sequenced to investigate the variability between the isolates as well as the variability between the South African isolates and GFLV sequences available in Genbank. Sequence identities between clones from different GFLV isolates from South Africa were between 86-99% and 94-99% at the nucleotide and amino acid levels, respectively. Phylogenetic analysis based on the coat protein gene sequences showed that the South African isolates form two distinct clades or sub-populations. No significant correlation was found between geographical origin and symptoms, nor between geographical origin and sequence variability or between grapevine cultivar and symptom expression. Of the 23 samples tested with all three tests, 21 tested positive with rapid-direct-one-tube-RT-PCR, 19 with the ImmunoStrips and 17 with an imported DAS-ELISA kit (Agdia). Rapid-direct-one-tube-RT-PCR was found to be the most reliable technique for GFLV detection. Although the establishment of a DAS-ELISA directed to the South African strain(s) of GFLV was not successful, an alternative PCR based diagnostic system was developed, and proved to be sensitive and reliable. RT-PCR based diagnostic assays are generally accepted to be more sensitive than DAS-ELISA, but the latter is still used as the diagnostic assay of choice for routine testing due to ease of use. This rapid-direct-one-tube-RT-PCR assay is a rapid, sensitive and reliable diagnostic test, reducing the prevalence of false negatives, contributing to a virus free viticulture industry. The rapid-direct-one-tube-RT-PCR assay is as easy to use as DAS-ELISA, faster and can be performed by semi skilled workers, thus providing all the advantages associated with DAS-ELISA.
164

Molecular genetic study of wheat rusts affecting cereal production in the Western Cape

Le Maitre, Nicholas Carlyle 03 1900 (has links)
Thesis (MSc (Genetics))--University of Stellenbosch, 2010. / ENGLISH ABSTRACT: Microsatellites were used to differentiate Leaf (Puccinia triticina Eriks.) and Yellow rust (Puccinia striiformis Westend. f. sp. tritici Eriks.) pathotypes. There was sufficient diversity in the Leaf rust microsatellite markers to differentiate the pathotypes and create a phylogenetic tree of Leaf rust. Three of the microsatellite markers were sufficient to differentiate all the Leaf rust pathotypes. Sufficient diversity in the Yellow rust microsatellite markers was also observed which made it possible to differentiate the pathotypes. Only three pathotypes were used so no phylogenetic inference was made. Two microsatellite markers were sufficient to differentiate all the yellow rust pathotypes. Microsatellite and Amplified Fragment Length Polymorphisms (AFLP) markers were used to differentiate Stem rust (Puccinia graminis f. sp. tritici Eriks. and Henn.) pathotypes, and the data was combined for phylogenetic analysis. AFLP bands unique to each Stem rust pathotype were converted to Sequence Characterised Amplified Region (SCAR) markers. A single specific SCAR marker was created for UVPgt52. A second SCAR marker amplified four of the eight pathotypes. None of the other SCAR markers were specific. A 270 basepair fragment of the ITS1 region of the rDNA gene of all the Puccinia spp. was also sequenced in order to develop pathotype specific primers that could be used in a Real Time-PCR to determine relative levels of pathogen inoculum in a sample. Unfortunately insufficient diversity in the sequences of the ITS1 region of the rDNA gene did not allow unique primers to be designed for each pathotype making it impossible to proceed with the relative quantification using Real Time-PCR. Following marker development ninety one field isolates were collected from eleven sites in the Overberg and Swartland regions during 2008 and 2009. In the field isolates, four different Leaf rust pathotypes were identifiable. UVPgt13 and UVPgt10 were most prevalent. The most prevalent Stem rust pathotypes were UVPgt50, UVPgt52, UVPgt54 and UVPgt57. Only 6E16A- was identifiable in the Yellow rust isolates. There were no apparent patterns in the distribution of Leaf, Stem or Yellow rust. Leaf and Stem rust were widely distributed, while Yellow rust was confined to three sites in the central South Cape, the only sites where climatic conditions were favourable for its development during the sampling period. The low levels of diversity found in the rust population when compared to international populations are probably due to the relatively small population size, the lack of a host for sexual reproduction, the small sample size, the effective monoculture and the strong selective pressure created by artificial control methods. / AFRIKAANSE OPSOMMING: Mikrosatellietmerkers is gebruik om Blaar- (Puccinia triticina Eriks.) en Geelroes-( Puccinia striiformis Westend. f. sp. tritici Eriks.) patotipes te onderskei. Daar was genoeg diversiteit in die Blaarroesmerkers om verskillende patotipes te kon onderskei en om „n filogenetiese-boom te kon saamstel. Met drie van die mikrosatellietmerkers was dit moontlik om al die Blaarroespatotipes te kon onderskei. Daar was genoeg diversiteit in die Geelroesmerkers om al die patotipes te kon skei en met twee van die mikrosatellietmerkers kon al drie Geelroespatotipes van mekaar onderskei word. Mikrosatelliet- en ge-Amplifiseerde-Fragment-Lengte-Polimorfismes (AFLP) is gebruik om die Stamroes- (Puccinia graminis f. sp. tritici Eriks. and Henn.) patotipes te skei. AFLP-fragmente uniek aan „n spesifieke patotipe is omgeskakel na Volgorde-Spesifieke-ge-Amplifiseerde-Streek (SCAR) merkers. „n Spesifieke SCAR-merker is gemaak vir UVPgt52. „n Tweede SCAR-merker het vier van die patotipes geidentifiseer. Nie een van die ander SCAR-merkers was spesifiek t.o.v. „n spesifieke patotipe nie. Die volgorde van „n 270 basispaar fragment van die ITS1-streek van die rDNS-geen van al die Puccinia spp. is bepaal om patotipe spesifieke inleiers te kon ontwerp. Hierdie inleiers kan gebruik word om „n Intydse-Polimerase-Ketting-Reaksie (RT-PCR) te ontwerp om sodoende die relatiewe vlakke van die patogeen besmetting in „n monster te bepaal. Daar was nie genoeg diversiteit in die bepaalde volgordes om die spes1fieke inleiers te kon identifiseer nie en dus is RT-PCR laat vaar. Na die ontwikkeling van die merkers was een-en-negentig veldmonsters ingesamel afkomstig van elf lokaliteite in die Overberg en Swartland gedurende 2008 en 2009. Vier Blaarroespatotipes was uitkenbaar. Blaarroespatotipes UVPrt10 en UVPrt13 was die mees algemeenste. UVPgt50, UVPgt52, UVPgt54 en UVPgt57 was die mees algemene Stamroespatotipes. Net 6E16A- is geidentifiseer by die Geelroes-isolate. Daar was geen patroon in die verspreiding van Blaar-, Stam- of Geelroes patotipes. Blaar- en Stamroes was die wydste versprei, maar Geelroes het net by drie lokale in die sentrale Suid-Kaap voorgekom. Die lokaliteite is die enigste waar die weersomstandighede gunstig was vir Geelroes ontwikkeling gedurende die periode van monsterneming. Die lae vlakke van diversiteit wat in die roespopulasie gevind was is in teenstelling met internasionale populasies. Dit mag moontlik wees as gevolg van die relatief beperkte populasie grootte, die afwesigheid van „n gasheer vir seksuele voortplanting, die beperkte hoeveelheid monsters wat ingesamel is en die sterk selektiewe druk weens kunsmatige beheer.
165

The molecular characterization of South African isolates of Grapevine Rupestris Stem Pitting-associated virus (GRSPaV)

Noach, Liesl Christine 12 1900 (has links)
Thesis (MSc (Genetics))--University of Stellenbosch, 2010. / Includes bibliography. / ENGLISH ABSTRACT: The first aim of this study was to reliably and rapidly detect Grapevine rupestris stem pittingassociated virus (GRSPaV) in grapevine. This was achieved by screening 94 grapevines using crude plant extracts in both quantitative and conventional reverse transcription polymerase chain reaction (RT-PCR). The second aim was to establish a technique capable of differentiating GRSPaV sequence variants. The application of this technique is for the largescale screening of diseased vines to associate sequence variants of GRSPaV with disease symptoms. Nested quantitative polymerase chain reaction and high resolution melting assays (qPCR-HRM) were developed for three regions of the GRSPaV genome (coat protein, RNAdependant RNA-polymerase and triple gene block movement protein). The qPCR-HRM technique using the high saturation dye, EvaGreen™, and the Rotor-Gene™ 6000 analyzer was validated with a panel of sixteen sequence-characterized viral isolates. Diluted RT-PCR products and cloned cDNA gave the most consistent amplification plots and dissociation profiles. RT-PCR products generated from total RNA extracts were used as template for qPCR-HRM assays and for direct sequencing of sixteen samples in the three aforementioned regions. The average amplification efficiency for qPCR was 1.52±0.04. Auto-calling of userdefine genotypes was performed at a confidence interval of 70%. Phylogenetic analysis of the three regions of the GRSPaV genome was performed with published GenBank sequences to confirm the HRM data. The dominant sequence variants found in the South African sample set radiated with Group II, reference full-length variant GRSPaV-SG1. GRSPaV-infected samples can in future be subjected to qPCR-HRM assays developed during this study. This can be performed to establish similarity to known genotypes and therefore phylogenetic groups. Mixed infection of sequence variants and quasi-species were a common occurrence. The assay will be useful in establishing correlation of specific genotypes to different phenotypical expression of viral disease. This could provide insight into the etiology of diseases associated with GRSPaV. / AFRIKAANSE OPSOMMING: Die eerste doel van hierdie studie was om die virus wat met Rupestris-stamverpitting (Grapevine rupestris stem pitting-associated virus of “GRSPaV”) in wingerd verbind is, vinnig en betroubaar op te spoor. Dit is bereik deur 94 wingerdstokke vir die teenwoordigheid van die virus te toets met beide kwantitatiewe en konvensionele trutranskripsie polimerase kettingreaksies (RT - PCR) vanaf ongesuiwerde plant-ekstraksies. Die tweede doel was die daarstelling van ’n tegniek om onderskeid te tref tussen variante van GRSPaV met verskillende nukleotiedvolgordes. Hierdie tegniek kan op groot skaal gebruik word om ge-affekteerde wingerdstokke te toets om sodoende siektesimptome met spesifieke variante van GRSPaV te verbind. Ge-neste kwantitatiewe polimerase-kettingreaksies (qPCR) en hoë-resolusie smelt-analises (HRM) is ontwikkel vir drie streke van die GRSPaV-genoom (mantelproteïen, RNS-afhanklike RNS-polimerase en trippelgeenblok bewegingsproteïen). Die tegniek van qPCR-HRM met die hoë-versadingingskleurstof EvaGreen™ en die Rotor- Gene™ 6000 ontleder se geldigheid is bevestig deur vergelyking met ’n paneel van sestien virus-isolate waarvan die volgorde reeds bepaal is. Verdunde RT-PCR-produkte en gekloneerde DNS het die mees konsekwente amplifikasie-uitstipping en dissosiasieprofiele opgelewer. RT-PCR-produkte wat vanuit totale RNS-ekstrakte verkry is, is as templaat vir qPCR-HRM-analises gebruik. Dieselfde produkte is ook gebruik, om die volgorde van sestien monsters in drie streke direk te bepaal. Die gemiddelde amplifikasiedoeltreffendheid van die qPCR was 1.52±0.04. Gebruiker-gedefinieerde genotipes is deur middel van outooproeping teen ’n vertroue-interval van 70% uitgevoer. Filogenetiese analises vir drie streke van die GRSPaV-genoom is uitgevoer met gepubliseerde GenBank-volgordes om die HRMdata te bevestig. Die dominante volgorde-variante in die stel Suid-Afrikaanse monsters het ooreengestem met Groep II, vollengte-verwysingsvariant GRSPaV-SG1. Monsters wat met GRSPaV besmet is kan in die toekoms onderwerp word aan die qPCR-HRM-analises wat in hierdie studie ontwikkel is. Dit kan uitgevoer word om ooreenkomste met bekende genotipes te bepaal, en dus ook met filogenetiese groepe. Die besmetting van plante met meer as een volgorde-variant het algemeen voorgekom. Die kwasi-spesies populasie-struktuur van die virus het ook gedurig na vore gekom. Die toets sal nuttig wees in die bepaling van korrelasies tussen spesifieke genotipes en verskillende fenotipiese voorkomste van virussiektes. Dit kan insig verleen in die etiologie van siektes wat met GRSPaV verbind word.
166

Molecular investigation of genetic and environmental factors contributing to obesity in adolescent learners residing in the semi-urban/rural areas of the Western Cape Province, South Africa

Yako, Yandiswa Yolanda 12 1900 (has links)
Thesis (PhD)--Stellenbosch University, 2012. / Includes bibliography / ENGLISH ABSTRACT: Background/Aims: Obesity has increased rapidly in South African children and adolescents with significant variability observed among racial groups. Genes that regulate appetite have been studied in different populations worldwide, but their role in obesity among South African adolescents is unknown. The present study aimed at investigating the role of these genes, and their combined effect with physical activity in the development of obesity among South African adolescents. Methods: A total of 1564 South African school learners of Caucasian (n= 146), Mixed Ancestry (n= 872) and Black African (n= 537) ethnic groups were recruited for a research project that aimed to elucidate diabetes and the metabolic syndrome in children and adolescents attending schools in periurban areas of the Western Cape. The present case-control study included 227 obese-overweight (115 Black Africans and 112 Mixed Ancestry), and 204 normal weight (94 Black Africans and 110 Mixed Ancestry) adolescents learners. The learners were genotyped for nine polymorphisms (LEP: 19G>A, Lys36Arg, Val94Met; LEPR: Lys109Arg; Gln223Arg, Lys656Asn; CART: c.160-33G>A, c.499delA, and c.517A>G; GHRL: Leu72Met; and MC3R: Thr6Lys, Val81Ile) using allele-specific restriction enzyme analysis and automated sequencing. Genotype and haplotype associations with anthropometric variables such as body mass index (BMI), waist, hip, and mid-upper-arm circumferences (WC, HC, MUAC), and metabolic traits (fasting blood glucose, high density lipoproteincholesterol, total cholesterol), and blood pressure were further conducted. Furthermore, the type and frequency of physical activity was assessed by means of structured questionnaires; and its effect on obesity-related variables investigated in learners that were genotyped for the MC3R Thr6Lys and Val81Ile polymorphisms. Results: In a stepwise backward logistic regression analysis (containing age, gender, and LEP, LEPR, CART and GHRL polymorphisms), CART c.517A>G was independently significantly associated with obesity (OR= 5.98; 95%CI= 2.02, 21.27). CART c.517G carriers had higher MUAC (b coefficient= 1.88; 95%CI= 0.31, 3.44) while the LEPR 109Arg allele was significantly associated with decreased BMI (b coefficient = -2.36; 95%CI= -4.24, -0.47), WC (b coefficient = -5.66; 95%CI= -9.89, -1.44) and MUAC (b coefficient = -1.61; 95%CI= -3.00, -0.22); after adjusting for age, gender, and ethnicity. The haplotype containing the three LEP polymorphisms (A-A-A compared to the reference G-A-G haplotype) increased BMI (p= 0.0155), MUAC (p= 0.0146), and HC (p= 0.0128). The minor alleles of the MC3R polymorphisms decreased BMI, HC, WC, MUAC and TC; whilst only the Thr6Lys was associated with systolic and diastolic blood pressure (p= 0.0047 and 0.0027, respectively) in Mixed Ancestry learners. Doing house chores was associated with lower total cholesterol, independently and in the presence of the 81Ile allele (b coefficient = -0.355; 95%CI= 0.148, 0.561). Conclusion: To our knowledge, this is the first study that reports CART c.517A>G polymorphism as a risk factor for obesity in adolescents. Furthermore, the present study demonstrated that the MC3R polymorphisms had a positive effect on total cholesterol, which was further enhanced in physically active individuals. Similar to other studies, LEPR Lys109Arg and LEP polymorphisms were associated with variations in obesity-related variables among Black African and Mixed Ancestry South African learners. / AFRIKAANSE OPSOMMING: Agtergrond/Doelwitte: Vetsug het drasties toegeneem in Suid-Afrikaanse kinders en adelossente met ‘n beduidende variasie opgemerk tussen verskillende rassegroepe. Gene verantwoordelik vir regulering van eetlus is reeds wêreldwyd in verskillende bevolkingsgroepe bestudeer, maar hul rol in oorgewig Suid-Afrikaanse adolessente is onbekend. Die huidige studie was daarop gerig om ondersoek in te stel na die rol van hierdie gene en hul gekombineerde effek met fisiese aktiwiteit in die ontwikkeling van vetsug onder Suid-Afrikaanse adolessente. Metodes: ‘n Totaal van 1564 Suid-Afrikaanse leerders van Kaukasiese Afkoms (n=146), Gemengde Afkoms (n=872) en Swart Afkoms (n= 537) was gewerf in die navorsingsprojek wat ten doel gehad het om kinders en adolosente met diabetes en die metaboliese sindroom te identifiseer wat skole bygewoon het in semi-voorstedelike gebiede van die Wes-Kaap. Die huidige gevalle studie het 227 vetsugtige-oorgewig (115 Swart Afkoms en 110 Gemengde Afkoms) en 204 normale gewig (94 Swart Afkoms en 110 Gemengde Afkoms) leerders ingesluit. Die leerders was gegenotipeer vir nege polimorfismes (LEP: 19G>A, Lys36Arg, Val94Met; LEPR: Lys109Arg; Gln223Arg, Lys656Asn; CART: c.160-33G>A, c.499delA, and c.517A>G; GHRL: Leu72Met; and MC3R: Thr6Lys, Val81Ile) met die gebruik van alleel-spesifieke restriksie ensiem analises en geoutomatiseerde DNA volgorde bepalings tegnieke. Genotipiese en haplotipiese assosiasies met antropometriese veranderlikes soos liggaamsmassa indeks (BMI), middel-, heup- en mid-boarm omtrek (WC, HC, MUAC), metaboliese tendense (vastende bloed glukose, hoë-digtheid lipoproteïen-cholesterol, totale cholesterol) en bloeddruk was ook uitgevoer. Die tipe en frekwensie fisiese aktiwiteit was geassesseer deur middel van gestruktureerde vraelyste; en die uitwerking daarvan op vetsugverwante veranderlikes ondersoek in leerders wat vir die MC3R Thr6Lys en Val81Ile polimorfismes gegenotipeer was. Resultate: Statistiese ontleding (‘‘stepwise backward logistic regression analysis”), wat ouderdom, geslag en polimorfismes (LEP, LEPR, CART GHRL) ingesluit het, het getoon dat CART c.517A>G betekenisvol onafhanklik geassosiasieer was met vetsug (OR= 5.98; 95% CI= 2.02, 21.27). CART c.517G draers het ‘n hoër MUAC waarde gehad (b koeffisient = 1.88; 95%CI= 0.31, 3.44), terwyl die LEPR 109Arg alleel betekenisvol geassosieer was met verlaagde BMI ((b koeffisient = -2.36; 95%CI= -4.24, -0.47), WC (b koeffisient = -5.66; 95%CI= -9.89, -1.44) en MUAC (b koeffisient = -1.61; 95%CI= -3.00, -0.22) na die aanpassing van ouderdom, geslag en etnisiteit. Die haplotipe met die drie LEP polimorfismes (A-A-A teenoor die G-A-G verwysingshaplotipe) het die BMI (p= 0.0155), MUAC (p= 0.0146) en HC (p= 0.0128) verhoog. Die mindere allele van die MC3R polimorfismes het die BMI, HC, WC, MUAC en TC verlaag; terwyl slegs die Thr6Lys polymorfisme met sistolies en diastolies bloeddruk (p= 0.0047 en p= 0.0027, onderskeidelik) geassosieer was in Gemengde Afkoms leerders. Die verrigting van algemene huistake was geassosieer met laer totale kolesterol vlakke, onafhanklik en in die teenwoordigheid van die 81lle alleel (b koeffisient= -0.355; 95%CI= 0.148, 0.561). Gevolgtrekking: Na ons wete is hierdie die eerste studie wat die CART c.517A>G polimorfisme as ‘n risikofaktor vir vetsug in adolessente aantoon. Die huidige studie toon ook dat die MC3R polimorfisme ‘n positiewe effek op totale kolesterol gehad het, wat ook verder versterk was in fisiese aktiewe individue. Soortgelyk aan ander studies, was die LEPR Lys109Arg en LEP polimorfismes geassosieer met variasies in vetsug-verwante veranderlikes onder Suid-Afrikaanse Swart en Gemengde Afkoms leerders. / This research was supported by a grant from the University Research Fund of the Cape Peninsula University of Technology, Harry Crossley, University of Stellenbosch, Faculty of Health Sciences, Medical Research Council, and the National Health Laboratory Services, South Africa.

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