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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.
191

The graminaceous rusts and smuts of Kansas

Haard, Richard Thomas. January 1963 (has links)
Call number: LD2668 .T4 1963 H32 / Master of Science
192

Environmental effects on Peronospora trifoliorum oospore production in seedlings of two alfalfa clones ; Attempts to germinate Peronospora trifoliorum oospores

Hodgden, L. D. January 1978 (has links)
Call number: LD2668 .T4 1978 H63 / Master of Science
193

Lignin as a mechanism of field resistance to Phytophthora rot in soybeans

Curry, Joseph Timothy. January 1984 (has links)
Call number: LD2668 .T4 1984 C87 / Master of Science
194

Some post-harvest physiological studies of potatoes and relation of some potato cultivars to incidence of internal brown spot

Lafta, Abbas Mubadar. January 1985 (has links)
Call number: LD2668 .T4 1985 L33 / Master of Science
195

A comparison of techniques for screening for resistance to the chinch bug, Blissus leucopterus leucopterus (Say), in sorghum

Meehan, Mitchell Elwin. January 1985 (has links)
Call number: LD2668 .T4 1985 M43 / Master of Science
196

Breeding for Smut Resistance in Arizona-Grown Wheat

Bryan, W. E. 15 March 1937 (has links)
This item was digitized as part of the Million Books Project led by Carnegie Mellon University and supported by grants from the National Science Foundation (NSF). Cornell University coordinated the participation of land-grant and agricultural libraries in providing historical agricultural information for the digitization project; the University of Arizona Libraries, the College of Agriculture and Life Sciences, and the Office of Arid Lands Studies collaborated in the selection and provision of material for the digitization project.
197

Assessment of inoculation techniques to evalute apple resistance to Phytophthora cactorum

Zondo, Patience Thembelihle 03 1900 (has links)
Thesis (MSc)--University of Stellenbosch, 2001. / ENGLISH ABSTRACT: Phytophthora cactorum (Lebert & Cohn) Schrot. is the primary cause of crown, collar and root rot diseases of apple (Malus domestica Borkh.) trees worldwide. This pathogen is most destructive in commercial apple orchards under waterlogged soil conditions and has recently been identified as causing serious disease in some South African apple orchards. Crown, collar and root diseases are difficult to control because of their unpredictability and catastrophic nature. The use of resistant cultivars and rootstocks is economical and environmentally considerate. Therefore the need to develop screening techniques that will enable the selection of desirable disease resistant traits as part of an apple-breeding program in South Africa was identified. The work undertaken in this study was aimed at optimizing different techniques to test resistance. Using two direct inoculation techniques (excised stem and intact stem) the aggressiveness of lO isolates of P. cactorum on apple rootstocks was determined. The susceptibilities of five apple rootstocks were also compared. Results have shown isolate by rootstock interaction which means isolate aggressiveness was influenced by rootstocks tested. The selectivity of isolates suggests that there may be several strains of the pathogen. Population studies of the pathogen might contribute valuable information that could lead to better interpretation of results. Rootstock susceptibility was monitored in vitro throughout the season by inoculating at monthly intervals for 26-months. It was observed that during winter, rootstock susceptibility was low compared to high susceptibility during summer. These results have revealed new information regarding changes in the relative resistance of the different rootstocks over the growing season, e.g. the susceptibility pattern of rootstock MMl06 occurred 1 to -2 months later than that of other rootstocks. This finding has important implications on the way in which resistance test results are interpreted, and emphasizes the importance of not relying on point sampling. Furthermore, useful information has been acquired regarding the epidemiology of the disease with regard to "windows of susceptibility". The phenomenon of a phase shift in susceptibility of different rootstocks needs to be tested on a broader scale to assess whether it has any practical application on resistance testing. Although different inoculation techniques are applied in breeding programs, up to now there is no consensus on which technique works best for seedling selections. Since large numbers of individuals must be tested to improve the chances of detecting resistant genotypes, mass inoculations of young seedlings is a rapid way of identifying resistant individuals. Two different screening methods were tested during this study. Using the sand-bran technique, seedlings were transplanted onto inoculated soil and the root mass was used as a measure of resistance. In a second method zoospore inoculum was applied to seedlings growing in a sand:bark mixture at different concentrations and the seedlings were subjected either to water drenching or not. In both trials the aggressiveness of isolates differed significantly from each other and only higher inoculum concentrations were effective in causing disease. The age of seedlings used in tests emerged as an important factor. Seedlings under five-months-old should not be used. Drenching inoculated seedlings enhanced disease development but the production of sufficiently high numbers of zoospores was a laborious task. Thus, it is recommended that the sand-bran inoculum technique be tested with the drenching treatment for mass selection. In conclusion this study confirms the importance of both choice of isolate and choice of inoculation intervals in determining susceptibility of rootstocks to infection. In spite of the fact that stem inoculation bioassays have limited resemblance to natural disease situations, these bioassays are useful for obtaining an indication as to whether genotypes have a degree of resistance and merit further testing. For this reason refinement of the stem inoculation bioassay is worthwhile pursuing. With regard to seedling trials, both the sand-bran and the zoospore technique appear promising but refinement of these techniques is necessary in order to present a more practical way of testing large volumes of seedlings. / AFRIKAANSE OPSOMMING: Evaluering van inokulasietegnieke om weerstand teen Phytophthora cactorum in appels te evalueer: Phytophthora cactorum (Lebert & Cohn) Schrot. is die primêre oorsaak van kroon-, kraag en wortelvrot van appelbome (Malus domestica Borkh.). Dit is die mees verwoestende patogeen in kommersiële appelboorde waar daar versuipte toestande grond voorkom. P. cactorum is onlangs identifiseer as die patogeen wat ernstige kroon- en kraag-verotting in Suid Afrikaanse appelboorde veroorsaak. Kroon-, kraag- en wortelvrot is moeilik om te beheer as gevolg van die onvoorspelbaarheid en rampspoedige aard van die siekte. Die gebruik van kultivars en onderstamme wat weerstandbiedend is teen siektes en plae is omgewingsvriendelik en is ekonomies van belang, dus het die behoefte ontstaan om inokulasietegnieke te ontwikkelom weerstandige saailinge te identifiseer en te selekteer as deel van 'n appelteelprogram in Suid Afrika. Die doelwit van hierdie studie is om verskillende inokulasietegnieke te toets en te verfyn om weerstand in appelsaailinge te identifiseer. Deur gebruik te maak van twee inokulasietegnieke (die afgesnyde loot- en intakte loot tegniek), is die relatiewe aggressiwiteit van 10 isolate van P. cactorum en die vatbaarheid van vyf appelonderstamme ondersoek. Resultate het aangetoon dat die aggressiwiteit van die isolate gevarieer het na aanleiding van die onderstam wat getoets is. Die selektiwiteit van die isolate is 'n aanduiding dat daar moontlik verskeie rasse van die patogeen voorkom. Toekomstige studies op die populasiestruktuur van P. cactorum sal 'n belangrike bydrae maak tot die interpretasie van resultate oor weerstand en weerstandsteling. Die vatbaarheid van onderstamme was ook in in vitro proewe ondersoek deur maandelikse inokulasies toe te pas oor 'n tydperk van 26 maande. Dit is opgemerk dat die onderstamvatbaarheid gedurende die winter laag was in vergelyking met die somer. Nie al die onderstamme het dieselfe gereageer gedurende verskillende toetstye nie. Hierdie resultate toon aan dat die relatiewe weerstand van verskillende onderstamme oor die groeiseisoen verskil, byvoorbeeld die vatbare reaksie van die onderstam 'l\.1MI06' het een tot twee maande later voorgekom in vergelyking met ander onderstamme wat getoets is. Hierdie bevinding het belangrike implikasies op die interpretasie van weerstandstoetsing en beklemtoon die moontlike tekortkominge in enkelproefwaarnemings. Bruikbare inligting ten opsigte van die epidemiologie van die siekte is versamel wat beskryf kan word in terme van vensters van vatbaarheid wat verskil van onderstam tot onderstam. Verdere ondersoeke in die verband word aanbeveel. Hoewel verskeie inokulasietegnieke bestaan om jong saailinge vir weerstand te toets, is daar tot op hierdie stadium nog nie ooreenstemming oor die beste tegniek wat toegepas moet word om saailingseleksie te doen nie. Omdat groot getalle saailinge getoets moet tydens die seleksieproses sal massa-inokulasie van saailinge die aangewese metode wees. Twee verskillende inokulasie tegnieke is getoets in die studie. Deur gebruik te maak van die sandsemel tegniek, is saailinge geplant in geinfesteerde plantmedium, waartydens die wortelmassa van saailinge gebruik is om die reaksie op infeksie te kwantifiseer. Die soëspoor inokulasietegniek was toegepas op saailinge wat in 'n sand en basmengsel geplant is teen verskillende inokulurnkonsentrasies. 'n Waterverdrenkingsbehandeling is ook getoets. In albei hierdie proewe het die aggressiwiteit van die isolate van mekaar verskil. Slegs die hoër inokulumkonsentrasies was effektief in die ontwikkeling van die siekte. Die ouderdom van saailinge is ook uitgewys as 'n belangrike faktor wat 'n rol speel in weerstandstoetsing. Saailinge jonger as 5 maande word nie aanbeveel vir hierdie toetse nie. Verdrenking van saailinge het die voorkoms van die siekte verhoog, maar die produksie van groot getalle soëspore was 'n beperkende faktor in die uitvoering van die proef Dit word aanbeveel dat die sand-semel inokulasietegniek verder evalueer moet word onder verskeie toestande, onder andere deur dit met verdrenkinghte kombineer. Die belang van die keuse van isolaat en inokulasiedatum in bepaling van relatiewe weerstand van onderstamme teen P. cactorum is tydens die studie bevestig. Afgesien van die beperking van die staminokulasietegnieke in soverre dit verwyderd is van natuurlike infeksie, word die tegnieke aanbeveel om 'n indikasie te kry van die relatiewe weerstand van onderstamme. Beide die sand-semel en soëspoor tegnieke kan gebruik word om weerstandige saailinge te identifiseer, maar tegniese verfyning van hierdie tegnieke is nodig om saailinge in massa te evalueer.
198

Invloed van doppenetrasieweerstand op die oesstadium van druiwe

Van Dyk, B. W. (Burger Wynand) January 1992 (has links)
Thesis (MScAgric)--Stellenbosch University, 1992. / One microfiche copy / ENGLISH ABSTRACT: The possibility of harvesting grapes at an earlier stage of maturity, based on differences in glucose and fructose concentration which influence the sweetness of grapes, was investigated. Although differences between cultivars were found the extent was not such that a specific cultivar could be selected in order to harvest at a lower sugar concentration, but with the same sweetness. Certain characteristics of table and wine grape cultivars with respect to anatomical composition and skin penetration resistance (SPR) were also investigated in order to ascertain the extent to which grapes would resist external damage, and to what extent turgor and skin thickness contributed to SPR. Daily variances in SPR confirm that not only skin strength, but also the turgor of the grape berry contributed to SPR. Skin penetration resistance seems to be a good criterion of the extent to which cultivars would resist external damage, because it is based on the toughness of the skin and the turgor of the berry. / AFRIKAANSE OPSOMMING: Die moontlikheid van vroeer oes op grond van verskille in die glukose- en fruktosekonsentrasie wat 'n invloed op die soetheid van druiwe mag he, is ondersoek. Daar is gevind dat die verskille wat tussen cultivars voorkom nie van so 'n grootte-orde is dat 'n spesifieke cultivar geselekteer kan word ten einde by 'n laer totale suiker, maar by dieselfde soetheidsgraad, te kan oes nie. Verder is sekere eienskappe van tafel- en wyndruifcultivars t.o.v. anatomiese samestelling en doppenetrasieweerstand (DP\V) ondersoek om die moontlike weerstand teen eksterne beserings en die mate waartoe turgor en dopdikte 'n invloed daarop mag uitoefen, _vas te stel. Daaglikse variasie in DPW het bevestig dat die DPW nie alleen afhanklik is van dopsterkte nie, maar ook van die turgor van die korrel. Doppenetrasieweerstand blyk 'n goeie maatstaf te wees vir die mate waartoe cultivars weerstand hied teen sekere eksterne beserings omdat dit gebaseer is op dopsterkte en turgor van die korrel.
199

Isolation and characterisation of a polygalacturonase-inhibiting protein (PGIP) and its encoding gene from Vitis vinifera L.

De Ascensao, Ana 12 1900 (has links)
Thesis (PhD)--Stellenbosch University, 2001. / ENGLISH ABSTRACT: Polygalacturonase-inhibiting proteins (PGIPs) are present in the cell walls of a variety of plant species. These proteins have been shown to specifically inhibit endopolygalacturonases (endo-PGs) secreted by invading fungal pathogens as part of the induced disease resistance mechanism of plants. This is the first report on the isolation and characterisation of a pgip gene from Vitis vinifera L., designated grapevine pgip1. A single open reading frame encoding a deduced polypeptide of 333 amino acids with a predicted molecular mass of 37.1 kOa and a calculated isoelectric point of 8.61 was identified from a 5.6 kb subgenomic fragment of V. vinifera cv Pinotage. Nucleotide and derived amino acid sequence analysis of grapevine pgip1 showed significant homology with other characterised PGIP encoding genes and revealed features characteristic of PGIPs found in several other plant families. Genomic DNA analysis showed that grapevine pgip1 belongs to a small multigene family in Vitis cultivars. From Northern blot analysis it was evident that expression of the PGIP family is both tissue- and developmental stage specific. The grapevine pgip1 was transiently expressed in Nicotiana benthamiana L. with potato virus X (PVX) as a vector. Grapevine PGIP1 isolated from crude protein extracts of PVX-infected N. benthamiana were tested and showed inhibitory activity against polygalacturonases (PGs) from Botrytis cinerea. Grapevine PGIPs have not previously been purified and characterised. Molecular analyses have confirmed that PGIPs are typically encoded by multigene families and that the inhibitor specificities and kinetics of the isolated proteins differ within and among species. In this study, two PGIP isomers from V. vinifera berries were isolated. The one isomer, designated PGIP-A, was partially purified and had a molecular mass of 39 kOa, whereas the other PGIP, designated PGIP-B, was purified and had a molecular mass of 42 kOa as determined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SOS-PAGE) and Western blot analysis. Both proteins were cell wall-bound. Enzymatic deglycosylation confirmed that PGIP-B is a glycosylated protein. Grapevine PGIP-A showed strong inhibitory activity against a homogeneous PG from Aspergillus niger and to a lesser extent against PG from Fusarium moniliforme, but was unable to interact with a crude PG preparation from B. cinerea. Grapevine PGIP-B was able to strongly inhibit PGs from B. cinerea as well as from Colletotrichum gleosporoides, yet showed no inhibition towards PG from A. niger. The grapevine pgip1 gene was expressed under the control of the Cauliflower mosaic virus (CaMV) 35S promoter in tobacco plants via Agrobacterium tumefaciensmediated transformation. Transgenic tobacco plants expressing the grapevine PGIP (gPGIP1) were used to demonstrate the effectiveness of this inhibitor against fungal PGs and to investigate whether gPGIP1 influences disease development. Northern blot analysis identified 19 transgenic plants expressing pgip1 transcript levels. Crude PGIP extracts from the transgenic tobacco plants inhibited PGs from B. cinerea and C. gleosporoides, but not PG from A. niger. Leaves from untransformed tobacco plants, from transgenic tobacco lines showing high and low PG inhibition, and from transgenic plants that did not express pgip1, were inoculated with B. cinerea. Transgenic leaves showed a reduction in the size of necrotic lesions of macerated tissues of approximately 45% relative to control and non-expressing transgenic leaves. The results from the heterologous expression of gPGIP1, together with the results from the protein purifications and inhibition studies, indicate that the isolated grapevine pgip1 gene encodes the isolated PGIP-B isomer. This work has ; established a good model system to study certain aspects of plant-pathogen interactions in grapevine. Heterologous expression of gPGIP1 has demonstrated that PGIP inhibition of fungal PGs slows disease development of B. cinerea in planta. / AFRIKAANSE OPSOMMING: Sien volteks vir opsomming
200

The development and characterisation of grapevine virus-based expression vectors

Du Preez, Jacques 03 1900 (has links)
Thesis (PhD (Genetics))--University of Stellenbosch, 2010. / ENGLISH ABSTRACT: Grapevine (Vitis vinifera L.) is a very important agricultural commodity that needs to be protected. To achieve this several in vivo tools are needed for the study of this crop and the pathogens that infect it. Recently the grapevine genome has been sequenced and the next important step will be gene annotation and function using these in vivo tools. In this study the use of Grapevine virus A (GVA), genus Vitivirus, family Flexiviridae, as transient expression and VIGS vector for heterologous protein expression and functional genomics in Nicotiana benthamiana and V. vinifera were evaluated. Full-length genomic sequences of three South African variants of the virus (GTR1-1, GTG11-1 and GTR1-2) were generated and used in a molecular sequence comparison study. Results confirmed the separation of GVA variants into three groups, with group III (mild variants) being the most distantly related. It showed the high molecular heterogeneity of the virus and that ORF 2 was the most diverse. The GVA variants GTG11-1, GTR1-2 and GTR1-1 were placed in molecular groups I, II and III respectively. A collaboration study investigating the molecular divergence of GVA variants linked to Shiraz disease (SD), described two interesting GVA variants of group II, namely GTR1-2 and P163-M5 (Goszczynski et al., 2008). The group II variants were found to be closely linked to the expression of SD. GTR1-2 was isolated from a susceptible grapevine plant that never showed SD symptoms (Goszczynski 2007). The P163-M5 variant that resulted in exceedingly severe symptoms in N. benthamiana and is that used as SD positive control by the grapevine industry, was found to contain a 119 nt insert within the native ORF2. Comparative analysis performed on the complete nt and aa sequences of group II GVA variants suggested that the components in the GVA genome that cause pathogenicity in V. vinifera are more complex (or different) to those that cause pathogenicity in N. benthamiana. The three South African variants (GTR1-1, GTG11-1 and GTR1-2) were assembled into fulllength cDNA clones under control of CaMV 35S promoters. After several strategies were attempted, including a population cloning strategy for GTR1-2, none of the clones generated were able to replicate in N. benthamiana plants. A single amino acid substitution at position 13 (Tyr/Y Cys/C) in ORF 5 of the GTR1-2 cDNA clone was shown to abolish or reduce replication of the virus to below a detectable level. Two infectious clones of Israeli variants of GVA (T7-GVA-GR5 and T7-GVA118, obtained from M. Mawassi) were brought under control of a CaMV 35S promoter (35S-GVA-GR5 and 35S-GVA118). Both clones were infectious, able to replicate, move systemically and induce typical GVA symptoms after agroinfiltration in N. benthamiana. These Israeli clones served as backbone for further experiments in characterisation of transient expression and VIGS vectors. The use of GVA as gene insertion vector (35S-GVA118) and gene exchange vector (35S-GVA-GR5- ORF2+sgMP) in N. benthamiana and V. vinifera was compared. The gene insertion vector, 35S-GVA118 was based on the full-length GVA genome. The gene exchange vector, 35SGVA- GR5- ORF2+sgMP, was constructed in this study by elimination of ORF 2 and insertion of a sgMP and unique restriction sites to facilitate transgene insertion. In N. benthamiana both vectors showed similar GUS expression levels and photobleaching symptoms upon virus-induced NbPDS silencing. In V. vinifera limited GUS expression levels and VIGS photobleaching symptoms were observed for the gene insertion vector, 35SGVA118. No GUS expression was observed for the gene exchange vector 35S-GVA-GR5- ORF2+sgMP in this host. As for silencing, one plant, agroinfiltrated with 35S-GVA-GR5- ORF2-VvPDS+sgMP, developed photobleaching symptoms in 3 systemic infected leaves after 4 months. This study showed that GVA can be used as gene insertion and gene exchange vector for expression and VIGS in N. benthamiana, but in grapevine its use is limited to expression and silencing of genes in the phloem tissue. It is also the first report that ORF 2 of GVA is not needed for long distance movement in grapevine. To investigate the possible role of the P163-M5 119 nt insertion and the GVA ORF 2 (of unknown function), in expression of symptoms in plants, ORF 2 of a 35S-GVA-GR5 cDNA clone was removed and subsequently substituted by the corresponding ORFs of four South African GVA variants. Upon agro-infiltration into N. benthamiana leaves, all chimaeric GVA constructs were able to move systemically through the plant. At this stage no correlation could be found between severity of symptoms, the presence of the P163-M5 insert and the specific GVA ORF 2 present in the chimaeras, indicating that other factors in the viral genome or the host plant probably play a crucial role. This study contributed to the pool of available in vivo tools for study and improvement of the valuable grapevine crop. It also opened several exciting research avenues to pursue in the near future. / AFRIKAANSE OPSOMMING: Wingerd (Vitis vinifera L.) is ‘n baie belangrike landboukundige gewas wat beskerm moet word. Om die rede word verskeie in vivo gereedskap vir die bestudering van die wingerdplant, en die patogene wat dit infekteer benodig. Die wingerd genoom se volgorde is bepaal en dus is die volgende logiese stap om die gene te annoteer en funksie daaraan toe te skryf. In hierdie studie is die gebruik van Grapevine virus A (GVA), genus Vitivirus, familie Flexiviridae, as tydelike uitdrukking- en virus-geinduseerde geenuitdowingsvektor vir heteroloë proteïen uitdrukking en funksionele genoomstudies in Nicotiana benthamiana en V. Vinifera getoets. Vollengte genoomvolgordes van drie Suid-Afrikaanse variante van die virus (GTR1-1, GTG11-1 en GTR1-2) is gegenereer en in ‘n molekulêre volgorde vergelyking studie gebruik. Resultate het die verdeling van GVA variante in drie groepe, waar groep III die verste verwant is, bevestig. Dit het ook gewys dat die virus ‘n baie hoë molekulêre heterogeniteit het en dat oopleesraam 2 (ORF 2) die mees divers is. ‘n Samewerking studie waar die molekulêre diversiteit van GVA variante, gekoppel aan Shiraz siekte (SD), ondersoek is, is twee interessante variante van groep II beskryf, naamlik GTR1-2 en P163-M5 (Goszczynski et al., 2008). Groep II variante is vooraf gevind om nou verwant te wees aan die ontwikkeling van SD in wingerd. Die GTR1-2 variant is uit ’n vatbare wingerd plant, wat nooit SD-simptome vertoon het nie, geïsoleer (Goszczynski et al., 2007). In die ORF 2 van die P163-M5 variant, wat simptome van die ergste graad in N. benthamiana geïnduseer het, en ook deur die industrie as betroubare SD-positiewe kontrole gebruik word, is ’n 119 nt invoeging gevind. Die vergelykende analise wat uitgevoer is, het daarop gedui dat die determinante van patogenisiteit in die GVA genoom moontlik meer kompleks kan wees in V. vinifera as in N. benthamiana. Die drie Suid-Afrikaanse variante (GTR1-1, GTG11-1 en GTR1-2) is in afsonderlike vollengte cDNA klone, onder beheer van CaMV 35S promotors, aanmekaargesit. Nadat verskeie kloneringstrategieë, insluitend ’n populasie kloneringstrategie vir die GTR1-2 kloon, gebruik is, het geen een van die cDNA klone die vermoë besit om in N. benthamiana te repliseer nie. ’n Enkele aminosuur substitusie in posisie 13 (Tyr/Y Cys/C) in ORF 5 van die GTR1-2 kloon, het die replisering van die virus tot laer as ’n opspoorbare vlak verlaag. Twee infektiewe klone van Israeliese GVA variante (T7-GVAGR5 en T7-GVA118, verkry van M. Mawassi) is onder beheer van ‘n CaMV 35S promotor geplaas (35S-GVA-GR5 and 35S-GVA118). Beide klone het na agro-infiltrasie in N. benthamiana plante gerepliseer, sistemies beweeg en tipiese GVA simptome geinduseer. Hierdie twee klone het as raamwerk gedien vir verdere eksperimente in karakterisering van tydelike uitdrukkings- en VIGS vektore. Die gebruik van GVA as geen-insvoegingsvektor (35S-GVA118) en geen-vervangingsvektor (35S-GVA-GR5- ORF2+sgMP) is in N. benthamiana en V. vinifera vergelyk. Die geen-invoegingsvektor 35S-GVA118, was op die vollengte GVA genoom gebasseer. Die geen-vervangingsvektor 35S-GVA-GR5- ORF2+sgMP, was in hierdie studie gekonstrueer. Dit is gemaak eerstens deur eliminasie van ORF 2 in die 35S-GVA-GR5 kloon, en tweedens deur die invoeging van ’n subgenomiese promotor van die beweginsproteïen (sgMP) en unieke beperkings-ensiemsetels om klonering van transgene te fasiliteer. Beide vektore het in N. benthamiana vergelykbare GUS uitdrukkingsvlakke en fotobleikende simptome getoon na virus-geinduseerde NbPDS uitdowing. In V. Vinifera is beperkte GUS uitdrukkingsvlakke en VIGS fotobleikende simptome opgemerk met die geen-invoegingsvektor, 35S-GVA118. Geen GUS uitdrukking is in hierdie gasheerplant met die geen-vervangingsvektor opgemerk nie. Slegs een wingerdplant het fotobleikende simptome, na 4 maande in 3 sistemies geïnfekteerde blare gewys, na agroinfiltrasie van die 35S-GVA-GR5- ORF2-VvPDS+sgMP konstruk. Hierdie studie het bevestig dat GVA as geen-invoeging en geen-vervangingsvektor, vir heteroloë proteïenuitdrukking en VIGS, in N. benthamiana gebruik kan word, maar dit blyk of die gebruik daarvan in wingerd meer tot die floeëm weefsel beperk is. Hierdie studie wys vir die eerste keer dat ORF 2 nie nodig is vir langafstand beweging van die virus in wingerd nie. Om die moontlike rol van die P163-M5 119 nt invoeging en die GVA ORF 2 (met onbekende funksie), in die uitdrukking van simptome in plante te ondersoek, is ORF 2 van die 35SGVA- GR5 cDNA kloon verwyder en daaropvolgens vervang met die ooreenstemmende ORFs van vier Suid-Afrikaanse GVA variante. Na agro-infiltrasie in N. benthamiana blare, het al die chimeras die vermoë gehad om te repliseer, sistemies te beweeg en simptome te induseer. Geen korrelasie kon gevind word tussen die graad van simptome, die teenwoordigheid van die P163-M5 insersie en die spesifieke GVA ORF 2 teenwoordig in die chimeras nie, wat dus daarop dui dat ander faktore in die virusgenoom of die gasheerplant `n moontlike belangrike rol kan speel. Hierdie studie het bygedrae tot die beskikbare poel van in vivo gereedskap vir die bestudering en verbetering van die kosbare wingerdgewas. Dit het ook talle interessante navorsingsgeleenthede oopgemaak om in die nabye toekoms te betree.

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