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The graminaceous rusts and smuts of KansasHaard, Richard Thomas. January 1963 (has links)
Call number: LD2668 .T4 1963 H32 / Master of Science
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Environmental effects on Peronospora trifoliorum oospore production in seedlings of two alfalfa clones ; Attempts to germinate Peronospora trifoliorum oosporesHodgden, L. D. January 1978 (has links)
Call number: LD2668 .T4 1978 H63 / Master of Science
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Lignin as a mechanism of field resistance to Phytophthora rot in soybeansCurry, Joseph Timothy. January 1984 (has links)
Call number: LD2668 .T4 1984 C87 / Master of Science
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Some post-harvest physiological studies of potatoes and relation of some potato cultivars to incidence of internal brown spotLafta, Abbas Mubadar. January 1985 (has links)
Call number: LD2668 .T4 1985 L33 / Master of Science
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A comparison of techniques for screening for resistance to the chinch bug, Blissus leucopterus leucopterus (Say), in sorghumMeehan, Mitchell Elwin. January 1985 (has links)
Call number: LD2668 .T4 1985 M43 / Master of Science
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Breeding for Smut Resistance in Arizona-Grown WheatBryan, 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.
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Assessment of inoculation techniques to evalute apple resistance to Phytophthora cactorumZondo, 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.
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Invloed van doppenetrasieweerstand op die oesstadium van druiweVan 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.
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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
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The development and characterisation of grapevine virus-based expression vectorsDu 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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