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Studies on Phakopsora pachyrhizi, the causal organism of soybean rust.Nunkumar, Archana. January 2006 (has links)
Phakopsora pachyrhizi H. Syd and P. Syd, the causal organism of soybean rust
(SBR) was first reported in Japan in 1902. In 1934 the pathogen was found in several
other Asian countries and as far south as Australia. In India, SBR was first reported
on soybeans in 1951. There have been several early reports of SBR in equatorial
Africa but the first confirmed report of P. pachyrhizi on the African continent was in
1996 from Kenya, Rwanda and Uganda. Since then, the pathogen has spread south
with reports from Zambia and Zimbabwe in 1998 and in Mozambique in 2000.
In February 2001, P. pachyrhizi was first detected on soybeans near Vryheid, in
Northern KwaZulu-Natal, South Africa (SA). As the season progressed, the disease
was observed in other parts of the province, and epidemic levels were found in the
Cedara, Greytown, Howick and Karkloof production regions. Soybean rust
subsequently spread to Amsterdam and Ermelo in the Highveld region of SA. The
disease reappeared in SA in March 2002. It is now established that the pathogen is a
threat to soybean production in the country with yield losses in the region of 10-80%.
A literature review on SBR investigating the taxonomy of the pathogen, its
morphology, symptoms, host range, infection process, epidemiology, control options
and the economic importance of P. pachyrhizi was complied to provide the necessary
background information to conduct research under local conditions and to assist in
interpretation of results of experiments.
Epidemiological trials were conducted at the University of KwaZulu-Natal under
controlled environmental conditions in a dew chamber and conviron. Development of
P. pachyrhizi on the susceptible cultivar (LS5995) was quantified in combinations of
seven temperatures (15,19,21,24,26,28 and 30°C) and five leaf wetness durations
(LWD) (6,9,12,14 and 16hrs) at three relative humidities (RH) (75%, 85% and 95%).
Studies indicate that optimum temperature for uredospore infection is 21-24°C with a
LWD greater than 12hrs and RH 85-95%. The number of pustules as well as lesion
size on the abaxial and adaxial leaf surface increased with increasing LWD at all the
RH values tested. Infection did not occur on plants incubated at 15°C and 30°C at
85% or 95%RH whereas at 75%RH infection did not occur on plants incubated at
15°C, 19°C and 30°C regardless of LWD. Number of pustules per lesion produced at
75%, 85% and 95%RH was highest at 24°C and showed a gradual increase with
increasing LWD. Lesion size on both leaf surfaces increased after 12hrs LWD at 24°C
at 75% and 85%RH whereas at 95%RH lesion size increased after 14hrs LWD at
24°C.
Exposure of uredospores to ultraviolet light which is equivalent to ultraviolet C
(sunlight) which is < 280nm, shows a decrease in germination (7%). Under
continuous darkness, the germination percentage was found to range from 58% after
48 hrs. Germination was found to peak at 16hrs in darkness with a gradual decrease
as time increased whereas germination under ultraviolet light was highest after 6hrs
with a gradual decrease with increased exposure to light. Germ tube lengths were
found to be shorter when exposed to ultraviolet light (107µm) compared to controls
kept in the dark (181µm). Results obtained clearly show a negative effect of ultraviolet
light on the germination and germ tube length of uredospores. A 0.1 ml suspension of
uredospores on 1.25% water agar Petri dishes was exposed to cycles of 14h
ultraviolet light and 10h darkness for 48h. Results indicate an increase in germination
percentage of uredospores when exposed to 10h of darkness following a 14h period
under ultraviolet light.
Controlled environmental studies were conducted to determine alternative hosts of P.
pachyrhizi in SA. The control used in this experiment was Prima 2000, a susceptible
cultivar to soybean rust. Seven legume plants [Cajanus cajan (L.) Huth, Glycine max
(L.) Merr, Lablab purpureus (L.) Sweet, Lupinus angustifolius (L.) Finnish, Phaseolus
vulgaris (L.), Pueraria lobata (M&S) Wild and Vigna unguiculata (L.) Walp] and three
dry bean lines (Bonus; OPS-RS2 and PAN 159) showed typical SBR symptoms when
rated after 21 days post inoculation with uredospores for percentage disease severity.
Disease severity was significantly different within the alternative hosts, but G. max,
P. vulgaris and P. lobata were not significantly different from Prima 2000 (control). A
uredospore suspension of 2.5 x 10(5) uredospores ml(-1) from plants that showed typical
SBR symptoms was made and inoculated on to Prima 2000, a susceptible soybean
cultivar. Uredospores from pustules on G. max, L. purpureus, L. angustifolius, P.
vulgaris, P. lobata, V. unguiculata, Bonus and PAN 159 produced viable uredospores
on PRIMA 2000. These plants are considered alternative hosts of P. pachyrhizi.
Effect of leaf age on susceptibility of soybean to SBR was tested under controlled
environmental conditions. Mean number of lesions as well as lesion size were greater
on younger leaves than on older leaves of plants at the same physiological age.
Plants at the early vegetative and reproductive stages had a significantly lower
number of lesions as well as a smaller lesion size. Plants at the V6 and R1 growth
stages were significantly more susceptible to P. pachyrhizi than plants at other
developmental stages.
Trichoderma harzianum Rifai, Eco-77® a commercial biological control product, was
evaluated for its efficacy as a biological control agent of P. pachyrhizi. Trichoderma
harzianum sprayed at the standard concentration on infected soybean plants was
significantly more effective in controlling P. pachyrhizi than plants sprayed at 1/2X
and 2x the standard concentration. This was noted in both Trial 1 and 2. Data indicate
that spraying the filtrate two days after inoculation produces less disease. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2006.
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Breeding and evaluation of cassava for high storage root yield and early bulking in Uganda.Tumuhimbise, Robooni. January 2013 (has links)
Cassava (Manihot esculenta Crantz), is the world’s most widely grown starch storage root crop. It is a principal food staple in sub-Saharan Africa where it accounts for approximately one-third of the total production of staple food crops. It plays a key role as a food security and an income-generating crop for millions of smallholder farmers. In Uganda, cassava ranks second to bananas (Musa spp.) in terms of area occupied, total production and per capita consumption; however, nearly 5% of the total population experiences hunger with the prevalence of food energy deficiency at the country level standing at 48%. Cassava is a crop with high potential to alleviate food shortages and energy deficiencies, owing to its unique advantages of producing acceptable yields and starch on infertile soils amidst erratic rainfall, when most other crops would fail. Hoewever, its yield potential has not been fully realised since most of the cassava cultivars grown are susceptible to pests and diseases, low yielding and late bulking. The main objective of the research was to develop high yielding, early bulking cassava genotypes that combine resistance to cassava brown streak disease (CBSD) and cassava mosaic disease (CMD) with farmer preferred traits for cultivation in Uganda. The specific objectives were to: (i) evaluate farmers’ attitudes to and/or perceptions of cassava early bulking, production constraints and cultivar preferences; (ii) determine the extent of genetic variability in storage root bulking and other important traits of selected cassava genotypes; (iii) assess the effects of genotype x environment interaction on early bulking and related traits of selected cassava genotypes; (iv) develop and evaluate cassava F1 families for early bulking in terms of the attainment of early, high fresh storage root yield (FSRY) and resistance to CBSD and CMD; and (v) determine the combining ability and gene action controlling early bulking and yield-related traits, as well as resistance to CBSD and CMD. Through the farmer participatory survey, a number of cassava production constraints were identified, key of which were: diseases, especially CBSD and CMD; lack of early bulking cultivars; rodents and insect pests. Farmers rated early bulking as the second most important preferred trait after FSRY, but suggested that early bulking should be complemented with high dry mass content (DMC), sweetness, high FSRY and resistance to pests and diseases. The analysis of variance of 12 cassava genotypes selected for evaluation in three diverse locations and at five different harvest times indicated significant variation among genotypes, harvest times, locations and their interactions for FSRY and most of the other traits evaluated. Fresh storage root yield and the other traits evaluated were predominantly under the control of genetic variation, indicating that genetic advance would be achieved through hybridisation of the test genotypes. Additive main effects and multiplicative interaction (AMMI) analysis of the data collected at nine months after planting (MAP) indicated a non-significant GEI for early FSRY, but significant GEI for other traits assessed. Eight of the 12 genotypes analysed had relatively low interaction with locations for early FSRY, signifying that these genotypes were relatively stable for early FSRY. Thirty-six F1 families were generated from a 9 x 9 diallel and exhibited a high degree of variation between and within families for all the traits assessed at the seedling evaluation stage. Diallel analysis at the seedling evaluation stage at 10 MAP indicated that additive gene effects were predominant in the expression of early FSRY and most of the other traits analysed. At the clonal evaluation stage, the 36 families were assessed for early FSRY at 8 MAP and this trait together with most of the other traits assessed were found to be predominantly under the control of non-additive gene effects. High mid- and better-parent heterosis for early FSRY was recorded in most families at the clonal evaluation stage with NASE3 x Nyara, Nyara x B11 and NASE3 x B11 recording the highest. Selection from the 36 families at the clonal evaluation stage based on farmers’ top two preferred traits, viz. early bulking for FSRY and DMC, plus resistance to CBSD and CMD identified 50 genotypes that had early FSRY of ≥25 t ha-1 at 8 MAP compared to the best parent, CT1 that had 15.9 t ha-1 at 8 MAP. The selected genotypes also had high DMC and dual resistance to CMD and CBSD. Advancement of the selected genotypes should go a long way towards increasing cassava yield per unit time, reducing food shortages and increasing the income of smallholder farmers in Uganda. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2013.
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Breeding investigations on utility of maize streak virus resistant germplasm for hybrid development in the tropics.Gichuru, Lilian Njeri. 12 May 2014 (has links)
Maize (Zea mays L.) supports millions of livelihoods in sub-Saharan Africa (SSA) in terms of
food and feed. Production of the crop is however limited by several factors, among these, maize
streak virus (MSV) disease. Although extensively studied, MSV remains a serious problem in
SSA due to several challenges in breeding MSV resistant maize varieties. These include
integration of MSV resistant germplasm from different backgrounds, reliance on a few resistant
sources, and genotype x environment interactions. This study was designed to assess the
breeding potential of several MSV resistant lines in hybrid combinations. Understanding
architecture of genetic divergence and background of these genotypes would greatly aid in
breeding high yielding and stable MSV resistant hybrids. Experiments were conducted during
2010 to 2012 seasons in Kenya. Diallel crosses and SSR markers were used to characterize
MSV resistant maize inbred lines from three programs of CIMMYT, KARI and IITA.
In general, this study revealed that MSV is still an important problem in Kenya with high
incidence and severity levels in the farmers’ fields. The levels of MSV resistance in locally
grown hybrids needs to be improved. Farmers challenged breeders to develop new hybrids that
combine early maturing, high yield potential and MSV resistance.
The study was successful in identifying the best eight inbred lines for use in breeding new maize
hybrids with MSV resistance. The nature of gene effects was established for the first time, in
particular the role of epistasis and G x E in conditioning MSV resistance in hybrids. Results
indicate serious implications for previous models that ignored epistasis in studying MSV
resistance in maize. The inbreds Z419, S558, CML509 and Osu23i, displayed high levels of
epistasis for MSV resistance. Unless strong sources of MSV resistance, such as MUL114 and
CML509, are used, breeding resistant hybrids will require parents that carry dominant
resistance genes. The additive-dominance model was adequate to explain northern leaf blight
(NLB) resistance in hybrids, indicating fewer complications in breeding NLB resistant hybrids.
The study also reveals that SSR genetic distance data can be used to predict hybrid
performance, especially when the correct set of markers is used. Many previous studies have
not found any significant relationship between genetic distance and heterosis, due to large
G x E and use of a wrong set of markers. The diallel analysis and SSR data established the
important heterotic groups, which will be exploited for efficient development of MSV resistant
maize hybrids. These strategies will be recommended to programs that emphasize MSV
resistance in maize hybrids. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2013.
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Interaction between root lesion nematode, Pratylenchus neglectus, and root-rotting fungi of wheat / by Abdolhossein Taheri.Taheri, Abdolhossein January 1996 (has links)
Bibliography: leaves 307-329. / xvi, 329 leaves, [21] leaves of plates : ill. (chiefly col.), map ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / This study concludes that in soils in South Australia where root-rotting fungi and P. neglectus exist together, root disease of wheat is caused by their combined effect. Evidence suggests that P. neglectus not only contributes to this interaction through mechanical wounding of roots, but also causes biochemical and physiological changes in plants, making them more prone to fungal infection. / Thesis (Ph.D.)--University of Adelaide, Dept. of Plant Science, 1996
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Species of Pythium associated with barley in South Australia / by J.I. BratoloveanuBratoloveanu, J. I. January 1985 (has links)
Bibliography: leaves 140-158 / ix, 158 leaves, [23] leaves of plates : ill., 1 map ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (Ph.D.1985) Dept. of Plant Pathology, University of Adelaide
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Studies upon the plant parasitic nematodes and their controlByars, Luther P. January 1900 (has links)
Thesis (Ph. D.)--University of Wisconsin--Madison, 1919. / Title from added collective thesis title page. Part 1 reprinted from Phytopathology, vol. 4, no. 4 (Aug. 1914), p. [323]-326, plate XXI ; Part 2 reprinted from Phytopathology, vol. IX, no. 2 (Feb. 1919), p. [93]-103, plate IX ; Part 3: Bulletin / United States Department of Agriculture, no. 818 (5 Jan. 1920) (see OCLC #16627505), 14 p., V p. of plates ; Part 4: Bulletin / United States Department of Agriculture, no. 842 (7 Sept. 1920) (see OCLC #16627722), 40 p., VI p. of plates. Includes bibliographical references.
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Real time PCR as a versatile tool for virus detection and transgenic plant analysisMalan, Stefanie 12 1900 (has links)
Thesis (MSc (Genetics))--University of Stellenbosch, 2009. / ENGLISH ABSTRACT: South Africa is regarded as one of the top wine producing countries in the world.
One of the threats to the sustainability of the wine industry is viral diseases of which
Grapevine leafroll-associated virus 3 (GLRaV-3) and Grapevine virus A (GVA) are
considered to be the most important and wide spread. Scion material is regularly
tested for viruses; however scion material is often grafted onto rootstocks that have
questionable phytosanitary status. Virus detection in rootstocks is challenging due to
low and varying titres, but is imperative as a viral control mechanism. An additional
viral control mechanism is the use of transgenic grapevine material which offers
resistance to grapevine infection.
The objective of this project was to establish a detection system using real time PCR
(qPCR) techniques, to accurately and routinely detect GLRaV-3 and GVA in
rootstock propagation material. qPCR would furthermore be used to perform
molecular characterisation of transgenic plants containing a GLRaV-3 antiviral
ΔHSP-Mut construct.
A severely infected vineyard (Nietvoorbij farm) in the Stellenbosch area was
screened throughout the grapevine growing season to investigate virus prevalence
throughout the season and to determine the optimal time for sensitive virus detection.
A large scale screening of nursery propagation material for GLRaV-3 infection was
also conducted. The qRT-PCR results were compared to DAS-ELISA results to
compare the efficacy and sensitivity of the two techniques. For the severely infected
vineyard, the ability to detect GLRaV-3 increased as the season progressed towards
winter. qRT-PCR was more sensitive and accurate in detecting GLRaV-3 than DASELISA,
as the latter technique delivered numerous false positive results later in the
season. The best time to screen for GLRaV-3 in the Western Cape region was from
the end of July to September. For the nursery screenings, our qRT-PCR results were
compared to the results of the DAS-ELISA performed by the specific nurseries. No
GLRaV-3 infection was detected in the specific samples received from the two
different nurseries. The results for all the samples correlated between the two techniques. This confirms that the propagation material of these nurseries has a
healthy phytosanitary status with regards to GLRaV-3.
However, the detection of GVA in the severely infected vineyard yielded inconsistent
results. Detection ability fluctuated throughout the season and no specific trend in
seasonal variation and virus titre fluctuation could be established. The highest
percentage of GVA infected samples were detected during September, April and the
end of July. Previously published universal primers were used for the detection of
GVA, but further investigation indicated that they might not be suitable for sensitive
detection of specific GVA variants present in South Africa.
Vitis vinifera was transformed with a GLRaV-3 antiviral construct, ΔHSP-Mut.
SYBR Green Real time PCR (qPCR) and qRT-PCR were utilised as alternative
methods for molecular characterisation of transgenic plants. The qPCR and Southern
blot results correlated for 76.5% of the samples. This illustrated the ability of qPCR
to accurately estimate transgene copy numbers. Various samples were identified
during qRT-PCR amplification that exhibited high mRNA expression levels of the
transgene. These samples are ideal for further viral resistance studies.
This study illustrated that the versatility of real time PCR renders it a valuable tool for
accurate virus detection as well as copy number determination. / AFRIKAANSE OPSOMMING: Suid Afrika word geag as een van die top wyn produserende lande ter wereld. Die volhoubaarheid van die wynbedryf word onder andere bedreig deur virus-infeksies.
Grapevine leafroll associated virus 3 (GLRaV-3) en Grapevine virus A (GVA) is van
die mees belangrike virusse wat siektes veroorsaak in Suid-Afrikaanse wingerde.
Wingerd bo-stok materiaal word gereeld getoets vir hierdie virusse, maar hierdie
materiaal word meestal geënt op onderstokmateriaal waarvan die virus status
onbekend is. Virus opsporing in onderstokke word egter gekompliseer deur baie lae
en variërende virus konsentrasies, maar opsporing in voortplantingsmateriaal is ‘n
noodsaaklike beheermeganisme vir virus-infeksie.
Die doel van die projek was om ‘n opsporingsisteem te ontwikkel via kwantitatiewe
PCR (qPCR) tegnieke vir akkurate en gereelde toetsing van GLRaV-3 en GVA in
onderstokmateriaal. qPCR sal ook verder gebruik word vir molekulêre
karakterisering van transgeniese plante wat ‘n GLRaV-3 antivirale ΔHSP-Mut
konstruk bevat.
‘n Hoogs geïnfekteerde wingerd was regdeur die seisoen getoets om seisoenale
fluktuasies in viruskonsentrasie te ondersoek en om die optimale tydstip vir
sensitiewe virus opsporing te bepaal. ‘n Grootskaalse toetsing van kwekery
voortplantingsmateriaal vir GLRaV-3 infeksie was ook uitgevoer. Die qRT-PCR
resultate is met die DAS-ELISA resultate vergelyk om die effektiwiteit en
sensitiwiteit van die twee tegnieke te vergelyk. Vir die hoogs geïnfekteerde wingerd
het die GLRaV-3 opsporing toegeneem met die verloop van die seisoen tot en met
winter. qRT-PCR was meer sensitief en akkuraat as DAS-ELISA in die opsporing
van GLRaV-3, weens verskeie vals positiewe resultate wat later in die seisoen deur
die laasgenoemde tegniek verkry is. Die beste tyd om vir GLRaV-3 te toets is vanaf
einde Julie tot September. Tydens die kwekery toetsings was qRT-PCR resultate met
die DAS-ELISA resultate van die spesifieke kwekerye vergelyk. Geen GLRaV-3
infeksie was waargeneem in die spesifieke monsters wat vanaf die kwekerye ontvang
is nie. Die resultate van die twee tegnieke het ooreengestem vir al die monsters wat
v
getoets is. Dit het bevestig dat die voortplantingsmateriaal van hierdie kwekerye
gesonde fitosanitêre status met betrekking tot GLRaV-3 gehad het.
Die opsporing van GVA in die geïnfekteerde wingerd het egter wisselvallige resultate
gelewer. Opsporing van die virus het ook regdeur die seisoen gefluktueer en geen
spesifieke neiging in seisoenale opsporingsvermoë kon gemaak word nie. Die
hoogste persentasie GVA geïnfekteerde monsters was waargeneem tydens
September, April en die einde van Julie. Voorheen gepubliseerde universele inleiers
was gebruik vir die opsporing van GVA, maar verdere ondersoeke het getoon dat
hierdie inleiers nie noodwendig geskik is vir sensitiewe opsporing van GVA variante
wat teenwoordig is in Suid-Afrika nie.
Vitis vinifera was getransformeer met ‘n GLRaV-3 antivirale konstruct, ΔHSP-Mut.
SYBR Green Real time PCR (qPCR) en qRT-PCR was ingespan as alternatiewe
metodes vir molekulêre karaterisering van transgeniese plante. Die qPCR en
Southern-klad resultate het ooreengestem vir 76.5% van die monsters. Dit illustreer
die vermoë van qPCR om akkurate kopie-getalle van transgene te bepaal. Verskeie
plante is geïdentifiseer tydens qRT-PCR amplifisering wat hoë vlakke van transgeen
mRNA uitdrukking getoon het. Hierdie monsters is ideaal vir verdere virus
weerstandbiedendheids studies.
Hierdie studie het die veelsydigheid van real time PCR bewys en getoon dat dit ‘n
kosbare tegniek is vir akkurate virus opsporing sowel as kopie-getal bepaling.
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A metagenomic approach using next-generation sequencing for viral profiling of a vineyard and genetic characterization of grapevine virus ECoetzee, Beatrix 12 1900 (has links)
Thesis (MSc (Genetics))--University of Stellenbosch, 2010. / Includes bibliography. / Title page: Dept. of Genetics, Faculty of Science / ENGLISH ABSTRACT: Next-generation sequencing technologies are increasingly used in metagenomic studies, largely
due to the high sequence data throughput capacity and unbiased approach in determining the
genetic composition of an unknown environmental sample. This study investigated the
applicability of the Illumina next-generation sequencing platform for metagenomic sequencing
of grapevine viruses to provide the first complete viral profile, or virome, of a diseased
vineyard.
Leaf material was harvested from 44 randomly selected vines in a leafroll-diseased vineyard in
South Africa. Sample material was pooled and double-stranded RNA extracted. The dsRNA was
sequenced as a paired-end sequencing run using the Illumina sequencing-by-synthesis
technique, and more than 19 million sequence reads, equivalent to approximately 837
megabases of metagenomic sequence data, were obtained. Of these data, approximately 400
megabases could be assembled into 449 scaffolds, using the de novo assembler Velvet. These
scaffolds were subjected to BLAST searches against the NCBI databases and top hit scores were
used for virus identification. Based on the BLAST results, suitable sequences were selected from
the NCBI database and used as reference sequence in MAQ mapping assemblies.
The bioinformatic analyses allowed for the determination of the virus species present, the most
prominent variants, and the relative abundance of each. Four known grapevine viral pathogens
were identified. Grapevine leafroll-associated virus 3, representing 59% of the analyzed short
read sequence data, was identified as the most prominent virus species. Three variants of this
virus were detected: GP18 was the most abundant, followed by a minor Cl766/NY1 variant and
a potential novel grapevine leafroll-associated ampelovirus. A single Grapevine rupestris stem
pitting ]associated virus variant, similar to SG1, and a Grapevine virus A variant, a member of
molecular group III, were identified. This study is also the first to report the presence of
Grapevine virus E (GVE) in South African vineyards. Grapevine virus E was further genetically characterized and the genome sequence of GVE
isolate SA94 determined. The GVE SA94 genome sequence, 7568 nucleotides in length, is the
first complete genome sequence for the virus species. The genome organization of GVE SA94 is
typical of vitiviruses, but in contrast to other RNA viruses, the AlkB domain is located within the
helicase domain in open reading frame 1 (ORF 1). Grapevine virus E SA94 shares nearly 100%
nucleotide identity with the Japanese TvP15 isolate and GVE 3404, a de novo scaffold generated
from the metagenomic sequence data.
Bioinformatic analysis of metagenomic sequence data further revealed the presence of three
fungus-infecting viral families, Chrysoviridae, Totiviridae and the unclassified dsRNA virus,
Fusarium graminearum dsRNA mycovirus 4. A virus from the family Chrysoviridae, similar to
Penicillium chrysogenum virus, was the second most abundant virus detected.
We demonstrated the successful application of a short read sequencing technology, such as the
Illumina platform, for viral profiling of an infected vineyard. To our knowledge this is the first
application of the Illumina technology for this purpose. / AFRIKAANSE OPSOMMING: Volgende-generasie tegnologie om basis volgordes van nukleiensure te bepaal, word al meer
gebruik in metagenomiese studies. Dit is veral weens die hoe data-omset kapasiteit en
onbevooroordeelde aanslag in die bepaling van die genetiese samestelling van onbekende
omgewingsmonsters. Hierdie studie het die aanwending van die Illumina volgende-generasie
volgorde-bepalingsplatform in 'n metagenomiese studie van wingerdvirusse, ondersoek. Dit het
ten doel gehad om die eerste volledige virus profiel, of viroom, van 'n geinfekteerde wingerd
saam te stel.
Blaarmateriaal is verkry vanaf 44 lukraak-gekose wingerdstokke in 'n rolblad-geinfekteerde
wingerd in Suid-Afrika. Monster materiaal is saamgevoeg en dubbelstring-RNS geekstraheer.
Die dubbelstring-RNS is onderwerp aan gepaarde-ent volgorde-bepaling deur gebruik te maak
van die Illumina volgorde-bepaling-deur-sintese tegniek. Meer as 19 miljoen volgorde reekse,
ekwivalent aan ongeveer 837 megabasisse volgorde data, is verkry. Van hierdie data kon
ongeveer 400 megabasisse saamgevoeg word in 449 konstrukte ("scaffolds"), deur gebruik te
maak van die de novo samesteller Velvet. Hierdie konstrukte is onderwerp aan BLAST soektogte
teen die NCBI databasisse en die hoogste trefslag-telling is gebruik vir virus identifikasie. Op
grond van die "BLAST" resultate is geskikte volgordes geselekteer vanaf die NCBI databasis en
gebruik as verwysingvolgordes in MAQ kartering-analises.
Met die bioinfomatika analises kon die virus spesies teenwoordig, asook die mees prominente
variante en relatiewe voorkoms van elk, bepaal word. Vier bekende virus wingerdpatogene is
geidentifiseer. Grapevine leafroll-associated virus 3, verteenwoordig deur 59% van die
geanaliseerde kort-reeks volgorde data, is identifiseer as die mees prominente virus spesie. Drie
variante van die virus is in die wingerdmonster opgespoor: GP18 kom die mees algemeen voor,
gevolg deur 'n CL-766/NY1 variant en 'n potensiele nuwe wingerd rolblad-geassosieerde
ampelovirus. 'n Enkele Grapevine rupestris stem pitting-associated virus variant, soortgelyk aan
SG1, en 'n Grapevine virus A variant, 'n lid van molekulere groep III, is geidentifiseer. Hierdie
studie is ook die eerste om die teenwoordigheid van Grapevine virus E (GVE) in Suid-Afrikaanse
wingerde te rapporteer. Grapevine virus E is verder geneties gekarakteriseer en die genoomvolgorde van GVE isolaat
SA94 is bepaal. Die GVE SA94 genoomvolgorde, 7568 nukleotiede lank, is die eerste volledige
genoomvolgorde vir hierdie virus spesie. Die genoomorganisasie is tipies van vitivirusse, maar
in kontras met ander RNA virusse is die AlkB domein binne-in die helikase domein van
oopleesraam 1 (ORF 1) geleë. Grapevine virus E SA94 deel byna 100% nukleotied identiteit met
die Japannese TvP15 isolaat en GVE 3404, 'n de novo konstruk gegenereer vanaf die
metagenomiese volgorde data.
Bioinformatika analises van die metagenomiese volgorde data het verder die teenwoordigheid
van drie swam-infekterende virus families, die Chrysoviridae, Totiviridae en ongeklassifiseerde
dubbelstring-RNS virus, Fusarium graminearum dsRNA mycovirus 4, aangetoon. 'n Virus van die
Chrysoviridae familie, soortgelyk aan Penicillium chrysogenum virus, het die tweede meeste
voorgekom in die wingerd monster.
Hierdie studie demonstreer die suksesvolle toepassing van 'n kort reeks volgorde-bepalingstegnologie
soos die Illumina platform, vir die opstel van 'n virusprofiel van 'n
geinfekteerde wingerd. Sover ons kennis strek is hierdie die eerste aanwending van die Illumina
tegnologie vir hierdie doel.
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Yield response of Fusarium infected maize seed treated with biological control agent formulationsGerber, Johan,1961- 11 1900 (has links)
Potential vegetative and reproductive increases in yield, as well as the biological efficacy against Fusarium verticillioides and F. proliferatum causing ear and stem rot in maize crops of commercially-formulated micro-organism formulation T-Gro (Trichoderma harzianum isolate DB103 WP) combined with Spartacus (Beauveria bassiana isolate DB 105 WP), T-Gro combined with Armenius (Bacillus subtilis isolate DB 109 WP), T-Gro combined with Maximus (Bacillus subtilis isolate DB 108 WP), T-Gro combined with Shelter (Bacillus subtilis isolate DB 101), T-Gro combined with Bismarck (Microbacterium maritypicum isolate DB 107 WP), as well as individual treatments of T-Gro, Armenius, Bismarck, Maximus and Shelter, were investigated when applied to maize seed and soil under field conditions. All the micro-organism treatments were compared with Thiram 750WP (750g/kg thiram WP) and an untreated control.
The micro-organism treatments showed an increase in vegetative as well as reproductive yields when compared to the reference product Thiram 750 WP and the untreated control. There were no observations of adverse effects on the germination of maize seed in all the treatments that were applied. The three isolates B. subtilis, T. harzianum, and M. maritypicum, showed a significant reduction in vascular tissue discolouration of the main and ear stems, indicating a potential to be used in the reduction and control of diseases caused by Fusarium spp. Results also showed poor to very good increases of stem and foliage biomass as well as cob yield per plant produced by the micro-organism treatments when compared to the untreated control. The highest cob yield per plant that differed significantly from the untreated control was produced by T-Gro and Shelter.
No phytotoxicity of any kind was observed with the application of the micro-organism formulations and they could therefore be deemed suitable to be used for the treatment of maize seed. The micro-organism formulations containing fungal and bacterial biological control agents have the potential to be used in commercial maize production to increase vegetative and reproductive yields and reduce the severity of ear and stem rot in maize. / Agriculture Animal Health and Human Ecology / M.Sc. (Agriculture)
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Potyvirus: caracterização parcial de espécies em plantas daninhas associadas a cultura do pimentão, avaliação de genótipos de alface e análise subcelular do eIF4E e de proteínas do Lettuce mosaic virusMoura, Mônika Fecury [UNESP] 18 April 2013 (has links) (PDF)
Made available in DSpace on 2014-06-11T19:34:59Z (GMT). No. of bitstreams: 0
Previous issue date: 2013-04-18Bitstream added on 2014-06-13T20:27:47Z : No. of bitstreams: 1
moura_mf_dr_botfca.pdf: 469933 bytes, checksum: eccaa27134acf7a66cc0606cbff1deaa (MD5) / Os potyvírus constituem cerca de 90% das espécies conhecidas da família Potyviridae. No Brasil ocasionam sérios entraves em alface (Lactuca sativa L.) e em pimentão (Capsicum annuum L.), onde se pode citar o Lettuce mosaic virus – LMV e o Pepper yellow mosaic virus (PepYMV), respectivamente. Com o intuito de melhor compreender o reservatório natural de potyvírus em plantas invasoras, amostras foram coletadas em áreas produtoras de pimentão e analisadas utilizando-se antissoro anti-potyvirus (Agdia). Entre estas plantas positivas, destacou-se Solanum americanum Mill, onde foi verificada infecção mista do Cucumber mosaic virus e do Potato virus Y, e Commelina benghalensis L. em que foi encontrado um possível novo potyvírus com a maior identidade de nucleotídeos da proteína capsidial (62%) com a espécie Hardenbergia mosaic virus. Este potyvírus não foi transmitido por extrato vegetal, bem como por afídeos para plantas de pimentão e Nicotiana tabaccum TNN. Na região codificadora para a proteína capsidial do potyvirus não foi encontrado o domínio DAG, relacionado a transmissão por afídeos. Visando encontrar possíveis fontes de resistência ao Lettuce mosaic virus - LMV, genótipos foram inoculados com o isolado LMV-AF-199 (LMV-Most) e o fator de iniciação de tradução eucariótico eIF4E destes genótipos analisado. Em Calona e Salinas-88, conhecidas previamente como portadoras dos genes recessivos mol1 e mol2 foram observados sintomas em todas as plantas inoculadas e verificado o padrão típico do eIF4E1 e eIF4E2, respectivamente... / The Potyvirus genus corresponds to 90% of known species of the Potyviridae family. In Brazil potyviruses causes serious problems in lettuce (Lactuca sativa L) and in pepper crops (Capsicum annuum L.), which we can highlight Lettuce mosaic virus – LMV and Pepper yellow mosaic virus (PepYMV), respectively. To increase knowledge about the natural reservoir of potyviruses in weeds, samples were collected from a pepper producer area and analyzed for potyvirus using antiserum anti-potyvirus (Agdia). Solanum americanum Mill was identified as a host for Cucumber mosaic virus and Potato virus Y. In Commelina benghalensis L. a possible new species of potyvirus was found with higher nucleotide identity of the coat protein (62%) with Hardenbergia mosaic virus. This potyvirus could not be transmitted by aphids to sweetpepper and... (Complete abstract click electronic access below)
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