Inter-row mechanical cultivation was proposed as a supplement to or substitute for conventional weed control methods currently used in snap bean, carrot and lettuce production. Several types of cultivators were assessed and compared. The effect of mechanical cultivation on crop yield and quality was studied by counting, weighing and grading bean pods, carrot roots and lettuce heads. The effect of mechanical cultivation on disease incidence was studied by surveying fields during the season and by determining the number and weight of diseased pods, roots and heads at harvest. The relationship between the level of Cercospora blight on carrots and potential impacts on yield was also investigated by measuring plant characteristics and the amount of force needed to separate carrot foliage from root. The effect of mechanical cultivation on the phenology of snap bean flowering was studied by determining how long it took for a plant to produce 50% of its flowers and counting how many flowers and pods a plant produced. In general, mechanical cultivation did not affect normal crop production and may be used to replace or complement conventional weed control methods. There was little variation among different cultivators within one season, but cultivator effects differed among crops and from one year to the next.

No description available.

Thesis (PhD (Agric))--University of Stellenbosch, 2003. / ENGLISH ABSTRACT: During a survey in the greater Cape Flats Heterodera schachtii was found to be widespread on cauliflower, Brussels sprouts, beetroot and cabbage. The numbers present were above two eggs and juveniles per gram of soil, generally regarded as the economic threshhold level of infestation and requiring control. The damage potential of H schachtii on vegetables, as well as the ability of certain weeds to serve as a source of infection on subsequent crop plantings was studied under greenhouse conditions and resulted in a reduction of yield and root weight of crops. Population densities of H schachtii increased significantly under favourable hosts like cabbage where densities of 198 eggs and juveniles per gram of soil were reached. The most commonly occurring weeds maintained nematode development and increased their population densities. They can thus serve as alternative hosts in the absence of susceptible hosts and should be routinely controlled. The life cycle and biology of H schachtii was also studied. Penetration of plant tissue and subsequent development on vegetables, weeds and trap crops were observed. Penetration was successful on all crops tested reaching 37% and 52% at inoculum levels of 22 and 11 juveniles per gram of soil, respectively. Subsequent development of H schachtii on weeds and vegetables was similar, but in the case of cauliflower and black nightshade as hosts, their life cycle was shorter in comparison to other crops. The possible existence of varying susceptibility of crops to different populations of H schachtii was examined by comparing the rates of penetration in crops and reproduction of geographically isolated populations of H schachtii in the greater Cape Flats. When root penetration, virulence and juvenile emergence were examined, populations from Lynedoch and Philippi were distinct from the other populations. Subsequently, representative individuals of these populations were subjected to PCR-RFLP, but with these techniques real differences between the various populations could not be adequately detected. The environmental parameters such as soil texture, temperature and pH on H schachtii were investigated as to their influence on the root weight and yield of crops. Reductions in the yield of beetroot and cabbage were observed with soil temperatures ranging between 15 to 30°C. Migration and penetration of H schachtii juveniles declined with an increase in clay and silt content of the soil. Above a 34% silt and clay content of soil, no migration and penetration took place. Root penetration levels of 30% and higher were reached with pH varying between 4.5 and 7.4. This resulted in a significant reduction in yield of crops. Crop rotation is an essential component of non-chemical control. In the case of H schachtii, it required one host crop in four non-host rotational cycles to maintain the population of the nematode in the soilless than three eggs and juveniles per gram of soil. The inclusion of a trap crop reduced the population densities to below two eggs and juveniles per gram of soil. It therefore also forms an integral part of a control strategy. Solarization proved successful as a physical control method. Best results were obtained in summer with clear polyethylene which led to a 97% reduction of infective juveniles. This method can be applied during the late summer in the greater Cape Flats, just before the onset of winter. This may safeguard future spring plantings. The need for effective control strategies in order to reduce the numbers of H schachtii is of the utmost importance to ensure vegetable production in the future. Small-scale farmers should therefore be educated in this respect. / AFRIKAANSE OPSOMMING: Heterodera schachtii het wydverspreid in 'n opname in die groter Kaapse Vlakte voorgekom op beet, blomkool, Brusselse spruite en kopkool. Die nematode getalle by alle lokaliteite was bo die algemeen aanvaarbare ekonomiese drempelwaarde van twee eiers en larwes per gram grond wat beheer regverdig. Die skadepotensiaal van H schachtii op groente, sowel as die vermoë van sekere onkruide om as infeksie bronne te dien vir opvolgende gewasse, is in glashuise ondersoek en het tot 'n verlaging in opbrengs en wortelmassa by gashere gelei. Die populasie digthede van H schachtii het met die aanplant van geskikte gashere tot vlakke van 198 eiers en larvae per gram grond gestyg. Die mees algemeen voorkomende gasheeronkruide het nematode ontwikkeling in stand gehou en selfs tot 'n populasie verhoging gelei. Hierdie onkruide is 'n beperkende faktor vir die verbouing van groente aangesien die onkruide as alternatiewe gasheer kan dien in die afwesigheid van gashere en onkruidbeheer moet dus op 'n gereelde basis toegepas word. Die lewenssiklus en biologie van H schachtii is ondersoek deurdat die penetrasie van gasheer wortels en die daaropvolgende ontwikkeling op groente, onkruide en vanggewasse vergelyk is. Penetrasie, vyf dae na inokulasie, is met alle gashere verkry met 37% en 52% penetrasie met inokulum vlakke van 22 en 11 larwes per gram grond onderskeidelik. Daaropvolgende ontwikkeling van H schachtii was soortgelyk op groente en onkruide, maar blomkool en nastergal het as gashere 'n verkorte lewenssiklus tot gevolg gehad. Die moontlikheid van verskille in die virulensie van H schachtii is ondersoek deur die penetrasie van gewasse en reproduksie vlakke van nematodes van nege verskillende geografies geskeide populasies in the groter Kaapse Vlakte te vergelyk. Die Lynedoch en Philippi populasies het onderskeibare resultate gelewer ten opsigte van die populasies uit die ander lokaliteite, maar geen verskille kon met PKR-RFLP aangetoon word nie. Die invloed van omgewings parameters, grondtekstuur, temperatuur en pH, is op H schachtii ondersoek ten opsigte van opbrengste en wortelmassa van gewasse. Grondtemperature tussen 15°C - 30°C het tot die grootste daling in opbrengs gelei op kopkool en beet. Migrasie en penetrasie het afgeneem met 'n toename in klei en slik inhoud tot en met 'n klei en slik inhoud van 34%, waarna geen penetrasie en migrasie voorgekom het nie. Wortelpenetrasie van 30% en hoër het voorgekom by pH vlakke van tussen 4.5 - 7.4 met die gepaardgaande verlaging in opbrengs van gewasse. Afwisseling van gewasse is 'n essensiële metode van nie-chemiese beheer van nematode getalle in die grond. Die mees optimale rotasie ten opsigte van H schachtii beheer is met die aanplanting van een gasheer gewas in vier gewas aanplantings verkry. Die insluiting van 'n vanggewas in die gewas rotasie siklus het die nematode populasievlakke tot onder twee per gram grond laat daal. Solarisasie is suksesvol uitgevoer met deurskynende poli-etileen in die groter Kaapse Vlakte gedurende die somer met gevolglik 'n 97% vermindering van die getalle infektiewe nematodes. Effektiewe beheermaatreëls ten opsigte van H schachtii moet in die groter Kaapse Vlakte ingestel word om groente-produksie in hierdie gebied te verseker. Kleinboere moet in hierdie tegnieke opgelei word.

Potato virus Y (PVY) is an economically important virus worldwide. In South Africa, PVY has been shown to be a major limiting factor in the production of important solanaceous crops, including potato (Solanum tuberosum L.), pepper (Capsicum annuum L.), tomato (Lycopersicon esculentum Mill.) and tobacco (Nicotiana spp). The variability that PVY displays, wherever the virus occurs, merits the study of the isolates occurring in KwaZulu-Natal (KZN) in the Republic of South Africa (RSA). This characterization will provide a clear understanding of strains/isolates from local vegetables and how they relate to the other PVY strains already identified, as well as information that can be used to manage the diseases they cause. Hence, the aim of this project was to study the biological and genetic properties of PVY isolates infecting potato, tomato and pepper in KZN. Enzyme-linked immunosorbent assay (ELISA) using monoclonal antibodies and reverse transcription polymerase chain reaction (RT-PCR) using primers specific to all PVY strains were used to detect the virus in plant material showing PVY-like symptoms collected from various locations in KZN. A total of 39 isolates (18 isolates infecting tomato, 12 infecting potato and 9 infecting pepper) were further differentiated into strains by means of ELISA using strain specific antibodies and RT-PCR using primers specific to the different strains of PVY identified around the world. All PVY isolates infecting tomato and pepper tested positive for the ordinary PVYO strain with both ELISA and RT-PCR. PVY isolates infecting potato were more diverse and comprised the PVYN, PVYNTN and PVYNWilga strains, with mixed infections noted in some cases. The biological properties were studied by mechanically inoculating Chenopodium quinoa, Nicotiana tabacum cv Xanthi, N. tabacum cv Samsun, N. glutinosa, and N. rustica with leaf extracts from plants infected with the different PVY strains detected in this study. All inoculated C. quinoa plants did not show symptoms. All tobacco plants showing symptoms were tested for the presence of PVY by means of ELISA using monoclonal antibodies targeting all strains and electron microscopy using the leaf dip technique. Not all the inoculated tobacco tested positive with ELISA. The symptoms observed were therefore divided into PVY-related and PVY non- related. PVY-related symptoms included vein clearing, mosaic chlorosis, stunting, and vein necrosis. PVY non-related symptoms included wrinkles and leaf distortions. Potyvirus-like particles of about 700 nm were observed under the transmission electron microscope (TEM) from plants showing PVY-related symptoms while rod shaped viral particles of sizes varying between 70 and 400 nm were observed from plants showing non-PVY related symptoms. A portion of the virus genome (1067 bp) covering part of the coat protein gene and the 3’ non-translated region (NTR) of three PVYO isolates infecting tomato, one PVYO isolate infecting pepper and one PVYNWilga isolate infecting potato were amplified, cloned and sequenced. The 5’ NTR, P1, HC-Pro and part of P3 regions (2559 bp) of a PVYN isolate infecting potato were also amplified, cloned and sequenced. Sequence data was compared with selected PVY sequences from different geographical locations around the world. These were available on the NCBI website and subsequently used for phylogenic analyses. The sequenced genomic regions of the PVYN isolate were found to be 99% similar to the New Zealand PVYN isolate (GenBank accession number: AM268435), the Swiss PVYN isolate CH605 (X97895) and the American PVYN isolate Mont (AY884983). Moreover, the deduced amino acid sequence comparison of the genomic regions of the PVYN isolate revealed the presence of five distinct amino acids residues. The three amino acid residues (D205, K400, and E419), which determine the vein necrosis phenotype in tobacco, were also identified. The coat protein and 3’ NTR sequences of all KZN PVYO isolates infecting pepper and tomato were closely similar to each other than to KZN PVYNWilga isolate infecting potato. The phylogenic analysis clustered the KZN PVYN isolate with the European sublineage N, PVYNWilga isolate infecting potato with the American PVYO isolate Oz (EF026074) in the O lineage and all PVYO isolates infecting tomato and pepper in a new sublineage within the O lineage. Taken together, these results point to the presence of PVY in solanaceous vegetables cultivated in KZN and they lay the foundation for the formulation of effective control measure against PVY diseases in KZN. / Thesis (M.Sc.) - University of KwaZulu-Natal, Pietermaritzburg, 2009.

Made available in DSpace on 2015-03-26T13:37:54Z (GMT). No. of bitstreams: 1 texto completo.pdf: 752482 bytes, checksum: a4ec60bad1c1315d52f2a91132509d98 (MD5) Previous issue date: 2014-03-11 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / Pepper yellow mosaic virus (PepYMV) naturally infects sweet pepper and tomato plants, leading to severe losses since its first report in Brazil (Brasília, DF, 2002). Molecular and serological methods can be used to detect the pathogen. Serological methods for viral detection require the use of a high quality antiserum, offering good sensitivity and specificity. Traditionally, purified viral particles are used as immunogens. However, the purification process is very laborious and the final preparation may have unsatisfactory purity and/or concentration. Thus, the aim of this work was to produce a polyclonal antiserum against the recombinant capsid protein (CP) of PepYMV to allow its use both for diagnosis and for studies of the interaction of the PepYMV CP with other viral proteins and host factors. The coding sequence of the capsid protein gene of PepYMV was cloned into an expression vector (pRSET-A) and transformed into Escherichia coli strain BL21::DE3 for in vitro expression. The recombinant protein, fused to a histidine tag, was purified under denaturing conditions by affinity chromatography using a Ni-NTA column. The purified recombinant protein was dialyzed under renaturing conditions. Its integrity and identity were confirmed by polyacrylamide gel electrophoresis and mass spectrometer analyses. New Zealand rabbits were immunized with increasing amounts of the recombinant protein. The sensitivity and specificity of the antisera were analyzed by Western blot and indirect ELISA assays. The antisera raised against the recombinant CP showed good specificity and sensibility, proving to be a reliable tool for the detection of PepYMV. / O Pepper yellow mosaic virus (PepYMV), agente causal do mosaico amarelo do pimentão e do tomateiro, desde seu primeiro relato no Brasil no ano de 2002 em Brasília, DF, vem se disseminando em regiões produtoras de pimentão e tomate causando perdas substanciais ao produtores dessas culturas. Para identificação dessa enfermidade algumas ferramentas são utilizadas, dentre elas, podemos destacar os métodos moleculares e sorológicos, sendo estes últimos mais utilizados por apresentarem alta especificidade e sensibilidade, além de possuir um custo relativamente baixo. Para a produção de anti-soro, tradicionalmente, utilizam-se partículas virais concentradas como imunógenos. No entanto, o processo de purificação é muito trabalhoso e pode apresentar pureza e concentrações insatisfatórias. Desta forma, o objetivo deste trabalho foi produzir um anti-soro policlonal, a partir da proteína capsidial recombinante do PepYMV, que permita sua utilização tanto para diagnose quanto para estudos de interação da proteína capsidial do PepYMV com outras proteínas virais e fatores do hospedeiro. A sequência do gene da proteína capsidial do PepYMV foi clonada em vetor de expressão (pRSET-A) e transformada em Escherichia coli, linhagem BL21::DE3, para expressão in vitro. A proteína expressa fusionada a uma cauda de histidina foi purificada sob condições desnaturantes por cromatografia de afinidade em coluna de resina Ni-NTA. Em seguida a proteína purificada foi dialisada sob condições renaturantes e sua integridade e identidade foram confirmadas por gel de poliacrilamida a 12% e análise de espectrometria de massa. Dois coelhos da raça Nova Zelândia foram imunizados com quantidades crescentes da proteína recombinante dialisada adicionados do adjuvante de Freud incompleto na proporção 1:1. A sensibilidade e a especificidade do anti-soro foram testados por Western blot e ELISA indireto. O anti-soro produzido apresentou boa especificidade e sensibilidade, provando ser uma ferramenta confiável para a diagnose do PepYMV.