Thesis (M. Sc. Agric.(Plant Production)) -- University of Limpopo, 2019 / Nightshade (Solanum retroflexum Dun.) is among the most important indigenous leafy vegetables in Vhembe District, Limpopo Province, South Africa, due to its high values of beta-carotene, vitamin E, folic acid, ascorbic acid, calcium, iron and protein. Vhembe District occurs in the tropical regions of Limpopo Province and the production of vegetables is dependent upon the availability of irrigation water. An Integrated Drip Irrigation System (IDIS) and a 3S planter were developed to save water by planting several plants/hole of drip irrigation system. The subsystems in IDIS allow for the production of different crops with different water requirements, whereas the 3S planter can be used for planting from one to nine plants/hole of drip irrigation system. Also, the subsystems could be used in assessing irrigation interval for crops under various planting densities. The interaction of irrigation interval and planting density of S. retroflexum had not been documented. The objective of this study, therefore, was to determine the interactive effects of irrigation interval and planting density on biomass yield and chemical nutrient elements (summer harvest only) of S. retroflexum under field conditions. The irrigation interval and planting density/hole were arranged in a split-plot experimental design, with eight replications. The main plot was irrigation interval and the subplot was the planting densities. Harvesting was done twice for both summer and winter experiments. The first harvest (H1) was done at 6 weeks after transplanting, with the second harvest (H2) being done at six weeks after the first harvest. Fresh shoots were oven-dried at 60°C for 72 h for the determination of dry matter. Mature leaves were powdered and analysed for mineral content (Ca, P, K, Mg, Na, Fe, Zn, Mn and Cu) using the ICPE-9000. Data were subjected to analysis of variance using SAS software. In the summer experiment, the interaction was significant (P ≤ 0.05) for dry shoot mass at H1 and H2. However, the contribution of xv the interaction in the total treatment variation (TTV) of the variable was negligent and therefore, only single factors were reported. Irrigation interval and planting density had highly significant (P ≤ 0.01) effects on plant variables during H1 and H2 in summer and winter. However, irrigation interval effects for dry shoot mass were not significant for summer H2. Interaction effects were significant for Ca, P, K, Mg, Mn and Cu in leaf tissues during summer H1, but were not significant for Na, Fe and Zn. Also, irrigation interval was significant for Ca, Mg, P, K, Na, Fe, Zn, Mn and Cu during summer H1, whereas planting density had no significant effects for all chemical nutrients except for Ca, P and K during summer H1. Dry shoot mass of S. retroflexum increased linearly with increasing irrigation interval and planting density. Results suggested that most nutrient elements increased with deficit irrigation water and higher planting density, whilst P decreased under high planting density. The study showed that there is a high potential for saving water through longer irrigation intervals and produce good high yields at a higher planting density. In conclusion, the use of IDIS and 3S planter to promote growth and accumulation of essential nutrient elements on S. retroflexum demonstrated that longer irrigation interval and higher plant density per drip irrigation hole could be suitable for cultivation of this indigenous vegetable. The recommendation of this study is that higher planting density and longer irrigation intervals are key determinants of higher biomass yield and water saving strategies for large-scale production of the crop. Further, the mineral composition of the crop was under the influence of higher planting density and irrigation intervals.

Thesis (M. A. Agriculture (Plant Protection)) -- University of Limpopo, 2019 / The successful cultivation of nightshade (Solanum retroflexum) as a leafy vegetable with the nutritional potential of contributing to food security in marginalised communities of Limpopo Province could be limited by high population densities of root-knot (Meloidogyne species) nematodes. However, the use of Nemarioc-AL/AG and Nemafric-BL/BG phytonematicides in suppressing nematodes and not being phytotoxic requires the empirically-developed non-phytotoxic concentration, technically referred to as Mean Concentration Stimulation Point (MCSP). The MCSP, developed using the Curve-fitting Allelochemical Response Data (CARD) computer-based model, is crop-specific, hence it should be developed for every crop. The objective of this study was to investigate the influence of Nemarioc-AL/AG and Nemafric-BL/BG phytonematicides on growth of nightshade, accumulation of essential nutrient elements and cucurbitacin residues in nightshade leaves. Microplots were established by inserting 20-cm-diameter plastic pots into 10-cm-deep holes at 0.6 m intra-row and 0.6 m inter-row spacing. Each pot was filled with 10 000 cm3 steam-pasteurised river sand and Hygromix at 3:1. After establishment, Nemarioc-AL and Nemafric-BL phytonematicides were applied at 7-day interval, whereas, Nemarioc-AG and Nemafric-BG phytonematicides were only applied at planting. Two separate experiments for Nemarioc-AL and Nemafric-BL phytonematicides were conducted in summer (November-January) 2017/2018 under microplot conditions with each comprising treatments namely; 0, 2, 4, 8, 16, 32 and 64%, similarly, two separate experiments for the following phytonematicides, Nemarioc-AG and Nemafric-BG comprised treatments namely; 0, 2, 4, 6, 8, 10 and 12 g arranged in a randomised complete block design (RCBD), with 12 replications. The nutrient elements in leaf tissues of nightshade were analysed using the inductively coupled plasma optical emission spectrometry (ICPE-9000) while, cucurbitacin A and B were xii each quantified using the isocratic elution Shimadzu HPLC Prominence with Shimadzu CTO-20A diode array detector. Plant growth and nutrient elements variables were subjected to the CARD computer-based model to generate biological indices to generate the curves, quadratic equations and the related biological indices (Dm, Rh, k) (Liu et al., 2003). The MCSP values were calculated using the biological indices of plant or nutrient element variables which, along with increasing concentration of Nemarioc-AL, Nemafric BL, Nemarioc-AG and Nemafric-BG phytonematicides, exhibited positive quadratic relations, with R2 ≥ 25. Using cucurbitacin A and B standards, residues of Nemarioc AL/AG and Nemafric-BL/BG phytonematicides, were not detected in nightshade leaves, respectively. Dry root mass and dry shoot mass of nightshade over increasing concentration of Nemarioc-AL phytonematicide each exhibited a quadratic relationship, with the models explained by 93 and 61%, respectively. Dry root mass, dry shoot mass, plant height, chlorophyll content and stem diameter against increasing concentration of Nemafric-BL phytonematicide each exhibited positive quadratic relationships with the models explained by 95, 72, 65, 78 and 62%, respectively. Plant height, stem diameter and dry root mass against increasing concentration of Nemarioc-AG phytonematicide each exhibited positive quadratic relationships with their models explained by 93, 88 and 91%, respectively. Dry shoot mass and stem diameter against increasing concentration of Nemafric-BG phytonematicide each exhibited positive quadratic relationships with their models explained by 94 and 84%, respectively. Na, Fe and K over increasing concentration of Nemarioc-AL phytonematicide each exhibited positive quadratic relationships with their associations explained by 96, 91 and 95%, respectively. Zn over increasing concentration of Nemafric-BL phytonematicide exhibited positive quadratic relationship with the model explained by 98%. Fe over increasing concentration of Nemarioc-AG phytonematicide exhibited positive quadratic xiii relationship with the association explained by 91%. Fe, Na, K and Zn over increasing concentration of Nemafric-BG phytonematicide each exhibited positive quadratic relationships with their associations explained by 81, 90, 80 and 89%, respectively, whereas, on the contrary, Zn over increasing concentration of Nemarioc-AG phytonematicide exhibited negative quadratic relationship with the association explained by 96%. Significant (P ≤ 0.05) plant variables were subjected to CARD, to generate biological indices which were used to compute the MCSP using the relation: MCSP = Dm + Rh/2 and the overall sensitivity value (∑k). In Nemarioc-AL phytonematicide trial, MCSP = 3.02% and ∑k = 1 for plant variables, whereas, MCSP and ∑k for nutrient elements were 12.09% and 1, respectively. In Nemafric-BL phytonematicide trial, MCSP = 3.08% and ∑k = 0 for plant variables, while MCSP = 2484.14% and ∑k = 0 for nutrient elements. In Nemarioc-AG phytonematicide trial, MCSP = 3.47 g and ∑k = 0 for plant variables, whereas, for nutrient elements MCSP = 8.49 g and ∑k = 1. In Nemafric-BG phytonematicide trial, MCSP = 4.70 g and ∑k = 0 for plant variables, whereas, MCSP =723.75 g and ∑k = 1 for nutrient elements. In conclusion, the application of Nemarioc-AL/AG and Nemafric-BL/BG phytonematicides had the ability to stimulate the growth of nightshade and enhance the accumulation of the selected nutrient elements without leaving cucurbitacin chemical residues in leaf tissues of nightshade. / National Research Foundation (NRF) and the Land Bank Chair of Agriculture

Orientador: Rogério Falleiros Carvalho / Resumo: O nitrogênio (N), elemento essencial ao crescimento e desenvolvimento vegetal, é absorvido principalmente como nitrato (NO3-). Esse elemento pode controlar a arquitetura de raízes e dessa forma favorecer a absorção de nutrientes pelas plantas. Para isso, tem sido mostrado que esses eventos estão relacionados ao controle hormonal, especialmente as auxinas, pois esse hormônio controla o crescimento das raízes de modo dependente de NO3-. Entretanto, esses mecanismos são complexos, pois dependem de várias moléculas, especialmente peróxido de hidrogênio (H2O2), o qual parece atuar nas sinalizações das auxinas em situações estressoras nas plantas, a exemplo da deficiência de N. Entretanto, a forma como auxinas e H2O2 interagem nas raízes sob deficiência de N foi pouco explorado. Assim, objetivou-se verificar se as auxinas dependem de H2O2 nas respostas das raízes de tomateiro à deficiência de NO3-. Para isso, plantas de da cultivar Micro-Tom de tomateiro foram cultivadas em hidroponia sob suficiência e deficiência de NO3-, ambas com presença e ou ausência de inibidor de transporte de auxinas, o (ácido 2,3,5-triiodo-benzóico- TIBA) e inibidor do acúmulo de H2O2 (Catalase). O delineamento experimental foi inteiramente casualizado, disposto em fatorial 2x4, com três repetições. Foi analisado comprimento e biomassa seca da parte aérea e biomassa seca, comprimento, área e volume das raízes. Além disso, foi quantificado conteúdo de MDA e H2O2. A deficiência de NO3- diminuiu o crescimento... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Nitrogen (N) is essential for plant growth and development, and is mainly absorbed as nitrate (NO3-). It also controls root architecture, improving plant nutritional status. These characteristics are related to hormonal control, especially by auxins. Auxins control root growth via NO3-dependent cell expansion, but these mechanisms are complex and depend on several molecules. Hydrogen peroxide (H2O2) appears to act on these auxins signalling in plant stress situations. However, how auxins and H2O2 interact in roots under N deficiency has not been well explored. Therefore, this study aimed to verify if auxins depend on H2O2 for responses of tomato roots to N deficiency. Micro-Tom plants were grown in hydroponic systems under sufficiency and deficiency of NO3-, both with and without auxin transport inhibitor, 2,3,5-triiodo-benzoic acid (TIBA), and the inhibitor of H2O2 accumulation (Catalase) in plants. The experiment was carried out in a completely randomized design in a factorial scheme (2x4) with three replications. Shoot length and dry biomass, as well as root dry biomass, length, area and volume were analysed. In addition, MDA and H2O2 content were also quantified. NO3- deficiency reduced shoot growth and, interestingly, promoted higher root growth. Furthermore, TIBA inhibited root growth under both NO3- conditions and the presence of catalase in NO3-deficiency resulted in lower root growth. These results show that auxins depend on H2O2 for tomato root responses to N defici... (Complete abstract click electronic access below) / Mestre

Meloiodgyne incognita, one of the most aggressive plant parasitic nematodes species on potato in South Africa, belongs to a group of plant parasitic nematodes commonly known as root knot nematodes (RKN). This group of nematodes is widely distributed throughout the world. Meloidogyne spp. cause major economic losses to important crops such as potato and therefore decrease their market value in many countries across the world. The second stage juveniles are the only mobile and infective phase of the root knot nematode. As they infect host roots, they create wounds that can be used by other plant pathogens to penetrate the host in large numbers. The most effective management strategy for root knot nematodes is the use of nematicides such as Temik and Methyl bromide. However, these have been banned due to adverse on the environment. Therefore, Meloidogyne spp. will inevitably become a big problem in the potato industry of many countries due to the lack of effective alternatives to banned chemicals. Pectobacterium carotovorum subsp. brasiliensis (Pcb) is one of the most important soft rot-causing agents in South Africa. This pathogen belongs to a group of pathogens commonly known as soft rot Enterobacteriaceae (SRE). Bacteria belonging to this group of pathogens are known to cause soft rot and blackleg diseases on potato and other crops. Pcb is known as an opportunistic pathogen that can only penetrate host root tissue through natural openings or wounds that result from a variety of agents. Post penetration, the bacteria will increase in number and cause soft rot and blackleg. As rotting plant tissue disintegrates the bacteria escapes into the soil where it serves as inoculum and can infect healthy hosts. Many interactions have been documented between Meloidogyne spp. and other plant pathogens but to our knowledge there are no interactions that have been reported between Meloidogyne spp. and Pectobacterium spp. Considering the life cycles of RKN and SRE, we hypothesised that there could be an interaction between the two pathogen groups. Since both RKN and SRE are potato pathogens, they share the same space in the rhizosphere. This likely can lead to synergies and complex formation between the two pathogens. Likely, the wounds created by RKN J2s as they penetrate plant tissue can potentially be used by opportunistic Pcb to infect various hosts. It is from these identified overlaps that the first part of this study focused on investigating the potential interaction between M. incognita and Pcb. The first objective was to determine whether Pcb can attach onto M. incognita J2s and, if this was the case, to determine whether the J2s can disseminate the bacteria as they move around in the environment. The second objective was to determine whether there is a synergistic interaction between RKN and SRE and the combined effect of the two pathogens on their host Solanum tuberosum cv Mondial. The results obtained in the first part of the study strongly suggested that Pcb can attach onto M. incognita J2s and can be disseminated as the J2s move around in the environment. Thus, this indicated that there is a synergistic interaction between M. incognita and Pcb as there was increased disease severity and incidence in plants inoculated with both pathogens compared to those inoculated with individual pathogens. Significantly higher Pcb concentrations were found in plants inoculated with both pathogens. There was no breakage of tolerance to Pcb-caused blackleg on an otherwise resistant cultivar, BP1. The second aim of this study was to determine whether the induction of natural resistance using environmentally friendly resistance inducing chemicals can potentially be used as an alternative to chemical control. To this end, the effect of three inducers at different concentrations, amongst DL-β-aminobutyric acid, Acibenzolar-s-methyl and Messenger on potato plants infected with RKN was compared. The most effective resistance inducer amongst the three was 20mM BABA as it was able to reduce the number of J2s that penetrated host tissue, the number of females in the roots and the rate of egg production. Furthermore, the galling index observed in potato roots was significantly lower when plants were treated with 20mM BABA. Additionally, the reduced rate of RKN infection in plants primed with 20mM led to a decrease in the rate of Pcb infection. / Dissertation (MSc)--University of Pretoria, 2013. / gm2013 / Microbiology and Plant Pathology / Unrestricted

Potato (Solanum tuberosum) is regarded as one of the single most important vegetable crops in South Africa, with an average annual production of 2 million metric tons. The potato industry contributes to an average of $ 0.37b worth of potatoes annually. Over the years, potato production in South Africa has been affected by, among other factors, diseases and plant parasitic nematodes particularly root-knot nematodes (Meloidogyne spp.). In infected potato fields, root-knot nematodes cause great damage to the crop leading to substantial losses in yield and compromised produce quality. The direct and indirect damage caused by Meloidogyne species results in revenue loss due to a high number of table and processing potatoes rejected in markets both locally and internationally. The presence of resistance breaking Meloidogyne populations, the withdrawal of methyl bromide and lack of commercially grown resistant cultivars suggests that growers are likely to experience more losses in the future. Furthermore, distribution of seed tubers harbouring root-knot nematodes, which may also be asymptomatic, inadvertently facilitates transmission of these parasites to new areas thus perpetuating the problem. Therefore, for the potato industry to adequately address the threat of root-knot nematodes, accurate identification and quantification of root knot nematode levels in the field as well as in seed tubers is of importance. Currently most methods of identifying Meloidogyne species largely rely on the use of morphological traits. However, it can be a challenge to accurately differentiate between closely related species using morphology and other classical methods. To resolve this, recent trends globally have focused on the development of DNA-based diagnostics to rapidly and accurately identify different Meloidogyne species. This study therefore sought to; (a) develop a PCR-based diagnostic tool for accurate detection and identification of various Meloidogyne species parasitizing potatoes in South Africa; (b) use this tool to map their distribution and; (c)develop real-time PCR (qPCR) techniques for accurate quantification and characterization of tropical Meloidogyne species from infected potato tubers. In this study, of the 78 composite potato tuber samples collected from various potato growing regions across seven provinces, 24% were found infected with M. javanica, 23% with M. incognita, 17% with M. arenaria, 14% with M. enterolobii, 3% M. chitwoodi, 1% M. hapla and 1% as M. artiellia. The identity of the remaining 17% could not be established. The three tropical species; M. javanica, M. incognita and M. arenaria were identified as the dominant species, occurring almost in every region sampled. Meloidogyne hapla and M. enterolobii occurred in Mpumalanga and KwaZulu–Natal respectively while M. chitwoodi was isolated from two growers located within the Free State. In the study the use of HRMC and real-time PCR was also developed for identification and quantification of tropical Meloidogyne species infesting potato tubers. Using these two techniques, we were able to show that Meloidogyne arenaria populations produced specific melting peaks (79.3183± 0.0295°C, P < 0.05) thus distinguishing themselves from M. incognita (79.5025± 0.0224°C, P < 0.05) and M. javanica (79.96 ± 0.0459°C, P < 0.05). Real-time PCR was also able to detect 1.53/100th of a nematode using second stage juveniles. / Dissertation (MSc)--University of Pretoria, 2012. / Microbiology and Plant Pathology / unrestricted

Orientador: João Domingos Rodrigues / Resumo: Dois experimentos foram conduzidos em 2018 e 2019 para avaliar os efeitos de diferentes fungicidas de dois importantes grupos: estrobilurinas e carboxamidas, sobre a eficácia fotossintética e produtividade de plantas de tomate, inoculadas com Alternaria sp., em ambiente protegido na Fazenda Ituaú, Salto - SP, Brasil. O delineamento experimental aplicado em ambos os experimentos foi de blocos ao acaso, com oito tratamentos e cinco repetições: testemunha inoculada (água + inóculo), testemunha absoluta (água), azoxystrobina (50,0 g ha-1), boscalida (50,0 g ha-1), azoxystrobina (50,0 g ha-1) + boscalida (50,0 g ha-1), mandestrobina (194 g ha-1), impirfluxam (30 g ha-1), mandestrobina (194 g ha-1) + impirfluxam (30 g ha-1), aplicados em intervalos de 15 dias, totalizando 7 a 8 aplicações no ciclo da cultura. O efeito no desenvolvimento das plantas foi medido pela altura das plantas e produção dos cinco primeiros cachos. A eficiência fotossintética foi calculada pela medição das trocas gasosas, fluorescência da clorofila a, teores de pigmentos fotossintéticos, e produtividade. Os fungicidas apresentaram algumas melhorias nos parâmetros analisados, como melhorando os componentes da eficiência fotossintética e da produção de frutos comerciais, porém sem apresentar diferenças significativas em relação às testemunhas. Os resultados demonstram eficácia no controle de Alternaria sp. aplicando os fungicidas em tomateiro, e também apontam para efeitos adicionais em componentes da eficiênci... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Two experiments were carried out in 2018 and 2019 to evaluate the effects of the application of different fungicides from two important groups: strobilurins and carboxamides, on the photosynthetic efficiency and yield of tomato plants, inoculated with Alternaria sp., grown in greenhouse at Fazenda Ituaú, Salto - SP, Brazil. The experimental design applied in both trials was randomized blocks with eight treatments and five replicates: inoculated control (water + inoculum), absolute control (water), azoxystrobin (50,0 g ha-1), boscalid (50,0 g ha-1), azoxystrobin (50,0 g ha-1) + boscalid (50,0 g ha-1), mandestrobin (194 g ha-1), inpyrfluxam (30 g ha-1), mandestrobin (194 g ha-1) + inpyrfluxam (30 g ha-1), applied at intervals of 15 days, totalizing 7 or 8 applications in the crop cycle. The effect on plant development was measured by plant height and the production of the five initial bunches. The photosynthetic efficiency was calculated from measurements of gas exchange, chlorophyll a fluorescence, content of photosynthetic pigments, and tomato productivity were evaluated. All fungicides positively affected some evaluated parameters, improving photosynthetic efficiency components and commercial fruit production, however without significant difference to the controls. The results demonstrate the efficacy of the use of these fungicides in controlling Alternaria sp. in tomato and also appoint to some additional effects on photosynthesis efficiency components and antioxidant enzym... (Complete abstract click electronic access below) / Doutor

Orientador: Leonardo Bianco / Resumo: Sendo a beldroega uma planta infestante da cultura do tomate e uma hospedeira natural de nematoides do gênero Meloidogyne, levanta-se a hipótese de que a ocorrência dessa planta daninha em áreas de plantios de tomate possa interferir no crescimento e produtividade do tomateiro, dependendo do período de convivência, e que existe uma relação da sua ocorrência com a infestação de M. incognita no tomateiro, e que é possível controlá-la com seletividade ao tomateiro. Dessa forma, com este estudo, objetivou-se: a) avaliar se há interferência da beldroega no crescimento e produtividade do tomateiro; b) determinar o período de convivência da planta daninha com a cultura que não afete o crescimento e a produtividade do tomateiro; c) avaliar se a beldroega infestando a cultura do tomate atua como hospedeira de Meloidogyne javanica e com isso reduz a sua infestação no tomateiro; d) estudar a seletividade de herbicidas ao tomateiro visando o controle da beldroega em pré e pós-emergência e) verificar se a velocidade de controle do metribuzim elimina de imediato a interferência dessa planta daninha no tomateiro. Para tanto, esse estudo constou de três experimentos. No primeiro, os tratamentos experimentais consistiram de três períodos de convivência do tomateiro „Dylla‟ com a beldroega (14, 21 e 60 dias após o transplantio), com um tratamento para tomate sem convivência com a planta daninha e outro tratamento para a beldroega sem convivência com a cultura, todos com ou sem a inoculação de ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Purslane is an infesting plant in tomato culture and a natural host of nematodes of the genus Meloidogyne, the hypothesis arises that the occurrence of this weed in tomato plantation areas may interfere in tomato growth and productivity depending on the period coexistence between them. Also that is a relationship between it ́s occurrence and the infestation of M . incognita in tomato and that it is possible to control it with tomato selective products. This study aimed to: a) assess whether purslane interferes with tomato growth and productivity; b) determine the period of coexistence of the weed with the crop that does not affect the growth and productivity of the tomato; c) to evaluate whether purslane infesting the tomato crop acts as host of Meloidogyne javanica and thereby reduces its infestation in tomato; d) to study the selectivity of herbicides for tomato aiming at the control of purslane in pre and postemergence e) to verify if the speed of control of the metribuzim immediately eliminates the interference of this weed in the tomato. This study consisted of three experiments. In the first, the experimental treatments consisted in three coexistence periods of tomato 'Dylla' with purslane (14, 21 and 60 days after transplanting), with a treatment for tomatoes without living with weeds and another treatment for purslane without coexistence with culture, all with or without Meloidogyne javanica inoculation. The treatments were arranged in a randomized block design, in 6 ... (Complete abstract click electronic access below) / Mestre

La producción de alcohol se encuentra relacionada directamente con la producción de Biocombustibles, debido al desplazamiento de importantes volúmenes de alcohol para su conversión en etanol o alcohol anhídrido; usado como complemento o sustituto de la gasolina. Esta situación ha generado que la caña de azúcar, principal materia prima para la elaboración de alcohol se convierta en un “bien escaso” y sea necesario buscar otras potenciales fuentes para su producción. En el Perú, existe también el potencial para producir el mencionado producto, usando los tubérculos como base, especialmente la papa; a través de la conversión de almidones en azúcares y su posterior fermentación y destilación en alcohol etílico. La presente investigación tiene por finalidad demostrar la viabilidad de la instalación de una planta de producción de alcohol de papa en la provincia de Chota, departamento de Cajamarca, con la intención de atender al mercado interno, con un producto de alta calidad, orientado al uso industrial, farmacéutico y cosmético. La metodología desarrollada comprende la determinación de viabilidad de Mercado, Organizacional, Técnico Operativa, Económica financiera y ambiental. Finalmente se concluye que es viable la instalación de una planta de producción de alcohol en la provincia de Chota.

Phytophthora erythroseptica Pethyb. is a devastating fungal pathogen of potato (Solanum tuberosum). The pathogen causes a disease known as pink rot. Pink rot results in necrosis and decay of tubers; and is responsible for major losses pre and post harvest. Attributes of the disease are progressing toward epidemic proportions. To help prevent such dramatic outbreaks of the disease, understanding the factors associated with incidence will provide opportunities to control the pathogen. A link between pH and disease severity has been observed. We studied the effects of pH and Ca to determine their influence on disease development. Low pH and Ca deficiencies are often inter-related factors that can be causal of one another. The pH effect could be due to either H+ or Ca ions. To separate their interactive effects we tested each variable individually using hydroponics and nutrient solutions. We assessed disease severity by assigning an Infection Coefficient (IC) to each root and stolon samples. The IC values were determined by quantifying DNA and then creating a ratio of host to pathogen DNA within root tissue. The DNA was measured by quantitative RT PCR. Statistical analysis showed significance in greater pathogen presence at more acidic pH and lower levels of available Ca. Significant reductions in IC values were observed when pH was elevated above pH 7. There was a notable increase in colonizing pathogen DNA at pH 5. Ca was significant, and as levels of soluble Ca increased the degree of disease severity became smaller. The effect of Ca was found not to be dependent upon pH. These results suggest immediate and cost effective applied management strategies to reduce incidence and disease severity outbreaks. Amending the soil with lime to increase soil pH and Ca content is one such potential method.

Pathogen derived resistance (PDR) is a recent breakthrough where plant hosts can be made to be resistant to viral infections through transformation with conserved viral genes. Given the severity of Tospovirus diseases worldwide (particularly in tomato), PDR has the potential to garner large yield returns where pathogen populations have overcome the established resistance. Tomato breeding lines FLA7804, FLA8044, and the research line MP1 were used in transformation experiments with potions of the Tomato spotted wilt virus (TSWV) N-gene, and two other chimerical viral nucleocapsid gene constructs from, Impatiens necrotic spot virus (INSV), and Groundnut ringspot virus (GRSV). We conducted 19 independent transformations consisting of 300 to 700 14-day old whole cotyledons each for a total number of approximately 9,000 potentially transformed explants. Of those, approximately 6,300 explants failed to produce regenerants, 2,419 explants underwent abnormal development on elongation media, 187 failed to root, and 215 plants to be characterized genetically. Of the 215 plants, 9 were from FLA 7804, 96 from FLA 8044, and 110 from MP1. Both PCR and Southern blot hybridization analysis later confirmed that none of the 215 plants were transgenic. Opposite to tomato, we were able to transform Arabidopsis thaliana ecotype wassilewskija (Ws) via floral dip with the above listed constructs demonstrating that constructs were not deleterious within a plant once fully introgressed. Sixteen independent transformants in the T0 generation resulted from 19,000 germinated seed from three dipped plants resulting in a total transformation rate of 0.08%. Of the 1,000 T1 seed germinated on kanamycin media from each of the 16 putative Arabidopsis plants transformed with the construct containing elements of the N-gene from all three of the aforementioned tospoviruses, four populations exhibited simple Mendelian inheritance of the transgene. DNA walking analysis yielded amplification of the unknown region outside the nptII region of the insert for three of the four remaining transformants, which was subsequently sequenced and mapped to chromosomes 1, 3, and 4. There were 25 T1 individuals selected from each population and transferred to soil for DNA extraction and zygosity determination. Homozygous T2 seed was collected for future resistance studies.