The nematodes from the genus Meloidogyne are the most important causal agents of plant damages considering that they have a large geographical distribution and they are difficult to be controlled. Considering that the actual information about species from the genus Meloidogyne infecting plants in agriculture areas are still short in the State of CearÃ, the present research was developed with the following objectives: identify Meloidogyne species and races which occur in the different micro-regions from the State of Cearà and identify the natural hosts for M. enterolobii in guava (Pisidium guajava) orchards. A hundred and twelve plants with gall symptoms in the roots were collected from producing areas of 29 counties including 13 micro-regions from the State of CearÃ. It was observed that 112 nematode populations collected from infected plants, 46 presented phenotype typical of M. enterolobii, 27 of M. incognita, 15 of M. javanica, five of M. arenaria and one of M. hapla. Six nematode populations presented esterase phenotype distinct from those known for the Meloidogyne species already described in Brazil. In the physiologic race determination it was found races 2 and 3 of M. incognita, and race 1 of M. arenaria and a race 2 of M. javanica. The following nematode associations found in the present paper were not yet mentioned which could demonstrate that they could be the first report in the State: M. incognita in Beta vulgaris var. cicla L., B. vulgaris L., Capsicum chinensi Jacq. and Spondia tuberosa x S. mombim; M. javanica in Sieges beckiaorientalis L., S. mombim L., Hybanthus ipecacuanha (L.) Oken. and Abelmoschus esculentus L.; M. arenaria in Impatiens walleriana L., Morinda citrifolia L. and Duranta repens L. var. aurea; and M. hapla in Rosa sp. Meloidogyne enterolobii was identified in all guava root samples collected in the orchards distributed in 13 counties from the State. Besides guava, M. enterolobii was also detected in Malpighia glabra L., Ipomoea batatas (L.) Lam, Solanummelongena L., Cactus sp., Emilia fosbergii Nicolson, Hypericum sp., Inga edulis Mart., Carica papaya L., Ocimum basilicum L., Solanum americanum Mill, S. paniculatum L., Gladiolus sp. and C. frutescens L. The present research is the first report about those plant nematode associations in the State of CearÃ. The specie M. enterolboii was present in 52% of the counties and in 77% of the micro-regions visited, while M. incognita and M. javanica were detected only in 35% and 31% of the visited counties, respectively, and in 46% of the micro-region visited. The present research will contribute to update the scientific information about the occurrence of Meloidogyne species in the main agriculture producing regions from the State of CearÃ. / Os nematoides pertencentes ao gÃnero Meloidogyne estÃo entre os maiores agentes causadores de danos em plantas, pois possuem ampla distribuiÃÃo geogrÃfica e sÃo de difÃcil controle. Considerando que as informaÃÃes atualizadas sobre as espÃcies de Meloidogyne afetando plantas em Ãreas de produÃÃo agrÃcola ainda sÃo escassas no Estado do CearÃ, conduziu-se a presente pesquisa com os seguintes objetivos: identificar espÃcies e raÃas de Meloidogyne que ocorrem nas diferentes microrregiÃes do Estado do Cearà e identificar hospedeiras de M. enterolobii em pomares de goiabeira. Cento e doze amostras de plantas infestadas foram coletadas em 29 municÃpios em Ãreas produtoras do estado pertencentes a 13 diferentes microrregiÃes. Verificou-se que das 112 populaÃÃes obtidas nas coletas, 46 apresentaram fenÃtipos tÃpicos de M. enterolobii, 27 de M. incognita, 15 de M. javanica, cinco de M. arenaria, uma de M. hapla. Seis populaÃÃes apresentaram fenÃtipos de esterase distintos daqueles conhecidos para as espÃcies de Meloidogyne jà relatadas no Brasil. Na determinaÃÃo das raÃas fisiolÃgicas, foram encontradas as raÃas 2 e 3 para M. incognita, a raÃa 1 para M. arenaria e a raÃa 2 para M. javanica. As seguintes associaÃÃes encontradas neste trabalho nÃo foram ainda mencionadas podendo ser os primeiros relatos no estado: M. incognita em acelga (Beta vulgaris var. cicla L.), beterraba (B. vulgaris L.), pimenta de cheiro (Capsicum chinensi Jacq.) e umbu-cajà (Spondia tuberosa x S. mombim); M. javanica em botÃo de ouro (Siegesbeckia orientalis L.), cajà (S. mombim L.), papaconha (Hybanthus ipecacuanha (L.) Oken.) e quiabo (Abelmoschus esculentus L.); M. arenaria em maria-sem-vergonha (Impatiens walleriana L.), noni (Morinda citrifolia L.) e pingo de ouro (Duranta repens L. var. aurea); M. hapla em roseira (Rosa sp.). A espÃcie M. enterolobii foi identificada em todas as amostras de raÃzes de goiabeira coletadas em pomares distribuÃdos em 13 municÃpios do estado. AlÃm da goiabaeira, M. enterolobii foi constatada tambÃm em acerola (Malpighia glabra L.), batata doce (Ipomoea batatas (L.) Lam), berinjela (Solanum melongena L.), cactos (Cactus sp.), falsa serralha (Emilia fosbergii Nicolson), Hypericum sp, ingà (Inga edulis Mart.), mamÃo (Carica papaya L.), manjericÃo (Ocimum basilicum L.), maria-pretinha (Solanum americanum Mill), jurubeba (S. paniculatum L.), palma (Gladiolus sp.) e pimenta tabasco (C. frutescens L.), associaÃÃes estas relatadas pela primeira vez no Estado do CearÃ. A espÃcie M. enterolboii estava presente em 52% dos municÃpios e em 77% das microrregiÃes visitadas enquanto que M. incognita e M. javanica foram constatadas em 35% e 31% dos municÃpios, respectivamente, ambas em 46% das microrregiÃes visitadas. Este estudo vem contribuir na atualizaÃÃo das informaÃÃes sobre a ocorrÃncia de espÃcies de Meloidogyne nas principais regiÃes produtoras do Estado do CearÃ.

Thesis (M.Sc. (Plant Production)) -- University of Limpopo, 2013 / Two phytonematicides were researched and developed from fermented crude extracts of wild watermelon (Cucumis africanus) and wild cucumber (Cucumis myriocarpus) fruits for use as alternatives to methyl bromide in managing root-knot (Meloidogyne species) nematodes in tomato (Solanum lycopersicum) production. Fruits of C. africanus contain cucurbitacin B (C32H48O8), while those of C. myriocarpus contain cucurbitacin A, which comprises cucumin (C27H40O9) and leptodermin (C27H38O8). Phytonematicides from C. africanus and C. myriocarpus fruits are referred to as nemafric-B and nemarioc-A, respectively. The two phytonematicides, due to their origin from plant species with allelochemicals, have high potential of being phytotoxic to crops. The use of the Curve-fitting Allelochemical Response Dosage (CARD) computer-based model assisted in the establishment of concentrations which were stimulatory to growth of tomato (Solanum lycopersicum) plants, while exhibiting nematoxic properties to Meloidogyne species. The two phytonematicides were developed from crude extracts of fruits dried at 52˚C in air-forced ovens and ground in a Wiley mill through 1-mm-opening sieves. However, equipment for drying and grinding fruits would not be accessible to smallholder farmers who wished to prepare their own products on-farm. The objective of this study therefore, was to determine whether nemafric-BL and nemarioc-AL produced from fresh fruit of the two Cucumis species would be suitable for use (i.e. non phytotoxic) in tomato production for managing population densities of M. incognita race 2. In order to distinguish the products of fresh (F) fruits from those of dried (D) fruits, they were code-named nemafricF-BL or nemariocF-BL and nemafricD-BL or nemariocD AL, respectively, where G and L denoted granular and liquid formulations, respectively. Tomato cv. ‘Floradade’ seedlings were infested with 3 000 eggs and second-stage xv juveniles of M. incognita race 2. An equivalent of 40 g and 80 g dried fruit mass of nemafric-B and nemarioc-A, namely, 284 g and 411 g fresh fruit mass for nemafric-B and nemarioc-A, respectively, were separately fermented using EMROSA effective micro-organisms mixed with 16 L chlorine-free tapwater in 20 L container for 14 days at ± 25˚C, allowing pH to gradually decline to ± 3.7. Separate experiments for each product run concurrently. Treatments, namely, 0, 2, 4, 8, 16, 32 and 64% concentrations, where for instance, 2% = 20 ml/1000 ml x 100, were arranged in a randomised complete block design, with 10 replications. Blocking in the greenhouse was done for wind direction which was regularly erected by fans for cooling down the greenhouse. At 56 days after weekly application of each treatment, flower number, fruit number, dry shoot mass, dry root mass, dry fruit mass, plant height, stem diameter and nematode numbers were each subjected to analysis of variance. Nematode data were, prior to analysis, transformed using log10(x + 1), but untransformed data were reported. Using the sum of squares, nemafric-BL and nemarioc-AL treatments affected dry root mass, dry shoot mass, flowers number, fruit number, plant height and stem diameter. Nemafric-BL contributed 67%, 78%, 58%, 43%, 60% and 26%, while nemarioc-AL contributed 71%, 61%, 19%, 35%, 34% and 24% to total treatment variation of the six respective variables. Plant variables with significant (P ≤ 0.05) treatment effects were further subjected to the CARD model to generate seven biological indices, with three distinct phases, namely, stimulation, neutral and inhibition phases. Using the quantified stimulation phase, the mean concentration stimulation range (MCSR) was computed for each variable using two biological indices, namely, threshold stimulation point (Dm) and saturation point (Rh). The CARD model explained 98%, 99%, 98% and 98% of the quadratic models of dry root mass, dry shoot mass, plant height and stem diameter, xvi respectively, against increasing concentrations of nemarioc-AL. Similarly, the CARD model explained 99%, 96%, 84% and 93% of total treatment variation in the respective plant variables. The integrated MCSR [MSCR = Dm + (Rh/2)] for nemafric-BL on tomato plants was 7%, while that for nemarioc-AL was 4%. In the CARD model, the overall sensitivities (∑k) of tomato plants exposed to nemafric-BL and nemarioc-AL were 3 units and 5 units, respectively. Tomato plants were therefore, less sensitive to nemarioc-AL since it had higher ∑k value than nemafric-BL. At 4% nemarioc-AL and at 7% nemafric-BL, the two phytonematicides were each highly suppressive to population densities of M. incognita race 2. In conclusion, on the basis of non-phytotoxicity of the computed MCSR values and their suppressive effects on population densities of M. incognita race 2, the smallholder farmers could produce nemafric-BL and nemarioc-AL phytonematicides on-farm. However, the production of the two products from fresh fruits would not be sustainable since fruits of the two Cucumis species are highly seasonal due to the high incidence of post-harvest decays. / The Land Bank Chair of Agriculture – University of Limpopo, Limpopo Agro-processing Technology Station,and the Flemish Interuniversity Council of Belgium

Thesis (MSc. (Horticulture)) -- University of Limpopo, 2016. / Pelargonium sidoides has numerous medicinal applications, with economic potential to serve as a future cultigen in smallholder farming systems. However, it is highly susceptible to the root-knot (Meloidogyne species) nematodes, without any identifiable nematode resistant genotypes. Nemarioc-AL and Nemafric-BL phytonematicides, with cucurbitacin A and cucurbitacin B active ingredients, respectively, are being researched and developed as an alternative to synthetic nematicides at the University of Limpopo. However, since active ingredients in phytonematicides are allelochemicals, the two phytonematicides have the potential of inducing phytotoxicity on crops protected against nematode damage. The objectives of the study, therefore, were (1) to determine the non-phytotoxic concentration of Nemarioc-AL phytonematicide on plant growth of P. sidoides, and (2) to determine the non-phytotoxic concentration of Nemafric-BL phytonematicide in plant growth of P. sidoides. Cuttings were raised in 30-cm-diameter plastic pots containing 10 000 ml steam-pasteurised river sand and Hygromix-T at 3:1 (v/v) under microplot conditions in autumn (March-May) and repeated in spring (August October) 2015. After establishment each plant was inoculated with 5 000 eggs and second-stage juveniles (J2s) of M. javanica. Six treatments, namely, 0, 2, 4, 6, 8 and 10% concentrations of each phytonematicide on separate trials were arranged in a randomised complete block design, with seven replicates. At 56 days after inoculation, in Experiment 1, Nemarioc-AL phytonematicide, treatment significantly (P ≤ 0.05) affected plant height, dry root mass and root galls, contributing 62, 69 and 70% to total treatment variation of the three variables, respectively. Relative to untreated control Nemarioc-AL phytonematicide increased plant height and dry root mass by 34 to 61% xxi and 20 to 76%, respectively, with a slight decrease by 5% in plant height at the highest concentration. However, the material decreased root galls by 5 to 50%. Significant (P ≤ 0.05) plant variables were subjected to Curve fitting-allelochemical respond dosage model, to generate biological indices which were used to compute the mean concentration stimulation point (MCSP) using the relation: MCSP = Dm + Rh/2 and the overall sensitivity value (∑k). In Experiment 1, MCSP = 6.18% and ∑k = 3. Plant variables and increasing concentration of phytonematicide exhibited quadratic relations. Treatments reduced nematode variables, at all levels including at the lowest, but the effect were not different. In Experiment 2, Nemarioc-AL phytonematicide treatment effects were not significant on plant variables except for root galls, but were significant for root nematodes except for eggs. Data for plant variables in Experiment 2 were not subjected to Curve fitting-allelochemical respond dosage model because they were not significant (P ≤ 0.05). In Experiment 1, Nemafric-BL phytonematicide treatment significantly (P ≤ 0.05) affected plant height and root galls, contributing 63 and 67% to total treatment variation of the two variables, respectively. Relatively to untreated control, plant height was increased by 10 to 36%, while root galls was reduced by 2.43 to 60%. In Experiment 1, MCSP = 2.87% and ∑k = 3. Concentrations of Nemafric-BL phytonematicide significantly (P ≤ 0.05) reduced eggs, juveniles and Pf at all levels including at the lowest, but the effect were not significant different, with treatments contributing 78, 72 and 90% to the total treatment variation. In Experiment 2, Nemafric BL phytonematicide treatment effects were not significant on plant variables except for root galls, but were significant for root. In conclusion, Nemarioc-AL and Nemafric-BL xxii phytonematicides could be applied at the lowest concentration of 2% where it was shown to be effective in suppressing population densities of M. javanica. / Agricultural Research Council (ARC), National Research Fund (NRF) , Flemish Inter university Council of Belgium and Land Bank Chair of Agriculture ─ University of Limpopo

Thesis (M.Sc. (Horticulture)) -- University of Limpopo, 2017. / Root-knot (Meloidogyne species) nematodes are economically destructive pathogens of over 3000 species, whereas others have resistance to Meloidogyne species. Wild watermelon (Cucumis africanus) and wild cucumber (Cucumis myriocarpus) are highly resistant to Meloidogyne species, particularly M. incognita and M. javanica. The two Cucumis species are used in inter-generic grafting with watermelon (Citrullus lanatus) as nematode resistant rootstocks. Also, the two Cucumis species are used in traditional medicine and in plant-parasitic nematode management as phytonematicides. The form of nematode resistance, which is essential in plant breeding, is not documented for the two Cucumis species. The objective of this study was to determine the form of nematode resistance in the two Cucumis species to M. incognita and M. javanica under greenhouse conditions. Four parallel experiments were each conducted under greenhouse conditions. Uniform six-week old Cucumis seedlings were transplanted into 250 ml polystyrene cups filled with 200 ml growing medium of steam-pasteurised fine sand. A week after transplanting, Cucumis seedlings were each infested by dispensing approximately 100 M. incognita second-stage juveniles (J2) or M. javanica J2 using a 20 ml plastic syringe by placing into 5-cm-deep furrow around the seedling stem and covered with growing medium. Treatments (periodic harvest intervals) were arranged in a randomised complete block design, replicated five times. Five seedlings from each experiment were harvested every second day, for 30 days, with stained roots being assessed for necrotic spot (suberised cells) number, giant cell number, proliferation of rootlet interference number and root gall number. Periodic harvest intervals were highly significant (P ≤ 0.01) on necrotic spot number, proliferation of rootlet interference number and root gall number in C. africanus-M. incognita relations, but were not significant for giant cell number. Treatments contributed 59, 64 and 50% in total treatment variation (TTV) of necrotic spot number, proliferation of rootlet interference number and root gall number, respectively. Harvest period had highly significant effects on necrotic spot number, giant cell number, proliferation of rootlet interference number and root gall number in C. africanus-M. javanica relations. Treatments contributed 55, 71, 63 and 59% in TTV of necrotic spot number, giant cell number, proliferation of rootlet interference number and root gall number, respectively. Periodic harvest intervals were significant (P ≤ 0.05) on giant cell number and highly significant on root gall number in C. myriocarpus-M. incognita relations. However, there were no significant treatment differences on necrotic spot number and proliferation of rootlet interference number. Treatments contributed 57 and 57% in TTV of root gall number and giant cell number, respectively. Harvest period had highly significant effects on giant cell number, proliferation of rootlet interference number and root gall number, but were not significant on necrotic spot number in C. myriocarpus-M. javanica relations. Treatments accounted for 67, 49 and 53% in TTV of giant cell number, proliferation of rootlet interference number and root gall number, respectively. In conclusion, the mechanism of resistance to M. incognita and M. javanica in both C. africanus and C. myriocarpus was post-infectional nematode resistance, which has attributes for introgression into commercial nematode-susceptible Cucumis cultivars. / Agricultural Research Council (ARC), National Research Foundation of South Africa; and the ARC-Universities Collaboration Centre for Smallholder Farmers

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

Thesis (M.Sc. Agriculture (Agronomy) -- University of Limpopo, 2019 / Most sweet potato-producing regions in South Africa are heavily infested by the root knot (Meloidogyne species) nematodes, which are difficult to manage since the withdrawal of the highly effective fumigant synthetic chemical nematicides. Prior to the withdrawal, the management of Meloidogyne species was not a priority in sweet potato (Ipomoea batatas L.) production since methyl bromide was highly effective in suppressing nematodes. The withdrawal resulted in the introduction of various alternative nematode management strategies, with nematode resistance being the most preferred. However, progress in the use of nematode resistance had been hindered by limited information on accurate species identification since Meloidogyne species have a wide host range and some biological races. The objectives of the study were (1) to determine the degree of nematode resistance in sweet potato cv. 'Mafutha' to M. javanica, M. incognita races 2 and M. incognita race 4 and (2) to investigate the mechanism of resistance in sweet potato cv. 'Mafutha' to M. javanica, M. incognita race 2 and M. incognita race 4. A total of six Experiments were conducted. In each, treatments comprised 0, 25, 50, 125, 250, 625, 1250, 3125 and 5250 eggs and second-stage juveniles (J2), arranged in a randomised complete block design (RCBD), with six replications. Uniform rooted sweet potato cuttings were transplanted in 20-cm-diameter plastic pots filled with steam pasteurised (300˚C for 1 hour) loam soil and Hygromix-T mixed at 3:1 (v/v) ratio. At 56 days after inoculation, plant variables and nematodes in roots were collected. Meloidogyne javanica inoculum levels in Experiment 1 had highly significant (P ≤ 0.01) effects on dry shoot mass and, stem diameter, contributing 74% and 50% in total treatment variation (TTV) of the respective variables, whereas under M. incognita race 2 inoculum levels contributed 70% and 56% in TTV of dry root mass and dry shoot mass, respectively. Meloidogyne incognita race 4 inoculum levels contributed 65% xx and 58% in TTV of stem diameter and dry shoot mass, respectively. In Experiment 2, M. javanica treatment levels contributed 56% in TTV of dry root mass, whereas M. incognita race 2 inoculum levels had no significant effect on any plant variable. In contrast, M. incognita race 4 contributed 51% in TTV of vine length. In Experiment 1, the nematode levels had significant effects on reproductive potential (RP) values, with treatments contributing 96%, 86% and 76% in TTV of RP values in M. javanica, M. incognita race 2 and M. incognita race 4, respectively. In Experiment 2, treatments contributed 79%, 46% and 61% in TTV of RP values in the respective Meloidogyne species. Results of the study suggested that growth of sweet potato cv. 'Mafutha' was affected by nematode infection, whereas the test nematodes were able to reproduce and develop on the test potato cultivar. In conclusion, sweet potato cv. 'Mafutha' was susceptible to M. javanica, M. incognita race 2 and M. incognita race 4 and therefore, the cultivar should not be included in crop rotation programmes intended to manage tropical Meloidogyne species and races in Limpopo Province, South Africa. Since the cultivar was susceptible to the test nematodes, the study did not evaluate the mechanism of resistance. / Agricultural Research Council (ARC), National Research Foundation (NRF) and the Land Bank Chair of Agriculture

Les nématodes à galles du genre Meloidogyne sont des parasites obligatoires des plantes capables d’infecter un large panel de plantes d’intérêt agronomique. Ces parasites ont la capacité d’induire la différenciation de cinq à sept cellules racinaires en cellules géantes, hypertrophiées, métaboliquement actives et multinucléées. Ces cellules géantes constituent le site nourricier indispensable au nématode, et sur lequel il va s’alimenter jusqu’à sa reproduction. Le développement de ces cellules entraine une déformation racinaire appelée « galle » qui va perturber l’absorption de nutriments de la plante et l’affaiblir. Des études transcriptomiques ont montré qu’une vaste reprogrammation transcriptionnelle a lieu lors de la formation de la galle. Cette thèse vise à caractériser le rôle des petits ARN, des ARN non codants, au cours de la formation des galles induites par M. incognita. Les petits ARN non codants sont des régulateurs clés de l’expression génique et comprennent deux grandes familles : les microARN (miARN) et les petits ARN interférents (siARN). Pour cela, les petits ARN de racines de la plante modèle Arabidopsis thaliana saines et infectées par le nématode à galle M. incognita ont été caractérisés par séquençage haut débit à 7 et 14 jours après infection, deux stades importants du développement des cellules géantes. Cette étude a permis d’identifier 24 miARN d’Arabidopsis différentiellement exprimés dans les galles en comparaison aux racines non infectées. L’analyse fonctionnelle de ces miARN a permis de valider le profil d’expression dans les galles de cinq miARN et de démontrer le rôle de miR159 dans la réponse de la plante à M. incognita. De plus, une approche pangénomique a été réalisée afin d’identifier les gènes susceptibles d’être régulés par les siARN lors de l’interaction. En conclusion, ce travail a contribué à démontrer d’une part l’implication des miARN dans l’interaction plante - nématode à galles et a permis l’identification des gènes potentiellement régulés par les siARN lors de l’interaction et impliqués dans la formation des cellules géantes induites par les nématodes à galles. / Root-Knot-Nematodes are obligate plant parasites able to infect a large panel of cultivated plants. These parasites have the ability to induce redifferentiation of five to seven root cells into specialized giant cells, hypertrophied, multinucleated and metabolically overactive. These giant cells form the feeding site upon which nematodes feed continuously until reproduction. Giant cells development leads to a root deformation, named gall, which disturbs plant nutrients absorption causing its weakening. Transcriptomic studies showed that a huge transcriptional reprogramming occurs during gall development. This project aims to characterize the role of small RNAs, non-coding RNAs, during gall development induced by M. incognita. Small non-coding RNAs are key regulators of gene expression and include two major families: microRNAs (miRNAs) and small interfering RNAs (siRNAs). Thus, small RNAs from roots of the model plant Arabidopsis thaliana healthy or infected by M. incognita were characterized by Next Generation Sequencing at 7 and 14 days after infection, two important stages of gall development. This study led to the identification of 24 plant microRNAs differentially expressed in galls compared to uninfected roots. Functional analysis of these miRNAs validated the expression pattern in galls of five miRNAs and demonstrated the role of miR159 in the plant response to M. incognita. In addition, a genome-wide approach was used to identify genes that could be regulated by siRNAs during the interaction. In conclusion, this work contributed todemonstrate, on one hand, the involvement of microRNAs in the plant - RKN interaction and allowed the identification of genes potentially regulated by small interfering and involved in the formation of giant cells induced by root-knot nematodes.

Thesis (M.A. Agriculture. (Plant Production)) -- University of Limpopo, 2021 / Root-knot (Meloidogyne species) nematodes are host to most plant species, with the success of most crops being dependent upon proper nematode management tactics. Sweet potato (Ipomoea batatas L.) is highly susceptible to root-knot nematodes, with physical damage being visible on roots. The withdrawal of highly effective fumigant synthetic nematicides from the agrochemical markets resulted in a need to investigate alternative strategies for managing high nematode population densities, with the use of nematode resistance being the most preferred strategy. The objectives of this study were (1) to establish whether sweet potato cv. 'Blesbok' would be resistant to M. javanica under greenhouse conditions, (2) to investigate whether cucurbitacin containing phytonematicides would be comparable to Velum synthetic nematicide in suppressing Meloidogyne species. For Objective 1, treatments comprised 0, 5, 25, 125, 625, 3125 and 15625 eggs and second-stage juveniles (J2), had six replications and validated in time. Uniform sweet potato cuttings were transplanted in 20-cm diameter plastic pots, filled with steam pasteurised (300°C for 1 hour) loam soil. At 56 days after inoculation, plant growth, plant nutrient and nematode variables were assessed using analysis of variance and subjected to lines of the best fit. Treatments had significant (P ≤ 0.05) effects on eggs and highly significant (P ≤ 0.01) effects on J2, final nematode population densities (Pf) and the reproductive factor (RF), contributing 39, 45, 42 and 92% in total treatment variation (TTV) of the respective variables. Treatments did not have significant effects on plant variables. Calcium, K, Mg and Fe versus M. javanica levels each exhibited negative quadratic relations, with the models being explained by associations from 59 to 96%. In contrast, Zn versus M. javanica levels exhibited positive quadratic relation, with the model being explained by 80 and 98% association and optimised at 125 M. javanica units. For Objective 2, four treatments, namely, untreated control, Nemarioc-AL phytonematicide, Nemafric-BL phytonematicide and Velum had 10 replications and also validated in time. The plantlets with well-developed root system were transplanted under field conditions. Data for Object 2 did not comply with the requirements for ANOVA and were therefore subjected to Principal Component Analysis (PCA). Nemafric-BL phytonematicide treatment in both experiments reduced eggs, J2 in roots and J2 in soil and RP of Meloidogyne species, with the results being comparable to those of Velum synthetic nematicide. Nemarioc-AL phytonematicide reduced J2 in roots and in soil of Meloidogyne species, without affecting eggs in roots and RP. Nemafric-BL phytonematicide and Velum each increased plant growth variables in Experiment 1 and Experiment 2, whereas Nemarioc-AL phytonematicide did not have significant effects on plant growth variables. Velum chemical nematicide stimulated the accumulation of most essential nutrient elements in leaf tissues of the test cultivar, followed by Nemafric-BL phytonematicide, whereas Nemarioc-AL phytonematicide had no significant effects on the accumulation of essential nutrient elements. The study had two major outcomes, namely, (1) that the efficacy of Nemafric-BL phytonematicide was comparable to that of Velum chemical nematicide in suppression of population densities of Meloidogyne species in cv. ′Blesbok′ under field conditions and (2) that cv. ′Blesbok′ was tolerant to M. javanica and therefore, it was not necessary to investigate the mechanisms of nematode resistance. / Agricultural Research Council (ARC) and National Research Foundation (NRF)

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