Thesis (M.Sc. Agriculture (Plant Protection)) -- University of Limpopo, 2021 / In use of phytonematicides as an alternative to synthetic chemical nematicides, the major challenge had been the development of appropriate application technologies, which are currently limited to the ground leaching technology (GLT) and botinemagation (BNT) systems. The former is labour-intensive, whereas the latter requires infrastructure that could be costly for smallholder farmers. The priming of seeds with hypogenous germination properties in phytonematicide solutions could serve as an alternative method of the application of phytonematicides, where the cotyledons would serve as carriers of the active ingredients that are leached into the rhizosphere for suppression of nematode numbers. However, since germination is a chemical process, it is not known whether the active ingredients in cucurbitacin containing phytonematicides would interfere with germination and the subsequent emergence of the seedlings through the incidence of phytotoxicity as observed in the use of the products in crop production. The objectives of the study, therefore, were (1) to investigate the sensitivity and overall sensitivity of pea (Pisum sativum L.) plants to Nemarioc-AL and Nemafric-BL phytonematicides, and (2) to determine the mean concentration point (MCSP) for pea-inoculated with Meloidogyne incognita under greenhouse and microplot conditions, where seeds were previously primed in phytonematicide solutions. Two separate trials were conducted with seven treatments, namely, 0, 2, 4, 8, 16, 32 and 64% Nemarioc-AL or Nemafric-BL phytonematicide, arranged in completely randomised design (CRD), with 8 replications each. Pea seeds were primed in Nemarioc-AL and Nemafric-BL phytonematicide solutions for two hours and shade dried prior to sowing. In vitro trial, 10 seeds were spread uniformly on a moistened filter paper in sterilised petri-dishes with lids and placed in an incubator at 25oC. In vivo trials were under greenhouse and micro-plot conditions, pea seeds were sown in 25-cm and 30-cm diameter plastic pots, respectively. Pots were filled with pasteurised loam soil. Seedlings were inoculated with 5 000 eggs + second-stage juveniles (J2) of M. incognita. Treatments in each case included priming seeds as explained earlier, arranged in a randomised complete block design (RCBD), with 6 replications under greenhouse conditions and 8 replications under micro-plot conditions. In all cases, plant growth variables were assessed using the Curve-fitting Allelochemical Response Dose (CARD) model to generate biological indices which were used to calculate MCSP and the overall sensitivity (Σk). Nematode variables in inoculated trials were assessed using the regression model. In vitro trials, germination variables had positive quadratic relation versus Nemafric-BL phytonematicide, with MCSP= 0.62 % and ∑k = 34 units. In contrast, tested germination variables exhibited negative quadratic relations versus Nemarioc-AL phytonematicide. In greenhouse trials, MCSP values for Nemarioc-AL and Nemafric-BL phytonematicides were 0.62 and 2.18 %, respectively, with ∑k = 0. Plant height (R2 = 0.86), stem diameter (R2 = 0.93) and chlorophyll content (R2 = 0.85), exhibited positive quadratic relationship against Nemarioc-AL phytonematicide, whereas, plant height (R2 = 0.95), stem diameter (R2 = 0.92), chlorophyll content (R2 = 0.89), number of flowers (R2 = 0.93) and dry shoot mass (R2 = 0.94), exhibited positive quadratic relationship against Nemafric-BL phytonematicide. In micro-plot trials, MCSP values for Nemarioc-AL and Nemafric-BL phytonematicides were 0.71 and 2.45 %, respectively, with ∑k = 0. Plant height (R2 = 0.95), stem diameter (R2 = 0.98), chlorophyll content (R2 = 0.98), and gall ratings (R2 = 0.98), exhibited positive quadratic relationships against Nemarioc-AL phytonematicide, while chlorophyll content (R2 = 0.97) and gall ratings (R2 = 0.96) exhibited positive quadratic relationships against Nemafric-BL phytonematicide. All degrees of Nemarioc-AL and Nemafric-BL phytonematicides profoundly reduced nematode numbers under greenhouse and micro-plot trials. In conclusion, both Nemarioc-AL and Nemafric-BL phytonematicides could be applied through the priming technology on pea seeds which have hypogenous germination properties in suppression of nematode population densities. / National Research Foundation (NRF)

In this work, different Rotylenchulus reniformis nematode population numbers affecting cotton plants were spectrally classified using Self-Organized Maps. The hyperspectral reflectance of cotton plants affected by different nematode population numbers were analyzed in order to extract information from the signal that would lead to a fieldworthy methodology for predicting nematode population numbers extant in a plant's rhizosphere. Hyperspectral reflectances from both control and field nematode infestations were used in this work. Various feature extraction and dimensionality reduction methods (e.g., PCA, DWT, and SOM-based methods) were used to extract a reduced set of features. These extracted features were then classified using a supervised SOM classification method. Additionally, this work explores the possibility of combining the standard feature extraction methods with self-organized maps to extract a reduced set of features in order to increase classification accuracies.

No description available.

No description available.

No description available.

No description available.

No description available.

The cyst nematode, Globodera tabacum tabacum Behrens, and the parasitic angiosperm, Egyptian broomrape, Orobanche aegyptiaca Pers., are obligate root parasites that cause severe yield and quality loss of many important crop hosts. Although these represent two diverse classes of parasites, they have significant similarities in the modes of parasitism and complex interactions with their hosts. Conventional control methods have had limited success in controlling these parasites. The overall objective of this research was to engineer resistance to the cyst nematode and Egyptian broomrape by expressing genes encoding parasite specific toxins under the control of parasite-responsive promoters using tobacco (Nicotiana tabacum L. cv. Xanthi). For nematode resistance, an anti-feeding strategy was employed utilizing the tomato proteinase inhibitor I (PI-I) gene as a nematode specific toxin. Transgenic tobacco plants were generated that expressed genes encoding an intracellarly retained or secreted form of tomato PI-I under the control of the nematode-inducible promoter, derived from tomato (Lycopersicon esculentum L.) Hmg2 gene. Our goals were to determine the effectiveness of local PI-I expression on nematode resistance and to determine if intracellular or extracellular PI-I deposition enhances resistance. Two constructs were generated that contained either the coding region of the tomato PI-I gene, lacking the signal sequence (EM1), or the coding region of PI-I including the signal sequence (EM2), fused to the nematode-responsive Hmg2 promoter. Transgenic PI-I plants were inoculated with G. t. tabacum cysts and evaluated for nematode interactions. Our results suggest that local expression of intercellular of PI-I significantly reduced cyst production when compared to the nontransformed controls. For broomrape resistance, a well characterized R/avr gene pair, the tobacco N resistance gene and the tobacco mosaic virus replicase (TMV) gene, was utilized to create novel gene-for-gene resistance via a N gene-mediated hypersensitive response (HR) to limit broomrape parasitism. The bean (Phaselous vulgaris L.) chalcone synthase 8 (CHS8) promoter has been characterized as a broomrape–responsive promoter. We introduced the CHS8:TMV replicase gene construct into tobacco plants that contains an endogenous N gene. Transgenic tobacco plants were inoculated with O. aegyptiaca seeds and monitored for parasite attachment and development. The expression of the TMV replicase leads to a significant reduction in broomrape parasitism. These genetic engineering strategies show promise in enhancing resistance to these destructive parasites. / Ph. D.

Field studies were conducted over two growing seasons to evaluate the effect of different levels of third instar European corn borer, Ostrinia nubilalis Hübner (Lepidoptera: Crambidae), on whole-plant dry matter in corn grown for silage. Mean (± SEM) whole-plant dry matter was significantly greater by 18.8% in uninfested control plants than in plants with an infestation level of 6 larvae/plant in 2004. Whole-plant dry matter in 2005 was significantly greater by 10.5% in control plants than in plants with an infestation level of 5 larvae/plant. Economic injury levels were calculated for each year using regression equations between whole-plant dry matter and European corn borer infestation level. Plant growth stage and infestation level had no effect on percent acid detergent fiber, neutral detergent fiber, and crude protein values for either year. Greenhouse studies were conducted to examine the relationship between aboveground herbivory by European corn borer and belowground herbivory by root knot nematode, Meloidogyne incognita Chitwood (Tylenchida: Heteroderidae), in corn. Two experiments were conducted to measure belowground herbivory by M. incognita in juvenile penetrations and eggs/root system. In the first experiment, the main effects interaction was not significant for either M. incognita juvenile penetrations or eggs/root system. Overall mean juvenile penetrations/root system across all three growth stages, at infestation levels of 1 and 3 larvae/plant were significantly less than in the non-infested control. In addition, overall mean eggs/root system at an infestation level of 3 larvae/plant were significantly less than in the control. In the second experiment, the main effects interaction was significant for both juvenile penetrations and eggs/root system. At the 8 and 10 leaf growth stages, juvenile penetrations/root system at infestation levels of 1 and 3 larvae/plant were significantly less than in the control. In addition, eggs/root system at an infestation level of 3 larvae/plant were significantly less than in the control, at all growth stages. In the reciprocal study, which examined the effect of different M. incognita inoculation levels on European corn borer stalk tunneling, no significant effect of inoculation level on European corn borer stalk tunneling was found. / Ph. D.

An investigation of the effect of Meloidogyne hapla, M. arenaria, M. arenaria thamesi, M. incognita acrita, and M. incognita on Ligustrum ovalifolium, L. japonicum, and Abelia floribunda was conducted. With the exception of M. hapla on L. ovalifolium and M. arenaria thamesi on A. floribunda, all 5 species of nematodes were pathogenic on the 3 host species. Striking differences in growth of the L. ovalifolium plants inoculated with the 4 root-knot species that caused significant reduction could easily be seen. Reduction in weight of the plants inoculated with the 4 root-knot species varied from 39.8% to 63.6% of the controls, and the reduction in total length of stems of these plants varied from 37.2% to 58.5% of the controls at the termination of the experiment. Reduction in weight of the L. japonicum plants inoculated with root-knot species varied from 29.2% to 51.7% of the controls. All plants inoculated with root-knot were significantly reduced in weight compared to controls. There were no visual differences in growth among the A. floribunda treatments; however, omitting the M. arenaria thamesi treatment which was not significantly different from control, the reduction in weight of the inoculated plants varied from 26.81 to 41.71 of the controls. / M.S.