Benard Ngwene

Influence of different mineral nitrogen sources (NO3−-N vs. NH4+-N) on arbuscular mycorrhiza development and N transfer in a Glomus intraradices–cowpea symbiosis

Labeled nitrogen (15 N) was applied to a soil-based substrate in order to study the uptake of N by Glomus intraradices extraradical mycelium (ERM) from different mineral N (NO3− vs. NH4+) sources and the subsequent transfer to cowpea plants. Fungal compartments (FCs) were placed within the plant growth substrate to simulate soil patches containing root-inaccessible, but mycorrhiza-accessible, N. The fungus was able to take up both N-forms, NO3− and NH4+. However, the amount of N transferred from the FC to the plant was higher when NO3− was applied to the FC. In contrast, analysis of ERM harvested from the FC showed a higher 15 N enrichment when the FC was supplied with 15NH4+ compared with 15NO3−. The 15 N shoot/root ratio of plants supplied with 15NO3− was much higher than that of plants supplied with 15NH4+, indicative of a faster transfer of 15NO3− from the root to the shoot and a higher accumulation of 15NH4+ in the root and/or intraradical mycelium. It is concluded that hyphae of the arbuscular mycorrhizal fungus may absorb NH4+ preferentially over NO3− but that export of N from the hyphae to the root and shoot may be greater following NO3− uptake. The need for NH4+ to be assimilated into organically bound N prior to transport into the plant is discussed.

Indigenous leafy vegetables of Eastern Africa — A source of extraordinary secondary plant metabolites

Indigenous African leafy vegetables vary enormously in their secondary plant metabolites whereat genus and the species have a great impact. In African nightshade (Solanum scabrum), spiderplant (Cleome gynandra), amaranth (Amaranthus cruentus), cowpea (Vigna unguiculata), Ethiopian kale (Brassica carinata) and common kale (Brassica oleracea) the specific secondary metabolite profile was elucidated and gained detailed data about carotenoids, chlorophylls, glucosinolates and phenolic compounds all having an appropriate contribution to health beneficial properties of indigenous African leafy vegetables. Exemplarily, various quercetin glycosides such as quercetin-3-rutinoside occur in high concentrations in African nightshade, spiderplant, and amaranth between ~1400-3300μg/g DW. Additionally the extraordinary hydroxycinnamic acid derivatives such as glucaric isomers and isocitric acid isomers are found especially in amaranth (up to ~1250μg/g DW) and spiderplant (up to 120μg/g DW). Carotenoids concentrations are high in amaranth (up to101.7μg/g DW) and spiderplants (up to 64.7μg/g DW) showing high concentrations of β-carotene, the pro-vitamin A. In contrast to the ubiquitous occurring phenolics and carotenoids, glucosinolates are only present in the Brassicales species Ethiopian kale, common kale and spiderplant characterized by diverse glucosinolate profiles. Generally, the consumption of a variety of these indigenous African leafy vegetables can be recommended to contribute to different benefits such as antioxidant activity, increase pro-vitamin A and anticancerogenic compounds in a healthy diet.

Phosphate utilization by the fungal root endophyte Piriformospora indica

Background and aimThe root endophytic fungus Piriformospora indica increases plant resistance and tolerance to stress and promotes plant growth, but its ability to support plant nutrition is still controversially discussed. Irrespective of a potential nutrient transport towards the plant, the fungus might release P from sources unavailable for plant usage by transformation to available forms.MethodsTo test this hypothesis, sterile solid and liquid in vitro cultures of P. indica supplied with different organic and inorganic P sources were established. Cultures were investigated for growth, solubilised P, enzyme activities, RNA accumulation of the four genes encoding phosphate transporters and the two genes for acid phosphatases and phytases respectively found in P. indica genome, and for pH values in the media.ResultsP. indica growth was higher in the presence of inorganic P than in organic P sources. Significant amounts of P were solubilised by P. indica from Ca3(PO4)2 and rock phosphate. However, no relevant intra- or extracellular enzymatic activity was detected despite RNA accumulation of related genes. In general, the genes were all repressed by higher amounts of inorganic P and were expressed the most when the fungus received phytate. We observed a decrease in medium pH in the presence of P. indica irrespective of the P source.ConclusionsP. indica is able to solubilise phosphate from inorganic, but not from organic P sources. This P solubilisation is not due to enzymatic activities but rather to the lowering of the medium pH.

Effects of single and mixed inoculation with two arbuscular mycorrhizal fungi in two different levels of phosphorus supply on β-carotene concentrations in sweet potato (Ipomoea batatas L.) tubers

AimsThis study aimed to determine the effect of arbuscular mycorrhizal (AM) fungi and phosphorus (P) supply levels on β-carotene concentrations in sweet potato (Ipomoea batatas L.) tubers.MethodsTwo commercial AM fungal isolates of Glomus intraradices (IFP Glintra) and Glomus mosseae (IFP Glm) which differ in their life cycles were used. Sweet potato plants were grown in a horizontal split-root system that consisted of two root compartments. A root-free fungal compartment that allowed the quantification of mycelial development was inserted into each root compartment. The two root compartments were inoculated either with the same or with different AM isolates, or remained free of mycorrhizal propagules. Each fungal treatment was carried out in two P supply levels.ResultsIn the low P supply level, mycorrhizal colonization significantly increased β-carotene concentrations in sweet potato tubers compared with the non-mycorrhizal plants. Glomus intraradices appeared to be more efficient in increasing β-carotene concentrations than G. mosseae. Dual inoculation of the root system with the two mycorrhizal fungi did not result in a higher increase in tuber β-carotene concentrations than inoculation with the single isolates. Improved P nutrition led to higher plant tuber biomass but was not associated with increased β-carotene concentrations.ConclusionsThe results indicate a remarkable potential of mycorrhizal fungi to improve β-carotene concentrations in sweet potato tubers in low P fertilized soils. These results also suggest that β-carotene metabolism in sweet potato tubers might be specifically activated by root mycorrhizal colonization.

The role of plant processing for the cancer preventive potential of Ethiopian kale ( Brassica carinata )

ABSTRACT Background: Ethiopian kale (Brassica carinata) is a horticulturally important crop used as leafy vegetable in large parts of East and Southern Africa. The leaves are reported to contain high concentrations of health-promoting secondary plant metabolites. However, scientific knowledge on their health benefits is scarce. Objective: This study aimed to determine the cancer preventive potential of B. carinata using a human liver in vitro model focusing on processing effects on the pattern of secondary plant metabolites and bioactivity. Design: B. carinata was cultivated under controlled conditions and differentially processed (raw, fermented, or cooked) after harvesting. Human liver cancer cells (HepG2) were treated with ethanolic extracts of raw or processed B. carinata leaves and analyzed for their anti-genotoxic, anti-oxidant, and cytostatic potential. Chemical analyses were carried out on glucosinolates including breakdown products, phenolic compounds, carotenoids, and chlorophyll content. Results: Pre-treatment with B. carinata extracts concentration dependently reduced aflatoxin-induced DNA damage in the Comet assay, reduced the production of reactive oxygen species as determined by electron paramagnetic resonance spectroscopy, and induced Nrf2-mediated gene expression. Increasing extract concentrations also promoted cytostasis. Processing had a significant effect on the content of secondary plant metabolites. However, different processing methodologies did not dramatically decrease bioactivity, but enhanced the protective effect in some of the endpoints studied. Conclusion: Our findings highlight the cancer preventive potential of B. carinata as indicated by the protection of human liver cells against aflatoxin in vitro. In general, consumption of B. carinata should be encouraged as part of chemopreventive measures to combat prevalence of aflatoxin-induced diseases.

Intercropping Induces Changes in Specific Secondary Metabolite Concentration in Ethiopian Kale (Brassica carinata) and African Nightshade (Solanum scabrum) under Controlled Conditions

Intercropping is widespread in small-holder farming systems in tropical regions and is also practiced in the cultivation of indigenous vegetables, to alleviate the multiple burdens of malnutrition. Due to interspecific competition and/or complementation between intercrops, intercropping may lead to changes in plants accumulation of minerals and secondary metabolites and hence, alter nutritional quality for consumers. Intercropping aims to intensify land productivity, while ensuring that nutritional quality is not compromised. This study aimed to investigate changes in minerals and secondary plant metabolites in intercropped Brassica carinata and Solanum scabrum, two important African indigenous vegetables, and evaluated the suitability of this combination for dryer areas. B. carinata and S. scabrum were grown for 6 weeks under controlled conditions in a greenhouse trial. Large rootboxes (8000 cm3 volume) were specifically designed for this experiment. Each rootbox was planted with two plants, either of the same plant species (mono) or one of each plant species (mixed). A quartz sand/soil substrate was used and fertilized adequately for optimal plant growth. During the last 4 weeks of the experiment, the plants were either supplied with optimal (65% WHC) or low (30% WHC) irrigation, to test the effect of a late-season drought. Intercropping increased total glucosinolate content in B. carinata, while maintaining biomass production and the contents of other health related minerals in both B. carinata and S. scabrum. Moreover, low irrigation led to an increase in carotene accumulation in both mono and intercropped S. scabrum, but not in B. carinata, while the majority of kaempferol glycosides and hydroxycinnamic acid derivatives of both species were decreased by intercropping and drought treatment. This study indicates that some health-related phytochemicals can be modified by intercropping or late-season drought, but field validation of these results is necessary before definite recommendation can be made to stakeholders.

African Nightshade (Solanum scabrum Mill.): Impact of Cultivation and Plant Processing on Its Health Promoting Potential as Determined in a Human Liver Cell Model

Plant cultivation and processing may impact nutrient and phytochemical content of vegetables. The present study aimed at determining the influence of cultivation and processing on the health promoting capacity of African nightshade (Solanum scabrum Mill.) leaves, an indigenous vegetable, rich in nutrients and phytochemicals. Anti-genotoxicity against the human liver carcinogen aflatoxin B1 (AFB1) as determined by the comet assay and radical oxygen species (ROS) scavenging capacity of ethanolic and aqueous extracts were investigated in human derived liver (HepG2) cells. ROS scavenging activity was assessed using electron paramagnetic spin resonance and quantification of ARE/Nrf2 mediated gene expression. The cultivation was done under different environmental conditions. The processing included fermentation and cooking; postharvest ultraviolet irradiation (UV-C) treatment was also investigated. Overall, S. scabrum extracts showed strong health promoting potential, the highest potential was observed with the fermented extract, which showed a 60% reduction of AFB1 induced DNA damage and a 38% reduction in FeSO4 induced oxidative stress. The content of total polyphenols, carotenoids and chlorophylls was indeed affected by cultivation and processing. Based on the present in vitro findings consumption of S. scabrum leaves could be further encouraged, preferentially after cooking or fermentation of the plant.